198 annales universitatis paedagogicae cracoviensis studia naturae, 1: 198–201, 2016, issn 2543-8832 on 15th of march 2016 in the main building of the pedagogical university (pu) of kraków, on podchorążych 2 street, the open day was organised. like every year, a lot of people took part in this event – starting with the guests, throughout the university authorities, lecturers and students. in this year the open day has been held under the patronage of the mayor of kraków, professor jacek majchrowski and magni�cence rector of pedagogical university, professor michał śliwa. �e aim of this event was to present the structure and functioning of the university to pupils in primary, junior high and secondary schools and to encourage them to undertake their future studies at our university. guests could read the latest o�er of studies for the academic year 2016/2017, see how the pedagogical university looks like, talk to the teaching sta� and ask about all the bothering issues which are related to studying in our alma mater. in the main hall of pu, each institutes prepared their demonstration stand, where students and lecturers encouraged candidates to choose particular courses. �ey presented the wide spectrum of scienti�c possibilities o�ered by the university. �e students and open day of pedagogical university of kraków (kraków, 15th march 2016) dzień otwarty uniwersytetu pedagogicznego w krakowie (kraków, 15 marca 2016 roku) w dniu 15 marca 2016 w budynku głównym uniwersytetu pedagogicznego (up) im. komisji edukacji narodowej w  krakowie, przy ul. podchorążych 2, zorganizowano dzień otwarty uczelni. jak co roku, udział w  nim wzięło wiele osób – poczynając od zaproszonych gości, poprzez władze uczelni oraz wykładowców i  studentów. tegoroczny dzień otwarty został objęty patronatem prezydenta miasta krakowa, prof. jacka majchrowskiego oraz jm rektora up, prof. michała śliwy. celem tego wydarzenia było przybliżenie struktury i funkcjonowania uniwersytetu uczniom szkół podstawowych, gimnazjalnych i  średnich oraz zachęcenie ich do podjęcia w przyszłości studiów na naszej uczelni. goście mogli zapoznać się z  najnowszą ofertą studiów na rok akademicki 2016/2017, zobaczyć jak wyglądają zakątki up, porozmawiać z  kadrą dydaktyczną i  zapytać o  wszystkie nurtujące ich zagadnienia związane ze studiowaniem w naszej alma mater. w auli głównej każdy z instytutów przygotował swoje stoisko pokazowe, przy którym studenci wraz z  wykładowcami zachęcali do wyboru poszczególnych kierunków, poprzez prezentację szerokiego spektrum możliwości naukowych, jakie daje uczel199 r eportsinstitutes employees showed the speci�city of each course using experiments, presentations and thematic exhibitions. institute of biology at the position for the bioinformatics and biological regeneration presented issues which are related to phylogenetic analysis of the nucleotide sequences and the study of evolutionary relatedness between organisms (fig. 1). at the stand for the biology and environmental protection the following issues were presented: chlorophyll �uorescence as an indicator of the vitality of plants, “fragrance memory” and the computer simulation of healthy eating. an additional attraction was the presentation prepared by assoc. prof. małgorzata kłyś, who showed the study of biology and population of common pest of warehouses called grain weevil (sitophilus granarius l.) moreover, there were various activities which were organised by: institute of special education – “sign language”; institute of geography – geological activities: demonstration of microand macrofossils, rock specimens, geological experiments; institute of mathematics – “the world of math games”; institute for occupational safety and citizenship education – medical rescue demonstration, and many others. there was also students council stand where students prepared a lot of interesting contests. in one of them, the main prise was a free trip to the adaptive camp called “adapciak 2016”. �ere were many people at the open day this year – including children (fig. 2) and youth and we hope that information obtained from students and lecturers showed how scienti�c work and teaching looks at nia. dzięki eksperymentom, prezentacjom, a  także wystawom tematycznym, studenci i  pracownicy instytutów przybliżali gościom specy�kę każdego z kierunków. na stanowisku dla kierunku bioinformatyka i  odnowa biologiczna, instytut biologii prezentował zagadnienia dotyczące m.in. analizy �logenetycznej sekwencji nukleotydowych oraz badania pokrewieństwa ewolucyjnego między organizmami (ryc. 1). z kolei na stanowisku dla kierunku biologia i  ochrona środowiska przedstawiano zagadnienia dotyczące �uorescencji chloro�lu jako wskaźnika witalności roślin, „pamięci zapachowej” oraz symulacji komputerowej zdrowego odżywiania. dodatkową atrakcją była prezentacja przygotowana przez dr hab. małgorzatę kłyś, obrazująca badania biologii i populacji pospolitego szkodnika magazynów – wołka zbożowego (sitophilus granarius l.). ponadto przeprowadzono różnego rodzaju warsztaty, które zorganizowały m.in.: instytut pedagogiki specjalnej – „język migowy”; instytut geogra�i – warsztaty geologiczne: pokaz mikroi makroskamieniałości, okazów skalnych, eksperymentów geologicznych; instytut matematyki – „świat gier matematycznych”; instytut bezpieczeństwa i  edukacji obywatelskiej – pokaz ratownictwa i inne. swoje stoisko miał również samorząd studencki, który przygotował wiele ciekawych konkursów. w jednym z nich główną nagrodą był bezpłatny wyjazd na obóz adaptacyjny, tzw. „adapciak 2016”. tegoroczny dzień otwarty zgromadził wiele osób – w tym dzieci (ryc. 2) i młodzież szkolną, a informacje uzyskane od studentów i  wykładowców miejmy nadzieję przybliżyły zainteresowanym, jak wygląda praca nauko200 fig. 1. �e youngest participants of the open day at the pedagogical university in kraków (photo. w. wojtaś) ryc. 1. najmłodsi uczestnicy dnia otwartego uniwersytetu pedagogicznego w krakowie (fot. w. wojtaś) fig. 2. �e presentation of the position of the bioinformatic conducted by msc k. możdżeń and phd g. migdałek (department of plant physiology pu) (photo. w. wojtaś) ryc. 2. prezentacja na stanowisku dla kierunku bioinformatyka prowadzona przez mgr k. możdżeń oraz dr g. migdałka (zakład fizjologii roślin up) (fot. w. wojtaś) r ep or ts 201 r eports ksenia strzeżoń department of botany, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, ksenia1922@gmail.com our university. may the motto of the pedagogical university: “prestige, professionalism, modernity” be the slogan of many new students in the next academic year. wa oraz dydaktyka na naszym uniwersytecie. oby motto uniwersytetu pedagogicznego: „prestiż, profesjonalizm, nowoczesność” było od nowego roku akademickiego hasłem wielu nowych studentów. 176 annales universitatis paedagogicae cracoviensis studia naturae, 2: 176–180, 2017, issn 2543-8832 reports open day of pedagogical university of cracow, kraków, 17th march 2017, poland dzień otwarty uniwersytetu pedagogicznego w krakowie, kraków, 17 marzec 2017, polska the open day of pedagogical university in cracow took place on march 17 2017 under the auspices of the president of cracow – prof. jacek majchrowski and the rector of the university – prof. kazimierz karolczak. as every year, the mission of this event was to approximate the structure and functioning of the school to students in primary schools, lower-secondary and secondary schools. potential students were able to get acquainted with the latest academic offer for the academic year 2017/2018 and they were able to participate in numerous shows and experiments organised by university employees, phd students and students of our university. the opening of this meeting was made by rector prof. kazimierz karolczak in the presence of vice-rectors, deans and other representatives of the university authorities (fig. 1). between the hours 9 a.m. and 5 p.m., in the main hall, each of the institutes presented a wide range of proposals for research and teaching. through presentations, experienc17 marca 2017 roku odbył się dzień otwarty uniwersytetu pedagogicznego w  krakowie pod patronatem prezydenta miasta krakowa – prof. jacka majchrowskiego, jak i  rektora uniwersytetu – prof. kazimierza karolczaka. jak co roku, misją tego wydarzenia było przybliżenie struktury i  funkcjonowania uczelni uczniom szkół podstawowych, gimnazjalnych i  średnich. potencjalni studenci mogli zapoznać się z  najnowszą ofertą studiów na rok akademicki 2017/2018 oraz wziąć udział w  licznych pokazach i  eksperymentach zorganizowanych przez pracowników, doktorantów i studentów naszej uczelni. uroczystego otwarcia niniejszego spotkania dokonał rektor prof. kazimierz karolczak w obecności prorektorów, dziekanów oraz innych przedstawicieli władz uczelni (ryc. 1). w  godzinach od 9 do 17, w  auli głównej każdy z  instytutów zaprezentował szeroki wachlarz propozycji naukowo-dydaktycznych. dzięki prezentacjom, doświadczeniom i  ekspozycjom tematycznym, przybliżona została odwiedzającym indywidu177 r eports es and thematic expositions, the individuality and excellence of each field of study has been approximated. institute of security and civic education organised a quiz on the basis of knowledge about geography and history of security of the third polish republic and the world, training of cardiopulmonary resuscitation on the phantom (bls, aed) and presented the rules of correct bandage. department of psychology and the scientific club of students of psychology up “sensorium” presented experiments in the field of cognitive psychology “expeditions into the depths of mind”, corsis blocks (monkey ladder), gorilla experiment (blindness of attention) and cambridge brain sciences experiments. in the institute of information science, phd małgorzata lebda gave a lecture entitled “pepsi or coca-cola? anatomy of the logo”, alność i  wyjątkowość każdego z  kierunków proponowanych studiów. instytut bezpieczeństwa i  edukacji obywatelskiej zorganizował quiz z  podstaw wiedzy o  geografii i  historii bezpieczeństwa iii rp i świata, szkolenie z resuscytacji krążeniowo-oddechowej na fantomie (bls, aed) oraz zaprezentował zasady prawidłowego bandażowania. katedra psychologii i koło naukowe studentów psychologii up „sensorium” zaprezentowali eksperymenty z zakresu psychologii poznawczej tzw. „ekspedycje w głąb umysłu”, klocki corsiego (monkey ladder), gorilla experminet (ślepota z  braku uwagi) oraz eksperymenty z cambridge brain sciences. w instytucie nauk o informacji dr małgorzata lebda wygłosiła referat pt. „pepsi czy coca-cola? o  anatomii logo”, a  dr stanisław skórka próbował odpowiedzieć na pytanie „architekt informacji: kto, co i  dlaczego?”. fig. 1. rector of the pedagogical university prof. kazimierz karolczak and vice-rector for development assoc. prof. robert stawarz among students representing the university (photo. m. kania) ryc. 1. rektor uniwersytetu pedagogicznego prof. kazimierz karolczak oraz prorektor ds. rozwoju dr hab. robert stawarz wśród studentów reprezentujących uczelnię (fot. m. kania) 178 r ep or ts and phd stanisław skórka tried to answer the question “architect of information: who, what and why?”. phd magdalena lubińska-bogacka from the institute of social work presented the issue on “social work to contemporary social problems”. at the department of art took place visiting photography and animation studio, workshop graphics, design and sculpture. prof. piotr jargusz organised a painting workshop entitled “around the still life” for secondary school. the staff of the up library allowed to visit the libraries magazines and to get acquainted with its functioning. in turn, the center for sport and recreation presented several minutes of fitness, and members of the student club “bakałarz” organised a theater workshop entitled “theater bakałarz”. in the institute of technology very popular were demonstrations of robots (fig. 2), print and 3d scans, paper-making workshops, electronics and microcontrollers labs dr magdalena lubińska-bogacka z instytutu pracy socjalnej zaprezentowała zagadnienia na temat „praca socjalna wobec współczesnych problemów społecznych”. na wydziale sztuki odbyło się m.in. zwiedzanie pracowni fotografii oraz animacji, grafiki warsztatowej, wzornictwa, rzeźby. dla szkół licealnych prof. piotr jargusz zorganizował warsztaty malarskie pt. „wokół martwej natury”. pracownicy biblioteki głównej up umożliwili zwiedzanie magazynów biblioteki i zapoznanie się z jej funkcjonowaniem. z kolei centrum sportu i rekreacji prezentowało kilkuminutowe układy fitness, a członkowie klubu studenckiego „bakałarz” zorganizowali warsztaty teatralne pt. teatr „bakałarz”. w instytucie techniki dużym zainteresowaniem cieszyły się pokazy robotów (ryc. 2), druku i  skanowania 3d, warsztaty papieroplastyczne, laboratorium elektroniki i mikrokontrolerów oraz wykład „0-ra i  1-nki czyli krótka wycieczka do wnętrz komputera”. fig. 2. demonstration of robots for the youngest participants of this event (photo. a. ciejka) ryc. 2. pokaz robotów dla najmłodszych uczestników tego wydarzenia (fot. a. ciejka) 179 r eports and the lecture “0 and 1 – a short trip to the interior of the computer”. in the institute of physics staff presented their experiences of electricity and electrostatics, presented levitating magnets and heat engines, discussed the rules governing natural phenomena and attempted to observe sunspots using the telescope. in the institute of biology there were lectures and practical exercises on the world of plants and fungi, workshops and chemical demonstrations based on catalytic reactions, visiting the spectrometry laboratory and zoological laboratory. in the glasshouse, the staff and students of the department of plant physiology presented simple experiences illustrating important processes of plant life. to make the visitors happy, at the stand there was prepared organic apples, which at the same time were meant to promote the pedagogical university, by attached the school logo to the fruit (fig. 3). w  instytucie fizyki pracownicy przedstawili doświadczenia z  elektryczności i  elektrostatyki, zaprezentowali lewitujące magnesy i silniki cieplne, omówili reguły rządzące zjawiskami przyrodniczymi oraz podjęli próbę obserwacji plam słonecznych za pomocą lunety. w  instytucie biologii odbyły się m.in. prelekcje i zajęcia praktyczne na temat świata roślin i  grzybów, warsztaty i  pokazy chemiczne w  oparciu o  reakcje katalityczne, zwiedzanie laboratorium spektrometrii oraz pracowni zoologicznej. w szklarni, pracownicy i studenci zakładu fizjologii roślin zaprezentowali gościom proste doświadczenia, obrazujące ważne procesy życiowe roślin. w  celu umilenia czasu zwiedzającym, na stoisku tym przygotowany został poczęstunek w  postaci ekologicznych jabłek, które równocześnie miały za zadanie promowanie uniwersytetu pedagogicznego, poprzez dołączone do owoców bileciki z  logiem uczelni (ryc. 3). fig. 3. organic apples with special ticket advertising the pedagogical university in cracow (photo. i. konieczna) ryc. 3. ekologiczne jabłka z specjalnym bilecikiem reklamującym uniwersytet pedagogiczny w krakowie (fot. i. konieczna) 180 r ep or ts the institute of geography organised demonstrations of processes taking place in the geographical environment and presented creative balloon modeling. for the youngest participants there was a face painting and for some older there was wall photo and flash mob. during the open days, potential students could talk to the students and lecturers of the university and get more information about each subject. particularly important was the individual dialogue at the student-student level which was an important experience for prospective students. they were able to find out what the study at the pedagogical university looks like from the perspective of older colleagues. certainly this year open day contributed to the popularization of studies at the pedagogical university. among the youth and children there was an interest in both, the university as a scientific and didactic unit as well as the particular fields of study that were presented during the event. we hope that the information gained in this way and the whole atmosphere of this meeting interested and inspired future students to take up education at the pedagogical university in krakow. we also hope that these activities will attract new students who want to develop their passions and interests in the next academic year. instytut geografii zorganizował pokazy procesów zachodzących w  środowisku geograficznym oraz zaprezentował kreatywne modelowanie balonów. dla najmłodszych uczestników przewidziano malowanie twarzy tzw. „face painting”, a dla nieco starszych zdjęcia na tłach przygotowanych przez studentów tzw. „foto ściana” oraz „flash mob” taneczny. podczas dni otwartych, potencjalni studenci mogli bezpośrednio porozmawiać ze studentami i  wykładowcami uczelni oraz zasięgnąć bliższych informacji na temat poszczególnych kierunków. szczególnie istotnym elementem był tutaj indywidualny dialog na szczeblu uczeń – uczeń, będący ważnym doświadczeniem dla przyszłych kandydatów na studia. mogli oni dowiedzieć się, jak z perspektywy starszych kolegów wygląda studiowanie na uniwersytecie pedagogicznym. z  pewnością tegoroczny dzień otwarty przyczynił się do popularyzacji studiów na uniwersytecie pedagogicznym. wśród młodzieży i dzieci widoczne było zainteresowanie, zarówno uczelnią jako jednostką naukowo-dydaktyczną, jak również poszczególnymi kierunkami, które zostały zaprezentowane podczas trwania tego wydarzenia. oby zdobyte w ten sposób informacje oraz cała atmosfera tego spotkania zaciekawiły i  zainspirowały przyszłych studentów do podjęcia edukacji właśnie na uniwersytecie pedagogicznym w krakowie. można mieć nadzieję, że poczynione działania zaprocentują już w następnym roku akademickim nowymi studentami, chcącymi rozwijać swoje pasje i  zainteresowania. iwona konieczna, karina wieczorek institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland, i.konieczna08@gmail.com 221 if you want to get to know more about science, participate in camps, research projects, scientific conferences, and also promote science through participation in the “scientists night”, “science festival”, and other popular science events, we invite you to join the interdisciplinary scientific group of the biology and geography phd students. the group works at the faculty of geography and biology of the pedagogical university of the national education commission in cracow. as a member of the group, you will meet interesting people, broaden your general knowledge by attending meetings with scientists, and gain new experience by participating in numerous trainings and workshops, and improve your teaching skills in working with youth and children. “do you want to be a scientist?” – we invite you to join the scientific group of the biology and geography phd students. here everyone can be a scientist! the first interdisciplinary scientific group of the biology and geography phd students of the faculty of geography and biology of the pedagogical university was founded in 2014. assoc. prof. magdalena the scientific group of biology and geography phd students of the faculty of geography and biology of the pedagogical university in cracow koło naukowe doktorantów biologii i geografii przy wydziale geograficzno-biologicznym uniwersytetu pedagogicznego w krakowie jeśli chcesz poznać bliżej naukę, wziąć udział w obozach, projektach badawczych, konferencjach naukowych, a  dodatkowo promować naukę poprzez udział w  “nocy naukowców”, “festiwalu nauki” i  innych wydarzeniach popularnonaukowych, zapraszamy do włączenia się w  działalność interdyscyplinarnego koła naukowego doktorantów biologii i  geografii. koło działa przy wydziale geograficzno-biologicznym uniwersytetu pedagogicznego im. komisji edukacji narodowej w  krakowie. będąc członkiem koła poznasz ciekawych ludzi, poszerzysz swoją wiedzę ogólną poprzez uczestnictwo w  spotkaniach z  ludźmi nauki, zdobędziesz nowe doświadczenie, biorąc udział w  licznych szkoleniach i  warsztatach oraz udoskonalisz swoje umiejętności dydaktyczne w  pracy z  młodzieżą i dziećmi. chcesz “być naukowcem?” – zapraszamy do koła naukowego doktorantów biologii i geografii tutaj każdy może nim być! pierwsze interdyscyplinarne koło naukowe doktorantów biologii i  geografii wydziału geograficzno-biologicznego uniwersytetu pedagogicznego zostało założone w 2014 roku. od tego czasu opiekunem koła annales universitatis paedagogicae cracoviensis studia naturae, 2: 221–224, 2017, issn 2543-8832 222 r ep or ts nowak-chmura has been the supervisor of the group since then. each year, the scientific group brings together new members, both biology and geography phd students. the main objectives of the scientific group are, first and foremost, the broadly understood scientific development of phd students, the creation of interdisciplinary research teams, popularisation of science, and integration of the phd students environment. members of the group actively and willingly participate in popular science classes for children at schools and other educational institutions, as well as conduct workshops and lectures at the children and parents university of the pedagogical university. doctoral students also participate in the promotion of science during the “science festival” (fig. 1) in cracow and the “scientists night”. in addition to popular science activity, phd every year, jest pani dr hab. magdalena nowak-chmura. z roku na rok koło naukowe zrzesza nowych członków, zarówno doktorantów biologii, jak i  geografii. głównymi celami działalności koła naukowego jest przede wszystkim: szeroko pojęty naukowy rozwój doktorantów, tworzenie interdyscyplinarnych zespołów badawczych, popularyzowanie nauki oraz integracja środowiska doktorantów. członkowie koła chętnie i  aktywnie biorą udział w  zajęciach popularnonaukowych dla dzieci w szkołach oraz innych placówkach oświatowych, jak również prowadzą warsztaty i wykłady na uniwersytecie dzieci i  rodziców uniwersytetu pedagogicznego. doktoranci biorą również udział w promocji nauki podczas “festiwalu nauki” (ryc. 1) w  krakowie oraz “nocy naukowców”. oprócz działalności popularnonaukowej, doktoranci włączają się co roku w organizację “akcji sos – uczelfig. 1. phd students from the scientific group of the biology and geography phd students and students from the institute of biology participating in the science festival 2017 (photo. b. zyśk) ryc. 1. doktoranci z koła naukowego doktorantów biologii i geografii oraz studenci instytutu biologii, biorący udział w festiwalu nauki 2017 (fot. b. zyśk) 223 r eports students join the organisation of the “sos action – universities to animal shelters”, during which they volunteer to collect food, money, and other items for homeless animals from the cracow animal shelter (fig. 2). one of the ideas of the members of the scientific group is the “scientists workshops”. during the first workshop of this type, one could gain knowledge about the organisation of scientific work, including, e.g., the following: is it worth using the bibliography manager? where does on start to write an article? what makes it easy to write a publication? how does one choose the appropriate scientific journal? both the phd students and scientifically-active employees participated in the “scientists workshops”. thanks to this, it was possible to exchange views and expenie schroniskom zwierząt”, podczas której charytatywnie pomagają zbierać jedzenie, pieniądze oraz inne przedmioty dla bezdomnych zwierząt z  krakowskiego schroniska zwierząt (ryc. 2). jednym z  pomysłów członków koła naukowego są “warsztaty naukowca”. na pierwszych tego rodzaju warsztatach można było zdobyć wiedzę na temat organizacji pracy naukowej w  tym, np.: czy warto używać menadżera bibliografii? od czego zacząć pisanie artykułu? co ułatwia pisanie publikacji? i  jak wybrać odpowiednie czasopismo naukowe. udział w  “warsztatach naukowca” wzięli, zarówno doktoranci, jak i pracownicy czynni naukowo. dało to możliwość bezpośredniej wymiany poglądów i  doświadczeń, pomocnych w  przygotowafig. 2. collection of food for homeless animals organised by the phd students of the scientific group within the “sos action – universities for animal shelters” (photo. ł. binkowski) ryc. 2. zbiórka żywności dla bezdomnych zwierząt zorganizowana przez doktorantów koła naukowego w ramach “akcji sos uczelnie schroniskom zwierząt” (fot. ł. binkowski) 224 r ep or ts anna kocoń phd student in department of invertebrate zoology and parasitology, pedagogical university of cracow, poland, a_kocon@wp.pl riences directly, helping to prepare scientific articles or other ways of presenting the research results. participants of the scientific group also participate in science camps, where they have the opportunity to collect the research material in the field, which will be subsequently appropriately developed, and the results of these studies will be published in scientific journals. the place and date of the meetings of the scientific group are adjusted to the free time of phd students, because many have to combine scientific work with other duties. everyone interested in the scientific group please contact: lkolo17@gmail.com. niu artykułów naukowych lub innych prezentacji wyników. uczestnicy koła naukowego biorą również udział w  obozach naukowych, dzięki którym mają możliwość zbioru w  terenie materiału badawczego, który później jest stosownie opracowywany, a  wyniki tych opracowań publikowane w  czasopismach naukowych. miejsce i  termin spotkań koła naukowego są dostosowywane do wolnego czasu doktorantów, ponieważ wielu musi godzić pracę naukową z innymi obowiązkami. zainteresowanych aktywnością w kole naukowym prosimy o  kontakt: lkolo17@ gmail.com. 187 national scientific conference “lichenology yesterday and today the tenth anniversary of the death of professor józef kiszka”, kraków, 24th march 2017, poland ogólnopolska konferencja naukowa „lichenologia wczoraj i dziś w dziesiątą rocznicę śmierci prof. dr hab. józefa kiszki”, kraków, 24 marzec 2017, polska on march 24, 2017 on the 10th anniversary of the death of prof. józef kiszka, at the institute of biology, pedagogical university of cracow, a lichenological conference took place. the purpose of the conference was to reminisce prof. józef kiszka, co-founder of the kraków school of lichenology, educator of several generations of polish lichenologists and tireless researcher of lichen biota of the polish carpathians. the main organiser of the conference was the department of botany of the institute of biology, where prof. j. kiszka was a long-term employee and manager. profesor józef kiszka (1939–2007) 24 marca 2017 w  10 rocznicą śmierci prof. dr hab. józefa kiszki, odbyła się w  instytucie biologii uniwersytetu pedagogicznego w  krakowie okolicznościowa konferencja lichenologiczna. celem konferencji było przypomnienie osoby prof. józefa kiszki, współtwórcy krakowskiej szkoły lichenologicznej, wychowawcy kilku pokoleń polskich lichenologów oraz niestrudzonego badacza bioty porostów polskich karpat. głównym organizatorem konferencji był zakład botaniki instytutu biologii up, którego wieloletnim pracownikiem i kierownikiem był prof. j. kiszka. annales universitatis paedagogicae cracoviensis studia naturae, 2: 187–190, 2017, issn 2543-8832 188 r ep or ts the conference was opened by prof. zbigniew szeląg, current head of the department of botany and former graduate student of prof. kiszka. the conference was attended by the following representatives of the pedagogical university: vice-rector for development assoc. prof. robert stawarz, dean of the faculty of geography and biology assoc. prof. grzegorz formicki and director of the institute of biology assoc. prof. andrzej rzepka. among the participants were former students and understudies of prof. kiszka, members of his family, numerous staff members of the institute of biology, students and representatives from various science centers in cracow, as well as specially invited guests from scientific institutions from all over poland. more than 100 people attended the conference. there were also many letters from people who could not participate in the conference for various reasons. the conference consisted of three parts: the oral (plenary), the reminiscent, and the outdoor, with the oral and reminiscent parts interlaced. in the plenary session, the participants of the conference had an opportunity to get acquainted with the issues of scientific research, which was conducted by professor kiszka. the introductory lecture, professor józef kiszka – researcher and teacher, was given by phd laura betleja, from the institute of biology, a student and a long-time collaborator of the professor. during this session, other guest speakers also delivered presentations: prof. barbara godzik from the institute of botany of the polish academy of sciences in cracow (“the impact of air pollution on biological diversity, or how lichens cannot be missed in environmental change konferencję uroczyście otwarł prof. dr hab. zbigniew szeląg, obecny kierownik zakładu botaniki, w  przeszłości magistrant prof. kiszki. w  otwarciu konferencji uczestniczyły władze uniwersytetu pedagogicznego w  osobach: prorektora ds. rozwoju dr hab. roberta stawarza, dziekana wydziału geograficzno-biologicznego dr hab. grzegorza formickiego oraz dyrektora instytutu biologii dr hab. andrzeja rzepki. wśród uczestników nie zabrakło dawnych studentów i wychowanków profesora kiszki, członków jego rodziny, licznie przybyłych pracowników instytutu biologii, studentów i  przedstawicieli nauki z  różnych krakowskich ośrodków oraz specjalnie zaproszonych gości z  jednostek naukowych z  całej polski. w konferencji uczestniczyło ponad sto osób. odczytano też wiele listów od osób, które z  różnych względów nie mogły uczestniczyć w konferencji. konferencja składała się z  trzech części: referatowej (plenarnej), wspomnieniowej i  plenerowej, przy czym części referatowa i wspomnieniowa wzajemnie się przeplatały. w  sesji plenarnej (referatowej), uczestnicy konferencji mieli okazję zapoznać się z problematyką badań naukowych, prowadzonych przez prof. kiszkę. referat wprowadzający pt. „profesor józef kiszka – badacz i  nauczyciel” wygłosiła dr laura betleja z  instytutu biologii, wychowanka i  wieloletnia współpracownica profesora. w  trakcie tej sesji, referaty wygłosili również inni zaproszeni goście: prof. dr hab. barbara godzik z  instytutu botaniki polskiej akademii nauk w krakowie („wpływ zanieczyszczeń powietrza na różnorodność biologiczną, czyli jak nie można pominąć porostów w  badaniach zmian śro189 r eportsresearch”). assoc. prof. martin kukwa of the university of gdańsk (“fungi in poland – history of research and state of knowledge”), assoc. prof. paweł czarnota, university of rzeszów (“special cases of taxonomic and chorological research of professor józef kiszka”), assoc. prof. beata krzewicka from the institute botany of the polish academy of sciences in cracow (“freshwater lichens of vistula sources, continued...”), assoc. prof. robert kościelniak from the pedagogical university of cracow (“contribution of professor józef kiszka in the knowledge of the lichen biota of the carpathians”) and prof. wiesław fałtynowicz from the university of wrocław (“current status of lichen biota in poland”). in the “reminiscent” part of the conference, co-workers, former students and friends shared their touching memories about prof. kiszka. there were also many letters from people who worked with the professor. of particular note was the letter from mr. andrzej sikorowski (a member of the cracow vocal and instrumental ensemble “pod budą”), a long-time friend of professor józef kiszka. in his letter, he described the specific relationship between the professor and sikorowski’s father, who, in the 1950s, being the founding editor of “echo of cracow”, funded a scholarship to a gifted student from sąspów primary school. this student turned out to be young józef kiszka from kalinów near cracow. professor kiszka never forgot his “benefactor”, and until his death, he regularly visited him with flowers on his name day. in addition, in this letter, sikorowski recalled how passionate professor kiszka was when talking about lichens and how he named every species he encountered during their walks together. dowiskowych”), prof. dr hab. martin kukwa z  uniwersytetu gdańskiego („grzyby naporostowe w polsce – historia badań i stan poznania”, dr hab. inż. paweł czarnota z  uniwersytetu rzeszowskiego („szczególne przypadki badań taksonomicznych i  chorologicznych profesora józefa kiszki”), dr hab. beata krzewicka z instytutu botaniki polskiej akademii nauk w  krakowie („porosty słodkowodne źródeł wisły, ciąg dalszy …”), dr hab. robert kościelniak z  uniwersytetu pedagogicznego w  krakowie („wkład profesora józefa kiszki w  poznanie bioty porostów karpat”) oraz prof. dr hab. wiesław fałtynowicz z uniwersytetu wrocławskiego („aktualny stan bioty porostów polski”). w  części „wspomnieniowej” konferencji współpracownicy, byli studenci i  przyjaciele podzielili się swoimi wzruszającymi wspomnieniami, dotyczącymi profesora kiszki. odczytano również wiele listów od osób, które współpracowały z  profesorem. szczególnie wzruszający był list od pana andrzeja sikorowskiego (członka krakowskiego zespołu wokalno-instrumentalnego „pod budą”), wieloletniego przyjaciela prof. józefa kiszki. w  liście opisał on specyficzną nić relacji między profesorem, a  swoim ojcem, który w  latach pięćdziesiątych jako redaktor naczelny „echa krakowa” ufundował stypendium dla zdolnego ucznia szkoły podstawowej w sąspowie. uczniem tym okazał się chłopiec józef kiszka, pochodzący z kalinowa k/krakowa. profesor kiszka nigdy nie zapomniał o  swoim „dobroczyńcy” i  do śmierci zjawiał się regularnie z  kwiatami na każdych jego imieninach. ponadto w liście tym sikorowski wspominał, z  jaką pasją profesor kiszka potrafił opowiadać o  porostach oraz jak podc190 r ep or ts finally, in the third part of the conference, participants drove to the cemetery of grębałów in cracow and laid flowers on the grave of professor józef kiszka, on the tenth anniversary of his death. professor józef kiszka was a man of great heart, incredibly hardworking, modest and kind. as an outstanding lichenologist, he will remain in our memory forever. zas wspólnych spacerów w  plenerze nazwał po polsku i  łacinie każdą napotkaną roślinę. na zakończenie, w  trzeciej części konferencji uczestnicy przejechali na cmentarz grębałów w  krakowie i  złożyli kwiaty na grobie profesora józefa kiszki, w  dziesiątą rocznice jego śmierci. profesor józef kiszka był człowiekiem o  wielkim sercu, nieprawdopodobnie pracowitym, skromnym i życzliwym. jako wybitny lichenolog pozostanie na zawsze w naszej pamięci. grzegorz rut, grzegorz migdałek, andrzej rzepka department of plant physiology, institute of biology, pedagogical university, podchorążych 2, 30-084 kraków, poland, grzegorz.rut@up.krakow.pl 191 on april 1st–2nd 2017, the 6th wrocław conference of students of technical and scientific studies “puzzel” took place at the biotechnology educational and research complex. the organisers, like every year, included the scientific groups of four wrocław universities: the student scientific group of biotechnologies of the university of wrocław, the scientific group of general building and the non-destructive research of the wrocław university of technology “etaksi” and the scientific society of materials science represented by prof. rudolf haimann from wrocław university of technology, student scientific group “bioenergia” and student scientific group “orgchem” from the university of life sciences and student scientific group “biomarkers in medical diagnostics” silesian piasts in wrocław. in 2017, the honorary patrons of the conference were the minister of science and higher education, the president of wrocław, rectors and deans of the faculties organising this conference. the partners and sponsors of the “puzzel” conference were polpharvith wrocław conference of students of technical and scientific studies “puzzel”, wrocław, 1st–2nd april 2017 vi wrocławska konferencja studentów nauk technicznych i ścisłych„puzzel”, wrocław, 1–2 kwietnia 2017 roku w  dniach 1–2 kwietnia 2017 w  budynku kompleksu edukacyjno-badawczego biotechnologii uniwersytetu wrocławskiego odbyła się vi edycja wrocławskiej konferencji studentów nauk technicznych i  ścisłych „puzzel”. jak co roku, organizatorami były koła naukowe czterech wrocławskich uczelni: studenckie koło naukowe biotechnologów „przybysz” przy uniwersytecie wrocławskim, koło naukowe budownictwa ogólnego i  badań nieniszczących politechniki wrocławskiej „etaksi”, koło naukowe materiałoznawstwa im. prof. rudolfa haimanna z  politechniki wrocławskiej, studenckie koło naukowe „bioenergia” i  studenckie koło naukowe „orgchem” z  uniwersytetu przyrodniczego oraz studenckie koło naukowe „biomarkery w diagnostyce medycznej” z uniwersytetu medycznego im. piastów śląskich we wrocławiu. w  bieżącym 2017 roku, patronami honorowymi konferencji „puzzel” byli, m.in. minister nauki i  szkolnictwa wyższego, prezydent miasta wrocławia oraz rektorzy i  dziekani wydziałów organizujących niannales universitatis paedagogicae cracoviensis studia naturae, 2: 191–195, 2017, issn 2543-8832 192 r ep or ts ma, cebris, eurx, wrocław research center eit+, root innovation, the manus and skillum foundations. the media patronage was taken by the studentews.pl portal, klaster.plusuj.pl, and biotechnologia.pl, the wrocław university of technology “żak” student monthly, “open innovation” and the foundation for development of science and business in the medical and exact sciences fields. the opening ceremony of the conference was inaugurated by a lecture by phd tomasz rożek, whose research interests focus on the physics of elementary particles. he leads the “sonda2” television program, works with “focus”, “national geographic”, “knowledge and life” and he is one of the co-founder of the scientific journalists association – naukowi.pl. on the same day, an honorary lecture entitled “biomonitoring genotoxicity of particulate air pollutants” was presented by assoc. prof. katarzyna piekarska from the faculty of environmental engineering, wroclaw university of technology. after a  brief coffee break, speakers discussed issues across various disciplines of science. they concerned, e.g., opportunities and threats of the revitalisation of multi-plate housing construction from the perspective of selected housing complexes, advertising in public spaces, problems of contemporary small cities on the context of selected cities with populations of ten thousand inhabitants in the lower silesian voivodship, the influence of unbalanced fertilisation on the soil environment, the influence of crystallisation properties of salt on the surface membranes, cleaning agents used in swimming pools as a  potential source of pool water pollutant niejsze przedsięwzięcie. wśród partnerów i  sponsorów konferencji znalazły się firmy: polpharma, cebris, eurx, wrocławskie centrum badań eit+, root innovation, fundacja manus i  skillum. patronat medialny nad tegoroczną edycją objęły: portal studentews.pl, klaster.plusuj.pl, biotechnologia.pl, miesięcznik studentów politechniki wrocławskiej „żak”, „otwarta innowacja” oraz fundacja rozwoju nauki i  biznesu w  obszarze nauk medycznych i  ścisłych. uroczyste otwarcie konferencji zainaugurował wykład dr tomasza rożka, którego zainteresowania naukowe dotyczą fizyki cząstek elementarnych. prowadzi on program telewizyjny „sonda2”, współpracuje z  czasopismami „focus”, „national geographic”, „wiedza i życie”, a także jest współzałożycielem stowarzyszenia dziennikarzy naukowych – naukowi.pl. w  tym samym dniu, wykład honorowy pt. „biomonitoring genotoksyczności pyłowych zanieczyszczeń powietrza atmosferycznego” wygłosiła dr hab. katarzyna piekarska z  wydziału inżynierii środowiska politechniki wrocławskiej. po krótkiej przerwie kawowej, prelegenci omawiali zagadnienia z  różnych dziedzin nauki. dotyczyły one, np.: szans i  zagrożeń rewitalizacji wielkopłytowego budownictwa mieszkaniowego na przykładzie wybranych zespołów osiedlowych, reklamy w przestrzeni publicznej, problemów współczesnego małego miasta na przykładzie wybranych miast do 10 tys. mieszkańców w  województwie dolnośląskim, wpływu niezrównoważonego nawożenia na środowisko glebowe, wpływu właściwości krystalizacyjnych soli na błony komórkowe, środków czystości stosowanych na pływalniach jako potencjalne źródło za193 r eportsalong with phosphorus compounds, and a  method of the selection of building materials in the reclamation of the opencast mine in cracow. on the same day (1st april 2017), a poster session was held in the hall of the education and research complex. during this session, participants presented, e.g., the phenomenon of hypoxia and acidification of the microenvironment, as mediators of cancer metastasis, celiac disease as a  life without gluten, the need for vitamin b12 in hemodialysis patients, montmorillonite as a catalyst in oxidation process of limonene with hydrogen peroxide, mounting mass based on silicone pressure-sensitive adhesives, the use of microwaves in organic chemistry, the extraction of anthocyanin pigments from cranberry fruits, the influence of different concentrations of cadmium on sweet maize (zea mays l. ‘landmark’) and others. they also tried to answer the following questions, e.g., “are the chia seeds superfood or superb business?”, “what are the emission spectra of plasma?” etc. it was easy to observe a very wide spectrum of research topics presented during the event. on sunday (2nd april 2017), lectures and sessions with students and young scientists discussed in biological, agricultural, medical, and environmental sciences. some of the topics included the following: the identification and marking of selected terpenes in thyme and rosemary plants using the lcms/ms technique, modern technological solutions used in the breeding dairy cows, the interactions of essential oils and entomopathogenic fungi, mechanisms of cancer metastasis, mixed cultures of bacteria and nieczyszczenia wody basenowej związkami fosforu, sposobu doboru materiału budowlanego w rekultywacji terenu kopalni odkrywkowej iłów w krakowie itd. w tym samym dniu (01. 04. 2017), w holu kompleksu edukacyjno-badawczego odbyła się sesja plakatowa. podczas tej sesji uczestnicy omawiali, np. zjawisko hipoksji i zakwaszenia mikrośrodowiska, jako mediatorów przerzutu nowotworowego, celiakii jako życia bez glutenu, zapotrzebowania na witaminę b12 u  pacjentów hemodializowanych, montmorylonit jako katalizator w  procesie utleniania limonenu z  nadtlenkiem wodoru, masa montażowa na bazie kleju silikonowego, stosowania mikrofal w  chemii organicznej, ekstrakcji barwników antocyjanowych z  owoców żurawiny, wpływu zróżnicowanej koncentracji kadmu na kukurydzę (zea mays l. ‘landmark’), i inne. próbowali także odpowiedzieć na pytania, np. – „czy nasiona chia to superfood czy superbiznes?”, „co widma emisyjne mówią o  plazmie?” itd. łatwo było zauważyć bardzo szerokie spektrum prezentowanych tu zagadnień badawczych. podczas niedzielnych (02. 04. 2017) sesji referatowych studenci i młodzi naukowcy poruszali tematy z zakresu nauk biologicznych, rolniczych, medycznych, ochrony środowiska, dotyczące, m.in.: identyfikacji i oznaczenia wybranych terpenów w  tymianku i  rozmarynie techniką lc-ms/ms, nowoczesnych rozwiązań technologicznych wykorzystywanych w  chowie krów mlecznych, interakcji olejków eterycznych i  grzybów entomopatogennych, mechanizmów przerzutowania nowotworów, hodowli mieszanych bakterii oraz szczepów brochothrix thermosphacta sneath & jones, wpływu substratu z alg mor194 r ep or ts strains brochothrix thermosphacta sneath & jones, the effect of marine algae substrate on the degree of protection of plants against stress of drought, the use of nanoparticle formulations to prolong the life of roses and narcissus flowers, tardigrades as organisms that will survive the end of the world, and the energy potential of agricultural biomass used for biogas in the area of the selected municipality. during the conference, the participants had the opportunity for in-depth discussions on their research projects. presenters of various posters and lectures answered the interesting questions raised by the audience, exchanged insights and shared their valuable comments. an additional attraction, which is rarely encountered during any conference, was the opportunity to present self-built robots, cultured crystals, rare plant species, and other research facilities. in this year’s edition of the conference “puzzel”, as many as 207 speakers took part in oral presentations, and 262 young scientists and researchers across the country and abroad presented their research during the poster session. the lecture topics were broad, from biological and chemical to architectural and the engineering of building construction. such diversity of topics helped in broadening the horizon of knowledge and in finding innovative solutions to various analysed problems. simultaneously, this demonstrates the great interest in research among young researchers. the organisers hope that this annual meeting will result in a  stronger integration of science and technology students, not only skich na stopień zabezpieczenia roślin przed stresem suszy, zastosowania preparatów zawierających nanocząstki do przedłużania trwałości kwiatów ciętych róż i  narcyzów, niesporczaków jako organizmów, które przetrwają koniec świata, czy potencjału energetycznego biomasy rolniczej wykorzystanej na cele biogazowe na terenie wybranej gminy. podczas całej konferencji w  kuluarach toczyły się zacięte dyskusje pomiędzy uczestnikami. autorzy referatów i  plakatów odpowiadali na zadawane im pytania, wymieniali się spostrzeżeniami i  cennymi komentarzami, przydatnymi w  dalszej pracy badawczej. dodatkową atrakcją, którą rzadko spotyka się podczas konferencji, była możliwość zaprezentowania własnoręcznie zbudowanych robotów, wyhodowanych kryształów, rzadkich gatunków roślin i  innych obiektów badawczych. w  tegorocznej edycji konferencji „puzzel”, aż 207 prelegentów zaprezentowało wyniki badań w formie ustnych wystąpień, a 262 młodych naukowców z kraju i zagranicy wzięło udział w  sesji plakatowej. tematyka prelekcji była bardzo szeroka, począwszy od zagadnień biologicznych, chemicznych, a  skończywszy na architekturze i  budownictwie. tak ogromna różnorodność tematyczna umożliwia poszerzenie horyzontów i  odnalezienie innowacyjnych rozwiązań analizowanych problemów. jednocześnie świadczy o  ogromnym zainteresowaniu problematyką badawczą wśród młodych naukowców. organizatorzy mają nadzieje, że to spotkanie z  roku na rok będzie owocowało coraz silniejszą integracją środowisk studentów kierunków ścisłych i  technicznych nie tylko 195 r eportsduring the conference, but also during daily work in scientific institutions throughout poland and abroad. w  trakcie konferencji, ale także podczas codziennej pracy w  jednostkach naukowych w całej polsce i poza jej granicami. katarzyna możdżeń1*, peiman zandi2, saikat kumar basu3 1department of plant physiology, institute of biology, pedagogical university, podchorążych 2, 30-084 kraków, poland, *katarzyna.mozdzen@ up. krakow. pl 2institute of crop science, chinese academy of agricultural sciences, beijing 100081, p. r. china, 3department of biological sciences, university of lethbridge, ab canada tik 3m4 207 annales universitatis paedagogicae cracoviensis studia naturae, 1: 207–210, 2016, issn 2543-8832 from 18th till 21st of may took place in kraków �e 16th festival of science, which was organised once again by pedagogical university (pu) of kraków. assoc. prof. robert stawarz, dean of the faculty of geography and biology at pedagogical university, presided the organising committee. it should be noted that also one year ago the pedagogical university took care of organisation of this festival. “time and space” – these words were the motto of this year’s edition, and as the patron was chosen stanisław lem – outstanding creator of science �ction, philosopher and futurologist. honorary patronage over the festival of science was assumed by the president of kraków, the voivodeship marshal of małopolska, the minister of education, the european commission representation in poland, the chairman of city council of kraków, the president of the college of rectors of kraków universities, national centre for research and development and the president of foundation for polish science. festival of science is widely popular among enthusiasts of science and innovation, as well as students from primary and secondary schools. every year a large number festival of science in institute of biology at pedagogical university (kraków, 18th–21st of may 2016) festiwal nauki w instytucie biologii uniwersytetu pedagogicznego (kraków, 18–21 maja 2016) w dniach 18–21 maja 2016 r. w krakowie odbywał się 16. festiwal nauki, którego głównym organizatorem został po raz kolejny uniwersytet pedagogiczny (up) im. komisji edukacji narodowej w  krakowie. komitetowi organizacyjnemu przewodniczył dziekan wydziału geogra�czno-biologicznego – dr hab. robert stawarz. należy wspomnieć, że również rok temu opieka nad festiwalem spoczywała w rękach uniwersytetu pedagogicznego. hasło przewodnie tegorocznej edycji to „czas i  przestrzeń”. patronem festiwalu w  tym roku został wybrany znakomity twórca literatury fantastyczno-naukowej, �lozof i  futurolog – stanisław lem. honorowy patronat tegorocznej edycji festiwalu objęli: prezydent miasta krakowa, wojewoda i  marszałek województwa małopolskiego, minister edukacji narodowej, przedstawicielstwo komisji europejskiej w  polsce, przewodniczący rady miejskiej krakowa, przewodniczący kolegium rektorów szkół wyższych krakowa, narodowe centrum badań i  rozwoju oraz prezes fundacji na rzecz nauki polskiej. festiwal nauki co roku cieszy się ogromnym zainteresowaniem, a  na krakowski rynek, jak również miejsca wydarzeń towa208 of people come not only to the main market square, but also to the various accompanying events in kraków. �e visitors were welcome in huge scienti�c town situated at the heart of the city, where each of higher education institutions in kraków and other research institutions prepared excellent attractions: curiosities, experiments and games from various �elds of science. at each of the university booths one could also see the o�er of education for future students (fig. 1). institute of biology, like every year, presented in his stands diversi�ed exposure and a wide range of physiological and biochemical experiments, referring to the keynote “time and space” – starting with exhibits of paleobiology up till “the evolution of the world”. department of invertebrate zoology and parasitology prepared an extremely current issue. here visitors were able to learn how to protect themselves against ticks during the upcoming holidays (fig. 2). rzyszących, przychodzą tłumy miłośników nauki i  innowacji oraz liczne grupy szkolne. przez trzy dni na rynku odwiedzić można było ogromne naukowe miasteczko, gdzie każda z krakowskich uczelni i inne placówki naukowo-badawcze przygotowały ciekawostki, nowatorskie eksperymenty oraz gry i  zabawy z  różnych dziedzin nauki. na każdym z uczelnianych stoisk można było również zapoznać się z ofertą edukacyjną dla przyszłych studentów (ryc. 1). instytut biologii, jak co roku, prezentował w  swoim namiocie urozmaiconą ekspozycję oraz bogatą ofertę doświadczeń biochemicznych i  �zjologicznych, nawiązujących do „czasu i  przestrzeni” – od eksponatów paleobiologii począwszy, na „ewolucji świata” skończywszy. zakład zoologii bezkręgowców i  parazytologii przygotował temat „na czasie”. odwiedzający mogli dowiedzieć się, jak podczas zbliżających się wakacji uchronić się przed kleszczami (ryc. 2). fig. 1. youth participating in the laboratory demonstrations prepared by the students of institute of biology of pedagogical university (photo. w. wojtaś) ryc. 1. młodzież uczestnicząca w przygotowanych przez studentów instytutu biologii up pokazach laboratoryjnych (fot. w. wojtaś) r ep or ts 209 r eports however, festival of science is not only the festival stalls, but also a number of sports and cultural events taking place in di�erent parts of kraków. in such a wide range everyone could found something for themselves. certainly all sport fans went to the tauron arena kraków to cheer for players of the ecological space tournament. it is the volleyball tournament intended for students from secondary schools. �is year the participants of tournament were graced by the presence of former polish representative – piotr gruszka – the best player of the european championships in 2009. �ere were also special cultural events: performances and concerts, for example awaited by many people concert of józef skrzek with the band and choirs. it is worth pointing out that the weather was nice again, and the weekend of the festival was one of the most beautiful in may. we hope that the next editions of the festival of science in kraków will be as successful jednak festiwal nauki to nie tylko festiwalowe miasteczko, ale także szereg imprez sportowo-kulturalnych, odbywających się w różnych częściach krakowa. w tak szerokiej ofercie każdy mógł znaleźć coś dla siebie. każdy miłośnik sportu z pewnością wybrał się na tauron arenę kraków, by dopingować zawodników ekologicznego turnieju kosmicznego. ten siatkarski turniej, ze specjalnym udziałem mistrza i  najlepszego zawodnika mistrzostw europy 2009, byłego reprezentanta polski – piotra gruszki, wyłonił najlepszą drużynę spośród zgłoszonych szkół ponadgimnazjalnych. nie zabrakło również wyjątkowych wydarzeń kulturalnych: spektakli i  koncertów, m.in. wyczekiwanego przez wielu koncertu józefa skrzeka wraz z  zespołem i  chórami. należy wspomnieć, że po raz kolejny pogoda okazała się bardzo łaskawa, a  festiwalowy weekend był jednym z najładniejszych w maju. liczymy, że kolejne edycje krakowskiego festiwalu nauki będą równie udane jak tefig. 2. children are observing specimens of ticks commonly occurring in poland (photo. w. wojtaś) ryc. 2. dzieci obserwujące okazy kleszczy występujących pospolicie w polsce (fot. w. wojtaś) 210 as this year’s (fig. 3). every year the university organisers put their heart and soul, a lot of energy and good ideas into providing an interesting program and creating a unique atmosphere for visitors. it is de�nitely worth to invite everyone to the festival of science in 2017. see you next year! jakub oliwa1, ksenia strzeżoń2 1 department of plant physiology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, jakuboliwa@gmail.com 2 department of botany, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland fig. 3. biology students who participated in this year’s festival of science (photo. w. wojtaś) ryc. 3. studenci biologii uczestniczący w tegorocznej edycji festiwalu nauki (fot. w. wojtaś) goroczna (ryc. 3). co roku każda jednostka organizująca pokazy wkłada mnóstwo energii, pomysłów i serca, by zapewnić interesujący program i  stworzyć wyjątkową atmosferę dla odwiedzających, dlatego już dziś warto zaprosić wszystkich na festiwal nauki 2017. do zobaczenia za rok! r ep or ts 170 annales universitatis paedagogicae cracoviensis studia naturae, 2: 170–175, 2017, issn 2543-8832 doi: 10.24917/25438832.2.13 s. suresh ramanan1, anatoliy a. khapugin2* 1kerala agricultural university kau, college of forestry, p.o vellanikkara, �rissur, 680656 kerala, india 2joint directorate of the mordovia state nature reserve and national park “smolny”, republic of mordovia, 430011 saransk, russia, *hapugin88@yandex.ru. activity of the intergovernmental science-policy platform on biodiversity and ecosystem services for biodiversity conservation �e e�orts of scientists to create environmental awareness among the policy makers as well as the public were started during the beginning of 19th century. globally, this awareness among people and subsequent changes in the climate and the natural phenomenon made policy makers and politicians more action oriented. one of such great e�ort taken is the foundation of intergovernmental panel on climate change in 1988 (herea�er – ipcc). it is an organisation that was awarded nobel prize in 2007 for its action and e�orts. furthermore, its reports and publications are well acknowledged and recognised politically as well as scienti�cally (smith et al., 2009; howarth et al., 2017). �e reports are mainly based on works that are already published. reports of this sort have great signi�cance and the e�orts behind them have to be appreciated. notwithstanding the due importance of climate change, biodiversity is not a lesser item on the table which needs great attention. besides, it is well-known that mitigation e�orts on climate change do help in biodiversity conservation as an indirect reaction (burch et al., 2014; viña et al., 2016). with the distance between the continents being getting reduced, societies have merged together. �is leads to an increased exchange of biological diversity between countries (van kleunen et al., 2015). for instance, the most of the vegetables on our dining table have reached us far from their centre of origin (vavilovs centres of origin) (vavilov, 1927). �is is a real example for exchange in local biodiversity. not only plants, even animals and insects (hilton, cuthbert, 2010; liebhold et al., 2016) are being exchanged, domesticated and introduced in places far from their centres of origin or its natural habitat. �e main motivation for this introduction is the bene�ts that are derived out of the introduced plants or animals. however, the impact of non-native organisms on the local biological diversity and the environment vary depending on species. history has many examples of the negative a ctivity of the intergovernm ental s cience-p olicy p latform on b iodiversity and ecosystem s ervices for biodiversity conservation 171 impacts caused by the introductions which became an invasive species. lantana camara l. (sharma et al., 2005), eichhornia crassipes (mart.) solms (villamagna, murphy, 2010), opuntia humifusa (raf.) raf. (dean, milton, 2000; fateryga, bagrikova, 2017) are some worldwide examples. globally, the list of invasive organisms is very exhaustive, which is prepared by the international union for conservation of nature invasive species specialist group. �e article in science journal (stokstad, 2017) begins with one such example of south american coatil (nasua nasua l.), which a species that was introduced as a pet in chile and has caused damage to a greater extent. �ese global examples mentioned above underline the need for understanding the biodiversity in order to avoid the catastrophic biodiversity loss. further, it is human nature to follow the path of successful personalities (reis et al., 2000). �e identical phenomenon did happen in the case of ipcc too. development of intergovernmental science-policy platform on biodiversity and ecosystem services (ipbes) �ere is clear scienti�c evidence that we are on the verge of a major biodiversity crisis. practically all facets of biodiversity are in sharp decline (johnson et al., 2017). to date, biodiversity decline is still being unrecognised, and this issue is not receiving proper focus in public resolutions. �is argument has been put forward by many scientists all over the world. yet still, loreau et al. (2006) brought out the paper titled as “diversity without representation”. it details the crux of the need for an international expert panel for biodiversity conservation, which should be more complex than existing one for climate change (ipcc). �e paper also details the activities that led to the formation of intergovernmental science-policy platform on biodiversity and ecosystem services (herea�er – ipbes). �e real development of the ipbes started once the biodiversity loss was highlighted in �e millennium ecosystem assessment report (larigauderie, mooney, 2010). in the meantime, a major conference of unesco was held in france during 2005 entitled as “biodiversity: science and governance” (barbault, le duc, 2005). during this conference, there was a call for an intergovernmental panel on biodiversity with a similar mechanism to that of the ipcc. �is conference produced two documents, the “paris declaration on biodiversity” and a conference statement. �e paris declaration proposed an international mechanism including all governmental and non-governmental institutions into an expert panel on biodiversity. as a follow-up, the french government launched the international mechanism of scienti�c expertise on biodiversity (herea�er – imoseb). �is was �nally revamped into an international science-policy interface, which was intergovernmental and also included many non-governmental stakeholders a�er the montpellier recommendation. s . s ur es h r am an an , a na to liy a . k ha pu gi n 172 in april 2008, representatives of the imoseb consultation and global strategy for the follow-up to the millennium ecosystem assessment met in paris (france). it was decided to merge these two initiatives in order to the support discussions on an “intergovernmental science-policy platform on biodiversity and ecosystem services”. shortly a�erwards, the conference of the parties was been held (may 2008, bonn, germany), where a joint concept on the goals and modalities for ipbes was presented. in meantime, 2008, the united nations environment programme (herea�er – unep) organised the �rst intergovernmental and multi-stakeholder meeting in putrajaya (malaysia), where the mechanism of interface for biodiversity and ecosystem services was discussed. �e next meeting was held in nairobi (kenya, 2009), which was attended by 225 delegates from 95 countries and numerous organisations. �us, the interest for ipbes in the science-policy community was clearly demonstrated. �e third intergovernmental and multi-stakeholder meeting was organised in south korea in 2010, followed by the �rst plenary meeting on ipbes in nairobi in 2011, where the platform’s modalities and institutional arrangements were framed. during the second plenary meeting in panama city (2012), a full-�edged platform was established (larigauderie, mooney, 2010). current achievements and non-recognition of ipbes �e current structure and functions of the ipbes are well detailed in their o�cial website (http://www.ipbes.net). �e output from this institution looks very remarkable. indeed, the �rst report on �ematic assessment of pollinators, pollination, and food production has incorporated more 3000 research papers. it has been highly appreciated by the policy makers as well as the scienti�c community (e.g., potts et al., 2016). �e second report of ipbes was the methodological assessment report on scenarios and models of biodiversity and ecosystem services (ipbes, 2016), which were also applauded globally. currently, the sphere of ipbes activity is still wider. besides keeping records and assessing the biodiversity trends, it also categorises real-world policy constraints in conservation issues. it also helps in the capacity building of governments and others. �e ipbes has employed over 1300 professionals to contribute in its two assessments reports. similar to ipcc and others international panels like international assessment of agricultural knowledge, science and technology for development, it does not conduct any research nor does it monitor climate related data or parameters. however, there were critical opinions on ipbes work in the scienti�c community. for example, turnhout et al. (2012) have indicated some omissions in the ipbes work and suggested nine recommendations for the improvement of ipbes e�ciency. �ey have suggested that the ipbes must focus on a much broader range of knowledge and stakeholders. but, as we see today, the ipbes uses the policy of attracting of external a ctivity of the intergovernm ental s cience-p olicy p latform on b iodiversity and ecosystem s ervices for biodiversity conservation 173 specialists to participate in the external review of the ipbes regional assessments of biodiversity and ecosystem services (larigauderie, watson, 2017). where is support at a difficult time? although ipbes performance is impressive, it has some problems. stokstad (2017) in his article “un biodiversity group confronts cash crunch” presents a clear picture on the current state of ipbes. �e author presents data that clearly indicates that the �nancial status of the ipbes is in an unfortunate condition. as a result, three major assessment reports have been delayed. one of the reports is on controlling invasive species; secondly, it is on the sustainable use of wild species; thirdly, it is on the cultures perspectives of ecosystem services and the quanti�cation of ecosystem services. �is delay is mainly due to the �nancial issues. furthermore, it resulted in the reduction in the ipbes annual budget by 8% (to $8.7 million) in 2017, and it has been decided to cut this budget to $5 million (by 30%) in 2018. as a result, some processes in the algorithm of biodiversity assessment and ecosystem services are delayed. as stokstad (2017) rightly noted, many members have re�ected on this situation. unfortunately, there are many cases with similar situations where impressive results are welcome and highly appreciated, but there is a lack or even the absence of appropriate �nancial support from a state (pei et al., 2012; rybakova, 2013; editorial, 2016). conclusions all these cases clearly indicate that governments, the private sector, and civil societies all over the world want more robust information on reasonable futures for biodiversity and ecosystem services. �ey want to know how the drivers are impacting biodiversity and ecosystem services that might undergo a dramatic change in the upcoming years. �ey also want to comprehend the repercussions of di�erent policy choices on biodiversity and ecosystem services, and how to achieve policy targets, e.g., the aichi targets. despite these numerous desires, they are all reluctant to fund it. �e concern about the economic situation in a country should not hide the problem of environmental quality and nature conservation. �e greatest threat is that the decline in public funding for scienti�c research and organisation, such as ipbes, will have an impact on the research quality. of course, we do not call for funding of every organisation. we say that appropriate funding must be provided to those who provide the relevant results. we would like to end by words of mahatma gandhi: �e world has enough for everyone’s need but not enough for everyone’s greed. s . s ur es h r am an an , a na to liy a . k ha pu gi n 174 acknowledgements �e authors would like to thank the two anonymous reviewers whose comments and recommendations have contributed to improving this paper. references barbault, r., le duc, j.p. 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(2016). e�ects of conservation policy on china’s forest recovery. science advances, 2(3), 1–8. p.e1500965. doi: 10.1126/sciadv.1500965 działalność międzyrządowej platformy naukowo-politycznej na rzecz różnorodności biologicznej i usług ekosystemowych dla zachowania różnorodności biologicznej streszczenie w artykule przedstawiono zwięzłe sprawozdanie dotyczące chronologii oraz działalności międzyrządowej platformy polityki naukowej w  sprawie różnorodności biologicznej i  usług ekosystemowych (ipbes). organizacja ta przywiązuje szczególną wagę do globalnego stanu różnorodności biologicznej, trendów i aktywności politycznej. artykuł prezentuje uogólniony obraz rozwoju i osiągnięć ipbes w trakcie jego istnienia. raporty ipbes ze względu na swoje znaczenie są uznawane i aprobowane na całym świecie. jednak rozpoznawalny sukces ipbes jest niweczony przez sytuację �nansową ipbes. w efekcie istnieje stan, która prowadzi ostatecznie do wniosku, że zawsze następuje hamowanie �nansowania wspólnego dobra. key words: biodiversity monitoring, ecological assessment, ecological policy, funding, nature conservation received: [2017.09.01] accepted: [2017.11.13] 211 annales universitatis paedagogicae cracoviensis studia naturae, 1: 211–215, 2016, issn 2543-8832 on september 30th 2016 took place the 10th edition of researchers night in małopolska. more than 30 di�erent research institutions, educational and cultural, took part in this scienti�c project. many activities in the �elds of chemistry, physics and astronomy, geology and geography, mathematics and computer science, bio-medical sciences, humanities, social and economic, art and technology, were prepared that night. in the bio-medical �eld, among others, institute of biology at the pedagogical university in kraków was presented. on this occasion, the employees, graduate students and students organised workshops, laboratories, lectures, games and fun. �ese presentations were not intended only for children, but also for adults. as part of researchers night many interesting scienti�c issues related to biological �eld were presented. each participant could learn something about the “secrets of the living and nonliving world” and see how complex scienti�c equipment works. during the chemistry workshops, entitled “salts in our environment; analytical reaction; chemiluminescence” the properties of chloride, sodium nitrate (v) of potassiresearchers night in małopolska at the institute of biology of the pu in kraków, september 30th 2016 małopolska noc naukowców w instytucie biologii up w krakowie, 30 września 2016 30. września 2016 roku odbyła się dziesiąta już edycja małopolskiej nocy naukowców. w przedsięwzięciu tym wzięło udział ponad 30 różnych instytucji naukowych, oświatowych i  kulturalnych. przygotowano na nią wiele atrakcji w  dziedzinach: chemii, �zyki i astronomii, geologii i geogra�i, matematyki i informatyki, nauk biologiczno-medycznych, humanistycznych, społecznych i  ekonomicznych, sztuki oraz techniki. w  dziedzinie nauk biologiczno-medycznych zaprezentował się m.in. instytut biologii uniwersytetu pedagogicznego w  krakowie. pracownicy instytutu, doktoranci oraz studenci zorganizowali z  tej okazji warsztaty, pokazy laboratoryjne, wykłady, gry oraz zabawy. przygotowano je nie tylko z przeznaczeniem dla dzieci, ale także dla dorosłych. w ramach wydarzenia zaprezentowano wiele ciekawych zagadnień naukowych, dotyczących szeroko rozumianej problematyki biologicznej. każdy uczestnik mógł zagłębić się przynajmniej w  niektóre „tajemnice świata ożywionego i  nieożywionego” oraz zapoznać z działaniem skomplikowanej aparatury badawczej. na warsztatach z  chemii, zatytułowanych „sole w naszym otoczeniu; reakcje ana212 um hydrogen carbonate and ammonium carbonate and calcium were demonstrated (fig. 1). presentations of precipitation of colourful salt deposits and decomposition of hydrogen peroxide were also conducted. additionally, during the course “colloids in cosmetics – receiving creams and lipsticks” participants tried to independently receive creams, lipsticks and lip glosses. department of biochemistry, biophysics and biotechnology, under the theme “elements of biochemistry in everyday life”, organised workshops for detection of proteins contained in milk, juice, tap water etc., and impact of alcohol and heavy metals salts on the protein was explained. �e experiments connected to surface tension within the framework of issues “elements of biophysics in everyday life” were also presented during that night (fig. 2). during the lecture “�e fruit �y as a model organism in genetic research” in the department of cell biology and genetics, lityczne; chemiluminescencja”, demonstrowano uczestnikom właściwości chlorku sodu, azotanu (v) potasu, wodorowęglanu sodu i amonu oraz węglanu wapnia (ryc. 1). przeprowadzono także pokazy strącania barwnych osadów soli oraz rozkładu perhydrolu. podczas zajęć „układy koloidalne w kosmetyce – otrzymywanie szminek i kremów” uczestnicy warsztatu podejmowali natomiast próby samodzielnego wykonywania kremów, szminek oraz błyszczyków. w  zakładzie biochemii, bio�zyki i  biotechnologii, w  ramach tematu „elementy biochemii w życiu codziennym”, zorganizowano warsztaty dotyczące wykrywania białek, m.in. w  mleku, soku, wodzie z  kranu itp., wpływu alkoholu oraz soli metali ciężkich na białka. demonstrowano również doświadczenia dotyczące napięcia powierzchniowego w  ramach problematyki „elementy bio�zyki w życiu codziennym” (ryc. 2). fig. 1. preparations for researchers night in małopolska in chemistry lab (photo. w. wojtaś) ryc. 1. przygotowania do małopolskiej nocy naukowców w laboratorium chemicznym (fot. w. wojtaś) r ep or ts 213 r eports on the example of the model organism were presented to participants, among other, the human problems induced by changes in the genetic material. �e genetic issues have been very popular for many years now. �e aim of the lecture and presentation titled “�e environmental detective on the trail” was for participants to see the laboratory equipment and methods used in various measurements on biological material. what is more, presentation entitled “mercury in food products” made participants aware of danger of this element for human health, and showed them how to protect themselves from its toxic e�ects. both lectures took place at the department of vertebrate zoology and human biology. �is department proposed also participation in a lecture about bats, their nutrition and interactions with plants, which was entitled “secrets of the bats”. �e department of invertebrate zoology prepared three interesting lectures and w  trakcie wykładu „muszka owocowa jako organizm modelowy w  badaniach genetycznych”, przygotowanego w  zakładzie biologii komórki i genetyki, na przykładzie modelowego organizmu, zaprezentowano uczestnikom m.in. zagadnienia induko wanych przez człowieka zmian w  materiale genetycznym. tematyka ta od lat cieszy się dużym zainteresowaniem. celem prelekcji i  pokazu „detektyw środowiskowy na tropie” było zapoznanie uczestników ze sprzętem laboratoryjnym oraz sposobami wykonywania różnorodnych pomiarów na materiale biologicznym, natomiast podczas prezentacji tematu „rtęć w  produktach spożywczych” uświadamiano zainteresowanym jak pierwiastek ten jest niebezpieczny dla zdrowia człowieka oraz w jaki sposób chronić się przed jego toksycznym działaniem. obydwie prelekcje zorganizowano w zakładzie zoologii kręgowców i biologii człowieka. zakład ten proponował rówfig. 2. young participants of researchers night in małopolska (photo. w. wojtaś) ryc. 2. młodzi uczestniczy małopolskiej nocy naukowców (fot. w. wojtaś) 214 demonstrations. during the lecture “insects and mites, pests of stored food products – a problem with which the world can not deal” were presented the live specimens of insects and mites living in barns and food warehouses. additionally, simple ways to detect and combat such insects were discussed. �e lectures entitled “attention! season for ticks! what each kraków inhabitant should know” and “ticks – dangerous mites. �e medical and veterinary importance of most common species of ticks in poland” were designed to present an overview of the epidemiological situation of tick-borne diseases in poland, with particular emphasis on signi�cance of ticks in medicine and veterinary. moreover, the �rst symptoms of tick-borne diseases, as well as areas of high risk of developing such diseases in poland (including recreational areas, parks, forest paths etc.) were indicated. basic knowledge of this subject plays a very important role in the prevention of these diseases. for fans of the plant world was prepared the lecture in dendrology �eld “what do wood grains say – the story stored in wood” (department of ecology and environmental protection). during the meeting the structure and secondary growth of trees was discussed. in addition, the lecture participants had the opportunity to carry out an independent dendrochronological analysis, consisting of determining the age of trees on the basis of annual growth rings paying attention to the conditions for their growth. �e lecture “light as inspiration for plants” and workshops prepared by sta� and phd students of the department of plant physiology were addressed to people internież uczestnictwo w  wykładzie o  nietoperzach, ich odżywianiu i  interakcjach z  roślinami, zatytułowanym „tajemnice nietoperzy”. w  zakładzie zoologii bezkręgowców przygotowano trzy interesujące prelekcje, połączone z  pokazami. w  prelekcji „owady i  roztocze, szkodniki magazynowanych produktów spożywczych – problem, z  którym nie może uporać się świat”, zaprezentowano żywe okazy owadów i  roztoczy żyjących w spichrzach oraz magazynach spożywczych. omawiano także proste sposoby ich wykrywania i  zwalczania. pozostałe wykłady: „uwaga! sezon na kleszcze! co każdy krakowianin o  kleszczach wiedzieć powinien” oraz „kleszcze – niebezpieczne roztocza. znaczenie medyczne i  weterynaryjne najczęściej występujących w  polsce gatunków kleszczy”, miały na celu przedstawić słuchaczom przegląd sytuacji epidemiologicznej chorób odkleszczowych na terenie polski, ze szczególnym uwzględnieniem kleszczy o  największym znaczeniu medycznym i  weterynaryjnym. ponadto przedstawiono pierwsze objawy chorób odkleszczowych, jak również obszary wysokiego ryzyka zachorowań na tego rodzaju choroby w  polsce (m.in. tereny rekreacyjne, parki, ścieżki leśne itp.). podstawowa wiedza na ten temat odgrywa bardzo ważną rolę w pro�laktyce tych chorób. dla miłośników świata roślin przygotowano prelekcję z  zakresu dendrologii „co mówią słoje historia zapisana w  drewnie” (zakład ekologii i  ochrony środowiska). podczas tej prelekcji omawiano budowę i przyrost wtórny drzew. dodatkowo uczestnicy prelekcji mieli możliwość przeprowadzenia samodzielnej analizy dendrochronologicznej, polegającej na określeniu wieku r ep or ts 215 r eportsested in laboratory experiments on plant material. during the workshop about plants, the role of the light intensity and colour during germination, etiolation and intensity of photosynthesis were discussed. based on the institute of biology of the pu example we can see how rich was the thematic range of this year’s researchers night. certainly it satis�ed both young and older “regulars” of the event. it is therefore not surprising that over the years researchers night in małopolska rises more and more interest among the inhabitants of the city and surrounding area. recent data shows that so far more than 70 000 people were interested in this event. it allows participants to have a close look at the work of scientists and ask specialists about things that every day are not very easy to understand. for scientists it gives the opportunity of direct contact with public, which might be an inspiration to take on new scienti�c topics as well. �erefore, the next editions of the researchers night in małopolska will be among the most liked and respected meetings popularising science in society. katarzyna możdżeń1, beata barabasz-krasny2 1 department of plant physiology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, kasiamozdzen@interia.pl 2 department of botany, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland drzew na podstawie słojów przyrostów rocznych, ze zwróceniem uwagi na warunki ich wzrostu. wykład „światło inspiracją dla roślin” oraz warsztaty przygotowane przez pracowników i doktorantów zakładu fizjologii roślin przeznaczono dla osób zainteresowanych eksperymentami laboratoryjnymi na materiale roślinnym. w  trakcie zajęć omawiano rolę natężenia i  barwy światła w  przebiegu kiełkowania, etiolacji oraz natężenia fotosyntezy u roślin. na przykładzie instytutu biologii up widać, jak bogata była oferta tematyczna tegorocznej edycji nocy naukowców. z  pewnością zadowoliła ona zarówno młodych, jak i  starszych „bywalców” tej imprezy. nie jest zatem dziwne, że z  upływem lat małopolska noc naukowców wzbudza coraz większe zainteresowanie wśród mieszkańców krakowa i  okolic. z  ostatnich danych wynika, że do tej pory przyciągnęła ponad 70 000 zainteresowanych. pozwala ona przyjrzeć się pracy naukowców z  bliska, zapytać o  to, co na co dzień nie sposób łatwo zrozumieć. naukowcom daje możliwość bezpośredniego kontaktu ze społeczeństwem, co może być również inspiracją do podjęcia nowej tematyki badawczej. dlatego kolejne edycje małopolskiej nocy naukowców wpisują się w  kalendarz lubianych i cenionych spotkań popularyzujących naukę w społeczeństwie. 196 annales universitatis paedagogicae cracoviensis studia naturae, 1: 196–197, 2016, issn 2543-8832 reports viiith interdisciplinary scientific conference tygiel 2016 “interdisciplinarity as a key to development”, lublin, 12th–13th march 2016 viii interdyscyplinarna konferencja naukowa tygiel 2016 „interdyscyplinarność kluczem do rozwoju”, lublin, 12–13 marca 2016 roku on 12th and 13th of march 2016 for the 8th time was organised interdisciplinary scienti�c conference “interdisciplinarity as a key to development” in lublin. �e main administrators of this conference were the foundation for promotion of science and development tygiel, as well as various students scientific clubs from university of life sciences in lublin and lublin university of technology. �e opening ceremony of the conference took place at the congress centre of the university of life sciences in lublin on akademicka 13 street. �e conference was inaugurated by associate professor andrzej łukasik presentation “ontological and epistemological aspects of the problem of measurement in quantum mechanics”, phd małgorzata goleman “behavioral disorders or ill-mannered dog?”, phd monika jach’s “microbiota, microbiont and superorganism” and the �ermolytix sp. z o.o. representative “developing innovative ways to increase the e�ciency of photovoltaic cells”. �e strictly scienti�c part of the conference, during which research in 31 various w  dniach 12–13 marca 2016 roku, już po raz ósmy odbyła się w  lublinie interdyscyplinarna konferencja naukowa „interdyscyplinarność kluczem do rozwoju”. głównymi organizatorami były fundacja na rzecz promocji nauki i rozwoju tygiel, a także różne koła naukowe uniwersytetu przyrodniczego w lublinie i politechniki lubelskiej. uroczyste otwarcie konferencji odbyło się w  centrum kongresowym uniwersytetu przyrodniczego w lublinie przy ulicy akademickiej 13. konferencję zainaugurowały wystąpienia dr hab. andrzeja łukasika „ontologiczne i epistemologiczne aspekty problemu pomiaru w  mechanice kwantowej”, dr inż. małgorzaty goleman „zaburzenia behawioralne czy pies źle wychowany?”, dr  moniki jach „mikrobiota, mikrobiom i  superorganizm” oraz przedstawiciela �rmy �ermolytix sp. z  o.o. „opracowanie innowacyjnej metody zwiększenia efektywności ogniw fotowoltaicznych”. po o�cjalnym otwarciu miała miejsce część ściśle naukowa, w  której zaprezentowano wystąpienia referatowe w  31 różnych 197 r eportsthematic sessions were organised a�er the of�cial opening ceremony. during the morning sessions, the participants raised the issue of cancer, health and human psychology. �ey discussed also issues in sections such as: religion and philosophy, politics and society, family and education, language and culture. �e results of research in the natural sciences and environment, tourism and spatial planning, biology of plants and animals were presented during the a�ernoon sessions. participants were focused also on the contemporary problems of lifestyle diseases and genetic issues which become more and more popular. �e problems of the medical, chemical, engineering and food industries sciences were referred during the tygiel conference. additionally, the poster session with 120 posters about di�erent research cases was organised. �e interdisciplinary scienti�c conference tygiel 2016 was without doubt a forum for the exchange of experiences and achievements in �elds of humanities, socio-economic, engineering, environmental and medical sciences. �e main aim of this interdisciplinary approach was to exchange scienti�c views, invite people to cooperation and organise the new research teams. �is conference brought together over 550 people from at least 60 universities in poland. sesjach tematycznych. podczas sesji przedpołudniowych poruszano problematykę chorób nowotworowych, ochrony zdrowia i psychologii człowieka. dyskutowano także na temat zagadnień w  sekcjach: religia i  �lozo�a, polityka i społeczeństwo, rodzina i edukacja, język i  kultura. w  sesjach popołudniowych prezentowano wyniki badań nauk przyrodniczych i środowiskowych, turystyki i gospodarki przestrzennej, biologii roślin i zwierząt. skupiono się także na problemie chorób cywilizacyjnych i  coraz bardziej popularnych ostatnio zagadnieniach z genetyki. referowano także tematy z zakresu nauk medycznych, chemicznych, inżynieryjnych oraz przemysłu spożywczego. w  ramach konferencji zorganizowano także sesję plakatową, podczas której zaprezentowano 120 plakatów o różnej problematyce badawczej. interdyscyplinarna konferencja naukowa tygiel 2016 bezsprzecznie stanowiła forum wymiany doświadczeń i  osiągnięć nauk humanistycznych, społeczno-ekonomicznych, inżynieryjnych, przyrodniczych i  medycznych. interdyscyplinarne podejście miało na celu wymianę poglądów, zachęcenie do współpracy czy tworzenia nowych zespołów badawczych. konferencja ta zgromadziła ponad 550 osób z  co najmniej 60 uczelni w polsce. edyta możdżeń independent researcher; kraków, poland, edytamozdzen@interia.pl 198 annales universitatis paedagogicae cracoviensis studia naturae, 1: 198–201, 2016, issn 2543-8832 on 15th of march 2016 in the main building of the pedagogical university (pu) of kraków, on podchorążych 2 street, the open day was organised. like every year, a lot of people took part in this event – starting with the guests, throughout the university authorities, lecturers and students. in this year the open day has been held under the patronage of the mayor of kraków, professor jacek majchrowski and magni�cence rector of pedagogical university, professor michał śliwa. �e aim of this event was to present the structure and functioning of the university to pupils in primary, junior high and secondary schools and to encourage them to undertake their future studies at our university. guests could read the latest o�er of studies for the academic year 2016/2017, see how the pedagogical university looks like, talk to the teaching sta� and ask about all the bothering issues which are related to studying in our alma mater. in the main hall of pu, each institutes prepared their demonstration stand, where students and lecturers encouraged candidates to choose particular courses. �ey presented the wide spectrum of scienti�c possibilities o�ered by the university. �e students and open day of pedagogical university of kraków (kraków, 15th march 2016) dzień otwarty uniwersytetu pedagogicznego w krakowie (kraków, 15 marca 2016 roku) w dniu 15 marca 2016 w budynku głównym uniwersytetu pedagogicznego (up) im. komisji edukacji narodowej w  krakowie, przy ul. podchorążych 2, zorganizowano dzień otwarty uczelni. jak co roku, udział w  nim wzięło wiele osób – poczynając od zaproszonych gości, poprzez władze uczelni oraz wykładowców i  studentów. tegoroczny dzień otwarty został objęty patronatem prezydenta miasta krakowa, prof. jacka majchrowskiego oraz jm rektora up, prof. michała śliwy. celem tego wydarzenia było przybliżenie struktury i funkcjonowania uniwersytetu uczniom szkół podstawowych, gimnazjalnych i  średnich oraz zachęcenie ich do podjęcia w przyszłości studiów na naszej uczelni. goście mogli zapoznać się z  najnowszą ofertą studiów na rok akademicki 2016/2017, zobaczyć jak wyglądają zakątki up, porozmawiać z  kadrą dydaktyczną i  zapytać o  wszystkie nurtujące ich zagadnienia związane ze studiowaniem w naszej alma mater. w auli głównej każdy z instytutów przygotował swoje stoisko pokazowe, przy którym studenci wraz z  wykładowcami zachęcali do wyboru poszczególnych kierunków, poprzez prezentację szerokiego spektrum możliwości naukowych, jakie daje uczel199 r eportsinstitutes employees showed the speci�city of each course using experiments, presentations and thematic exhibitions. institute of biology at the position for the bioinformatics and biological regeneration presented issues which are related to phylogenetic analysis of the nucleotide sequences and the study of evolutionary relatedness between organisms (fig. 1). at the stand for the biology and environmental protection the following issues were presented: chlorophyll �uorescence as an indicator of the vitality of plants, “fragrance memory” and the computer simulation of healthy eating. an additional attraction was the presentation prepared by assoc. prof. małgorzata kłyś, who showed the study of biology and population of common pest of warehouses called grain weevil (sitophilus granarius l.) moreover, there were various activities which were organised by: institute of special education – “sign language”; institute of geography – geological activities: demonstration of microand macrofossils, rock specimens, geological experiments; institute of mathematics – “the world of math games”; institute for occupational safety and citizenship education – medical rescue demonstration, and many others. there was also students council stand where students prepared a lot of interesting contests. in one of them, the main prise was a free trip to the adaptive camp called “adapciak 2016”. �ere were many people at the open day this year – including children (fig. 2) and youth and we hope that information obtained from students and lecturers showed how scienti�c work and teaching looks at nia. dzięki eksperymentom, prezentacjom, a  także wystawom tematycznym, studenci i  pracownicy instytutów przybliżali gościom specy�kę każdego z kierunków. na stanowisku dla kierunku bioinformatyka i  odnowa biologiczna, instytut biologii prezentował zagadnienia dotyczące m.in. analizy �logenetycznej sekwencji nukleotydowych oraz badania pokrewieństwa ewolucyjnego między organizmami (ryc. 1). z kolei na stanowisku dla kierunku biologia i  ochrona środowiska przedstawiano zagadnienia dotyczące �uorescencji chloro�lu jako wskaźnika witalności roślin, „pamięci zapachowej” oraz symulacji komputerowej zdrowego odżywiania. dodatkową atrakcją była prezentacja przygotowana przez dr hab. małgorzatę kłyś, obrazująca badania biologii i populacji pospolitego szkodnika magazynów – wołka zbożowego (sitophilus granarius l.). ponadto przeprowadzono różnego rodzaju warsztaty, które zorganizowały m.in.: instytut pedagogiki specjalnej – „język migowy”; instytut geogra�i – warsztaty geologiczne: pokaz mikroi makroskamieniałości, okazów skalnych, eksperymentów geologicznych; instytut matematyki – „świat gier matematycznych”; instytut bezpieczeństwa i  edukacji obywatelskiej – pokaz ratownictwa i inne. swoje stoisko miał również samorząd studencki, który przygotował wiele ciekawych konkursów. w jednym z nich główną nagrodą był bezpłatny wyjazd na obóz adaptacyjny, tzw. „adapciak 2016”. tegoroczny dzień otwarty zgromadził wiele osób – w tym dzieci (ryc. 2) i młodzież szkolną, a informacje uzyskane od studentów i  wykładowców miejmy nadzieję przybliżyły zainteresowanym, jak wygląda praca nauko200 fig. 1. �e youngest participants of the open day at the pedagogical university in kraków (photo. w. wojtaś) ryc. 1. najmłodsi uczestnicy dnia otwartego uniwersytetu pedagogicznego w krakowie (fot. w. wojtaś) fig. 2. �e presentation of the position of the bioinformatic conducted by msc k. możdżeń and phd g. migdałek (department of plant physiology pu) (photo. w. wojtaś) ryc. 2. prezentacja na stanowisku dla kierunku bioinformatyka prowadzona przez mgr k. możdżeń oraz dr g. migdałka (zakład fizjologii roślin up) (fot. w. wojtaś) r ep or ts 201 r eports ksenia strzeżoń department of botany, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, ksenia1922@gmail.com our university. may the motto of the pedagogical university: “prestige, professionalism, modernity” be the slogan of many new students in the next academic year. wa oraz dydaktyka na naszym uniwersytecie. oby motto uniwersytetu pedagogicznego: „prestiż, profesjonalizm, nowoczesność” było od nowego roku akademickiego hasłem wielu nowych studentów. 204 annales universitatis paedagogicae cracoviensis studia naturae, 2: 204–207, 2017, issn 2543-8832 iii international interdisciplinary conference “nature-culture-man”, kraków, 23rd–25th june, 2017, poland iii międzynarodowa interdyscyplinarna konferencja „natura-kultura-człowiek”, kraków, 23–25 czerwca 2017, polska the iii international, and viii interdisciplinary, scientific conference “nature-culture-man” took place in kraków on june 23– 25, 2017. the conference was organised by the self-government of phd students of the pedagogical university in cracow and the association of creators of science and culture “episteme”. this year, as in previous years, the pedagogical university in cracow and the ministry of science and higher education held the honorary patronage of this event. on the 23rd of june, on the first day of the conference, before its official opening, the participants took part in the exhibition entitled “nature/man/culture”, which presented works about different visions of contact, coexistence, and the relationship between man and reality. one could admire both the photographs and the artwork on this subject. for example, msc magdalena jędrzejko presented photographs from the “mirror error” series, and msc aleksandra kolobius presented a relief – “destruction of the electromagnetic field in 3 phases”. the next day, june 24, the rector of the pedagogical university of cracow, prof. kazimierz karolczak, officially opened the conference. the inaugural lectures were w krakowie, w dniach 23–25 czerwca 2017, odbyła się iii międzynarodowa, a  viii interdyscyplinarna konferencja naukowa „natura-kultura-człowiek”. organizatorami konferencji byli samorząd doktorantów uniwersytetu pedagogicznego w  krakowie oraz stowarzyszenie twórców nauki i  kultury „episteme”. w  tym roku, podobnie jak w  ubiegłych latach, honorowy patronat nad tym wydarzeniem objął uniwersytet pedagogiczny w krakowie oraz ministerstwo nauki i szkolnictwa wyższego. 23 czerwca, w  pierwszym dniu konferencji jeszcze przed jej oficjalnym otwarciem, uczestnicy wzięli udział w wystawie pt. „natura/człowiek/kultura”, która prezentowała prace opowiadające o  różnych wizjach kontaktu, współistnienia i  relacji pomiędzy człowiekiem, a  rzeczywistością. można było podziwiać, zarówno fotografie, jak i  grafiki o tej tematyce. na przykład, mgr magdalena jędrzejko zaprezentowała fotografie z  serii „mirror error”, a  mgr aleksandra kolobius relief – „zniszczenie pola elektromagnetycznego w 3 fazach”. następnego dnia, tj. 24 czerwca, rektor uniwersytetu pedagogicznego w  krakowie prof. dr hab. kazimierz karolczak dokonał 205 r eportsdelivered by the invited guests: assoc. prof. katarzyna potyrała from the pedagogical university in cracow (“the penetration of science into a wide public area – on communication and scientific mediation”), prof. suman suvedi bhattarai from the tribhuwan university in nepal (“the influence of the environment on the tourism of nepal”) and assoc. prof. hoa kim ngan nhu-tarnawska from the pedagogical university in cracow (“nuclear energy – was this discovery made by a woman?”). after a short break, the organisers invited the participants to take part in the planned thematic sessions of the conference. in the field of humanities and social sciences, the following, among others, appeared: błażej grysztar from the university of zielona góra (“krosno odrzańskie in the period of the third reich”) and izabella janda from the jagiellonian university (“culture as a rational human development in the view of jacques maritain”). natural sciences were presented by: renata rettinger (“slow tourism versus all inclusive tourism: the case of seaside regions of the dominican republic”), msc jakub oliwa (“photosynthetic activity of the white mustard plants in response to the effects of alcoholic extracts from sunflower roots”) and msc vo van thiep (“the growth characteristics of konosirus punctatus (schlegel, 1846) in tam giang – cau hai lagoons”) from the pedagogical university in cracow. the following made presentations within the technology sciences, e.g., patrycja marczewska from the silesian medical university (“the use of chemometric methods in the context of quality control of plant protection products”), and patrycja stasiak and adam krawiec from the technical oficjalnego otwarcia konferencji. wykłady inauguracyjne wygłosili zaproszeni goście: dr hab. katarzyna potyrała z  uniwersytetu pedagogicznego w  krakowie („przenikanie nauki do szerokiej przestrzeni publicznej – o komunikacji i mediacji naukowej”), prof. dr hab. suman suvedi bhattarai z uniwersytetu tribhuwan w  nepalu („wpływ środowiska na turystykę nepalu”) oraz dr hab. hoa kim ngan nhu-tarnawska z  uniwersytetu pedagogicznego w krakowie („energia jądrowa – czy tego odkrycia dokonała kobieta?”). po krótkiej przerwie organizatorzy zaprosili uczestników do udziału w  zaplanowanych sesjach tematycznych konferencji. w  zakresie nauk humanistyczno-społecznych wystąpili, m.in.: błażej grysztar z  uniwersytetu zielonogórskiego („krosno odrzańskie w  okresie iii rzeszy”) oraz izabella janda z  uniwersytetu jagiellońskiego („kultura, jako rozumny rozwój człowieka w  ujęciu jacquesa maritaina”). nauki przyrodnicze prezentowali: renata rettinger („slow tourism versus all incluslive tourism: przypadek nadmorskich regionów republiki dominikany”), mgr jakub oliwa („aktywność fotosyntetyczna roślin gorczycy białej w  odpowiedzi na działanie alkoholowych ekstraktów z  korzeni słonecznika ogrodowego”) oraz mgr vo van thiep („the growth characteristics of konosirus punctatus (schlegel, 1846) in tam giang – cau hai lagoons”) z uniwersytetu pedagogicznego w krakowie. w  obrębie nauk ścisło-technicznych wystąpili, np.: patrycja marczewska ze śląskiego uniwersytetu medycznego („wykorzystanie metod chemometrycznych w  kontekście kontroli jakości środków ochrony roślin”), patrycja stasiak i adam krawiec z politechniki łódz206 r ep or ts university of lodz (“pro-ecological solutions in urban development and their influence on society”). in the field of art, the lecturers included, among others, the following: msc małgorzata budlewska from the wroclaw university of technology (“from exhibit to recipient”), karolina dąbek from the academy of music in cracow (“the notion of nature in the works of iannis xenakis”), and magdalena stawska from the leon koźmiński academy in warsaw (“passion, theatre, culture as perpetual motion of the contemporary society”). the representatives of scientific centres from ukraine and employees, phd students, and students of the pedagogical university in cracow took part in the special panel entitled “poland and ukraine on the road to democracy”. as part of this panel, the researchers from the nieżyński state university presented various issues, e.g., “the complex diagnostics of the political manipulation as a natural part of the psychological defense” – prof. oleksandr boyko, and “the role of propaganda in countries with antidemocratic regimes (namely in socialist countries in terms of the second wave of the cold war)” – phd volodymyr martynenko. while “the neo-colonial discourse of presenting the ukrainian-polish relations in the polish internet media in 2014–2016” was presented by msc mateusz kamionka from the pedagogical university in cracow. participants of this session, “studies on animals” included, among others, msc damian winczewski from the university of szczecin (“animals in the social thought of rosa luxemburg”), msc dariusz gzyra from the pedagogical university in cracow (“modern community and animals – a sketch of crikiej („proekologiczne rozwiązania w zabudowie miejskiej i ich wpływ na społeczeństwo”). w  obszarze sztuki prelegentami byli, m.in.: mgr małgorzata budlewska z  politechniki wrocławskiej („od eksponatu do odbiorcy”), karolina dąbek z  akademii muzycznej w krakowie („wątki natury w twórczości iannisa xenakisa”) oraz magdalena stawska z akademii leona koźmińskiego w warszawie („pasja, teatr, kultura jako perpetuum mobile współczesnego społeczeństwa”). w  panelu specjalnym zatytułowanym „polska i  ukraina na drodze do demokracji” uczestniczyli przedstawiciele ośrodków naukowych z ukrainy oraz pracownicy, doktoranci i  studenci uniwersytetu pedagogicznego w  krakowie. w  ramach tego panelu naukowcy z nieżyńskiego państwowego uniwersytetu prezentowali różnorodne zagadnienia, np.: „the complex diagnostics of the politicial manipulation as a natural part of the psychological defense” – prof. dr hab. oleksandr boyko, “the role of propaganda in countries with antidemocratic regimes (namely in socialist countries in terms of the second wave of the cold war)” – dr volodymyr martynenko. natomiast „neokolonialny dyskurs przedstawiania stosunków ukraińsko-polskich w  polskich mediach interentowych w  latach 2014– 2016” zreferował mgr mateusz kamionka z uniwersytetu pedagogicznego w krakowie. uczestnikami sesji „studia nad zwierzętami” byli, m.in. mgr damian winczewski z  uniwersytetu szczecińskiego („zwierzęta w  myśli społecznej róży luksemburg”), mgr dariusz gzyra z  uniwersytetu pedagogicznego w  krakowie („nowoczesna wspólnota i  zwierzęta – szkic krytyki”) oraz dr sylwia dudek-mańkowska z  uniwersytetu war207 r eportsszawskiego („animal geography – zwierzę w badaniach geograficznych”). w godzinach popołudniowych odbyła się sesja plakatowa, w której wyniki badań zaprezentowali, m.in. mgr ksenia strzeżoń („ocena składu gatunkowego runi łąk użytkowych gminy wadowice„) oraz mgr anna kocoń („niebezpieczeństwo czyha w…trawie. czy turysta w beskidzie śląskim i żywieckim jest bezpieczny?”) z  uniwersytetu pedagogicznego w  krakowie, mgr. monika hanáková z  uniwersytetu konstantyna filozofa w  nitrze („physics olympiad in selected central european countries with focus on assessment methods”), dr andrii ratsilevych z uniwersytetu iwana franka na ukrainie („the social and political life of the poles and ukrainians in volyn under democratic process in 1917”) i  inni. po całodniowych „owocnych” obradach organizatorzy zaprosili uczestników konferencji do udziału w spotkaniu integracyjnym „doktorant party” w klubie „pub & club prestige”. następnego dnia dla zainteresowanych zaplanowano wycieczkę „magiczny kraków”. jej trasa obejmowała zwiedzanie nieznanych zakątków starego miasta oraz krakowskiego kazimierza, dawnej żydowskiej dzielnicy krakowa. tegoroczna interdyscyplinarna konferencja naukowa „natura-kultura-człowiek” zgromadziła ponad 150 osób z całego świata. różnorodność problematyki, podobnie jak w  ubiegłych latach, tak i  w  tym roku, miała na celu integracje naukowców z  różnych dziedzin nauki, co w  tegorocznej edycji na pewno zostało zrealizowane. tique”), and phd sylwia dudek-mańkowska from the university of warsaw (“animal geography – animals in geographic research”). a poster session was held in the afternoon, during which the results were presented by msc ksenia strzeżoń (“assessment of the species composition of the utility meadow grasslands of the municipality of wadowice”), and msc anna kocoń (“danger is lurking in… the grass. is the tourist in the silesian and żywiec beskids safe?”) from the pedagogical university in cracow, msc monika hanáková from the constantine the philosopher university in nitra (“physics olympiad the in selected central european countries with focus on assessment methods”), phd andrii ratsilevych from the ivan frank university in ukraine (“the social and political life of the poles and ukrainians in volyn under democratic process in 1917”), and others. after all-day ‘fruitful’ deliberations, the organizers invited the participants of the conference to take part in the integration meeting “doctoral party” in the “pub & club prestige” club. on the next day, the “magical kraków” trip was planned for those interested. its route included visiting the unknown nooks of the old town and kazimierz in kraków, the former jewish district of kraków. this year’s interdisciplinary scientific conference “nature-culture-man” gathered over 150 people from all over the world. the diversity of issues, as in previous years, was also aimed at the integration of scientists from various fields of science, which was certainly accomplished in this year’s edition. anna kocoń institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland, anna.kocon@up.krakow.pl 181 on march 17–20th, 2017, at the nanotechnology center of the gdańsk university of technology, was the ii interdisciplinary academic conference on environmental protection – iakoś. the organisers were representatives of the faculty of technical physics and applied mathematics, the faculty of civil and environmental engineering, and the faculty of chemistry, gdańsk university of technology. the members of scientific groups were also involved in the project: scientific group of students chemists of gdańsk university of technology, scientific group of engineering and water management “konfuzor”, student scientific group of gdańsk university of technology “microbiology in environmental engineering” and scientific group of physics students of gdańsk university of technology. on the first day of the conference, about 5 in the afternoon, the opening ceremony of the conference took place with the inaugural lecture “what is the impact of the diverse (local and global) anthropogenic activities on the antarctic environment?” presented by prof. żaneta polkowska with co-authorship msc eng. małgorzata szopińska (gdańsk ii interdisciplinary academic conference on environmental protection gdańsk, 17–20th march 2017, poland ii interdyscyplinarna akademicka konferencja ochrony środowiska, gdańsk, 17–20 marzec 2017, polska w dniach 17–20 marca 2017 w centrum nanotechnologii politechniki gdańskiej odbyła się ii interdyscyplinarna akademicka konferencja ochrony środowiska – iakoś. jej organizatorami byli przedstawiciele wydziału fizyki technicznej i matematyki stosowanej, wydziału inżynierii lądowej i środowiska oraz wydziału chemicznego politechniki gdańskiej. w  przedsięwzięcie zaangażowali się także członkowie kół naukowych: naukowe koło chemików studentów politechniki gdańskiej, koło naukowe inżynierii i  gospodarki wodnej „konfuzor”, koło naukowe studentów politechniki gdańskiej „mikrobiologia w  inżynierii środowiska” oraz koło naukowe studentów fizyki politechniki gdańskiej. w  pierwszym dniu konferencji, około godziny 17 nastąpiło uroczyste otwarcie tego spotkania, na którym wykład inaugurujący pt. „jaki wpływ ma różnorodna (lokalna i  globalna) działalność antropogeniczna na środowisko antarktyki?” wygłosiła prof. dr hab. inż. żaneta polkowska we współautorstwie z  mgr inż. małgorzatą szopińską (politechnika gdańska). jeszcze tego samego dnia w  godzinach wieczornych, organannales universitatis paedagogicae cracoviensis studia naturae, 2: 181–186, 2017, issn 2543-8832 182 r ep or ts university of technology). on the same day, in the evening, the organisers invited all participants to an integration meeting held at the manhattan mk bowling shopping centre. the purpose of this meeting was to establish contacts and initiate informal scientific discussions. just like last year, on the second day of the conference (18th march 2017), the event was attended by lecturers, both by participants and sponsors. the deliberations took place simultaneously in two groups. during the lecture session, before lunch in the ia and iia groups presentations were made by msc emil żukowski from the university of natural sciences and humanities in siedlce (“improvement of the physical and chemical properties of rust soils”), and msc eng. michał kropkowski from the university of technology and life sciences in bydgoszcz (“assessment of the influence of thermal conditions on the growth and development of arable crops in poland”), msc eng. justyna jonik from the military academy of technology in warsaw (“detection and determination of harmful organic compounds in industrial waste”), and msc eng. karolina matej-łukowicz from gdańsk university of technology (“location of pollutants in urban catchment basin, based on oliwski stream survey”). in the ib and iib groups presentations were made by, for example, msc beata belica from warsaw university (“protection of middle eastern water resources in conflicts”), msc eng. joanna jaskuła (“modeling transport of rubble in two-stage retention tank – kowalskie lake on the main river”), and msc eng. aleksandra simińska (“comprehensive monitoring of the retention izatorzy zaprosili wszystkich uczestników na spotkanie integracyjne, które odbyło się w mk bowling centrum handlowe manhattan. celem tego spotkania było nawiązanie kontaktów i  zainicjowanie nieformalnych dyskusji naukowych. podobnie jak w ubiegłym roku, w drugim dniu konferencji (18. 03. 2017) wydarzenie uświetniły wykłady prowadzone, zarówno przez uczestników, jak i  przedstawicieli firm sponsorskich. obrady toczyły się jednocześnie w  dwóch grupach. w  sesji referatowej przed obiadem w  grupie ia i  iia wystąpienia mieli, m.in. mgr emil żukowski z  uniwersytetu przyrodniczo-humanistycznego w  siedlcach („poprawa właściwości fizycznych i  chemicznych gleb rdzawych”), mgr inż. michał kropkowski z  uniwersytetu technologiczno-przyrodniczego w  bydgoszczy („ocena stanu nawodnień roślin uprawnych w  polsce na tle globalnym”), mgr inż. justyna jonik z  wojskowej akademii technicznej w  warszawie („wykrywanie i  oznaczanie szkodliwych związków organicznych w ściekach przemysłowych”) oraz mgr inż. karolina matej-łukowicz z  politechniki gdańskiej („rozmieszczenie zanieczyszczeń w zlewni zurbanizowanej, na podstawie badania potoku oliwskiego”). natomiast w grupie ib i  iib referowali, m.in.: mgr beata belica z  uniwersytetu warszawskiego („ochrona zasobów wodnych bliskiego wschodu podczas konfliktów”), mgr inż. joanna jaskuła („modelowanie transportu rumowiska w dwu stopniowym zbiorniku retencyjnym – jezioro kowalskie na rzece głównej”) i  mgr inż. aleksandra simińska („kompleksowy monitoring zbiornika retencyjnego na przykładzie zbiornika rydzyna”) z  uniwersytetu przy183 r eportstank on the example of the rydzyna reservoir”) from the poznań university of life sciences and msc eng. nikole nawrot from gdańsk university of technology (“quality of sediments deposited in the urban drainage system of urban drainage basin”). after the lunch break came the time for sponsorship lectures, which are resulted from studies commissioned by various companies, for example, the klimatherm sponsors lecture whose the speaker presented a subject of “using solar energy to improve the efficiency of refrigeration systems”, green water solutions presented “application of recycling of grey water and recycling rainwater in modern and ecological housing, commercial and public buildings: economy = ecology”, tece “tece floor – eco-friendly, efficient management!” and bimet presented “irox – is a polymer technology that is an ecological alternative to electroplating”. on the same day, in the afternoon, the first of two poster sessions took place in the hall of the building, where the 38 young scientists presented their research, e.g., msc eng. kamila lewicka from the jan długosz university in częstochowa prepared a poster titled “the possibilities of using biodegradable polymer matrices in controlled release of plant protection products”, luiza markowicz from gdańsk university of technology discussed of “cosmetics and personal hygiene as an environmental problem and analytical challenge”, and msc artur eichmann from gdańsk university presented results of “impact of alkaline diesel emission on flora and vegetation change on wejherowo cement plant”. after the whole day of the conference, the organisers invited all the parrodniczego w poznaniu oraz mgr inż. nikole nawrot z  politechniki gdańskiej („jakość osadów deponowanych w systemie kanalizacji deszczowej zlewni zurbanizowanej”). po przerwie obiadowej przyszedł czas na wykłady sponsorskie, będące efektem badań wykonywanych na zlecenie różnych firm, np: klimatherm – „wykorzystanie energii słonecznej do poprawy efektywności układów chłodniczych”, green water solutions – „zastosowanie recyklingu wody szarej i  odzysku deszczówki w  nowoczesnym i  ekologicznym budownictwie mieszkaniowym, komercyjnym oraz obiektach publicznych: ekonomia = ekologia”, tece – „tece floor – ekologicznie ogrzewaj, efektywnie zarządzaj!”, czy bimet – „irox – jest to technologia polimerowa stanowiąca ekologiczną alternatywę dla procesów galwanotechnicznych”. tego samego dnia, w  godzinach popołudniowych, w  holu budynku odbyła się pierwsza z  dwóch sesji plakatowych, w  której wyniki swoich badań zaprezentowało 38 młodych naukowców. na przykład, mgr inż. kamila lewicka z  uniwersytetu jana długosza w  częstochowie przygotowała poster na temat „możliwości zastosowania biodegradowalnych matryc polimerowych w kontrolowanym uwalnianiu środków ochrony roślin”, luiza markowicz z  politechniki gdańskiej omówiła „środki kosmetyczne i higieny osobistej jako problem środowiskowy i wyzwanie analityczne”, natomiast mgr artur eichmann z  uniwersytetu gdańskiego zaprezentował „wpływ emisji pyłów alkalicznych na zmianę flory i  roślinności na przykładzie cementowni wejherowo”. po całym dniu obrad organizatorzy zaprosili wszystkich uczestników do ud184 r ep or ts ticipants to take part in the next integration meeting at pub bifor. this gave us the opportunity to further discuss the topics presented during the second busy day. on the third day of the conference (19th march 2017), before dinner in the iiia and iva groups, speeches and communications were delivered, among others: msc eng. natalia zawrotna from the university of warmia and mazury in olsztyn (“synthesis of polyhydroxyacids with the use of aeromonas bacteria”) and msc agnieszka godela (“effect of wastewater treatment plant “warta” on the overall abundance of bacteria and abundance of metal-resistant bacteria in the warta river waters”), and msc monika lewańska (“rivers as a vector of the spread of sticks of the genus listeria”) from the jan długosz university in częstochowa. in the iiib and ivb groups: sara defratyka from the agh university of science and technology in cracow (“do cigarettes only affect smokers? a study on the impact of cigarettes on the environment”), msc jakub knurek from maria curie-skłodowska university in lublin (“genetic diversity of ash (fraxinus sp.) and its importance in determining susceptibility to diseases”), msc eng. piotr winiarz (“fuel cells as the future of power engineering”) and mikołaj żak from gdańsk university of technology (“high-pressure fuel cells in distributed generation”). in the pre-dinner session, there were more than 20 speakers. after the lunch break, the following invited guests from the gdańsk university of technology delivered speeches: assoc. prof. eng. krzysztof czerwionka (“wastewater treatment plants – from antiquity to the ziału w  kolejnym już spotkaniu integracyjnym w klubie pub bifor. dało to możliwość dalszych dyskusji na tematy prezentowane podczas drugiego pracowitego dnia obrad. w trzecim dniu konferencji (19.03.2017), przed obiadem w  grupach iiia i  iva referaty i  komunikaty wygłosili, m.in.: mgr inż. natalia zawrotna z  uniwersytetu warmińsko-mazurskiego w  olsztynie („synteza polihydroksykwasów z  wykorzystaniem bakterii z  rodzaju aeromonas”), mgr agnieszka godela („wpływ oczyszczalni ścieków „warta” na ogólną liczebność bakterii oraz liczebność bakterii metaloopornych w  wodach rzeki warty”) i  mgr monika lewańska („rzeki jako wektor rozprzestrzeniania się pałeczek z  rodzaju listeria”) z  uniwersytetu jana długosza w  częstochowie. w  grupach iiib i  ivb referowali, m.in.: sara defratyka z akademii górniczo-hutniczej w krakowie („czy papierosy trują tylko palaczy? badanie wpływu papierosów na środowisko naturalne”), mgr jakub knurek z  uniwersytetu marii curie-skłodowskiej z lublina („różnorodność genetyczna jesionu (fraxinus sp.) i jej znaczenie w  określaniu podatności na choroby”), a  także mgr inż. piotr winiarz („ogniwa paliwowe jako przyszłość energetyki”) oraz mikołaj żak („wysokotemperaturowe ogniwa paliwowe w  rozproszonej generacji energii elektrycznej”) z politechniki gdańskiej. łącznie w  przedobiedniej sesji wystąpiło ponad 20 prelegentów. po przerwie obiadowej referaty wygłosili zaproszeni goście z  politechniki gdańskiej: dr hab. inż. krzysztof czerwionka („oczyszczalnie ścieków – od starożytności do przyszłości”), dr hab. inż. jacek gębicki („czy sztuczny nos może zastąpić zmysł węchu?”) 185 r eportsfuture”), assoc. prof. eng. jacek gębicki (“can the artificial nose replace the sense of smell?”), and phd katarzyna jankowska (“polish polar station hornsund of stanislaw siedlecki patronate name – 40 years of polish research in the arctic”), participant of the polish polar expedition of the polish academy of sciences 2013/14. after the lectures and discussions, the organisers invited the participants to take part in the second poster session. the subjects of posters were very different and addressed both general and specific issues, e.g., msc sandra górska from warsaw medical university prepared a poster of “research of medicinal fungal (lentinula edodes) polysaccharide immunoactive structure fraction”, msc mateusz sapieja from maria curie-skłodowska university in lublin discussed on “winter study of rallus aquaticus water” and jakub maculewicz from university of gdańsk discussed issues related to “determination of allelopathic interaction occurring between synechococcus sp. species of baltic filamentous cyanobacteria”. in this session, the 33 young scientists presented the results of their research. at the end of the conference, thanks to the support of the sponsors, the authors of the best posters and speeches were rewarded. one of the special awards was an internship at green water solutions. for the most persistent, the following training was organised: “demonstration of the reaxys chemical base of elsevier”. there was also a tour around the city of gdańsk organised for the most persistent trainees. as the conference, organisers emphasize that the iakoś aim was, and still is, oraz dr katarzyna jankowska („polska stacja polarna hornsund im. stanisława siedleckiego – 40 lat polskich badań w  arktyce”), uczestniczka polskiej wyprawy polarnej pan 2013/14. po części referatowej i  dyskusjach, organizatorzy zaprosili do udziału w  drugiej sesji plakatowej. tematyka prezentowanych posterów była bardzo różnorodna i  dotyczyła, zarówno zagadnień ogólnych, jak i  szczegółowych. na przykład, mgr sandra górska z  warszawskiego uniwersytetu medycznego przygotowała plakat pt. „research of medicinal fungal (lentinula edodes) polysaccharide immunoactive structure fraction”, mgr mateusz sapieja z  uniwersytetu marii curie-skłodowskiej z lublina przedstawił temat „zimowe badania wodnika rallus aquaticus”, a  jakub maculewicz z  uniwersytetu gdańskiego omówił zagadnienia dotyczące „określenia oddziaływania allelopatycznego występującego pomiędzy synechococcus sp. a  wybranymi gatunkami bałtyckich nitkowatych sinic”. w  sesji tej wyniki badań zaprezentowało aż 33 młodych naukowców. na zakończenie konferencji, dzięki wsparciu sponsorów nagrodzono twórców najlepszych plakatów i wystąpień. jedną z nagród specjalnych był staż w  firmie green water solutions. dla najbardziej wytrwałych zorganizowano szkolenie: „demonstracja bazy chemicznej reaxys firmy elsevier”, a następnie wycieczkę po gdańsku. jak podkreślają organizatorzy konferencji, celem iakoś była i nadal jest dyskusja w przyjaznej, naukowej atmosferze, zarówno przedstawicieli nauki, jak i przemysłu. podczas tych paru dni starano się, aby każdy mógł znaleźć temat, który go zainteresuje i  zain186 r ep or ts for representatives of both science and industry to discuss issues in a friendly and scientific atmosphere. during those few days, people were trying to find a topic that would be interesting for them. the conference was attended by over 120 people from polish universities, who presented 72 posters and 48 oral presentations. spiruje do dalszych badań. w  konferencji wzięło udział ponad 120 osób z  polskich uczelni, którzy zaprezentowali 72 plakaty i 48 wystąpień ustnych. sylwia śliwińska-wilczewska1, peiman zandi2, katarzyna możdżeń3 1institute of oceanography university of gdańsk, piłsudzkiego 46, 81-378 gdynia, poland, ocessl@ug.edu.pl 2institute of environment and sustainable development in agriculture, chinese academy of agricultural sciences, p.o. box 10081 beijing, p. r. china 3department of plant physiology, institute of biology, pedagogical university, podchorążych 2, 30-084 kraków, poland 202 annales universitatis paedagogicae cracoviensis studia naturae, 1: 202–206, 2016, issn 2543-8832 vth intercollegiate symposium of biotechnology “symbiosis” 22nd–24th april 2016 v międzyuczelniane sympozjum biotechnologiczne „symbioza” 22–24 kwietnia 2016 roku from 22nd till 24th of april 2016 for the 5th time took place in warsaw (poland) intercollegiate symposium of biotechnology “symbiosis” in name of prof. krzysztof włodzimierz szewczyk. �e main organiser of this symposium was the warsaw association of biotechnology “symbiosis” with the support granted from other institutions, such as: warsaw university of technology, warsaw agricultural university, department of biology university of warsaw. honorary patronage took capital city of warsaw, the academy of young scientists and the university of warsaw. in the �rst plenary session “structural biotechnology” lectures were presented by matthias bochtler of the international institute of molecular and cell biology in warsaw, poland (“dna methylation and hydroxymethylation”) and wanda kukliński of the mrc laboratory of molecular biology in cambridge, england (“correlative light and electron microscopy as an approach to structural biology cell”). �en, a�er the co�ee break began oral presentations in two thematic sessions: “biotechnology of cancer” and “nucleic acid biotechnology”. among more interesting topics discussed during these meetings we w  dniach 22–24 kwietnia 2016 roku po raz piąty odbyło się w  warszawie międzyuczelniane sympozjum biotechnologiczne „symbioza” im. prof. krzysztofa włodzimierza szewczyka. jak co roku, głównym organizatorem sympozjum było warszawskie stowarzyszenie biotechnologiczne „symbioza”. wsparcia udzieliły także m.in. takie instytucje, jak: politechnika warszawska, szkoła główna gospodarstwa wiejskiego czy wydział biologii uniwersytetu warszawskiego. honorowy patronat nad sympozjum objęli: stołeczne miasto warszawa, akademia młodych uczonych oraz uniwersytet warszawski. w  pierwszej sesji plenarnej „biotechnologia strukturalna” referaty wygłosili matthias bochtler z  międzynarodowego instytutu biologii molekularnej i  komórkowej w  warszawie, polska („metylacja i  hydroksymetylacja dna„) oraz wanda kukliński z  mrc laboratorium biologii molekularnej w cambridge, anglia („korelacja światła i  mikroskopii elektronowej jako podejście do biologii strukturalnej komórki”). następnie po przerwie kawowej rozpoczęły się prezentacje ustne w dwóch sekcjach tematycznych: „biotechnologia nowotworów” oraz „biotechnologia kwasu nukleinowego”. z  ciekawszych 203 r eportscan mention “modern methods of diagnosis of acute myeloid leukemia” presented by anna konturek from the jagiellonian university in kraków and “studies on the plant 5’ mrna cap surveillance” referenced by aleksandra kwaśnik from university of warsaw. a�er another break took place the second plenary session about “microand nanobiotechnology”. within its framework, lectures were presented by julian �iele from the leibniz institute of polymer research in dresden, germany (“droplet micro�uidics – from design to material microbioreactors”) and salvador borrós from the universitat ramon llull in barcelona, spain (“design of polymeric bionanomaterials”). a�er busy day of the symposium participants could relax in the evening taking part in the city game – “mokotów field”. �e next day, on april 23rd at 9.30 am the third plenary session “plant biotechnology” was inaugurated. �e �rst speaker was stanisław karpiński from sggw in warsaw, poland (“�e role of energy dissipation and reactive oxygen species in retrograde cell death signaling in arabidopsis”) and the second one brendan davies of the university of leeds in england (“transcriptional repression in plant development and evolution”). a�er this session there was a brand presentation of industry polpharma biologics by piotr zień and edyta dawidczyk. a�er the break began oral presentations in the two following sessions: “medical biotechnology” and “protein biotechnology”. as examples of the topics covered by those discussions we can mention “breeding myocardial cells in microsystems �ow” presented by ewelina tomecka from warsaw university of tematów omawianych podczas tych obrad można wymienić „współczesne metody diagnostyczne ostrych białaczek szpikowych” prezentowane przez annę konturek z  uniwersytetu jagiellońskiego w  krakowie, czy „badania mody�kacji występujących na końcu 5’ mrna u  roślin” zreferowane przez aleksandrę kwaśnik z uniwersytetu warszawskiego. po kolejnej przerwie odbyła się druga tego dnia sesja plenarna, dotycząca „mikro i  nanobiotechnologii”. w  jej ramach referaty wygłosili julian �iele z instytutu badań polimerów leibniza w dreznie, niemcy („mikroprzepływ kropelkowy – od projektowania materiału do mikrobioreaktorów”) oraz salvador borrós z  uniwersytetu ramona llulla w  barcelonie, hiszpania („projekt nanomateriałów polimerowych”). po tak pracowitym dniu obrad uczestnicy sympozjum mogli zrelaksować się wieczorem biorąc udział w grze miejskiej – „pole mokotowskie”. kolejnego dnia, 23 kwietnia o 9.30, zainaugurowano trzecią sesję plenarną pt. „biotechnologia roślin”. pierwszym z referujących był stanisław karpiński ze szkoły głównej gospodarstwa wiejskiego w  warszawie, polska („rola rozpraszania energii i reaktywnych form tlenu w uwstecznianiu sygnalizacji śmierci komórkowej u  arabidopsis”), a  drugim brendan davies z  university of leeds, anglia („represja transkrypcji w  rozwoju i  ewolucji roślin”). po zakończeniu tej sesji miała miejsce prezentacja branżowa �rmy polpharma biologics, którą poprowadzili piotr zień oraz edyta dawidczyk. następnie po przerwie rozpoczęły się prezentacje ustne w  dwóch kolejnych sekcjach tematycznych: „biotechnologia medyczna” i  „biotechnologia białka”. jako przykładowe 204 technology or “investigation of mitochondrial activity in mammaia cell lines” described by jakub dominowski from the international institute of molecular and cell biology in warsaw. a�er a short co�ee break the fourth plenary session was held, entitled “cell genesis and development”. among the speakers were toni gabaldón estevan from the centre for genomic regulation in barcelona (“mitochondria and the origin of the eukaryotic cell), and michał pasternak from the university of cambridge (“live imaging rnai screen for genes essential for meiosis in mammalian oocytes”). a�er dinner deliberations were held in sessions “plant biotechnology” and “medical biotechnology”. among many interesting topics were discussed: “identi�cation and characterization of the coding sequences of homologues spl in lupinus luteus” presented by krzysztof michalski of the nicolaus copernicus university in toruń, poland and “curcumin as a potential tool in combined mitoxantrone-based chemotherapy of carcinosarma” presented by marcin luty from the jagiellonian university in kraków. immediately a�er those sessions there was the �rst part of the poster session. on 39 posters were presented the results of various studies of young scientists who had the opportunity to discuss and exchange experiences gained during the implementation of their own research projects. �ose discussions could continue during the evening integration meeting in the club “medic”. on the last day of the symposium (april 24th) began sessions of oral presentations in the �elds of “microbiology” and “nanobiotechnology”. among the examples of the tematy poruszane w ramach tych obrad można wymienić m.in.: „hodowla komórek mięśnia sercowego w  mikrosystemie przepływowym” prezentowany przez ewelinę tomecką z  politechniki warszawskiej, czy „badanie aktywności mitochondrialnej w liniach komórkowych ssaków” omawiane przez jakuba dominowskiego z międzynarodowego instytutu biologii molekularnej i  komórkowej w  warszawie. po krótkiej przerwie kawowej odbyła się druga tego dnia, a  czwarta w  kolejności sesja plenarna „geneza i  rozwój komórki”. wśród referujących znaleźli się: toni gabaldón estevan z  centrum regulacji genomu w barcelonie, hiszpania („mitochondria i pochodzenie komórek eukariotycznych”), a także michał pasternak z uniwersytetu w cambridge, anglia („techniki obrazowania cząsteczek rnai i genów istotnych dla mejozy zachodzącej w komórkach jajowych ssaków”). poobiednie obrady odbywały się w  sekcjach tematycznych: „biotechnologia roślin” oraz „biotechnologia medyczna”. wśród wielu interesujących tematów poruszanych w ramach tych sesji znalazły się m.in.: „identy�kacja i  charakterystyka sekwencji kodujących homologów spl u  lupinus luteus” referowana przez krzysztofa michalskiego z uniwersytetu mikołaja kopernika w toruniu, polska oraz „kurkumina jako potencjalne narzędzie dla opartej na mitoksantronie chemioterapii stosowanej w leczeniu nowotworów złośliwych” zaprezentowana przez marcina luty z  uniwersytetu jagiellońskiego w  krakowie. bezpośrednio po zakończeniu obrad w  sekcjach tematycznych nastąpiła pierwsza część sesji plakatowej. na 39 plakatach przedstawiono wyniki różnorodnych badań, a młodzi naukowcy mieli okazję podyskutor ep or ts 205 r eportstopics we can enumerate: “evaluation of the possibility of using waste from the �sh factories in the synthesis of microbial oil by yarrowia lipolytica” given by bartłomiej zieniuk from sggw in warsaw, poland and “how quantum mechanics limits our knowledge about biological systems?” presented by wojciech rządkowski from the university of warsaw. a�er completion of the oral presentations the second part of the poster session was held. �is time, the results of research were presented on 40 posters. during this part of the symposium, participants discussed also scienti�c issues of their interest and exchanged their experiences. �en the meeting was continued in the ��h plenary session on “cell biotechnology”. here lectures were delivered by maciej kurpisz from the institute of human genetics, academy of sciences in poznań, poland (“stem cells and �eir application in clinical practice”) and devrim kilinc from university college dublin, ireland (“combining mechanical and chemical stimuli that induce cell growth and death”). �e sixth plenary session took place a�er the lunch break and referred to the “rna biotechnology”. �e speakers in this session were artur jarmołowski from adam mickiewicz university in poznań, poland (“post-transcriptional coordination of splicing and mirna biogenesis in plants”) and andrzej dziembowski from the institute of biochemistry and biophysics in warsaw, poland (“human exosome complex in health and disease”). about 4 pm there was a summary and the o�cial closing of this year’s edition of the interuniversity biotechnology symposium. during those three days, scienti�c problems were referred and discussed in six plewać i wymienić się doświadczeniami zdobytymi w trakcie realizacji własnych projektów badawczych. dyskusje te można było kontynuować podczas wieczornego spotkania integracyjnego w klubie „medyk”. ostatniego dnia sympozjum (24 kwietnia) obrady rozpoczęto sesjami prezentacji ustnych z  zakresu „mikrobiologii” i  „nano-biotechnologii”. wśród przykładowych poruszanych tematów można wymienić: „ocena możliwości zastosowania odpadów z  rybnych zakładów produkcyjnych w  syntezie oleju mikrobiologicznego przez drożdże yarrowia lipolytica” wygłoszony przez bartłomieja zieniuka ze szkoły głównej gospodarstwa wiejskiego w  warszawie, czy „jak mechanika kwantowa ogranicza naszą wiedzę o  systemach biologicznych?” przedstawiony przez wojciecha rządkowskiego z  uniwersytetu warszawskiego. po zakończeniu prezentacji ustnych odbyła się druga część sesji plakatowej. tym razem zaprezentowano wyniki badań na 40 plakatach. również w tej części sympozjum uczestnicy dyskutowali na interesujące ich tematy naukowe i  wymieniali swoje doświadczenia. następnie obrady kontynuowano w  ramach piątej z  kolei sesji plenarnej, dotyczącej „biotechnologii komórki”. tutaj referaty wygłosili: maciej kurpisz z  instytutu genetyki człowieka pan w  poznaniu, polska („komórki macierzyste i  ich zastosowanie w  praktyce klinicznej”) oraz devrim kilinc z  wyższej szkoły w  dublinie, irlandia („połączenie mechaniczne i chemiczne stymulujące do wywołania wzrostu oraz śmierci komórek”). ostatnia, szósta sesja plenarna, miała miejsce po przerwie obiadowej i odnosiła się do „biotechnologii rna”. referującymi w  tej 206 nary sessions. �e sessions were attended by professors from barcelona, cambridge, dresden, dublin, poznan, warsaw and young scientists from various polish research centers. more than 30 young scientists gave oral presentations. students and graduate students from all over poland showed their study results on 79 posters. in total, this years symposium brought together about 200 participants, which proves the great interest in the exchange of knowledge and experience and above all the desire to establish cooperation and the need for integration among students, graduate students and young scientists. sesji byli artur jarmołowski z  uniwersytetu adama mickiewicza w  poznaniu, polska („post-transkrypcyjna koordynacja splatania i  biogenezy mirna u  roślin”) i  andrzej dziembowski z  instytutu biochemii i  bio�zyki pan w  warszawie, polska („ludzki kompleks egzosomu w  zdrowiu i  chorobie”). około godziny 16:00 odbyło się podsumowanie i  o�cjalne zamknięcie tegorocznej edycji międzyuczelnianego sympozjum biotechnologicznego. w ciągu trzech dni referowano i dyskutowano w ramach 6 sesji plenarnych. w obradach udział wzięli profesorowie, m.in. z  barcelony, cambridge, drezna, dublina, poznania, warszawy oraz młodzi naukowcy z  wielu polskich ośrodków badawczych. ponad 30 młodych naukowców wygłosiło prezentacje ustne, a  studenci i  doktoranci z  całej polski pokazali swoje wyniki badań na 79 plakatach. ogółem tegoroczne sympozjum zgromadziło około 200 uczestników, co świadczy o  dużym zainteresowaniu wymianą wiedzy i  doświadczeń, a  przede wszystkim chęci nawiązania współpracy oraz potrzebie integracji wśród studentów, doktorantów i młodych naukowców. katarzyna możdżeń1, anna sołtys-lelek2 1 department of plant physiology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, kasiamozdzen@interia.pl 2 ojców national park, 32-045 sułoszowa, ojców 9 r ep or ts 214 on september 29, 2017, the lesser poland night of scientists was held for the 11th time. this year, the partners of this event included, just like in the previous years, the cracow universities, the polish academy of sciences, as well as the nokia technology centre, comarch s.a., the klaster life science kraków foundation and the jagiellonian centre of innovations, the abb corporate research centre, the cracow specialised hospital of john paul ii, museum of archaeology in cracow, the museum of urban engineering in cracow, the solaris national synchrotron radiation centre, as well as libraries and schools in andrychów, skawina, chrzanów, nowy sącz, niepołomice and tarnów. the media patrons of the night of scientists were, among others, “knowledge and life”, tvp3 kraków, rmf maxxx, miastodzieci.pl, “polish diary”, czasdzieci.pl and the lesser poland network of digital cinemas along with the cinema development foundation. as part of this year meetings with science, the maria curie-sklodowska high school in andrychów organised workshops in chemistry “magical chemistry”, physics “physical riddles”, geology and geography researcher night in małopolska, 29th september 2017, poland małopolska noc naukowców, 29 wrzesień 2017, polska 29 września 2017 roku już po raz 11 odbyła się małopolska noc naukowców. w tym roku partnerami tego wydarzenia były, podobnie jak w  ubiegłych latach, krakowskie uczelnie wyższe, polska akademia nauk, a  także centrum technologiczne nokia, comarch s.a., fundacja klaster life science kraków i  jagiellońskie centrum innowacji, korporacyjne centrum badawcze abb, krakowski szpital specjalistyczny im. jana pawła ii, muzeum archeologiczne w krakowie, muzeum inżynierii miejskiej w  krakowie, narodowe centrum promieniowania synchrotronowego solaris oraz biblioteki i  szkoły w  andrychowie, skawinie, chrzanowie, nowym sączu, niepołomicach i  tarnowie. patronami medialnymi nocy naukowców byli m.in. wiedza i  życie, tvp3 kraków, rmf maxxx, miastodzieci.pl, dziennik polski, czasdzieci.pl oraz małopolska sieć kin cyfrowych wraz z  fundacją rozwoju kina. w ramach tegorocznych spotkań z nauką liceum marii curie-sklodowskiej w andrychowie zorganizowało warsztaty z  chemii „magiczna chemia”, z  fizyki „zagadki fizyczne”, z geologii i geografii „smaki i kolory świata oraz atrakcje turystyczne w  polsce”, mateannales universitatis paedagogicae cracoviensis studia naturae, 2: 214–220, 2017, issn 2543-8832 215 r eports“tastes and colours of the world and tourist attractions in poland”, and mathematics and computer science “programming is not hard”. the demonstrations of first aid and modern technology in sports took place in biology and medicine. the participants also explored the mysteries of the world of plants and animals “the kingdom of plants and animals”. they got acquainted with the theatre “theatre – modernity and tradition,” took part in the workshops titled “theatrical workshops – movement, word, props”, as well as saw a performance of a theatre group c’est la vie entitled “iwona, princess of burgundy”. art lovers explored the mysteries of thinking “what is visual thinking and how can we learn it?” and whether “creativity gives us wings”. in the public library in chrzanów, the visitors learned who maria skłodowska-curie was during the meeting “be like maria”, and took part in the workshops “the laws of physics in nature”. they also explored the secrets of modern information technologies during the lecture “the expanded reality is not only informative, but also entertaining”. they took part in classes on programming bases and learned about educational robots created contemporarily. in the laboratory of the wastewater treatment plant, they learned about microbiological and physicochemical methods of testing drinking water and physicochemical methods of treating wastewater and sediments. during the lecture, “start where you are. use what you have. do what you can!” the listeners became acquainted with ways to analyse the body composition and obtained advice from a nutritionist. participants also learned the secrets of the little-known field of knowledge in poland – matyki i informatyki „programowanie nie jest trudne”. z biologii i medycyny odbyły się pokazy pierwszej pomocy i nowoczesnych technologii w  sporcie. uczestnicy zgłębiali także tajemnice świata roślin i zwierząt „królestwo roślin i zwierząt”. zapoznali się z teatrem „teatr – nowoczesność i  tradycja”, wzięli udział w warsztatach pt. „warsztaty teatralne – ruch, słowo, rekwizyt”, a  także obejrzeli spektakl grupy teatralnej c’est la vie pt. „iwona, księżniczka burgunda”. miłośnicy sztuki zgłębiali tajemnice myślenia „czym jest myślenie wizualne i  jak można się go nauczyć?” i  czy „twórczość dodaje nam skrzydeł”. w  miejskiej bibliotece publicznej w  chrzanowie odwiedzający dowiedzieli się kim była maria skłodowska-curie podczas spotkania „bądź jak maria”, wzięli udział w  warsztatach „prawa fizyki w  przyrodzie”. zgłębiali także tajemnice nowoczesnych technologii informacyjnych podczas wykładu „rzeczywistość rozszerzona to nie tylko wartość informacyjna, ale również rozrywkowa”. uczestniczyli w  zajęciach z  podstaw programowania i  poznali współcześnie konstruowane roboty edukacyjne. w  laboratorium oczyszczalni ścieków dowiedzieli o  mikrobiologicznych i  fizykochemicznych sposobach badania wody pitnej i  fizykochemicznych metodach oznaczania ścieków i  osadów. podczas wykładu „zacznij tam gdzie jesteś. użyj tego, co masz. zrób to, co możesz!” słuchacze zaznajomili się ze sposobami analizy składu ciała i  uzyskali porady dietetyka. uczestnicy poznali również tajemnice mało znanej w polsce dziedziny wiedzy – trychologii. w  czasie warsztatów „badanie pod włos!” analizowali stan skóry głowy i włosów. wykład „rzeczywistość rozszerzona to nie 216 r ep or ts trychology. during the workshops, “examination against the grain!” they analysed the condition of the scalp and hair. the lecture “the expanded reality is not only informative, but also entertaining” aimed at combining the real world in real time, in the form of an image recorded by a camera lens with a digitally generated image. in cracow, the institute of forensic expertise, during the workshops, “murder mystery”, presented the methods of securing fingerprints and biological traces, comparative analyses of writings, and unknown substances, as well as genetic material. the university of agriculture organised chemical demonstrations of volcanoes, elephant pastes, locomotives, non-newtonian liquids, milk rainbows, foam, and giant bubbles. the scientists also explored the secrets of the microbiology of food products, and presented the ichtyofauna of the rivers of southern poland on the example of fish from the vistula river. during the chemistry classes, the researchers presented, among others, the explosive “kmicic sausage”, “smelly cocktail”, and answered the following questions: “what connects odysseus and the mamrot wine?”, “how to destroy a tank with a hot-dog skin and candies?”, “does 1+1 = 2?” and “does kitchen salt contain cyanide?” they also discussed the facts and myths about fruit and vegetable juices. they presented the biology and behaviour of laboratory animals. the personnel of the university of agriculture also organised lectures on “fish glowing in the depths”, “why should you clean after your dog, that is, parasites for the young and not only”, “biotechnology – history, division and colours”, and others. tylko wartość informacyjna, ale również rozrywkowa” miał na celu połączenie w  czasie rzeczywistym prawdziwego świata, w formie obrazu rejestrowanego przez obiektyw aparatu, z generowanym cyfrowo obrazem. w  krakowie, instytut ekspertyz sądowych podczas warsztatów „mordercza zagadka” zaprezentował metody zabezpieczania śladów daktyloskopijnych i  biologicznych, analizy porównawcze pisma i  nieznanych substancji oraz materiału genetycznego. uniwersytet rolniczy zorganizował pokazy chemiczne wulkanów, pasty dla słonia, lokomotywy, cieczy nienewtonowskiej, mlecznej tęczy, piany i  gigabańki. naukowcy zgłębiali także tajemnice mikroświata produktów spożywczych, zaprezentowali ichtiofaunę rzek południowej polski na przykładzie ryb z  wisły. podczas zajęć z  chemii, naukowcy zaprezentowali m.in. wybuchową „kiełbasę kmicica”, „cuchnący koktajl” oraz odpowiadali na pytania: „co łączy odyseusza i wino mamrot?”, „jak zniszczyć czołg za pomocą skórki z  parówki i  cukierków?”, „czy 1+1 = 2?”, „czy w soli kuchennej jest cyjanek?” omówili także fakty i mity o sokach owocowych i  warzywnych. przedstawiali biologię i  behawioryzm zwierząt laboratoryjnych. pracownicy uniwersytetu rolniczego zorganizowali również wykłady „ryby świecące w  głębinach”, „dlaczego należy sprzątać po swoim psie, czyli o  pasożytach dla najmłodszych i  nie tylko”, „biotechnologia – historia i podział i kolory” i inne. naukowcy z  politechniki krakowskiej zaprezentowali zagadnienia dotyczące produkcji miodu „dary z  pasieki – superfoods”. ujawnili tajemnice nanotechnologii „sekrety współczesnej nanotechnologii – wizja rozwoju 217 r eports scientists from the cracow university of technology presented the issue “superfoods – gifts from the apiary”. they revealed the secrets of nanotechnology “secrets of modern nanotechnology – the vision of development and awareness of threats”, sought answers to the question “if you are born to swim you will never drown – from the brazilian nut to the ‘boiling’ sand” and organised pyrotechnic shows. the jagiellonian university prepared popular science lectures “from an idea to the pharmacy, that is, new drugs for photodynamic therapy”, “chemist on the trail of crime”, “skin colours under the magnifying glass”, “molecules at your reach, that is, how computers help examine the micro world”, “molecular lego blocks, or serious scientists fun”. university employees invited people to visit i  świadomość zagrożeń”, poszukiwali odpowiedzi na pytanie „co ma pływać nie utonie – od brazylijskiego orzecha po ‘wrzący’ piasek” oraz zorganizowali pokazy pirotechniczne. uniwersytet jagielloński przygotował wykłady popularno-naukowe „od pomysłu do apteki, czyli o nowych lekach do terapii fotodynamicznej”, „chemik na tropie zbrodni”, „kolory skóry pod lupą”, „molekuły na wyciągnięcie ręki, czyli jak komputery pomagają badań mikroświat”, „molekularne klocki lego, czyli poważne zabawy naukowców”. pracownicy uniwersytetu zapraszali do zwiedzania pracowni laboratoryjnych: mikroskopii sił atomowych, mikroskopii konfokalnej, chemii środowiska, spektroskopii ramanowskiej, chemii sądowej. podczas warsztatów „magiczny świat małego biochemika” uczestnicy brali czynny udział w  przygotowaniu coca fig. 1. participants of the researcher night in małopolska during the workshops at the institute of biology of the pedagogical university (photo. w. wojtaś) ryc. 1. uczestnicy małopolskiej nocy naukowców podczas warsztatów w instytucie biologii uniwersytetu pedagogicznego (fot. w. wojtaś) 218 r ep or ts the laboratories of atomic power microscopy, confocal microscopy, environmental chemistry, raman spectroscopy, and judicial chemistry. during the “magical world of a small bio-chemist” workshops, participants took active part in preparing coca cola of tea, a volcanic lamp, or elephant foam. the cracow specialised hospital of john paul ii organised, among others lectures “psyche and soma” and the “cardiological prevention”. the visitors from cracow took part in stress management workshops, in laboratory experiments in labelling genetic mutations, and also learned methods of cellular cultures. the pedagogical university offered numerous lectures, demonstrations, and workshops in the field of biological sciences. during this year’s night of scientists, the visitors coli z  herbaty, lampy wulkanicznej czy słoniowej piany. krakowski szpital specjalistyczny im. jana pawła ii zorganizował m.in. wykłady „psyche i soma” oraz „prewencja kardiologiczna”. odwiedzający krakowianie wzięli udział w  warsztatach panowania nad stresem, w  eksperymentach laboratoryjnych z  oznaczania genetycznych mutacji, a także poznali metody hodowli komórkowych. uniwersytet pedagogiczny w  dziedzinie nauk biologicznych zaoferował liczne wykłady, pokazy i  warsztaty. podczas tegorocznej nocy naukowców odwiedzający poznali m.in. przystosowania owadów i roztoczy do życia w środowisku spichrzów i magazynów spożywczych. przybyli goście poznali różne aspekty szkodliwości owadów i roztoczy i jak wykrywać szkodniki w produktach spożywfig. 2. high school students participating in the lecture and workshops “fruit fly as a model organism in genetic research” (photo. w. wojtaś) ryc. 2. licealiści uczestniczący w wykładzie i warsztatach „muszka owocowa jako organizm modelowy w badaniach genetycznych” (fot. w. wojtaś) 219 r eportslearned, among others, the adaptation of insects and mites to life in the environment of granaries and food stores. the visitors learned about various harmful aspects of insects and mites, and how to detect pests in food products. the presentations “ticks – dangerous mites” and “attention! tick season! what each citizen of cracow should know about ticks” brought the problem of parasitic diseases closer to the listeners, especially in relation to tick-borne diseases (fig. 1). in addition, they provided practical information on the principles of prevention and protection against dangerous parasites. the shows of lipstick and cream formulation enjoyed the biggest interest, which included “how to do it” sessions organised within workshops, including “colloidal systems in cosmetics – obtaining creams, lipsticks, lip gloss”. participants also took part in the classes about “light as an inspiration for plants”, in experiments on surface tension, the detection of organic compounds in milk, juices, and various food products. during the workshops, “mercury in food products” the scientists explored the secrets of mercury damage. the participants could learn about genetic modifications during lectures and workshops “fruit fly as a model organism in genetic research” (fig. 2). the participants learned about the secrets of the biological laboratory during the meeting “environmental detective on the track”. it is impossible to list here all organisers and present the presentations, workshops, and lectures prepared by them, which took part in different parts of the city and beyond its borders. over 1200 scientists, 2000 students, and almost 250 scientific groups from 44 various institutions were engaged in this czych. prezentacje „kleszcze – niebezpieczne roztocza” i „uwaga! sezon na kleszcze! czyli co każdy krakowianin o  kleszczach wiedzieć powinien” przybliżyły słuchaczom problem chorób pasożytniczych a  zwłaszcza chorób odkleszczowych (ryc. 1). ponadto dostarczyły one praktycznych informacji o  zasadach profilaktyki i zabezpieczenia przed groźnymi pasożytami. największym zainteresowaniem cieszyły się pokazy otrzymywania szminek i kremów – jak to jest zrobione zorganizowane w ramach warsztatów „układy koloidalne w  kosmetyce – otrzymywanie kremów, szminek, błyszczyków”. uczestnicy dodatkowo wzięli udział w zajęciach o tematyce „światło inspiracją dla roślin”, w doświadczeniach dotyczących napięcia powierzchniowego, wykrywania związków organicznych w  mleku, sokach i  różnych produktach spożywczych. podczas warsztatów „rtęć w  produktach spożywczych” naukowcy zgłębiali tajemnice szkodliwości rtęci. o  modyfikacjach genetycznych uczestnicy mogli dowiedzieć się podczas prelekcji i  warsztatów „muszka owocowa jako organizm modelowy w  badaniach genetycznych” (ryc. 2). podczas spotkania „detektyw środowiskowy na tropie” słuchacze poznali tajemnice laboratorium biologicznego. nie sposób wymienić tutaj wszystkich organizatorów i  zaprezentować przygotowane przez nich pokazy, warsztaty i  prelekcje odbywające się w  różnych częściach miasta i  poza jego granicami. w  tegoroczną noc naukowców zaangażowało się ponad 1200 naukowców, 2000 studentów oraz prawie 250 kół naukowych z  44 różnych instytucji na czele z  urzędem marszałkowskim województwa małopolskiego. 220 r ep or ts anna kocoń1, katarzyna możdżeń2 1department of invertebrate zoology and parasitology, institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland, anna.kocon@up.krakow.pl 2department of plant physiology, institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland year’s night of scientists, led by the marshals office of the lesser poland voivodeship. year after year, the growing number of participants confirms the increasing interest in the lesser poland night of scientists. this type of event allows us not only to become familiar with science from behind the scene, but primarily provide direct contact with the scientists. we hope to meet again in a year to uncover new secrets of the world of science, admire exotic plants and animals, cars of the future, or dancing and talking robots. the number of people interested in these meetings shows that science fascinates both young and old. z roku na rok wzrastająca liczba uczestników potwierdza coraz większe zainteresowanie małopolską nocą naukowców. tego typu wydarzenie pozwala nie tylko na poznawanie nauki od kuchni, ale przede wszystkim na bezpośredni kontakt z  naukowcami. mamy nadzieję, że za rok znowu się spotkamy, aby odkrywać nowe tajemnice świata nauki, podziwiać egzotyczne rośliny i  zwierzęta, samochody przyszłości, czy tańczące i  mówiące roboty. po liczbie zainteresowanych udziałem w  tych spotkaniach jest pewne, że nauka fascynuje nie tylko młodych, ale i tych starszych. 190 annales universitatis paedagogicae cracoviensis studia naturae, 1: 190–195, 2016, issn 2543-8832 magdalena greczek-stachura*, patrycja zagata-leśnicka, mateusz ślęczka institute of biology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, *magresta@wp.pl analysis of the coliform in the wilga river (southern poland) introduction �e progressive development of industry, as well as the cities suburbanisation cause inevitable damages to the environment. �ese damages are related to water and soil pollution. only 2% of surface waters in poland belong to the �rst class purity. �e rest of water bodies is named as waters of poor quality. �e water pollution is caused usually by industrial wastewater and municipal sewage, which are directly discharged into rivers. furthermore, pre-treated wastewater in industrial facilities is also the factor leading to the environmental pollution (rozporządzenie ministra środowiska…, 2004). �e pollutions of drinking water are caused by the occurrence of leaky septic tanks or by direct sewage discharging into watercourses and the surface areas. �e sewages seep through the ground and a�er that they are present in groundwater. �e groundwater should be pure, however, results of the monitoring carried out in the nineties of the twentieth century revealed, that water bodies are not included in the �rst class purity. �e microbiological contamination of the surface waters can cause food poisoning of water consumers. moreover, it is responsible for �sh diseases, which in result are not suitable for consumption. �us, the monitoring of drinking water intakes, as well as smaller watercourses is indispensable (george et al., 2002; pant, mittal, 2007; hendricks, pool, 2012; budzińska et al., 2014). �e wilga river is the right tributary of the vistula river �owing through kraków with the length of 11.5 km. �e total length of the river is 21.4 km and its spring is located in the north-eastern part of the wieliczka foothills in pawlikowice (list of names…). �e middle course of the river is a part which strongly meanders and the �nal stretch has been dug up and straightened. in the part from swoszowice to kraków, wilga �ows within the built-up area. �e catchment of the river is located in the rural area. �e banks in the upper and middle courses are covered with alluvial 191 a nalysis of the coliform in the w ilga r iver (southern p oland) forests. �ese forests give way to grassy �ora in kraków. �e wilga river is relatively short comparing to other rivers of wieliczka foothills, but it is characterised by trough characteristic to mountain rivers with swi� stream and rocky riverbed and by deep trough which occurs in lowland rivers with slow stream. wilga belongs to the most polluted rivers in kraków (ocena jakości wód…, 2004). �e contaminations were mainly due to the occurrence of former industrial facilities in the ix district – łagiewniki (soda solvay factory). �e microbiological tests of water coliforms (the smallest volume of water with detected escherichia coli t. escherich) in di�erent locations along wilga. �is test is a basic method to indicate the presence of sewage in watercourses based on the appearance of a bubble in durham tube. �e bubble is the e�ect of the fermentation of lactose by e. coli. �e samples were collected from 16 locations along the wilga river from the spring in raciborsko up to podgórze in kraków (fig. 1). in places of sampling we measured water parameters like temperature and ph. �e samples were initially diluted tenfold with physiological saline and a�er that the 1 cm3 of water was transferred to the test tube (with durham tubes inside) with broth enriched with 1% lactose. experiments were conducted for samples with di�erent water dilutions (1:10, 1:100, 1:1000, 1:10 000, 1:100 000). a�er 24 hours of incubafig. 1. map of sampling locations along the wilga river 192 m ag da le na g re cz ek -s ta ch ur a, p at ry cj a za ga ta -l eś ni ck a, m at eu sz ś lę cz ka tion at the temperature of 37°c the occurrence of broth turbidity and the presence of bubble in durham tubes were observed. experiments were conducted during the hot summer of 2014. air temperatures were above 30°c and water temperatures ranged from 15 to 21°c. �e temperature of upper river course reached 19°c. �e low temperature of water is usually due to the swi� stream in upper course. in the middle course, with slower stream, the temperatures reached 20°c. �e highest temperature of water we measured in the mouth of the wilga river was 21°c which was correlated to slow stream high exposure to solar radiation as well as vistula backwaters. we detected the purest water in the middle course of the wilga river (in janowice and sygneczów) (fig. 2). �ere are no farm buildings near the river trough and river �ows fast. �e coliform was 1 which indicates that the water is not suitable to drink according to whipple scale. even at the spring of the wilga river there was no microbiologically pure water. �e value of coliform reached 0.1 and the contamination is caused by the presence of public toilet for inhabitants of the neighbouring plot near the river. �e coliform of 0.1 was measured in 6 samples collected from: pawlikowice, raciborsko (ponds), wrząsowice (near the bar “wilga”), swoszowice (near the boundaries of kraków), opatkowice and łagiewniki (near the sanctuary of divine mercy). �ere are courses of swi� stream in each of mentioned locations. �erefore, these courses should be well fig. 2. �e values of coliform of water collected from di�erent locations: 1 – river spring, 2 – raciborsko pounds, 3 – raciborsko 2, 4 – pawlikowice, 5 – janowice, 6 – sygneczów, 7 – gołkowice, 8 – wilga bar, 9 – swoszowice g.m., 10 – opatkowice, 11 – swoszowice, 12 – sanctuary of divine mercy, 13 – zakopiańska street, 14 – kopty forest, 15 – mateczny roundabout, 16 – river mouth 193 a nalysis of the coliform in the w ilga r iver (southern p oland) oxygenated and relatively cool (even a few degrees colder) (fig. 3). �ese locations are characterised by scattered buildings, the blanks are covered with meadows or even trees. �ere are neither farms near the river nor sewage discharge into the river. �us, the river reveals better microbiological parameters. �e wilga river tends to self-clean. it can be observed at the stretch: janowice – sygneczów – gołkowice as well as at the stretch: swoszowice – łagiewniki. we can suppose that if there were neither direct nor indirect discharges of the sewage into water, the river would reveal a signi�cant tendency to self-clean. �e rivers self-cleaning process is long-term, therefore, if there is a large amount of sewage discharge, the process is inhibited and hard to observe. �e wilga river is arti�cially regulated to avoid �ood of the river at the stretch from kraków up to the river mouth. �e most polluted part of the wilga river is located near to the mateczny roundabout – coliform of 1:10 000. �e second most polluted area was located at the mouth of the river: 1:100. �ese two stretches are characterised by slow stream and high temperature of water reaching 21°c. �ere is a sewer pipe near to the health resort with residue of the sewage sludge deposited on the grid protecting the river banks against lateral erosion. we also observed a lot of water birds – mainly ducks looking for food. �e ph of water samples collected from the wilga river is alkaline and it oscillates within 8.0–8.2, which is correlated with the type of substrate that river �ows through. fig. 3. �e temperature of water [°c] measured in 16 locations of sampling along the wilga river: 1 – river spring, 2 – raciborsko pounds, 3 – raciborsko 2, 4 – pawlikowice, 5 – janowice, 6 – sygneczów, 7 – gołkowice, 8 – wilga bar, 9 – swoszowice g.m., 10 – opatkowice, 11 – swoszowice, 12 – sanctuary of divine mercy, 13 – zakopiańska street, 14 – kopty forest, 15 – mateczny roundabout, 16 – river mouth 194 m ag da le na g re cz ek -s ta ch ur a, p at ry cj a za ga ta -l eś ni ck a, m at eu sz ś lę cz ka �e wilga river valley is cut in miocene schists and covered with quaternary sands (łojan, 2008). up to the mouth of the krzywica stream in swoszowice, the wilga river catchment is covered with loess soil and the tertiary �ysch layers beneath them consisting of shale, marl, sandstone and chalk layer. �e ph of water has an in�uence on water microorganism variety. �ese organisms require the water of neutral ph, but they also have an ability to adapt to the new environmental conditions, when water changes ph into weakly acid or alkaline (pawlaczyk-szpilowa, 1978). rapid ph changes of surface waters are mainly due to discharge of some industrial sewage without their pre-neutralisation. �e main cause of high values of coliform is discharge of large amount of sewage into the river, as well as the fertilisation of farmlands with manure which get into the soil and are washed down by rain into the river. �e surface waters contaminations by pathogenic bacteria, originating from both the sewage discharge and the sewage in�ltration into the ground waters are serious problems. �e purity of surface waters should be considered as poor. �ere is only 2% of rivers and lakes waters that ful�l the requirements for the �rst class purity (libudzisz, kowal, 2000). microbiological quality of waters a�ects the quality of raw materials and the food products with aquatic originating. waters that are polluted by bacteria belonging to the coli group are neither suitable to drink nor even to recreational purposes use. even minor contaminations of water by coli bacteria poses a risk of causing illnesses or even epidemics among people (langergraber, muellegger, 2005). references budzińska, k., szejniuk, b., jurek, a., traczykowski, a., michalska, m., berleć, k. (2014). e�ectiveness of removing microbial pollutants from wastewater by the activated sludge method. environment protection engineering, 40, 53–67. doi: 10.5277/epe140405 george, i., crop, p., servai, p. (2002). faecal removal in wastewater treatment plants studied by plate counts and enzymatic methods. water research, 36, 2601–2617. doi: 10.1016/s0043-1354(01)00475-4 hendricks, r., pool, e.j. (2012). �e e�ectiveness of sewage treatment processes to remove faecal pathogens and antibiotic residues. journal of environmental science and health, 47, 289–297. doi: 10.1080/10934529.2012.637432 langergraber, g., muellegger, e. (2005). ecological sanitation – a way to solve global sanitation problems?, evironmental international, 31, 433–444. doi: 10,1016/j.envint.2004.08.006 libudzisz, z., kowal, k. (2000). mikrobiologia techniczna. tom 1. łódź: wydawnictwo politechniki łódzkiej. [in polish] list of names of �owing waters. nazewnictwo geogra�czne polski. tom 1, hydronimy. część 1. wody płynące, źródła, wodospady, główny urząd geodezji i kartogra�i. http://ksng.gugik.gov.pl/pliki/ hydronimy1.pdf. [in polish] łojan, e. (2000). wpływ składników mineralnych na geochemię metali ciężkich w osadach dennych rzeki wilgi. kraków: akademia górniczo-hutnicza im. stanisława staszica. [in polish] pant, a., mittal, a.k. (2007). disinfection of wastewater: comparative evaluation of chlorination and dhs-biotower. journal of environmental biology, 28(4), 717–722. 195 a nalysis of the coliform in the w ilga r iver (southern p oland) pawlaczyk-szpilowa, m. (1978). mikrobiologia wody i ścieków. warszawa: wydawnictwo naukowe pwn. [in polish] rozporządzenie ministra środowiska z dnia 11 lutego 2004 r. w sprawie klasy�kacji dla prezentowania stanu wód powierzchniowych i podziemnych, sposobu prowadzenia monitoringu oraz sposobu interpretacji wyników i prezentacji stanu tych wód. dz. u. z 2004 r. nr 32, poz. 284. [in polish] wojewódzki inspektorat ochrony środowiska w krakowie. (2004) ocena jakości wód powierzchniowych w województwie małopolskim w 2004 roku. [in polish] analiza form coli w rzece wildze (południowa polska) streszczenie rzeka wilga należy do najbardziej zanieczyszczonych rzek w krakowie. zanieczyszczenie tej rzeki wynika głównie z obecności obiektów przemysłowych w jej pobliżu. analizy miana coli w próbkach wody pobranych w 16 miejscach (począwszy od źródła rzeki w raciborsku, aż do podgórza w krakowie) wzdłuż rzeki przeprowadzone zostały w 2014 roku. analiza ta jest podstawową metodą służącą do wykazania obecności ścieków w ciekach wodnych i jest oparta na pojawieniu się pęcherzyka powietrza w rurce durhama, co z kolei jest efektem fermentacji laktozy prowadzonej przez bakterie escherichia coli. wodę o największej czystości wykazano w środkowym biegu rzeki, gdzie zaobserwowano szybki nurt oraz brak zabudowań gospodarczych w pobliżu rzeki. wodę złej jakości stwierdzono nawet przy źródle rzeki, gdzie miano coli wynosiło 0,1. wilga wykazuje tendencję do samooczyszczania się, co można było zaobserwować na odcinkach: janowice – sygneczów – gołkowice oraz swoszowice – łagiewniki. rzeka ta wykazałaby sporą tendencję do samooczyszczania się w przypadku braku ścieków odprowadzanych w sposób pośredni i bezpośredni. samooczyszczanie się rzek jest procesem długotrwałym, dlatego przy znacznym dopływie ścieków proces ten jest hamowany i przez to trudny do zaobserwowania. key words: coliform, escherichia coli, water contaminations, wilga river received: [2016.09.02] accepted: [2016.10.25] 208 the xli national scientific conference on “zoning of arable weeds in poland” was held on 3rd and 4th of july 2017 in kraków (poland). the main organisers were scientists from department of agrotechnology and agricultural ecology at faculty of agriculture and economics (university of agriculture in kraków). the president of the organising committee (oc) was prof. teresa dąbkowska, the member of the oc was prof. teofil łabza, and the secretary of the oc was assoc. prof. joanna puła. the following scientists from polish universities and research centres were the part of a science committee: assoc. prof. julian chmiel (adam mickiewicz university in poznań), prof. teresa dąbkowska (university of agriculture in kraków), prof. krzysztof domaradzki (institute of soil science and plant cultivation-state research institute in puławy), prof. czesław hołdyński (university of warmia and mazury in olsztyn), prof. bogdan jackowiak (adam mickiewicz university in poznań), assoc. prof. zygmunt kącki (university of wrocław), prof. andrzej lepiarczyk (university of agriculture in kraków) – fig. 1, prof. teofil łabza xli national scientific conference on “zoning of arable weeds in poland”, kraków, 3rd–4th july 2017 xli krajowa konferencja naukowa z cyklu „rejonizacja chwastów segetalnych w polsce”, kraków, 3–4 lipiec 2017 w  krakowie w  dniach 3–4 lipca 2017 roku odbyła się już xli krajowa konferencja naukowa z  cyklu „rejonizacja chwastów segetalnych w  polsce”. głównymi organizatorami konferencji byli pracownicy katedry agrotechniki i ekologii rolniczej wydziału rolniczo-ekonomicznego uniwersytetu rolniczego im. hugona kołłątaja w  krakowie. przewodniczącą komitetu organizacyjnego była prof. dr hab. teresa dąbkowska, członkiem prof. dr hab. teofil łabza, a  sekretarzem dr hab. joanna puła. w  składzie komitetu naukowego byli przedstawiciele uczelni i jednostek naukowych z całej polski: dr hab. julian chmiel (uniwersytet im. adama mickiewicza w  poznaniu), prof. dr hab. teresa dąbkowska (uniwersytet rolniczy w krakowie), prof. dr hab. krzysztof domaradzki (instytut uprawy nawożenia i  gleboznawstwa – pib w  puławach), prof. dr hab. czesław hołdyński (uniwersytet warmińsko-mazurski w  olsztynie), prof. dr hab. bogdan jackowiak (uniwersytet im. adama mickiewicza w  poznaniu), dr hab. zygmunt kącki (uniwersytet wrocławski), prof. dr hab. andrzej lepiarczyk (uniwersytet rolniczy w  kraannales universitatis paedagogicae cracoviensis studia naturae, 2: 208–213, 2017, issn 2543-8832 209 r eports (university of agriculture in kraków), prof. janina skrzyczyńska (siedlce university of natural sciences humanities), assoc. prof. zbigniew sobisz (pomeranian academy in słupsk), and prof. czesława trąba (university of rzeszów). over 50 participants from 18 research centres have made their way to kraków to experience this conference first-hand. during the sessions of the conference, 6 plenary papers were presented, and 21 short scientific reports were delivered. the speeches were divided into 5 thematic sessions, as follows: i – communities of arable weeds and their biodiversity, ii – disappear and invasive alien species  – dynamic of weed-grown, iii – the biology and harmfulness of weeds, iv – allelopathy and non-chemical methods for the kowie) – ryc. 1, prof. dr hab. teofil łabza (uniwersytet rolniczy w krakowie), prof. dr hab. janina skrzyczyńska (uniwersytet przyrodniczo-humanistyczny w  siedlcach), dr hab. zbigniew sobisz (akademia pomorska w słupsku) oraz prof. dr hab. czesława trąba (uniwersytet rzeszowski). na konferencję zgłosiło się ponad 50 osób z 18 ośrodków naukowych z polski. w czasie obrad wygłoszono 6 referatów plenarnych oraz przedstawiono 21 krótkich doniesień. tematykę obrad podzielono na 5 sekcji tematycznych: i – zbiorowiska segetalne i ich bioróżnorodność, ii – gatunki zanikające i  inwazyjne oraz dynamika zachwaszczenia, iii – biologia i szkodliwość chwastów, iv – allelopatia i niechemiczne sposoby ograniczania wzrostu i  rozwoju chwastów, v – badania fig. 1. prof. andrzej lepiarczyk during the opening of the conference (photo. k. domaradzki) ryc. 1. wystąpienie prof. andrzeja lepiarczyka w trakcie otwarcia konferencji (fot. k. domaradzki) 210 r ep or ts limiting growth and development of weeds, v – research on arable weed infestation as a source of information on the diversity of flora and segetal plant communities in poland. during the forum and discussions, participants of the conference were able to share their observations concerning the current state of knowledge about the vegetation growing on agricultural land in various agricultural regions of poland. as every year, research on monocotyledonous weeds in crops, as well as in orchards and gardens, was an issue of concern for herbologists. much attention has been devoted to invasive alien species, which are increasingly a threat to native vegetation. plants that dominated not only the topics presented during the conference, but also the short reports, were species of the genus reynoutria sp. it turns out that they have already dominated many local biocenes. in poland, the presence of all three most expansive species of this type has been recorded: reynoutria japonica houtt, r. sachalinensis (f. schmidt) nakai and r. ×bohemica chrtek et chrtkova. in speeches, detailed morphology of these plants and the methods of limiting their spread by various methods of mechanical weed control combined with herbicides were presented. other plants, as invasive as reynoutria species, which occupy ever larger areas of wasteland in our country, are solidago canadiensis l. and s. gigantea aiton. a lot of attention during the conference speeches was also dedicated to them. these species reproduce with high efficiency, both vegetatively and generatively, which makes it easier for them to get control over new areas, especially those that are excluded from agricultural use. nad zachwaszczeniem jako źródło informacji o różnorodności flory i roślinności segetalnej w polsce. w  trakcie obrad oraz dyskusji uczestnicy konferencji mieli możliwość dzielenia się swoimi spostrzeżeniami, dotyczącymi obecnego stanu wiedzy na temat roślinności porastającej użytki rolne w  różnych rejonach rolniczych polski. jak co roku, w kręgu zainteresowania herbologów znalazły się badania nad diasporami chwastów jednoliściennych w uprawach, jak również w sadach i ogrodach. dużo uwagi poświęcono gatunkom inwazyjnym, będącym coraz częściej zagrożeniem dla rodzinnej bioróżnorodności. roślinami, które zdominowały nie tylko tematy prezentowanych w  czasie konferencji referatów, ale i doniesień były gatunki z rodzaju reynoutria sp. okazuje się, że opanowały one już wiele lokalnych biocenoz. na terenie polski odnotowano występowanie wszystkich trzech najbardziej ekspansywnych gatunków z tego rodzaju: reynoutria japonica houtt, r. sachalinensis (f. schmidt) nakai oraz r. ×bohemica chrtek et chrtkova. w wystąpieniach przedstawiano szczegółową morfologię tych roślin oraz sposoby ograniczania ich rozprzestrzeniania, poprzez różne zabiegi mechaniczne połączone z  opryskami herbicydowymi. innymi, równie inwazyjnymi jak rdestowce gatunkami, które zajmują coraz to większe powierzchnie nieużytków w naszym kraju, są nawłocie: solidago canadiensis l. i s. gigantea aiton. im również poświęcono wiele uwagi w  wystąpieniach konferencyjnych. gatunki te z  dużą wydajnością rozmnażają się, zarówno wegetatywnie, jak i  generatywnie, co ułatwia im opanowywanie nowych terenów, zwłaszcza wyłączonych z  użytkowania rol211 r eportsin the contents of the papers, attention was paid to the phenotypic plasticity of invasive solidago species and the possibility of limiting their occurrence through the use of various methods of liquidation of fallows, which have become notoriously a mainstay to these species. participants in the conference undertook a wide-ranging discussion within the framework of section v – “research on arable weed infestation as a source of information on the diversity of flora and segetal plant communities in poland”. the discussion concerned databases for relevés, their documentation, digitisation, and access to a wider range of scientists from across europe and the world. here, the characteristics of polish vegetation database (pvd), the european vegetative archive (eva), and the global database of vegetation-plot databases (givd) were presented. the need for digitisation of very rich sets of relevés, which are stored in the form of typography, has been pointed out. this is extremely valuable information, especially for the assessment of changes in plant communities in poland, which are often the scientific achievements of many research teams – botanists and herbologists. according to assoc. prof. zygmunt kącki from the university of wrocław, these resources constitute a considerable part of the accumulated phytosociological material on a european scale. a discussion was also held on the weed infestation of agricultural, horticultural, and vegetable crops. the focus was mainly on non-chemical methods of the elimination of undesirable species, which can be used in crops as alternative methods to reduce weed niczego. w  treści referatów zwrócono uwagę na plastyczność fenotypową inwazyjnych gatunków nawłoci oraz możliwości ograniczenia ich występowania poprzez stosowanie różnych sposobów likwidacji odłogów, które stały się ostoją dla tych gatunków. uczestnicy konferencji podjęli szeroko zakrojoną dyskusję w  ramach tematów v sekcji – „badania nad zachwaszczeniem jako źródło informacji o różnorodności flory i roślinności segetalnej w  polsce”. dyskusja ta dotyczyła baz danych dla zdjęć fitosocjologicznych, ich dokumentowania, digitalizacji i  udostępniania szerszemu gronu naukowców z całej europy i ze świata. przedstawiono tu, min. charakterystykę baz polskich (polish vegetation database-polska baza danych o roślinności, krajowe centrum roślinnych zasobów genowych), europejskich (european vegetative archive-eva) oraz globalnych komputerowych baz danych zbiorowisk roślinnych, występujących na polach uprawnych (global index of vegetation-plot databases-givd). wskazano na potrzebę zapisu elektronicznego bardzo bogatych zbiorów zdjęć fitosocjologicznych, które są przechowywane w  postaci maszynopisów. są to niezwykle cenne informacje, zwłaszcza dla oceny zmian w  zbiorowiskach roślinnych polski, stanowiące często wieloletni dorobek naukowy wielu zespołów badaczy – botaników i herbologów. w opinii dr hab. zygmunta kąckiego z  uniwersytetu wrocławskiego w  skali europejskiej zasoby te stanowią pokaźną część zgromadzonego materiału fitosocjologicznego. w  czasie obrad podjęto także dyskusję związaną z  zachwaszczeniem upraw rolniczych, sadowniczych i  warzywniczych. sku212 r ep or ts infestation. the example was given of the introduction of living mulch, among others, with festuca ovina agg. l. there were also examples of the potential use of products of plant origin in plant protection, which seems to be the right direction of research, not only in reducing weed infestation but also in the protection of the environment. attention has also been paid to the use of allelopathy in reducing weed infestation as one of non-chemical methods. the second day of the conference continued with a joint field session, during which all participants had a chance to acquaint themselves with the flora of segetal weed communities on calcareous soil, and generally with the flora of the cracow-czestochowa upland, especially the buffer zone of the ojcowski national park (fig. 2). the rare weeds infesting crop fields, such as piono się głównie nad metodami niechemicznymi eliminacji gatunków niepożądanych, które mogą być wykorzystane w  uprawach jako alternatywne metody do ograniczenia zachwaszczenia. jako przykład podano wprowadzanie żywej ściółki, m.in. z  festuca ovina agg. l. przedstawiono także przykłady wykorzystania w produkcji środków ochrony roślin preparatów pochodzenia roślinnego, co wydaje się właściwym kierunkiem badań, nie tylko w przypadku ograniczenia zachwaszczenia, ale także ochrony środowiska naturalnego. zwrócono również uwagę na wykorzystanie zjawiska allelopatii w ograniczeniu zachwaszczenia pól uprawnych, jako jednej z  metod niechemicznych. w  drugim dniu konferencji uczestnicy wzięli udział w sesji terenowej, podczas której mieli okazję zapoznać się z  florą zbiorowisk chwastów segetalnych gleb wapiennych fig. 2. participants of the conference during the field session (photo. k. domaradzki) ryc. 2. uczestnicy konferencji w trakcie sesji terenowej (fot. k. domaradzki) 213 r eportsscarlet pimpernel (anagallis arvensis l.), blue pimpernel (a. foemina mill.), yellow ball-mustard (neslia paniculata l), field larkspur (consolida regalis gray), blue fieldmadder (sherardia arvensis l.), and others were found in places with the segetal flora near miechów (niedźwiedź village) and in sułoszowa. during the session, scientists also visited a farm in which strawberries are grown (sułoszowa, poland), which is located in the buffer zone and the ecological corridor of the ojcowski national park. the farm specialises in both the production and sale of fruit and seedlings of various varieties of strawberries. the field session and the whole conference ended with a joint dinner at the “herbowa” restaurant in pieskowa skała castle. summaries of the presentations and posters presented during the conference were published in a special issue (book of abstracts) published by the university of agriculture in kraków. xli national scientific conference on “zoning of arable weeds in poland, kraków 3–4 july 2017” – pp. 115, isbn 978-83-64758-58-4, edited by t. dąbkowska and j. puła. oraz ogólnie z florą wyżyny krakowsko-częstochowskiej, zwłaszcza ojcowskiego parku narodowego (ryc. 2). na stanowiskach z florą segetalną w okolicach miechowa (wieś niedźwiedź) oraz w  sułoszowej odszukano, takie rzadkie chwasty segetalne, jak: kurzyślad polny (anagallis arvensis l.), k. błękitny (a. foemina mill.), ożędka groniasta (neslia paniculata l.), ostróżka polna (consolida regalis gray), rolnica pospolita (sherardia arvensis l.) i  inne. w  trakcie sesji odwiedzono również gospodarstwo rolne i szkółkę truskawek w sułoszowej, położone w otulinie oraz korytarzu ekologicznym ojcowskiego parku narodowego. gospodarstwo to specjalizuje się, w uprawie i produkcji sadzonek różnych odmian truskawek. sesja terenowa i  cała konferencja zakończyła się wspólnym, uroczystym obiadem w  restauracji „herbowa” na zamku w pieskowej skale. streszczenia wystąpień i  posterów prezentowanych w  trakcie konferencji zostały opublikowane w  specjalnie wydanym przez wydawnictwo uniwersytetu rolniczego w  krakowie opracowaniu pt. „xli krajowa konferencja naukowa z  cyklu rejonizacja chwastów segetalnych w  polsce, kraków 3–4 lipca 2017” – ss. 115, isbn 978-83-64758-584, pod redakcją t. dąbkowskiej oraz j. puły. joanna puła, angelika kliszcz department of agrotechnology and agricultural ecology, university of agriculture in kraków, mickiewicz ave. 21, 31-120 kraków, poland, rrpula@cyf-kr.edu.pl 27 annales universitatis paedagogicae cracoviensis studia naturae, 2: 27–35, 2017, issn 2543-8832 doi: 10.24917/25438832.2.2 klaudia świacka, alicja michnowska*, jakub maculewicz, izabela przednowek, iga ogrodowczyk, sebastian kozic institute of oceanography, university of gdańsk, gdynia, poland; *alicja.michnowska@ug.edu.pl composition of phytoplankton in the puck bay and the open baltic sea introduction �e biocenosis of the baltic sea is an unusual combination of freshwater and ocean �ora and fauna. �e number of �ora and fauna adapted to life in the brackish water is small, but there may be large quantities of individual species. compared to the oceans, the food chains in the baltic sea are simple. �e number of species reduces from the southern baltic sea to the north. �e low salinity of the northern baltic sea, the cold winters, and the sea freezing over set challenges for the adaptation of organisms. due to the slow water turnover, environmental toxins and eutrophying nutrients remain in the baltic sea and cause long-term e�ects (olli et al., 2011). �e puck bay (54°40′00″n; 18°35′00″e) is an isolated part of gulf of gdańsk, separated by hel peninsula. �e bay has frequently been the subject of marine biological and hydrological research. �is area has speci�c hydrological conditions. due to the fact that the most of the puck bay is shallow, a greater e�ect of wind on the water dynamics is observed in this area. �us anemobaric conditions in particular contribute to the movement of water masses. in the puck bay, a greater correlation between air and surface water temperatures is also observed, where rapid changes of surface water are triggered by changes in air temperature. �erefore, the composition and distribution of phytoplankton in this area di�er signi�cantly during the year (klekot, 1980; pliński, picińska, 1985). phytoplankton blooms are a poorly studied phenomenon. it is in general connected with seasonal changes of sun irradiance a�ecting water temperature thermocline forming and some other environmental or anthropogenic factors, as local hydrography, and the increases in�ow of pollutants (żmijewska et al., 2000). phytoplankton constitutes an elementary component in aquatic ecosystems. representing the base of the pyramid of productivity, the understanding and modelling of the aquatic ecok la ud ia ś w ia ck a, a lic ja m ic hn ow sk a, j ak ub m ac ul ew ic z, iz ab el a p rz ed no w ek , i ga o gr od ow cz yk , s eb as tia n k oz ic 28 system is not possible without knowledge of the species composition, productivity, and the biomass of phytoplankton. �ese changes in phytoplankton may re�ect major shi�s in environmental conditions (olenina et al., 2006). in order to better understand the algal blooms, seasonal monitoring is necessary. due to natural and methodological reasons, the composition and abundance data on phytoplankton species are highly variable, and consequently, their use as water quality indicators is usually restricted, and mostly chlorophyll a concentrations and primary production are monitored (e.g., satbałtyk). moreover, due to development in taxonomical classi�cation and methodology, su�ciently long comparable time series of phytoplankton are rare (suikkanen et al., 2007). �e aim of this study was to investigate and compare the composition of phytoplankton and the concentration of photosynthetic pigments on four selected stands located in the inside part of �e puck bay and open water of the baltic sea. in the present research, quantitative and qualitative methods were used. for qualitative analysis, samples were analysed by microscope (nicon eclipse 80i). every observed individual of phytoplankton was identi�ed to the genus or species level using speci�ed keys for baltic species of phytoplankton (pliński, 1980) and noted. fig. 1. location of the study area: the open sea (stands: 1, 2) and the gulf of gdańsk (stands: 3, 4) material and methods �e main parameters investigated in the present study were the composition of phytoplankton expressed as the presence of the identi�ed genera or species on each stand c om position of phytoplankton in the p uck b ay and the open b altic s ea 29 and the concentration of photosynthetic pigments: chlorophyll a, b, and c expressed in μg∙l-1. to investigate the composition of phytoplankton, a qualitative method was used. to analyse the concentration of photosynthetic pigments, the quantitative method was used. �e investigation was conducted on a phytoplankton community collected once from the coastal zone in poland of the open sea (stands: 1, 2) and the gulf of gdańsk (stands: 3, 4), in july 2017 (fig. 1). �e plankton samples for qualitative analysis were collected from the surface layer using a phytoplankton net with a diameter of 24.5 cm and 50 µm mesh size. for quantitative analysis, samples were also collected from the surface layer using 5 plastic containers which were �lled with 1 l of water. �e concentration of photosynthetic pigments was measured using a spectrophotometric method. for quantitative analyses the samples were �ltered (v = 1 l) through mn gf-5 glass �bre �lters and extracted with 5 ml cold 90% acetone and then frozen at a temperature of -20°c for 30 minutes. to remove cell debris and �lter particles, the pigment extract was centrifuged at 10000 rpm for 5 minutes. �e extinction was determined at λm 630, 647, 664, and 750 nm with a du530 uv-vis spectrophotometer (beckman) using 1 cm glass cuvette. �e concentration of pigment content was calculated in accordance with je�rey and humphrey (1975). �e measurement of photosynthetic pigment concentration was conducted in triplicate. results and discussion �e studied phytoplankton community was composed of di�erent representatives of cyanobacteria, bacillariophyta, pyrrophyta, and chlorophyta (tab. 1). in total, 17 taxa were observed on all stands, of which 14 were as a genus rank and 3 as a species rank (fig. 2). �e highest number of phytoplankton representatives was observed in stand 1, where 12 generas were noted. aphanizomenon �os-aquae, chlorella vulgaris, and navicula sp. were the only taxa noted on every stand. leptolyngbya sp. and chlorococcum sp. were observed only in stand 1, while coscinodiscus sp. and oscillatoria sp. were noted exclusively in stand 3. �e concentration of chlorophyll a was highest in stand 4 and reached 2.01 μg∙l-1 (fig. 3). in the other stands, chlorophyll a content was similar, and reached around 1.5 μg∙l-1. both concentrations of chlorophyll b and c were the lowest in stand 2. �e highest concentration of chlorophyll b was in stand 1 and reached 0.28 μg∙l-1, while the highest concentration of chlorophyll c was in stand 3, reaching 0.45 μg∙l-1. however, the concentration of chlorophyll c in stands 1, 3, and 4 was similar. in all investigated stands, the concentration of chlorophyll a was much higher than the concentration of other types of chlorophyll. k la ud ia ś w ia ck a, a lic ja m ic hn ow sk a, j ak ub m ac ul ew ic z, iz ab el a p rz ed no w ek , i ga o gr od ow cz yk , s eb as tia n k oz ic 30 tab. 1. individual genera and species of phytoplankton observed on the examined stands: the open sea (stands: 1, 2) and the gulf of gdańsk (stands: 3, 4) no. species/divisions* stands 1 2 3 4 *cyanobacteria 1. aphanizomenon �os-aquae ralfs ex bornet & fla-hault + + + + 2. dolichospermum sp. + + 3. leptolyngbya sp. + 4. microcystis sp. + + 5. nodularia spumigena mertens ex bornet & flahault + + 6. oscillatoria sp. + *pyrrophyta 7. peridinium sp. + + *bacillariophyta 8. chaetoceros sp. + 9. coscinodiscus sp. + 10. licmophora sp. + + 11. navicula sp. + + + + 12. nitzschia sp. + + 13. skeletonema sp. + + + *chlorophyta 14. chlorella vulgaris mertens ex bornet & flahault + + + + 15. chlorococcum sp. + 16. coelastrella sp. + 17. pediastrum sp. + total number of taxa 12 9 9 4 in the baltic sea during the summer months, cyclically observed phenomenon are the blooms of cyanobacteria, mainly of the order nostocales. nodularia spumigena and aphanizomenon �os-aquae are the most o�en noted species which are the main components of summer blooms (mazur-marzec et al., 2012). diatom blooms during spring and the huge uptake of biogenic compounds cause the decrease of those substances during summer, which makes the environmental conditions favourable for cyanobacteria. during the de�ciency of biogenic compounds, some species of cyanobacteria are capable of �xing atmospheric nitrogen. �e ability of �xing atmospheric nitrogen by some species of cyanobacteria gives them predominance in the competition for space and environmental resources. �erefore, the heterocytous organisms can rapidly increase their biomass in short periods of time even under conditions of inorganic nitrogen de�ciency (mazur-marzec et al., 2012; wasmund et al., 1998). in stand 2, the lowest concentration of chlorophyll c and b was observed, which may indicate the lower abundance of bacillariophyta and chlorophyta. out of the c om position of phytoplankton in the p uck b ay and the open b altic s ea 31 groups of algae that were identi�ed, only chlorophyta contains chlorophyll b in chloroplasts, while chlorophyll c is characteristic for bacillariophyta (je�rey, vesk, 1997; kuczyńska et al., 2015). a high concentration of chlorophyll a and a relatively low concentration of chlorophyll b and c may also indicate the domination of cyanobacteria and simultaneously a decrease in diatoms in the gulf of gdansk. moreover, sampling was taken in june when the measured temperature of surface water was of approximately 20°c, which induce blooms of blue-green algae (fleming, kaitala, 2006). �e di�erences in the concentration of chlorophyll b between all stands may be connected with the di�erences in the composition of phytoplankton, which is also indicated by present results (fig. 3; tab.1). a low concentration of chlorophyll c in stand 2 may suggest the lower biomass of bacillariophyta on this site. fig. 2. some species of baltic phytoplankton: a – aphanizomenon �osaquae ralfs ex bornet & flahault, b – nodularia spumigena mertens ex bornet & flahault, c – chlorella vulgaris beyerinck [beijerinck] (source: algaebase – guiry, guiry, 2017) k la ud ia ś w ia ck a, a lic ja m ic hn ow sk a, j ak ub m ac ul ew ic z, iz ab el a p rz ed no w ek , i ga o gr od ow cz yk , s eb as tia n k oz ic 32 unfavourable weather conditions (strong wind) probably contributed to the lower number of observed phytoplankton species. in the �rst stand on the end of the hel peninsula, the highest numbers of phytoplankton species were observed, which can be connected with the stable conditions during sampling. �e species observed in all stands were a. �os-aquae and c. vulgaris. �ese species are widespread and o�en noted in the gulf of gdańsk and also in the open water of the baltic sea (mazur-marzec et al., 2012). large quantities of aphanizomenon �os-aquae may contribute to the high concentration of chlorophyll a, even in stands characterised by a lower number of observed species of phytoplankton. many observations suggest that a. �os-aquae produce some compounds that may alter plankton composition and activity, although each strain of this species may have di�erent properties (mazur-marzec et al., 2012). �ere are also reports on the ability of these algae to a�ect various aquatic organisms, e.g., microalgae, but every target species may respond di�erently to the allelopathic activity (keating, 1978; ikawa et al., 1994; kearns, hunter, 2000). taking into the account potentially harmful e�ects of algal blooms, persistent monitoring programs must be taken into consideration (żmijewska et al., 2000). fig. 3. �e concentration of chlorophylls a, b, c on the examined stands: the open sea (stands: 1, 2) and the gulf of gdańsk (stands: 3, 4) c om position of phytoplankton in the p uck b ay and the open b altic s ea 33 references fleming, v., kaitala, s. (2006). phytoplankton spring bloom intensity index for the baltic sea estimated for the years 1992 to 2004. hydrobiologia, 554(1), 57–65. doi: 10.1007/s10750-005-1006-7 guiry, m.d., guiry, g.m. (2017). algaebase. world-wide electronic publication. aphanizomenon �osaquae ralfs ex bornet & flahault, nodularia spumigena mertens ex bornet & flahault, chlorella vulgaris beyerinck [beijerinck]. galway, ireland: national university of ireland, http://www.algaebase.org. ikawa, m., haney, j.f., sasner, j.j. (1994). inhibition of chlorella growth by the lipids of cyanobacterium microcystis aeruginosa. hydrobiologia, 331, 167–170. doi: 10.1007/bf00025418 je�rey, s., vesk, m. (1997). introduction to marine phytoplankton and their pigment signatures. in: s. je�rey, r. mantoura, s. wright (eds.). phytoplankton pigments in oceanography: guidelines to modern methods. paris, france: unesco publ., pp. 37–84. je�rey, s.t., humphrey, g.f. (1975). new spectrophotometric equations for determining chlorophylls a, b, c1 and c2 in higher plants, algae and natural phytoplankton. biochemie und physiologie der p�anzen, 167(2), 191–194. doi: 10.1016/s0015-3796(17)30778-3 kearns, k.d., hunter, m.d. (2000). green algal extracellular products regulate antialgal toxin production in a cyanobacterium. environmental microbiology, 2, 291–297. keating, k.i. (1978). blue-green algal inhibition of diatom growth: transition from mesotrophic to eutrophic community structure. science, 199, 971–973. doi: 10.1046/j.1462-2920.2000.00104.x klekot, l. (1980). zatoka pucka osobliwością hydrologiczną bałtyku. oceanologia, 12, 109–123. [in polish] kuczyńska, p., jemioła-rzemińska, m., strzałka, k. (2015). photosynthetic pigments in diatoms. marine drugs, 13(9), 5847–5881. doi:10.3390/md13095847 mazur-marzec, h., sutryk, k., kobos, j., hebel, a., hohlfeld, n., błaszczyk, a., toruńska, a., kaczkowska, m.j., łysiak-pastuszak, e., kraśniewski, w., jasser, i. (2012). occurrence of cyanobacteria and cyanotoxin in the southern baltic proper. filamentous cyanobacteria versus single-celled picocyanobacteria. hydrobiologia, 701, 235–252. doi: 10.1007/s10750-012-1278-7 olenina, i., hajdu, s., edler, l., andersson, a., wasmund, n., busch, s., göbel, j., gromisz, s., huseby, s., huttunen, m., jaanus, a., kokkonen, p., ledaine, i. niemkiewicz, e. (2006). biovolumes and size-classes of phytoplankton in the baltic sea helcom. baltic sea environmental programme, 106. olli, k., klais, r., tamminen, t., ptacnik, r., andersen, t. (2011). long term changes in the baltic sea phytoplankton community. boreal environmental resources, 16, 3–14. pliński, m. (1980). glony zatoki gdańskiej: klucz do oznaczania gatunkow. gdańsk: wydawnictwo uniwersytetu gdańskiego. [in polish] pliński, m., picińska, j. (1985). �e dynamics of seasonal change of the phytoplankton biomass in the gulf of gdańsk, oceanologia, 23, 77–83. suikkanen, s., laamanen, m., huttunen, m. (2007). long-term changes in summer phytoplankton communities of the open northern baltic sea. estuarine, coastal and shelf science, 71, 580–592. doi: 10.1016/j.ecss.2006.09.004 wasmund, n., nausch, g., matthäus, w. (1998). phytoplankton spring blooms in the southern baltic sea-spatio-temporal development and long-term trends. journal of plankton research, 20, 1099–1117. doi: 10.1093/plankt/20.6.1099 żmijewska, m.i., niemkiewicz, e., bielecka, l. (2000). abundance and species composition of plankton in the gulf of gdansk near the planned underwater outfall of the gdansk-wschod (gdansk-east) sewage treatment plant. oceanologia, 42, 335–357. k la ud ia ś w ia ck a, a lic ja m ic hn ow sk a, j ak ub m ac ul ew ic z, iz ab el a p rz ed no w ek , i ga o gr od ow cz yk , s eb as tia n k oz ic 34 abstract �e puck bay is an area characterised by speci�c hydrodynamic conditions that determine the distribution and composition of phytoplankton. �e aim of this study was to investigate the di�erences in the phytoplankton composition and the content of photosynthetic pigments between the puck bay and open baltic sea. �e material was collected from four stands which were localised in the inner and outer part of hel peninsula. in this study, it has been demonstrated that the composition of individual species of phytoplankton di�ered between stands in the inner and outer part of the puck bay. �is investigation has also shown that the number of phytoplankton taxa was similar in three stands and it was much lower on the last stand (stand 4). �e di�erences in the concentration of photosynthetic pigments between all stands have also been observed. key words: phytoplankton, puck bay, southern baltic sea, blooms received: [2017.07.04] accepted: [2017.10.26] struktura fitoplanktonu w zatoce puckiej oraz na otwartym morzu bałtyckim streszczenie zatoka pucka jest obszarem charakteryzującym się specy�cznymi warunkami hydrodynamicznymi, które determinują rozmieszczenie i skład �toplanktonu. głównym celem niniejszej pracy było zbadanie różnic w składzie taksonomicznym �toplanktonu oraz zawartości barwników fotosyntetycznych pomiędzy zatoką pucką, a otwartymi wodami morza bałtyckiego. materiał zebrano z czterech miejsc zlokalizowanych po wewnętrznej, jak i zewnętrznej, części półwyspu helskiego. w pracy wykazano, że istnieją różnice w składzie taksonomicznym �toplanktonu między wewnętrzną i zewnętrzną częścią zatoki puckiej. niniejsza praca pokazała, że liczba taksonów �toplanktonu na stanowiskach 1, 2 i 3 była zbliżona i znacznie wyższa niż na stanowisku 4. zaobserwowano również różnicę w stężeniach barwników fotosyntetycznych między badanymi stanowiskami. słowa kluczowe: �toplankton, zatoka pucka, morze bałtyckie, zakwity information on the authors klaudia świacka in her studies, she focuses on the impact of the non-steroidal anti-in�ammatory drugs on the baltic benthic fauna. in current laboratory research, she is investigating the bioaccumulation of diclofenac in mytilus trossulus tissues. moreover, she determined the occurrence and concentration of nsaids in the gulf of gdańsk in the water, sediment, and in the tissues of the organisms collected from di�erent stations. she is also interested in the taxonomy of marine organisms and would like to improve her knowledge and skills during future investigations. alicja michnowska in her research, she is interested in marine ecotoxicology, especially in the matter of marine invertebrates. her main interests include the calibration and modi�cation of methods for the determination of biochemical compounds that can serve as potential biomarkers for environmental condition including genotoxicity, metal contamination, and climate changes. �e aim of her work is to determine a suitable methodology for a multiple biomarker approach usable in ecotoxicologic studies as an environmental diagnostic tool. jakub maculewicz �e �eld of his interest is allelopathic interactions of phytoplankton, in particular, of baltic �lamentous cyanobacteria. he is investigating what in�uences of allelopathic compounds have on those organisms. in c om position of phytoplankton in the p uck b ay and the open b altic s ea 35 his studies, he uses the following innovative methods: analysing pigment content, chlorophyll a �uorescence, and measuring the rate of photosynthesis to determine what impact allelochemicals have on algae. izabela przednowek �e main area of her interest is the biology and ecology of zooplankton. currently, her research focuses on comparing the species composition of phyoplankton before and a�er the in�ow of water from the north sea. such studies have not been conducted previously in the baltic sea. iga ogrodowczyk she is a marine biology student, and the main research objective of her bachelor’s thesis was to test gst as a marker of neoplasia diseases in bivalve molluscs. currently, she is trying to expand her knowledge on this subject. she also investigates the spread of neoplasia in bivalve molluscs from the gulf of gdańsk. sebastian kozic he studies marine biology. from the beginning of these studies, he has been interested in investigating zoobenthos. he wrote his undergraduate thesis about polychaetes, and his master thesis will also be about polychaetes. he believes that, in a few years, he will be an expert in polychaetes. 85 annales universitatis paedagogicae cracoviensis studia naturae, 1: 85–94, 2016, issn 2543-8832 larysa m. makhynia department of pharmacognosy and botany, bogomolets national medical university, pushkinska 22 st., 01601 kyiv, ukraine, larisa_2015@ukr.net bidens frondosa l. resource evaluation in the dnipro river bottomland (in range of the forest steppes of ukraine) introduction modern globalization threats and the current trends of the climatic changes over particular areas contribute to the acceleration of spreading of invasive species and create conditions for their quicker rooting under new conditions (protopopova, 1991; bondar et al., 2011). in the study area two indigenous species (bidens tripartitа l., b. cernua l.) and two adventive ones (b. frondosa l., b. connata muehl.) are met. during the last 10–15 years a  sharp resource decrease of a  pharmacopoeic species of в. tripartita is viewed due to the fast spreading of an invasive one – b. frondosa. consequently, in the near future it will cause serious losses of biological variety and economic signi�cance of ecosystems in which this species has been spreading (protopopova, 1973, 1991; bortnyak, 1976; horbyk, husak, 1983; mosyakin, 1988a; chorna, 2001, 2006). b. frondosa was �rst found on the ukrainian territory in the town of kаniv in 1970 by a famous polish researcher j. kornaś (1971). also b. frondosa spreading was pointed out in the research of protopopova (1973, 1991), bortnyak (1976), kotov (1979), horbyk and husak (1983). mosyakin (1988b) noticed that this species is much more frequently met in kyiv than b. tripartitа, o�en creating dense undergrowth. minarchenko and timchenko (2002) indicated the fast spreading of b. frondosa throughout ukraine. danylyk and danylyk (2009) showed the habitat of two new species (b. frondosa, b. connata) as a part of shatsky national park �ora though they haven’t been met there before. �e authors explain this fast spreading of invasive species throughout ukraine as the increase of the vegetation anthropogenic transformation. some of makhynia’s publications (2005, 2009, 2012, 2015), as well as makhynya and strumens’ka’s ones (2010), are devoted to the issues of spreading and fast exploration of new, especially transformed areas, and extruding other indigenous species – b. tripartitа. 86 la ry sa m . m ak hy ni a �e conditions prevailing on the study area, i.e. the intensive development of the riverside territories for the management (creating private areas, �sh farms, treatment plants etc.) contribute to more and more successful occupation of transformed territories by the invasive species. �e next 10–15 years will have a signi�cant impact on b. tripartitа spreading and resources. due to this, topical is the search of objects with the analogous or similar chemical composition, su�cient and accessible resource base. it is obvious that a successful candidate for such research is b. frondosa, industrial storing of which may negatively in�uence the species further expansion. b. frondosa is not used in ukraine, but due to the research of foreign authors it may be rather a promising species. in the usa it refers to the pharmacopeia is used in gynecology and otolaryngology (morton, 1962; mitich, 1994). in northern america infusions from its roots and leaves are used in arrhythmias, bronchitis and laryngitis, and its fruits – in metrorrhagia (kolla, 1985). �e researches of japanese scientists show the antioxidant activity of tetrahydroxyаuronе of b. frondosa. it supports the issue on the prospects of the use of the raw material of this species and the topicality of its resource (venkateswarlu et al., 2004). �e aim of the study was to investigate the b. frondosa resources in the dnipro river bottomland as a promising substitute of b. tripartitа. study area �e bottomland of the medium forest steppe of the dnipro is located between kyiv and kremenchuk (fig. 1) and di�ers signi�cantly from those parts of the dnipro which are in the north in forest zone and in the south in the steppe zone by the nature of its bottomland, stream �ow and �oodplains with its plant and ground coating (afanas’yev, 1950). �e dnipro bottomland which occupies accumulative terrace is 120 km wide near kyiv and consists of �oodplains, upland, and loess terraces up to 35–40 m height above the river level (marynych, shyshchenko, 2006). due to its layering relief the bottomland belongs to the bug–dnipro level, the heights of which reach 200–300 m. it is located within the ukrainian shield. its surface looks like a  slightly wavy plain dissected by valleys and gullies. �ere are not many lowland inter�uves. bedrock exfoliates on the slopes of river valleys and gullies (marynych, shyshchenko, 2006). in accordance with agro-ground zoning of ukraine, the dnipro bottomland is in the northern sub-province of the right bank central high province of forest steppe zone of typical black earth and gray ashed soils (vernander, 1986). floodplain soils and sandy terraces lie on eluvium, the rest are formed on loess. riverine �oodplain is characterized as consisting of little turf, turf sand, rarely – sandy soils. soils of central �oodplain are wet meadow, sandy, and loamy soils. among soils of pre-terrace �oodplain there prevail meadow marsh and swamp loam ones. on 87 the major part of pre-terrace �oodplain, alluvial deposits of organic–mineral character are concentrated (afanas’yev, 1950). upland terraces of the psel and sula bottomlands, as well as the one of the dnipro, are covered with sod low-podzolic sandy soils. �e duration of spring �ood varies widely from the beginning of april till july. �e long spring �ood determines considerable dynamics of �ood plains, the development of which is fully linked with the dnipro �oodwaters. �e dnipro signi�cant solid �ow is of great importance for �oodplain formation (poryvkyna, 1986). �e vegetation of the study area is formed under the in�uence of the arti�cial reservoirs (kаniv and kremenchuk), created in 60s–70s as a result of the dnipro channel regulation and the construction of hydroelectric power plants. total water area constitutes 292 ha. reservoirs are characterized by a variable hydro regime due to which huge areas fig. 1. map scheme of resource areas placement of the dnipro bottomland (within the forest steppe of the ukraine). resource areas: i – kyiv, ii – protsivsko–kozynsriy, iii – pereyaslav–khmelnitsky, iv – kаniv, v – zolotonosha, vi – cherkasy, vii – chygyryn, viii – protsenky, ix – kremenchuk bidens frondosa l. resource evaluation in the d nipro r iver bottom land (in range of the forest steppes of u kraine) 88 la ry sa m . m ak hy ni a with periodic �ooding water are formed. �ey became biotopes of mass distribution of b. frondosa. vegetation is represented by �oodplain forest, meadow, psamophytic, marsh, higher aquatic, and ruderal types (afanas’yev, 1950; voytyuk, 1999). floodplain forests are spread fragmentary and occupy in average 10–15%, their distribution is determined by the �oodplain regime. meadow vegetation in respect of the �oristic is the richest and most widely spread, having an average of 70–75% of the total area. it includes steppe, real and marshy meadows. marshy meadows account for 12–15%. �ey occur sporadically, mainly in central and pre-terrace �oodplains. psammothytic vegetation occupies small areas and is represented by few communities (about 3–5%). �e vegetation of new ecotypes, among which the most frequently spread are, as it is already been mentioned, b. frondosa and b. tripartita, occupies rather signi�cant territories (about 20–35%) and is di�erentiated in relation to humidity on aquatic, riverside and ruderal communities with lemna minor l., salvinia natans all., trapa natans l. s. str., nuphar lutea (l.) sibth. et sm., which belong to higher aquatic vegetation which occurs fragmentarily (about 25–30%). �e vegetation of riverside ecotypes (5–15%) consists of typha angustifolia l., scirpus lacustris l., glyceria maxima (hartm.) holmb., phragmites australis (cav.) trin. ex steud., sagittaria sagittifolia l., bidens tripartita, b. cernua, b. connata and b. frondosa. ruderal vegetation is formed from violations, azotized areas and occupies 7–15%. in relation to the increased anthropogenic in�uence (recreational in particular) the communities of ruderal vegetation rather quickly supersede the natural ones and its areas increase. �e dominant communities were formed by xanthium albinum h. scholz., chenopodium album l., tussilago farfara l., polygonum hydropiper l. and bidens frondosa. material and methods resource research was carried out on the basis of the method of registration plots (borisova, shreter, 1966; borisova et al., 1982; kryilova, shreter, 1971; kryilova, 1973, 1981; shreter, 1986; kryilova et al., 1989; kryilova, kaporova, 1992). �e size of registration plots depends on the location of masses on the study area. registration plots were diagonally located on the area occupied by the community of species (if its size was not more than 0.5 ha) or were placed as parallel or separated transects on the extended areas (riverside territories). �e number of registration plots depended on the sample size (kryilova, shreter, 1971; kryilova, 1973, 1981; kryilova, kaporova, 1992). in all, 347 plots were laid. raw material collected on every sample plot was weighed. weighed data was entered into the associations description form. freshly gathered raw material was put in separate paper-bags with weight, place and date of collection being �xed on them. raw material drying was performed in accordance with conventional methods as required (gosudarstvennaya farmakopeya sssr, 89 1989). a�er drying the re-weighting was carried out. data on dry weight was entered into the form. main calculations were performed under camera conditions. recalculating, we received data about raw material weight averages of every investigated species in g/m2. drying coe�cient was determined by the formula с = 100×(а–в)/а, where а – freshly gathered raw material weight, в – dry weight (salo, 1972). biological reserve of raw material was determined as the sum of the area and phytomass from g/ m2. operational reserve of raw material was determined as the rate of 50% to the biological reserve since bidens frondosa is a herbaceous annual, the raw material of which is the aboveground organs. �is very amount of single use provides the minimal population ability to recover a�er raw material collection (kovalev, zhuravlev, 1989; yakovlevа, blinovoy, 2004; minarchenko, minarchenko, 2004; minarchenko, 2012). �e rotation of the possible volume of collection for these species must be carried out every two years (kovalev, zhuravlev, 1989; yakovlevа, blinovoy, 2004). statistical data processing was performed by the applied computer program excel for windows o�ce 2007 and windows xp, as well as the methods of variation statistics. a true average (м) and the error of the average (m) with an acceptable accuracy coe�cient (p) to 15% were calculated (lakin, 1990; tsarenko et al., 2000). results and discussion �e main reserve of bidens frondosa is located on low riverside areas of �oodplain and riverine parts of rivers, lakes, ponds, oxbow lakes, marshy forests, islands, riverside areas of water reservoirs mainly of the le� bank bottomland and less of the right bank. �ey create continuous tracks of undergrowth, occur sporadically, but also can be found in small groups. as to spreading b. frondosa occupies about 70% of growing areas of all representatives of the genus (tab. 1). �e largest areas are concentrated in cherkasy region (іv, v, vii resource areas), kyiv region (іі resource area) and on islands in poltava region (іх resource area). �e fewest ones are in kyiv region (і, ііi resource areas) and on the outskirts of villages of poltava region (viiі, іх resource areas) (fig. 1). much rarer spreading and signi�cantly smaller reserve are characteristic of b. tripartita (tab. 2). firstly, it is caused by its lower ecologic amplitude (vinogradova, 2003; vinogradova et al., 2009; vasileva, papchenkov, 2011; makhynya, 2009, 2011); secondly, by the conditions which have developed on the study area, that is, active conquering of riverside territories for management needs (the creation of private areas, �sh farms, treatment plants etc.) and excessive �uctuations in water levels of arti�cial reservoirs. another aspect which in�uences the intensity of b. tripartita extrusion is the speed of seed germination of b. frondosa in comparison with the aboriginal type and the tempos of development under the juvenile condition (vinogradova et al., 2009; makhynya, 2011). in the region b. tripartita occupies about 20% of growing bidens frondosa l. resource evaluation in the d nipro r iver bottom land (in range of the forest steppes of u kraine) 90 la ry sa m . m ak hy ni a area. its largest areas are concentrated in cherkasy region (v, vi resource areas) and in kyiv region (іi resource area). �e smallest are in kyiv region (і, ііі resource areas) and poltava region (vііі resource area) (fig. 1). �e peculiarities of territorial and ecological reserve di�erentiation of b. frondosa and b. tripartitа are established. �e main reasons for the reduction of this species are its narrow ecological amplitude. �ey are intensi�ed by the condition changes prevailing on the study area and being negative for this species. it is found that the largest raw material reserves are peculiar of b. frondosa in cherkasy region (іv, v, vii resource areas), kyiv region (іі resource area) and on the islands in poltava region (іх resource area). according to the features of annual use and the reserve accessibility of the areas b. frondosa are concentrated in cherkasy region (v resource area). total area constitutes 32 648 ha; biological reserve is correspondingly between 1130–1190 t, operational – between 565–595 t of dry raw material (tab. 1). b. tripartita occupies only 25 573 ha; biological reserve is 108–124 t, operational – 54–62 t of dry raw material (tab. 2). �e relevance of further resource research in other parts of the bottomland and monitoring by the resources dynamics of b. frondosa under the conditions of arti�cial reservoirs is obvious. �is species harvesting will assist its negative in�uence reduction on natural reproduction of b. tripartita. for more successful conservation of resources b. tripartita needs some passive protection measures of its reserves creating natural reserves objects of both state and regional levels. main objects of protection have to be herb resources, the reserves of which are sharply decreasing in ukraine. tab. 1. bidens frondosa raw material reserves resource area total spreading area [ha] total thickets area [ha] raw material phytomass [g/m2]; ±sd biological reserves of dry raw material [t] operational reserves of dry raw material [t] amount of possible annual use of dry raw material [t] kyiv 448 2.48 868.90 ±59.80 6.02–6.91 3.01–3.45 1.50–1.73 protsivsko– kozynsriy 5200 56.22 882.96 ±22.05 145.30–151.74 72.65–75.87 36.32–37.94 pereyaslav– khmelnitsky 1300 20.45 856.50 ±13.70 51.70–53.39 25.85–26.69 12.93–13.35 kаniv 4150 78.13 862.70 ±22.80 198.55–208.32 99.27–104.16 49.64–52.08 zolotonosha 4500 117.09 837.24 ±26.12 289.91–308.41 144.95–154.21 72.48–77.10 cherkasy 3900 42.89 779.70 ±30.03 103.44–109.83 51.72–54.92 25.86–27.46 chygyryn 8000 57.95 879.20 ±20.08 149.03–155.93 74.52–77.97 37.26–38.98 protsenkiv 1550 29.23 892.50 ±14.70 76.97–79.55 38.49–39.78 19.24–19.89 kremenchuk 3600 48.44 829.54 ±28.43 109.50–116.29 54.75–58.14 27.37–29.07 total 32648 452.88 854.36 ±26.41 1130.43–1190.39 565.21–595.19 282.61–297.60 91 tab. 2. bidens tripartita raw material reserves resource area total spreading area [ha] total thickets area [ha] raw material phytomass [g/m2]; ±sd biological reserves of dry raw material [t] operational reserves of dry raw material [t] amount of possible annual use of dry raw material [t] kyiv 248 1.05 428.80 ±48.50 0.59–0.75 0.30–0.38 0.15–0.19 protsivsko– kozynsriy 4825 42.24 266.60 ±21.30 15.28–18.05 7.64–9.03 3.82–4.51 pereyaslav– khmelnitsky 1300 14.85 243.20 ±12.60 5.14–5.70 2.57–2.85 1.28–1.42 kаniv 3750 45.25 230.00 ±14.60 14.15–15.93 7.08–7.96 3.54–3.98 zolotonosha 3920 53.50 327.23 ±25.50 23.16–27.09 11.58–13.55 5.79–6.77 cherkasy 3440 40.07 307.40 ±18.80 17.09–19.37 8.54–9.69 4.27–4.84 chygyryn 4200 28.62 311.70 ±12.70 12.93–14.04 6.47–7.02 3.23–3.51 protsenkiv 1550 7.14 282.20 ±22.50 2.78–3.26 1.39–1.63 0.69–0.81 kremenchuk 2340 31.17 396.40 ±34.20 16.95–20.18 8.47–10.09 4.24–5.04 total 25573 263.89 310.40 ±23.40 108.08–124.38 54.04–62.19 27.02–31.09 references afanas’yev, d.y. 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[in russian] abstract �e article concentrates on the investigation of bidens frondosa l. resources in the dnipro river bottomland as a promising substitute of b. tripartitа l. due to its phyto-resources deterioration on the territory of ukraine. �e features of their distribution, ecological and cenotic components, and these species reserve at the most perspective raw material signi�cant territories are identi�ed. �reat factors for b. tripartitа resources are clari�ed. �e research was performed on the basis of the method of registration plots with the subsequent calculation of the biological and operational reserves and the possible volume of the annual collection of dry raw material. registration plots were diagonally located on the area occupied by the community of species. in the extended areas they are placed as parallel or separated transects. b. frondosa plant raw material reserve is established and the decrease for b. tripartitа is proved. in connection with the increase of b. frondosa resource base, the prospects of using its raw material as a successful substitute of b. tripartitа are justi�ed. corresponding proposals are submitted to the ministry of ecology and natural resources of ukraine. key words: bidens frondosa, b. tripartita, bottomland, dnipro river, raw material received: [2016.05.29] accepted: [2016.09.04] bidens frondosa l. resource evaluation in the d nipro r iver bottom land (in range of the forest steppes of u kraine) 94 la ry sa m . m ak hy ni a uczep amerykański bidens frondosa l. ocena zasobów w terasie rzecznej dniepru (w zasięgu lasostepu ukrainy) streszczenie artykuł dotyczy badań zasobów uczepu amerykańskiego bidens frondosa l. w terasie rzecznej dniepru jako ekspansywnego następcy uczepu trójlistkowego b. tripartitа l., w połączeniu z pogorszeniem �to-zasobów drugiego z wymienionych gatunków na terenie ukrainy. określono cechy ich rozmieszczenia, ekologiczne i cenotyczne składniki oraz ich rezerwuar gatunkowy na znacznej części badanego terenu. sprecyzowano również czynniki zagrożenia zasobów dla b. tripartitа. badania przeprowadzono w oparciu o metodykę poletek z późniejszym wyznaczeniem biologicznych, operatywnych zasobów oraz wielkości pozyskania rocznego surowca suchej masy. stałe poletka były rozmieszczone diagonalnie na obszarze zajmowanym przez zbiorowiska z tymi gatunkami. na rozległych powierzchniach były one umiejscowione w równoległych lub odrębnych transektach. zasoby surowca roślinnego b. frondosa okazały się być stabilne, a  ich obniżanie udowodniono u b. tripartitа. w połączeniu ze wzrostem bazy surowcowej b. frondosa, perspektywy wykorzystania jego surowca jako substytutu b. tripartitа są uzasadnione. stosowne wnioski złożono do ministerstwa ekologii i zasobów naturalnych ukrainy. słowa kluczowe: bidens frondosa, b. tripartita, terasa rzeczna, dniepr, surowiec zielarski information on the author larysa м. makhynia currently she is researching the species of the genus bidens l. resource evaluation in the dnipro river bottomland (in range of the forest steppes of ukraine). also, she investigated the distribution, ecological and cenological features of new association communities of bidenti frondosae – bidentetum connatae. she has previously examined distribution, ontogeny, ecology, cenology, ecolo-cenotic strategy of the genus bidens (b. tripartita l., b. cernua l., b. frondosa l., b. connata muehl.). 179 annales universitatis paedagogicae cracoviensis studia naturae, 1: 179–189, 2016, issn 2543-8832 ksenia strzeżoń department of botany, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, ksenia1922@gmail.com herbaceous flora of village chocznia (southern poland) with particular focus on medicinal plants �e chemistry development in the nineteenth century caused that herbal preparations were replaced by synthetic medicines (sarwa, 2001). however, currently people began to appreciate the power of natural medicines again. herbal preparations have a weaker e�ect, which makes them safer. �e quantity of active ingredient in the capsule prepared in the laboratory is strictly determined. �e plants regulate themselves in terms of production of the amount of the active ingredient. �e revival of popularity of herb-derivative medicines is due to the fact that they cause fewer or none side effects. obviously one should not assume that any plant, in any amount will not cause side e�ects. common sense and knowledge of herbs should be enough to ensure their safe use (duke, 2011). in poland, many medicinal plants are still coming from the natural environment. however, in some regions of the country, especially in the vicinity of the large urban areas, the anthropogenic impact and the state of environmental pollution make it impossible to exploit medicinal plants. hence, there is a constant need to �nd new areas where the concentration of medicinal species is relatively large and the state of the environment is favorable for the harvest. so far, in the vicinity of the village chocznia (małopolska voivodeship) no �oristic exploration has been carried out. �erefore, the �oristic studies were undertaken with the aim to analyse herbaceous plants composition, with particular emphasis on medicinal species. chocznia (49°51’51” n, 19°26’39” e) is a village located in the western part of the małopolska voivodeship in wadowice commune (fig. 1). it is situated at the altitude of 300–380 m above sea level on the border of wieliczka foothills (pogórze wielickie) and the eastern part of small beskid (beskid mały) near wadowice, zawadka, kaczyna, inwałd and ponikiew (siemionow, 1984; zinkow, 2001). �e soils in this area are rather poor and shallow; pseudopodsolic and brown acidic soils are dominant. chocznia is located in a temperate warm climate, where annual 180 k se ni a s tr ze żo ń rainfall ranges from 1000 to 1400 mm. �e strongest winds are from the west and north-west and in spring and autumn warm mountain winds are causing thaws or rain, characteristic for this area. by the chocznia village two rivers �ow – choczenka and kanówka. choczenka begins its �ow on the gancarz mount (góra gancarz) slopes and �ows into the river skawa from right tributary of the vistula (wisła). �e second river is the kanówka with its source on the foothills under bliźniaki (siemionow, 1984; zinkow, 2001). chocznia area is mainly occupied by agriculture crops, to a lesser extent forests and forest land. in this area, as in the small beskid, dominant are spruce forests of beech (fagus sylvatica l.) and �r (abies alba mill.) considered as contaminant species classi�ed to the vaccinio–abietenion oberda. 1962. alliance. �ere can also be found small birch forests mainly in the admixture of pine forests querco roboris–pinetum (w. mat. 1981) j. mat. 1988. �e riverine riversides appear above the streams and rivers with alder (alnus sp.) and willows (salix sp.) in the tree stand. �ere are also preserved fragments of natural acid beech forest from the alliance luzulo–fagenion (lohm. ex r. tx. 1954) oberda. 1957 and fertile beech forests with the association dentario glandulose–fagetum w. mat. 1964 ex guzikowa et kornaś 1969 (siemionow, 1984; szafer, zarzycki, 1972). fig. 1. study area – distribution of research stands in the village of chocznia 181 h erbaceous flora of village c hocznia (southern p oland) w ith particular focus on m edicinal plants plants materials were collected in years 2014–2015. collections were held starting from march 2014 until the end of the growing season and in the spring on the beginning of the growing season of 2015. in the area of the analysed village 10 research stands with di�erent habitat conditions were determined (fig. 1). �e plants materials were collected several times during the season noting the phenology of species. �e most frequent two specimens of the species were collected. plants were identi�ed both in the area and a�er drying in the laboratory using the available keys, atlases and guides (broda, mowszowicz, 2000; macků, krejča, 1989; schönfelder, schönfelder, 1997; szafer et al., 1986). nomenclature of species was given from the online atlas of polish vascular plants (http://www.atlas-roslin.pl/). �e herbarium is a documentation of the study and it is deposited in the department of botany at the pedagogical university of kraków. �e available publications and atlases were used to determine which plants have medicinal properties and contain active substances (broda, mowszowicz, 2000; kawałko, 1986; krejča, kresanek, 1983; macků, krejča, 1989; nowiński, 1980; ożarowski, jaroniewski, 1987; polakowska, 1982; rausch, lotz, 2006; rumińska, 1983; rumińska, ożarowski, 1990; sarwa, 2001; schönfelder, schönfelder, 1997; senderski, 2007; starý, jirásek, 1976; volåk, stodola, 1987). moreover, all recorded species are classi�ed based on their frequency scale in the analysed area. for this aim, the following frequency scale was accepted: rare species – from 1 to 3 stands, frequent species – from 4 to 6 stands, common species – 7 and more stands. among the species recognised as common and frequent are medicinal plants potentially suitable for harvesting from their natural state. a total of 99 plant species belonging to 37 families were found in the chocznia village. of all the taxons, 60 plants had medicinal properties (appendix 1 – tab. 1), which is 60.61% of all species observed. �e families most abundant in medicinal species are the following: asteraceae (9 species), lamiaceae (7 species), rosaceae (5 species), fabaceae and brassicaceae (4 species), plantaginaceae (3 species). other families were represented by one or two species (fig. 2). all medicinal plants from chocznia stands belong to 27 families. �is is consistent with the described trends in �oristic literature, because these families are most strongly represented in the �ora of poland (szafer, zarzycki, 1972; szafer et al., 1986). sun�ower family is the largest not only in the native �ora of poland, but in the �ora of the whole world – includes as many as 25 000 species (kochanowska, nowak, 2003), whereas the deadnettle family is a reservoir of medicinal, spice and cosmetics species. �e plants belonging to this family are characterised by a high content of aromatic essential oils, which are widely used in herbal medicine, cosmetics and gastronomy. �eir properties are known and appreciated since ancient times (broda, mowszowicz, 2000; nowiński, 1980; rumińska, 1983; sarwa, 2001). 182 k se ni a s tr ze żo ń on the studied area, among the medicinal plants the frequent status had 27 species and common status had 11 species. potentially for the purpose of harvest of herbal medicine 38 of frequent and common species were suitable. each area has a speci�c microclimate and the habitat conditions, which is re�ected in the composition and species richness of its �ora. regularity is that the �oras of di�erent areas, more or less transformed, are dominated by rare species – generally common species are the least. although common species occupy the largest surfaces, their amount in comparison to the total number of taxons on that area is relatively small (szafer, zarzycki, 1972). �is is also con�rmed in the �ora of the analysed village (fig. 3, appendix 1 –tab. 1). �e greatest diversity of medicinal plants were found on the 2, 7, 8 stands, which are generally characterised by high species diversity (fig. 4). in all three cases, these are places characterised by a diversity of habitats. hence, species with di�erent spectra of environmental requirements are present on these stands. however, a large number of medicinal taxons does not mean that these species are there in quantities suitable for harvest. species diversity is generally connected with a greater mosaic of area and very o�en a lack of dominant species in plots. dispersion of medicinal plants is not conducive to harvesting them from their natural habitats. �erefore, an important element is the frequency of occurrence of medicinal species. �ey need to be characterised by the appropriate concentration on stands so that their harvest is not associated with a lot of e�ort to �nd them in the area. fig. 2. comparison of the total number of species and the number of medicinal plants belonging to individual families; others – include the families represented by only one species 18 14 9 8 7 5 5 4 3 3 3 3 3 2 2 2 2 2 2 2 12 9 7 4 4 2 5 2 1 3 0 2 0 2 1 2 1 2 0 1 0 5 10 15 20 25 30 35 ot he rs a st er ac ea e la m ia ce ae b ra ss ic ac ea e fa ba ce ae r an un cu la ce ae r os ac ea e b or ag in ac ea e c ar yo ph yl la ce ae pl an ta gi na ce ae c am pa nu la ce ae c on va lla ri ac ea e po ac ea e pa pa ve ra ce ae v io la ce ae pr im ul ac ea e g er an ia ce ae a po cy na ce ae r ub ia ce ae sc ro ph ul ar ia ce ae n um be r of s pe ci es plant families medicinal species all species 183 h erbaceous flora of village c hocznia (southern p oland) w ith particular focus on m edicinal plants �e results of the study showed that the possibilities of harvesting medicinal species in chocznia and the surrounding area are satisfactory (appendix 1 – tab. 1, fig. 3). on the analysed stands, among the therapeutic species commonly occur: achillea millefolium, aegopodium podagraria, capsella bursa-pastoris, cirsium rivulare, lychnis �os-cuculi, plantago lanceolata, p. maior, ranunculus acris, rumex acetosa, taraxacum o�cinale, urtica dioica. �e majority of these species are used in conventional medicine, but some of them were used so far mainly in folk medicine, e.g. cirsium rivulare, lychnis �os-cuculi or ranunculus acris. �ese plants are well known to local residents, but now very few people know about their medicinal properties. currently, people are less interested in collecting herbs for their own needs, because it requires a certain expertise knowledge. additionally, availability in sales of prepared herbs substrates are large and generally they are not too expensive (duke, 2011; ożarowski, jaroniewski, 1987). however, in many polish regions especially where there is relatively high unemployment due to the lack of large industrial plants, people engage in harvesting growing wild herbs for sale. �e herbal substrate, well collected from the natural habitat, is better in terms of the content of the di�erent active substances than that grown in arti�cial conditions (duke, 2011; nowiński, 1980; rumińska, 1983; sarwa, 2001; skarżyński, 1994). �erefore, throughout poland there are companies involved in the purchase of herbal raw materials. obviously, in companies buying herbal raw materials it is necessary to specify the place where plants were harvested. in this context, chocznia is an appropriate location for harvesting medicinal plants, both for industrial and personal consumption, because it is located far away from big cities and industry. �e state of the environment here is relatively good and agriculture rather extensive in nature. �ese factors provide a high quality herbal raw materials from this area. fig. 3. comparison of frequency of occurrence of all species and medicinal species on the studied area 41 40 18 22 27 11 0 5 10 15 20 25 30 35 40 45 rare frequent common c at eg or y of o cc ur re nc e number of species all species medicinal species 184 k se ni a s tr ze żo ń references atlas of polish vascular plants – atlas roślin naczyniowych polski. http://www.atlas-roslin.pl/ [in polish] broda, b., mowszowicz, j. (2000). przewodnik do oznaczania roślin leczniczych, trujących i użytkowych. warszawa: państwowe zakłady wydawnictw lekarskich. [in polish] duke, j.a. (2011). księga zdrowia. ziołowa apteka domowa. poznań: publicat. [in polish] kawałko, j.m. (1986). historie ziołowe. lublin: krajowa agencja wydawnicza. [in polish] kochanowska, j., nowak, t. (2003). pomoce dydaktyczne dla nauczycieli odwiedzających ogród botaniczny we wrocławiu. wrocław: u.t.r. interior. [in polish] krejča, j., kresanek, j. (1983). rośliny lecznicze. warszawa: wydawnictwo sport i turystyka. [in polish] macků, j., krejča, j. (1989). atlas roślin leczniczych. wrocław: zakład narodowy im. ossolińskich. [in polish] nowiński, m. (1980). dzieje upraw i roślin leczniczych. warszawa: państwowe wydawnictwo rolnicze i leśne. [in polish] ożarowski, a., jaroniewski, w. (1987). rośliny lecznicze i ich praktyczne zastosowanie. warszawa: instytut wydawniczy związków zawodowych. [in polish] polakowska, m. (1982). leśne rośliny zielarskie. warszawa: państwowe wydawnictwo rolnicze i leśne. [in polish] rausch, a., lotz, b. (2006). zioła. leksykon. warszawa: elipsa. [in polish] rumińska, a. (1983). rośliny lecznicze: podstawy biologii i agrotechniki. warszawa: państwowe wydawnictwo naukowe. [in polish] rumińska, a., ożarowski, a. (1990). leksykon roślin leczniczych. warszawa: państwowe wydawnictwo rolnicze i leśne. [in polish] sarwa, a.j. (2001). wielki leksykon roślin leczniczych. warszawa: wydawnictwo książka i wiedza. [in polish] schönfelder, i., schönfelder, p. (1997). rośliny lecznicze – rozpoznawanie, zbiór, stosowanie. warszawa: multico. [in polish] fig. 4. comparison of the participation of all taxons and medicinal species on distinguished research stands 40 45 25 39 42 34 51 52 40 41 26 29 19 28 24 16 29 33 26 23 0 20 40 60 80 100 1 2 3 4 5 6 7 8 9 10 n um be r of s pe ci es stands total number of species on stand number of medicinal species on stand 185 h erbaceous flora of village c hocznia (southern p oland) w ith particular focus on m edicinal plants senderski, m. (2007). prawie wszystko o ziołach. podkowa leśna: wydawnictwo senderski m. [in polish] siemionow, a. (1984). ziemia wadowicka. kraków: anczyca. [in polish] skarżyński, a. (1994). zioła czynią cuda. warszawa: comes. [in polish] skowron, c., ruła, m., targosz, e. (2006). wadowice koło choczni. wadowice: gra�kon. [in polish] starý, f., jirásek, v. (1976). rośliny lecznicze. warszawa: państwowe wydawnictwo rolnicze i leśne. [in polish] szafer, w., kulczyński, s., pawłowski, b. (1986). rośliny polskie. opisy i klucze do oznaczania wszystkich gatunków roślin naczyniowych rosnących w polsce bądź dziko, bądź też zdziczałych lub częściej hodowanych. warszawa: państwowe wydawnictwo naukowe. [in polish] szafer, w., zarzycki, k. (1972). szata roślinna polski. warszawa: państwowe wydawnictwo naukowe. [in polish] volåk, j., stodola, j. (1987). rośliny lecznicze. warszawa: państwowe wydawnictwo rolnicze i leśne. [in polish] zinkow, j. (2001). wadowice i okolice: monogra�czny przewodnik turystyczny i krajoznawczy po zachodniej części pogórza wielickiego oraz po wschodnich częściach pogórza śląskiego, kotliny oświęcimskiej i beskidu małego. wadowice: wydawnictwo gra�kon. [in polish] 186 k se ni a s tr ze żo ń appendix 1 tab. 1. list of herbaceous species found on the study area; category of frequency on study area: r – rare species, f – frequent species, c – common species; 1–10 – numbers of stands according to �gure 1, m – medicinal species no. name of species plant families stands frequency category medical status 1. achillea millefolium l. asteraceae 1, 2, 3, 4, 5, 7, 8, 9, 10 c m 2. aegopodium podagraria l. apiaceae 1, 2, 3, 4, 5, 6, 7, 8, 9 c m 3. agrimonia eupatoria l. rosaceae 2, 5, 6, 7, 9 f m 4. ajuga reptans l. lamiaceae 1, 4, 5, 7, 10 f – 5. alliaria petiolata (m. bieb.) cavara et grande brassicaceae 6, 8, 9, 10 f m 6. allium ursinum l. alliaceae 7 r m 7. anemone nemorosa l. ranunculaceae 2, 5, 6, 7, 8, 9, 10 c – 8. arabidopsis thaliana l. brassicaceae 1, 2, 3, 4, 5, 6, 7 c – 9. asarum europaeum l. aristolochiaceae 6, 7, 8, 9 f m 10. bellis perinnis l. asteraceae 2, 4, 8, 10 f m 11. calendula o�cinalis l. asteraceae 1 r m 12. caltha palustris l. ranunculaceae 2, 5, 7, 8, 9 f – 13. campanula patula l. campanulaceae 2, 4, 5, 7, 8, 9, 10 c – 14. c. rapunculoides l. campanulaceae 6, 10 r – 15. c. trachelium l. campanulaceae 6, 10 r – 16. capsella bursa-pastoris (l.) medik. brassicaceae 1, 2, 3, 4, 5, 7, 8 c m 17. cardamine amara l. brassicaceae 8, 9, 10 r – 18. c. pratensis l. brassicaceae 2, 4, 5, 7, 10 f m 19. centurea cyanus l. asteraceae 2, 3, 4, 5, 8 f m 20. cerastium holosteoides fr. em. hyl. caryophyllaceae 1, 3, 4, 5, 6, 7, 8, 9 c – 21. chamomilla recutita (l.) rauschert asteraceae 1, 3, 4 r m 22. chamomilla suaveolens (pursh) rydb. asteraceae 1, 3, 4, 6 f m 23. chelidonium majus l. papaveraceae 1, 10 r m 24. cichorium intybus l. asteraceae 1, 3, 4 r m 25. cirsium rivulare (jacq.) all. asteraceae 1, 3, 4, 5, 6, 7, 8, 9 c m 26. convallaria majalis l. convallariaceae 7, 8, 9, 10 f m 27. coronilla varia l. fabaceae 1, 4, 7, 8 f m 28. dactylis glomerata l. poaceae 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 c – 29. dentaria glandulosa waldst. et kitt. brassicaceae 5, 6, 7 r – 30. echium vulgare l. boraginaceae 1, 3, 4 r m 31. equitesum arvense l. equisetaceae 1, 3, 4, 5, 6, 7 f m 187 h erbaceous flora of village c hocznia (southern p oland) w ith particular focus on m edicinal plants 32. euphorbia cyparissias l. euphorbiaceae 3, 4, 5, 7 f m 33. ficaria verna huds. ranunculaceae 2, 5, 7, 8, 9 f m 34. galathus nivalis l. amaryllidaceae 7, 10 r – 35. fragaria vesca l. rosaceae 2, 6, 10 r m 36. galeobdolon luteum huds. lamiaceae 2, 6, 8, 10 f – 37. galium aparine l. rubiaceae 4, 5, 7, 8, 9 f – 38. g. mollugo l. rubiaceae 6, 7, 10 r – 39. geranium robertianum l. geraniaceae 7, 9 r m 40. g. phaeum l. geraniaceae 7, 10 r – 41. geum rivale l. rosaceae 1, 8 r m 42. glechoma hederacea l. lamiaceae 2, 3, 4, 10 f m 43. hesperis matronalis l. brassicaceae 1, 2 r – 44. hieracium murorum l. asteraceae 5, 6, 7, 10 f – 45. humulus lupulus l. cannabaceae 4, 9 r m 46. hypericum perforatum l. hypericaceae 1, 3, 4, 10 f m 47. iris pseudacorus l. iridaceae 1 r – 48. lamium album l. lamiaceae 2, 7, 9, 10 f m 49. lathyrus pratensis l. fabaceae 2, 8 r – 50. leucathemum vulgare lamm. asteraceae 3, 4, 7, 8 f – 51. luzula campestris (l.) dc juncaceae 1, 2, 4, 7, 10 f – 52. lychnis �os-cuculi l. caryophyllaceae 1, 2, 4, 5, 7, 8, 9, 10 c m 53. lysimachia vulgaris l. primulaceae 2, 6, 8, 9 f m 54. lythrum salicaria l. lythraceae 1, 2 r m 55. maianthemum bifolium (l.) f. w. schmidt convallariaceae 5, 6, 7, 10 f – 56. medicago lupulina l. fabaceae 1, 6, 8, 9 f – 57. mentha longifolia (l.) l. lamiaceae 1, 5, 8, 9 f m 58. myosotis palustris (l.) l. em. rchb. boraginaceae 2, 7, 8, 9 f – 59. ononis arvensis l. lamiaceae 4 r m 60. origanum vulgare l. lamiaceae 4 r m 61. oxalis acetosella l. oxalidaceae 6, 7, 8, 9, 10 f m 62. papaver rhoeas l. papaveraceae 2, 3, 4, 8 f m 63. petasites albus l. asteraceae 6, 8 r – 64. phalaris arundinacea l. poaceae 1 r – 65. plantago lanceolata l. plantaginaceae 1, 2, 4, 5, 7, 8, 10 c m 66. p. maior l. plantaginaceae 1, 2, 4, 5, 7, 8, 10 c m 67. p. media l. plantaginaceae 7, 8 r m 68. platanthera bifolia (l.) rich. orchidaceae 9 r – 69. poa pratensis l. poaceae 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 c – 70. polygonatum multi�orum (l.) all. convallariaceae 1, 5, 10 r m 71. potentilla anserina l. rosaceae 1, 2, 7, 8, 9 f m 72. p. erecta (l.) raeusch. rosaceae 8, 9 r m 188 k se ni a s tr ze żo ń 73. primula elatior (l.) hill primulaceae 2, 5, 8, 9, 10 f m 74. prunella vulgaris l. lamiaceae 2, 5, 6, 8 f m 75. ranunculus acris l. ranunculaceae 2, 3, 4, 5, 7, 8, 9, 10 c m 76. r. bulbosus l. ranunculaceae 2, 7, 8 r – 77. rudbeckia laciniata l. asteraceae 1 r – 78. rumex acetosa l. polygonaceae 2, 4, 5, 6, 7, 8, 9, 10 c m 79. salvia pratensis l. lamiaceae 1 r m 80. senecio jacobaea l. asteraceae 3, 4, 5, 6, 7, 10 f – 81. symphytum o�cinale l. boraginaceae 8, 9 r m 82. s. tuberosum l. boraginaceae 5, 6, 10 r – 83. taraxacum o�cinale f.h. wigg asteraceae 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 c m 84. �laspi arvense l. brassicaceae 3, 4, 5, 8 f m 85. �ymus serphyllum l. lamiaceae 1 r m 86. trifolium pratense l. fabaceae 2, 7, 8, 10 f m 87. t. repens l. fabaceae 2, 7, 8, 9, 10 f m 88. typha latifolia l. typhaceae 1 r – 89. urtica dioica l. urticaceae 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 c m 90. vaccinium myrtillus l. ericaceae 6, 7, 9 r m 91. valeriana simplicifolia (rchb.) kabath valerianaceae 2, 9 r – 92. veronica chamaedrys l. scrophulariace-ae 1, 2, 4, 5, 6, 7, 8, 9, 10 c – 93. v. o�cinalis l. scrophulariace-ae 2, 5, 6, 7 f m 94. vicia sepium l. fabaceae 2, 5, 7, 8, 9 f – 95. vinca minor l. apocynaceae 9 r m 96. vincetoxinum hirudinaria medik. apocynaceae 1, 2, 5, 10 f m 97. viola arvensis murray. violaceae 2, 3, 8 r m 98. v. reichenbachiana boreau violaceae 2, 5, 6, 8 f – 99. viscaria vulgaris rohl. caryophyllaceae 1, 5, 8 r – 189 h erbaceous flora of village c hocznia (southern p oland) w ith particular focus on m edicinal plants flora zielna miejscowości chocznia (południowa polska), ze szczególnym uwzględnieniem roślin leczniczych streszczenie w polsce większość roślin leczniczych nadal pozyskuje się z naturalnych siedlisk. jednak w niektórych regionach, zwłaszcza w okolicach dużych miast, antropogeniczne oddziaływania i zanieczyszczenia powodują, że zbiory tych roślin są niemożliwe. dlatego bardzo ważne jest, aby szukać miejsc, w których koncentracja roślin leczniczych będzie wystarczająca do pozyskiwania. celem niniejszej pracy było zbadanie składu �ory zielnej w obrębie wsi chocznia (województwo małopolskie, polska południowa), ze szczególnym zwróceniem uwagi na gatunki lecznicze. w różnych warunkach siedliskowych wyznaczono na badanym obszarze 10 stanowisk, z których zbierano materiał roślinny. zbiory wykonano w okresach wegetacyjnych w 2014 i 2015 roku. materiały zbierano kilka razy w ciągu sezonu, zwracając uwagę na fenologię gatunków. dokumentacją badań był zielnik, zdeponowany na uniwersytecie pedagogicznym w krakowie w zakładzie botaniki. analiza �orystyczna stanowisk wykazała łącznie 99 gatunków roślin zielnych z 37 rodzin. ponad połowa z nich (60 gatunków) posiadała właściwości lecznicze. najwięcej gatunków leczniczych reprezentowało rodziny: asteraceae (9 gatunków), lamiaceae (7 gatunków), rosaceae (5 gatunków), fabaceae i brassicaceae (po 4 gatunki), plantaginaceae (3 gatunki). wśród roślin leczniczych, gatunków częstych w skali terenu stwierdzono 27, a pospolitych 11. potencjalnie do zbioru na potrzeby ziołolecznictwa nadaje się tu łącznie 38 gatunków. badane stanowiska położone są z dala od strefy zanieczyszczeń, dużych miast i autostrad. stąd miejscowość ta wydaje się być dobrym obszarem do zbierania surowców zielarskich na potrzeby przemysłu farmaceutycznego i na użytek własny. key words: �ora, herbaceous, plant, medicinal plant, southern poland received: [2016.09.28] accepted: [2016.10.17] 107 annales universitatis paedagogicae cracoviensis studia naturae, 2: 107–113, 2017, issn 2543-8832 doi: 10.24917/25438832.2.8 anatoliy a. khapugin joint directorate of the mordovia state nature reserve and national park “smolny”, republic of mordovia, saransk, russia, *hapugin88@yandex.ru. a record of silene viscaria (l.) jess. (caryophyllaceae) with achromatic flowers in the mordovia state nature reserve (central russia) introduction silene viscaria (l.) jess. (syn.: lychnis viscaria l., steris viscaria (l.) ra�n., viscaria viscosa asch., v. vulgaris rohl.) is a perennial 25–80 cm high herb: stem erect, green, not branching in lower portion, glabrous, upper portion of the upper internodes glutinous, with two to �ve distinct internodes (clapham et al., 1981; gubanov et al., 2003). it inhabits dry grasslands, open forests, forest clearings, and ledges (kurtto, wesenberg, 2001; gubanov et al., 2003). s. viscaria is distributed in most of europe excluding the iberian peninsula, northern scandinavia, northern russia, most of south italy, and southern greece (jalas, suominen, 1986). moreover, it is an occasional and alien garden species in eastern north america (morton, 2005). in�orescences are compound dichasia, lax or slightly congested. each of them bear about 20–25 �owers. �e �owers are pollinated by insects, mainly bumblebees and butter�ies (jennersten, 1988). �e seeds are dispersed by gravity. in most literature, the colour of s. viscaria �owers is indicated as purple, purple-red, pink, or crimson (clapham et al., 1981; gubanov et al., 2003; morton, 2005; frajman et al., 2013). only few authors indicate cases of achromatism for s. viscaria �owers (gubanov et al., 2003; frajman et al., 2013). moreover, there is a lack of data on di�erences of white-�ower, and plants with normal coloured �owers. in particular, we cannot evaluate the frequency of their registering of the existence/absence of morphological di�erences between whiteand purple-�ower plants. nevertheless, the �owers colours play a signi�cant role both for plants and their insect pollinators (miller et al., 2011), because insects discriminate a wide range of patterns and shapes (dyer et al., 2008; hempel de ibarra et al., 2015), and entomophillous plants are highly dependent of insects. �at is why, in this study, we aimed to compare some morphological parameters of generative plants with achromatic �owers and individuals with purple ones in the same coenopopulations in conditions of the mordovia state nature reserve (central russia). a na to liy a . k ha pu gi n 108 material and methods �e �eld investigations were carried out at the southern border of the federal protected area, mordovia state nature reserve (54°7504ʹn; 43°4016ʹe), in june 2017. population studies were conducted according to aleksandrova (1964) and khapugin et al. (2014). �ree study plots (1×1 m) were established in silene viscaria coenopopulations where individuals with achromatic �owers were found. �e composition of accompanying �ora was recorded within square plots 10×10 m situated around small (1 m2) study plots established for the study of s. viscaria individuals. assessment of the s. viscaria individuals status was carried out on the basis of the morphological parameters of generative plants (20 plants with achromatic �owers vs. 57 plant with normally coloured �owers): height of plants, number of whorls of lateral branches per an in�orescence, number of �owers per an in�orescence, length of an in�orescence, and the percentage of an in�orescence length from a plants height. �e nomenclature and the taxonomy of the plant taxa of the accompanying �ora (appendix 1 – tab. 1) are presented in accordance with �e plant list… (2013) and euro+med plantbase (2006–2011). statistical analysis was performed in ms excel and past (hammer et al., 2001). �erefore, we used principal component analysis (pca) to de�ne the main morphometric parameters that di�erentiate both groups of plants in studied habitats. for interpretation of the ordination axes, values of measured morphometric parameters were plotted onto a pca ordination diagram as supplementary environmental data. results and discussion among the many species of colourful �owers, sometimes the specimens lacking this colour are referred to as albinotic forms. examples of 64 species of this type of plant were reported by czarna (2006) and others. a more detailed description of albinism in orchids was given, e.g., griesbach (1979), kohns and schneider (1993), selosse et al. (2004), and jakubska and schmidt (2005). examples of albinotic plants are also mentioned by trudell et al. (2003), and śliwiński and jakubska-busse (2010). however, there has been no analysis of the albinotic population of silene viscaria, which was the subject of the investigatation undertaken in this study. as a result of investigations, several records of white-�ower s. viscaria plants were found at the south of the mordovia state nature reserve (fig. 1). �ey occur within the same plant communities with s. viscaria individuals with �owers normally coloured. �e composition of �ora accompanying to s. viscaria in studied locations is presented by 37 species belonging to 34 genera and 22 families (appendix 1 – tab. 1). of these, 109 poaceae (6 species), plantaginaceae (4 species), rosaceae (3 species), asteraceae (3 species), and caryophyllaceae (3 species) contain the highest numbers of representatives. �e mean values of morphological parameters were determined for s. viscaria �owering individuals within three study plots. as a result, we found that these parameters are similar between plants with achromatic �owers and plants with typically coloured �owers (tab. 2). similarly, binkenstein and schaefer (2015) compared �ower colours of di�erent habitats and chromatic / achromatic components of �ower colours from the honeybees’ point of view. �ey found that �ower colours do not di�er between closed forest and open grassland habitats in any chromatic or achromatic aspect both from the bees’ perspective and without any model bias. fig. 1. examples of silene viscaria (l.) jess. �owering plants with achromatic (a) and normal �owers (b) at the southern border of the mordovia state nature reserve (photo. a.a. khapugin) tab. 2. morphological traits of silene viscaria (l.) jess. �owering plants with di�erent colouring of �owers parameters morphological traits height of individuals [cm] number of whorls of lateral branches per an in�orescence number of �owers per an in�orescence length of an in�orescence [cm] percent of an in�orescence length to a plant height [%] a n a n a n a n a n m 67.3 55.4 4.3 4.4 40.5 51.0 16.9 16.4 24.9 29.8 m 2.9 1.8 0.2 0.1 2.9 2.8 1.5 0.9 1.8 1.4 min 55.0 35.0 3.0 3.0 26.0 19.0 9.0 8.0 16.1 17.0 max 78.0 71.0 5.0 5.0 56.0 67.0 24.0 25.0 33.6 50.0 notes: m – mean value, m – error of the mean, min–max – minimal–maximal values; a – plants with achromatic �owers; n – plants with normally coloured �owers a record of silene viscaria (l.) jess. (c aryophyllaceae) w ith achrom atic flow ers in the m ordovia s tate n ature r eserve (c entral r ussia) a b a na to liy a . k ha pu gi n 110 in order to determine the presence/absence between both study plots and two groups of s. viscaria plants, we carried out conjoint principal component analysis (pca) with the involvement of averaged values of all measured morphometric traits in three study plots (fig. 2). despite of high similarity in morphological traits, plants with achromatic �owers are slightly higher than �owering individuals of s. viscaria with normally coloured �owers. although, on the basis of all obtained results, we may conclude that there are not signi�cant di�erences between s. viscaria plants with achromatic �owers and individuals with coloured �owers. we suggest accumulating data on a phenomenon of �owers achromatism amongst di�erent groups of plants in di�erent areas of the world. it will allow the understanding of the frequency of this phenomenon and, perhaps, its relationship with the biological traits of the plants. references aleksandrova, v.d. (1964). �e study of changes of vegetation cover. in: e.m. lavrenko, a.a. korchagina (eds.), field geobotany. moscow–leningrad: nauka, 399–447. [in russian] binkenstein, j., schaefer, h.m. (2015). flower colours in temperate forest and grassland habitats: a comparative study. arthropod-plant interactions, 9, 289–299. doi: 10.1007/s11829-015-9369-9 clapham, a.r., tutin, t.g., warburg, e.f. (1981). flora of the british isles. london: cambridge university press. czarna, a. (2006). albinotic �owers in plants widly growing on the area of the wielkopolska lowland. rocznik akademii rolniczej w poznaniu, 378, 37–43. fig. 2. principal component analysis (pca) ordination diagram of �owering silene viscaria individuals with achromatic (grey symbols) and normally coloured (black symbols) �owers for �rst (triangles), second (circles), and third (squares) study plots; all groups are arranged in accordance to morphologic traits: h – height of individuals, n – number of whorls of lateral branches per an in�orescence, f – number of �owers per an in�orescence, l – length of an in�orescence, p – percentage of an in�orescence length from a plants height 111 dyer, a.g., rosa, m.g., reser, d.h. (2008). honeybees can recognise images of complex natural scenes for use as potential landmarks. journal of experimental biology, 211(8), 1180–1186. doi: 10.1242/ jeb.016683. euro+med. (2006–2011). euro+med plantbase – the information resource for euro-mediterranean plant diversity. http://ww2.bgbm.org/europlusmed/ frajman, b., �ollesson, m., oxelman, b. (2013). taxonomic revision of atocion and viscaria (sileneae, caryophyllaceae). botanical journal of the linnean society, 173(2), 194–210. doi: 10.1111/boj.12090 griesbach, r.j. (1979). �e albino form of epipactis helleborine. american orchid society bulletin, 48(8), 808–809. gubanov, i.a., kiseleva, k.v., novikov, v.s., tikhomirov, v.n. (2003). illustrated determinants of plants of the middle russia 2: angiosperms (dicotyledons: dialypetalous). moscow: kmk scienti�c press ltd., institute of technological researches. [in russian] hammer, ø., harper, d.a.t., ryan, p.d. (2001). past: paleontological statistics so�ware pack-age for education and data analysis. palaeontologia electronica, 4(1), 1–9. http://palaeo-electronica.org/2001_1/ past/past.pdf hempel de ibarra, n., langridge, k.v, vorobyev, m. (2015). more than colour attraction: behavioural functions of �ower patterns. current opinion in insect science, 12, 64–70. doi: 10.1016/j. cois.2015.09.005 jakubska, a., schmidt, i. (2005). chlorophyll-free form of epipactis albensis novkov et rydlo (orchidaceae, neottieae) in the skarpa storczyków nature reserve near orsk (lover silesia, poland). acta botanica silesiaca, 2, 151–154. jalas, j., suominen, j. (1986). atlas florae europaeae, 7. caryophyllaceae (silenoideae). helsinki: committee for mapping the flora of europe and societas biologica fennica vanamo. jennersten, o. (1988). pollination of viscaria vulgaris (caryophyllaceae): the contributions of diurnal and nocturnal insects to seed set and seed predation. oikos, 52, 319–327. doi: 10.2307/3565205 khapugin, a.a., vargot, e.v., chugunov, g.g. (2014). research methods of vegetative cover of terrestrial ecosystems. in: l.v. egorov, a.b. ruchin, a.a. khapugin, o.n. artaev (eds.), methods of �eld environmental research. chapter 1. saransk: pushta, 4–42. [in russian] kohns, p., schneider, p. (1993). epipactis muelleri godfery var. chlorotica. die orchidee, 44(2), 104–105. [in german] kurtto, a., wesenberg, j. (2001). viscaria bernh. in: b. jonsell (ed.), flora nordica, 2. stockholm: �e bergius foundation, �e royal swedish academy of sciences, 172–175. miller, r., owens, s.j., rørslett, b. (2011). plants and colour: flowers and pollination. optics & lasertechnology, 43, 282–294. doi: 10.1016/j.optlastec.2008.12.018 morton, j.k. (2005). silene. in: flora of north america editorial committee (ed.), flora of north america north of mexico, 5. new york & oxford: oxford university press, 166–214. selosse, m.a., faccio, a., scappaticci, g., bonfante, p. (2004). chlorophyllous and achlorophyllous specimens of epipactis microphylla (neottieae, orchidaceae) are associated with ectomycorrhizal septomycetes, including tru�es. microbial ecology, 47(4), 416–426. doi: 10.1007/s00248-003-2034-3 śliwiński, m., jakubska-busse, a. (2010). albinotyczne kwiaty i formy roślin. zielona planeta, 2(89), 9–10. http://www.ekoklub.ehost.pl/pke_od/images/zp89.pdf [in polish] �e plant list. a working list of plant species (2013). �e plant list. version 1.1. http://www.theplantlist.org/ trudell, s.a., rygiewicz, p.t., edmonds, r.l. (2003). nitrogen and carbon stable isotope abundances support the mycoheterotrophic nature and host-speci�city of certain achlorophyllous plants. new phytologist, 160, 391–401. doi: 10.1046/j.1469-8137.2003.00876.x. a record of silene viscaria (l.) jess. (c aryophyllaceae) w ith achrom atic flow ers in the m ordovia s tate n ature r eserve (c entral r ussia) a na to liy a . k ha pu gi n 112 appendix 1 tab. 1. list of plant taxa growing together with silene viscaria in studied plant communities at the southern border of the mordovia state nature reserve no. species family plot 1 plot 2 plot 3 1. achillea millefolium l. asteraceae + + + 2. anthoxanthum odoratum l. poaceae + + + 3. betula pendula roth betulaceae + + 4. calamagrostis epigejos (l.) roth poaceae + 5. campanula patula l. campanulaceae + + 6. convallaria majalis l. asparagaceae + 7. festuca valesiaca schleich. ex gaudin poaceae + 8. fragaria vesca l. rosaceae + + 9. frangula alnus mill. rhamnaceae + 10. galium mollugo l. rubiaceae + + + 11. hypericum perforatum l. hypericaceae + + + 12. leucanthemum vulgare l. asteraceae + 13. linaria vulgaris mill. plantaginaceae + 14. luzula pilosa (l.) willd. juncaceae + + + 15. malus domestica borkh. rosaceae + 16. melampyrum nemorosum l. orobanchaceae + 17. m. pratense l. orobanchaceae + + + 18. melica nutans l. poaceae + + 19. orthilia secunda (l.) house ericaceae + 20. phleum pratense l. poaceae + 21. pilosella o cinarum vaill. asteraceae + + + 22. pimpinella saxifraga l. apiaceae + 23. pinus sylvestris l. pinaceae + + + 24. plantago lanceolata l. plantaginaceae + 25. platanthera bifolia (l.) rich. orchidaceae + 26. poa pratensis l. poaceae + + 27. prunella vulgaris l. lamiaceae + 28. ranunculus polyanthemos l. ranunculaceae + 29. rumex acetosella l. polygonaceae + 30. sorbus aucuparia l. rosaceae + + 31. stellaria graminea l. caryophyllaceae + 32. s. holostea l. caryophyllaceae + 33. silene viscaria (l.) jess. caryophyllaceae + + + 34. veronica chamaedrys l. plantaginaceae + + + 35. v. o cinalis l. plantaginaceae + 36. vicia sylvatica l. fabaceae + 37. viola mirabilis l. violaceae + 113 abstract silene viscaria (l.) jess. is a common species of central russian �ora. it has attractive purple, crimson, or dark-pink in�orescences. some literature sources indicate the possibility of white-colour �owers. however, there are no reliable published evidences of these cases. in this report, a record of s. viscaria plants with achromatic, white, �owers at the southern border of the mordovia state nature reserve is presented. some morphological traits of �owering individuals were measured: the height of reproductive individuals, the number of whorls of lateral branches per an in�orescence, the number of �owers per an in�orescence, the length of an in�orescence, and the percentage of an in�orescence length from a �owering plants height. data on the �ora accompanying s. viscaria are presented. as a result, no signi�cant di�erences between plants with achromatic �owers and plants with coloured �owers have been found. however, the height of white-�ower individuals was slightly higher. we suggest accumulating data on a phenomenon of the �ower’s achromatism amongst di�erent groups of plants in order to try to understand the frequency of this phenomenon and perhaps its impact on plants biology. key words: achromatism, �ower colour, morphological features, plant population, russia received: [2017.07.09] accepted: [2017.11.14] stanowisko silene viscaria (l.) jess. (caryophyllaceae) z kwiatami achromatycznymi w rezerwacie przyrody mordovia (rosja centralna) streszczenie silene viscaria (l.) jess. jest gatunkiem pospolitym we �orze centralnej rosji. ma atrakcyjne �oletowe, szkarłatne lub ciemnoróżowe kwiatostany. niektóre źródła literatury wskazywały na możliwość występowania kwiatów w kolorze białym u tego gatunku. jednak dotychczas nie było wiarygodnych opublikowanych danych na ten temat. w niniejszym opracowaniu zaprezentowano stanowisko s. viscaria z achromatycznymi, białymi kwiatami, zlokalizowane na południowej granicy rezerwatu przyrody mordovia. w trakcie badań zmierzono niektóre cechy morfologiczne osobników kwitnących: wysokość osobników reprodukcyjnych, liczbę okółków odgałęzień bocznych na kwiatostan, liczbę kwiatów na kwiatostan, długość kwiatostanu, procent długości kwiatostanu z długości rośliny kwitnącej. przedstawiono również dane dotyczące �ory towarzyszącej s. viscaria. w rezultacie nie stwierdzono różnic istotnych między roślinami o achromatycznych kwiatach i roślinami o kolorowych kwiatach. jednak długość osobników z białymi kwiatami była nieco większa. byłoby wskazane gromadzenie dalszych danych dotyczących zjawiska achromatyzmu kwiatów wśród różnych grup roślin, aby spróbować zrozumieć częstotliwość tego zjawiska i być może jego wpływ na biologię roślin. słowa kluczowe: achromatyzm, kwiaty barwne, cechy morfologiczne, populacje roślin, rosja information on the author anatoliy a. khapugin phd, senior researcher of the joint directorate of the mordovia state nature reserve and national park “smolny”. at present, his research and professional interests cover the following topics: �oristic and geobotanical studies, biodiversity and conservation, rare plants, population-based studies, forestry, and invasive plant species. a record of silene viscaria (l.) jess. (c aryophyllaceae) w ith achrom atic flow ers in the m ordovia s tate n ature r eserve (c entral r ussia) 165 annales universitatis paedagogicae cracoviensis studia naturae, 1: 165–178, 2016, issn 2543-8832 edgar mó1,2, william cetzal-ix3*, eliana noguera-savelli4, saikat kumar basu5, peiman zandi6, katarzyna możdżeń7 1 orquideario agronomía-cunor-usac, centro universitario del norte, universidad de san carlos de guatemala, cobán, alta verapaz, guatemala 2 orquigonia, centro de rescate y conservación de orquídeas, cobán, alta verapaz, guatemala 3 instituto tecnológico de chiná, calle 11 entre 22 y 28, colonia centro chiná 24 050, campeche, méxico, *rolito22@hotmail.com 4 catedrático conacyt, colegio de postgraduados campus campeche. carretera haltunchén-edzná km. 17.5, sichochac, champotón, campeche, méxico, c.p. 24450 5 department of biological sciences, university of lethbridge, lethbridge, alberta, canada 6 institute of crop science, chinese academy of agricultural sciences, beijing 100081, people’s republic of china 7 department of plant physiology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland cyperus canus j. presl & c. presl – a traditional source of fiber, its uses, products and cultural significances among ethnic communities of central america �e genus cyperus l. (cyperaceae) includes between 750 and 950 species (mostly annuals and perennials), the exact number depends on the accepted generic or unresolved names (larridon et al., 2011, �e plant list, 2013). most species of cyperus are predominantly aquatic to sub-aquatic in habitat (marshes, swamps, bogs, lakes, ponds, fens and grassland) and are characterised by their distinct hollow, circular or triangular stems with distinct ridges and furrows predominantly present in the tropics, sub-tropics, temperate and the sub-temperate regions (adams, 1994; tucker, 1983, 1994). �e genus is found in close association and ecological assemblages with different species of grasses (e.g. scleria secans (l.) urb., echinochloa colona (l.) link, pennisetum purpureum schumach., urochloa platyphylla (munro ex c. wright) r. d. webster and zea mays l.). both families have close evolutionary a�nities. �e species within the genus vary greatly in height from 3–5 cm at one end of the spectrum, to as high as over 4 or 5 m on the other. �e plants usually thrive well in water less than 1 m in depth (adams, 1994; tucker, 1983, 1994). �e hollow stems usually bear the slender leaves towards the base of the plant, compactly arranged on a whorl at the apex of the �oral items (castro-ramírez et al., 1991; castro-ramírez, 1994). �e �owers vary in color between greenish to pale whitish or greenish white, �owers are mostly anemophilous (wild pollinated) and �owers are aggregated in di�erent clusters between the leaves at the apex. �e aggre166 ed ga r m ó, w ill ia m c et za l-i x, e lia na n og ue ra -s av el li, s ai ka t k um ar b as u, p ei m an z an di , k at ar zy na m oż dż eń gation of the �owers on the apex of the stem is arranged in the form of an umbrella (standley, 1931; castro-ramírez et al., 1991; castro-ramírez, 1994). cyperus species have cosmopolitan distribution, some of these are considered as noxious weeds, while a large number of others have di�erent agricultural, horticultural, medicinal, pharmaceutical and industrial uses (jansen, 1993, 1998; magaña-alejandro et al., 2014). �e most emblematic species of the genus by its historical and utilitarian context is c. papyrus l., a plant used by the ancient egyptians to make paper. other species of economic importance are: c. articulatus l., c. canus j. presl. & c. presl., c. esculentus l. and c. rotundus l. (standley, steyermark, 1958; eshbaugh, 1983; tucker, 1983, 1994). in central america, c. canus is a plant widely used by the indigenous communities of mexico, guatemala, el salvador, honduras and nicaragua for the principal elaboration of mats (‘petates’) (cajas et al., 2009). secondarily for other objects such as baby carrier, briefcases, folders and occasionally hats; also for all types of moorings, as a cords or ropes. at home it is used to start �re in the kitchen (ludlow-wiechers, diego-pérez, 2002; cajas et al., 2009; magaña-alejandro et al., 2014). �e stems of this plant are used as an excellent �ber for weaving, in making high quality ropes and also for producing blowers, backpacks, hats and sandals used commonly by the local people (castro-ramírez et al., 1991; castro-ramírez, 1994; cajas et al., 2009). �e �bers are also applied by local people to make handicra�s such as baskets and bags. it is also used in preparing traditional food products of guatemala mostly for tying up sausages and food bags (‘pita’) (fig. 2 – appendix). c. canus appears in the local culture historically, as mentioned in the popol vuh. �is plant is commonly used in districts of alta verapaz, chimaltenango, escuintla, guatemala, quiché, sacatepéquez, santa rosa, retalhuleu and zacapa (castro-ramírez et al., 1991; adams, 1994; castro-ramírez, 1994; cajas et al., 2009). �e use and traditional knowledge of c. canus among the indigenous communities of central america have been transferred through successive generations as part of the local tradition and culture. however, these traditional activities are now being slowly abandoned; since the plant products are being replaced by modern chemistry-based synthetic products (hardy nelson, 1986; jansen, 1993, 1998; roquas, 1994). in this study we describe the uses and production processes of some of the most important products made from c. canus. systematics and distribution class: equisetopsida c. agardh subclass: magnoliidae novák ex takht. superorder: lilianae takht. order: poales small family: cyperaceae juss. genus: cyperus l. species: cyperus canus j. presl. & c. presl. 167 c. canus (fig. 3 – appendix 1) was collected by haenke in ‘regno mexicano’ (currently mexico) and described by presl and presl in 1828 in the work reliquiae haenkeanae (reliq. haenk.) 1(3): 179. �is species is a native plant of tropical america, distributed from southern mexico and guatemala to colombia (fig. 3 – appendix 1). it grows along the shores of lakes, streams and marshy places and is sometimes found on rocks, at altitudes of 200–1500 m a.m.s.l. (standley, 1931; castro-ramírez et al., 1991; castro-ramírez, 1994). however, it is quite rare to �nd the plants in the wild since locals treat them as a common weed (fig. 4 – appendix 1). common names of cyperus canus (tule) �e species is commonly referred to as ‘tule’. �e word originated from the nahuatl word ‘tullin’ or ‘tolin’ and is commonly used to mean herbs growing in the lakes and marshy creeks, that are applied in the manufacture of chairs, baskets, mats, and several other handicra�s (castro-ramírez et al., 1991; castro-ramírez, 1994). however, according to del paso y troncoso (1986), the word ‘tullin’ corresponds to the generic name used for several species of cyperaceae, typhaceae, poaceae and ponteridaceae (ludlow-wiechers, diego-pérez, 2002). from this perspective, martínez (1979) associated the name ‘tule’ to several di�erent species, such as: c. articulatus l., c. tenerrimus j. presl & c. presl., c. canus, pontederia cordata l., schoenoplectus americanus (pers.) volkart ex schinz & r. keller, s. validus (vahl) á. löve & löve, typha domingensis pers., and t. latifolia l. �e plant c. canus is called ‘sak’ in local mayan language fig. 1. distribution of cyperus canus in central america (based on herbarium records of tropicos, 2015) cyperus canus j. p resl & c . p resl – a traditional source of fiber, its uses, products and cultural significances am ong ethnic com m unities of c entral a m erica 168 ed ga r m ó, w ill ia m c et za l-i x, e lia na n og ue ra -s av el li, s ai ka t k um ar b as u, p ei m an z an di , k at ar zy na m oż dż eń due to the similarity of the stems to sugar cane (diego-pérez, 2010). it is also known in spanish as ‘tule negro’, ‘tule de petate’ (gonzález-mayorga, 2009), ‘junquillo’, ‘sivate’, ‘ca�ita’; and ‘say’ in q’eqchi’ language (standley, steyermark, 1958). harvesting process of tule in central america the harvesting season of tule steams starts a�er the blooming period (otherwise these are not well dried). �e stems are all cut down and a�er harvest is completed, the separation of the stems is conducted depending on the quality of the harvested stem. �e decayed or rotten stems are discarded and only the healthy ones are carefully scrutinised and selected for use downstream (gonzález-mayorca, 2009). �e harvest period is from february to may, tiller plants produce 75–200 stems per plant, in each crop growing up to 400–500 plants, but in the drying step, 400 stems are equivalent to one-pound weight. preparation of tule �e selected stems are placed under the sun for proper drying depending upon the criteria of the producer and based on local customs. it takes 4–5 days for drying in salvador, but even for 8 days in guatemala. once dried, stems are classi�ed according to the length of the rods since the longest stems enable the producers to develop high quality parts without �aws in the quality of the �bers. �e classi�ed stem is properly stored in high and dry places for thorough ventilation and also for protection against insects. �ey are usually wrapped in nylon bags and under proper storage conditions can be successfully stored for a year or so. �e triangular shape of the stem helps in producing three strands of �bers that are extracted using an indigenous tool known as ‘güiscoyol’ in guatemala. �is is locally made from palms (bactris sp.) or by using speci�c parts of bone as part of the traditional knowledge and technology used in the production process. in salvador, the tool used is called ‘rajador’. in both cases these tools are made by the same cra�smen or artisans (gonzález-mayorga, 2009). in guatemala, the central portion of the stem is known as the ‘heart of tule’ or ‘gut of tule’ (spanish: corazón del tule or tripa de tule) or ‘cibaque’. in salvador, it is called ‘rope or heart of tule’ (spanish: mecate o corazón de tule). �is �ber is also used in the manufacture of handbags, sandals, blowers, hats, side bags, sacks etc. fiber cra�s basketry is one of the oldest human activities from the dawn of human civilisation across the planet and constitutes an indispensable part of the daily tasks of several indigenous communities (e.g. maya and chontal maya people in mexico, q’eqchi’, xinca, poqomchi’, ch’orti’ and kaqchikel people in guatemala). local women historically have developed exceptional skills and knowledge over several generations in the art 169 of basketry and related textiles trades by judicious use of locally available resources. �ey have been solely responsible for transferring this traditional knowledge of manufacturing di�erent basketry products to their future generations (bustos, 1994). one of the most commonly and historically known traditional local products made from c. canus is the ‘petate’. in central america (mesoamerica), the mayas used basketry for making di�erent ceremonial utensils and for their daily use, that is demonstrated in the codices, steles and ceramics; where di�erent products such as blowers, belts, baskets and mats were presented (cajas et al., 2009). mats ‘petates’ tab. 1. comparative analysis of use and prices of di�erent cra�swomen using tule in preparing traditional �ber-based products and handicra�s cra�swomen comments uses prices carmen chocooj people only buy stems for mooring food mooring tamale ‘tamales’ 140 stems or strands (us$ 3.93) 4–5 stems or strands (us$ 0.66) elisa cú tule blowers do not sell much because it is comparatively costlier than the plastic blowers blowers and ropes for mooring candles blowers (us$ 1.31) eulalia teyul young generations do not practice this activity since they are not interested in learning this traditional cra�. she makes 6 blowers per day (5 am – 6 pm); and a mat of 60×40 cm (3 hours), 2×1.25–1.30 m (1 day), 5×1.25–1.30 m (3 days) mats and blowers 25 stems for a blower (us$ 1.31), mats of 60×40 cm (us$ 1.31), 2×1.25–1.30 m (us$ 1.97), 5×1.25–1.30 m (us$ 2.62) for the preparation of a mat (depending on size) ~80–210 stem barks of c. canus are commonly used depending on the target size or dimension of the �nished mat. generally, weaving is conducted on the �oor, on the other ‘petate’, bed sheet or nylon material. in some cases it is also done directly on the dirt �oor. �e bark is initially extended in a vertical arrangement and a�erwards the interweaving of the barks are made on a horizontal direction one a�er another. as the work proceeds, more stems are added vertically and interwoven horizontally; leaving a fraction non-interlaced to make a moored that �nally serves as the basic frame or contour (fig. 5 – appendix 1). blowers or fans �e blower or fan is generally prepared by using ~25 stems of c. canus that are carefully selected, having equal length and �exible consistency to facilitate the interlacing process. alternatively, water is repeatedly applied to appropriately so�en the stems to cyperus canus j. p resl & c . p resl – a traditional source of fiber, its uses, products and cultural significances am ong ethnic com m unities of c entral a m erica 170 ed ga r m ó, w ill ia m c et za l-i x, e lia na n og ue ra -s av el li, s ai ka t k um ar b as u, p ei m an z an di , k at ar zy na m oż dż eń make them more manageable and easy for the trade. �e process of weaving starts by mooring a stem in half on the wooden benches; where 6–10 stems are placed, next doubled in half and are then woven together. as they get interweaving other stems are added until the blower takes the shape. before making the middle blower, the base is made because it is the point of attachment of the 25 stems, then the base or gripper will be thick and resistant to movement (fig. 6 – appendix 1). we conducted personal interviews with women artisans in alta verapaz, guatemala, and we share some interesting facts about the production and costs of c. canus cra�s (tab. 1). market scenario of the tule products studies of the situation of tule �ber artisans in central america (mesoamerica) indicate recurrent problems in the ethnic communities studied across di�erent countries. although there is a demand for traditional tule �ber-based handicra�s and products in the local and regional markets, as well as with foreign tourists, the economic conditions of the artisans are deplorable since this is a poorly organised sector. as a result traditional artisans and cra�smen and women are heavily dependent on middlemen who have better knowledge of the markets, gross product prices and the distribution systems. �ese middlemen make the most of the existing system retaining greater share of the pro�t to themselves compared to the artisans and cra�smen and women. based on a case study on tule artisans from guatemala, lemus herrera et al. (2009) observed the importance of gender equity in the community development programs; particularly in areas where women do majority of cra�s work for their livelihoods and barter. in these rural areas majority of development programs unfortunately exclude women due to cultural practices in which most power is exercised and enjoyed by men in the traditional societies. �e researchers therefore suggested that it will be important to improve the quality of life of local indigenous womenfolk by empowering them in their communities and societies. another case study from central america that is worth mentioning is that of the masatepe municipality from nicaragua (santana et al., 2004). in this community too, the indigenous �ber-based cra�s work on tule for preparation of mats and related products are conducted mostly by women (86%). �eir average weekly pay varies between $us 22–29 (300–400 córdobas). several of the local �bre products are regularly exported to costa rica, mexico, the us and the european union. �is is an unorganised cottage industry sector and hence exact data for the trade is not available or published. however, based on general estimates it could be mentioned that the trade is quite small in scale compared to other organised production sectors in central america. �e price of mat generated with one or two bunches of tule (each constituting on average ~150 leaves of cyperus canus) is ~us$ 22 (300 córdobas). monthly production is between 6–12 medium size mats, single, without decoration with pricing of 171 us$ 4–11 (50–150 córdobas) each. �e best quality mats with drawings and decorations vary in price range between us$ 11–22 (150–300 córdobas) each. however, the latter is made in smaller quantities only, around 6 mats per workshop unit (santana et al., 2004). in general, tule artisans in central america need consultancies in terms of education, training, empowerment and funding to help them develop into an organised sector for earning better economic return for their traditional artisan work and cra�smanship. �is will allow them to slowly establish an e�cient and e�ective production system in terms of better technology, diversi�cation and improving the quality of their products based on current market demands, better product distribution network and �nancial management. references adams, c.d. 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[in spanish] santana, r., montagnini, f., louman, b., villalobos, r., gómez, m. (2004). la industria de artesanías de masaya y masatepe, nicaragua: demanda por materia prima de bosques tropicales. recursos naturales y ambiente, 42, 77–85. [in spanish] standley, p.c. (1931). �e cyperaceae of central america. field museum of natural history, botanical series, 8, 237–292. standley, p.c., steyermark, j. (1958). cyperaceae, en flora of guatemala. fieldiana: botany, 24(1), 90–196. �e plant list. (2013). a working list of all plant species (version 1.1). http://www.theplantlist.org tropicos. (2015). missouri botanical garden. http://www.tropicos.org/name/9904590 tucker, g.c. (1983). �e taxonomy of cyperus (cyperaceae) in costa rica and panama. systematic botany monographs, 2, 1–85. tucker, g.c. (1994). revision of the mexican species of cyperus (cyperaceae). systematic botany monographs, 43, 1–213. 173 fig. 2. di�erent products and cra�s made with tule; a – blower, b – a plastic blower for comparison; processed �ber strands: c–d – ropes for tying up sausages, e – ropes for tying up tamales, f – ropes for hanging candles, g – ropes for tying up food bags, h – �nished high quality mats (photo. e. mó) appendix 1 cyperus canus j. p resl & c . p resl – a traditional source of fiber, its uses, products and cultural significances am ong ethnic com m unities of c entral a m erica 174 ed ga r m ó, w ill ia m c et za l-i x, e lia na n og ue ra -s av el li, s ai ka t k um ar b as u, p ei m an z an di , k at ar zy na m oż dż eń fig. 3. cyperus canus j. presl & c. presl; a – plant habit, b–c – young stems, d – mature stems, e – leaves and in�orescence, f – in�orescence enlarged, g – �owers (photo. e. mó) 175 fig. 4. cyperus canus j. presl & c. presl; a – plants growing in a �eld crop of corn, b – juvenile plants, c – plants growing in sub-aquatic habitat, d – plants mixed with corn (photo. e. mó) cyperus canus j. p resl & c . p resl – a traditional source of fiber, its uses, products and cultural significances am ong ethnic com m unities of c entral a m erica 176 ed ga r m ó, w ill ia m c et za l-i x, e lia na n og ue ra -s av el li, s ai ka t k um ar b as u, p ei m an z an di , k at ar zy na m oż dż eń fig. 5. production of mats ‘petates’ with tule; a–e – selection and preparation of stem barks, f–g – stem barks of di�erent dimensions, h–i – interlacing process of the stems, j – �nished mats (photo. e. mó) 177 fig. 6. production of a blower or fan with tule; a – selections of stems, b – traditional operating wooden bench, c – stems moored to the wooden bench, d–f – interlacing process with the stems, g – middle blower, h–i – �nished blower (photo. e. mó) cyperus canus j. p resl & c . p resl – a traditional source of fiber, its uses, products and cultural significances am ong ethnic com m unities of c entral a m erica 178 ed ga r m ó, w ill ia m c et za l-i x, e lia na n og ue ra -s av el li, s ai ka t k um ar b as u, p ei m an z an di , k at ar zy na m oż dż eń cyperus canus j. presl & c. presl – tradycyjne źródło błonnika, zastosowanie, produkty i znaczenie kulturowe wśród społeczności etnicznych streszczenie rodzaj cyperus l. (cyperaceae) obejmuje od 750 do 950 gatunków. występuje w ścisłym powiązaniu z ekologicznymi ugrupowaniami różnych gatunków traw. gatunki z rodzaju cyperus posiadają rozmieszczenie kosmopolityczne. niektóre z nich są uważane za szkodliwe chwasty uprawowe, jednak duża ich liczba znalazła zastosowanie w rolnictwie, ogrodnictwie oraz w przemyśle farmaceutycznym. cyperus canus j. presl & c. presl (cyperaceae) jest rośliną naturalną dla tropikalnej ameryki, rosnącą od południowego meksyku oraz gwatemali do kolumbii. gatunek ten występuje wzdłuż brzegów jezior, strumieni i bagien, a czasami na skałach, na wysokości 200–1500 m n.p.m. w naturalnym środowisku jest rośliną dość rzadko spotykaną, ponieważ traktowany jest jako tępiony chwast. w  ameryce środkowej c. canus jest rośliną powszechnie stosowaną do produkcji dywanów, nosidełek dla dzieci, portfeli, teczek, kapeluszy i lin. w gwatemali, pędy tego gatunku są używane do wyplatania koszy i  torebek, do produktów codziennego użytku (śpiworów, sandałów, wachlarzy) oraz przygotowywania tradycyjnych potraw. w artykule tym opisano zastosowanie i procesy produkcji niektórych ważnych wyrobów wykonywanych z dzikorosnącego c. canus. słowa kluczowe: central america, cyperus canus, stem, practical use received: [2016.07.13] accepted: [2016.09.06] 42 annales universitatis paedagogicae cracoviensis studia naturae, 1: 42–51, 2016, issn 2543-8832 svetlana gáperová1*, ján gáper2,3, terézia gašparcová1, kateřina náplavová3,5, peter pristaš4 1 faculty of natural sciences, matej bel university, tajovského 40, 974 01 banská bystrica, slovak republic, *svetlana.gaperova@umb.sk 2 faculty of ecology and environmental sciences, technical university in zvolen, t.g. masaryka 24, 960 63 zvolen, slovak republic 3 faculty of sciences, university of ostrava, chittussiho 10, 710 00 ostrava, czech republic 4 institute of biology and ecology, faculty of natural sciences, pavol josef šafárik university, moyzesova 11, 040 01 košice, slovak republic 5 ceb-centre of biological engineering, university of minho, campus de gualtar, braga, portugal morphological variability of fomes fomentarius basidiomata based on literature data introduction �e genus fomes is a taxon accepted in the polyporaceae family of polyporales order in class agaricomycetes, subphylum agaricomycotina and phylum basidiomycota. fomes fomentarius is the type species of the genus (donk, 1960). �ere are two morphological species in the genus fomes recognized at present: f. fomentarius (l.) fr. and f. fasciatus (sw.) cooke (ryvarden, 1991). although only mean basidiospore size is a helpful morphological characteristic in identi�cation of the respective species, its and rpb2 sequence data and optimum temperature for hyphal growth in vitro, support separation of these two distinct species (mccormick et al., 2013a; gáper et al., 2016). f. fomentarius is an ecologically and economically important polypore wood decay macrofungus with major roles not only in nutrient cycling in forest ecosystems as decomposer of dead wood and plant litter, but also as a source of medicinal and nutraceutical products (tello et al., 2005; collado et al., 2007; neifar et al., 2013; dresch et al., 2015). �ere have also been reports that it can persist as an endophyte in healthy plants for many years (baum et al., 2003; par�tt et al., 2010). �e species has been reported from africa, asia, europe, south america (chile) and north america (mccormick et al., 2013a, b; gáper, gáperová, 2014), but a revision of the f. fomentarius data from chile is necessary (gáper et al., 2016). in recent years, the existence of three distinct its lineages/sublineages among f. fomentarius strains has been clearly established. �e sublineage a1 consists of strains isolated from north america, whereas the sublineage a2 consists of strains recently 43 isolated only from europe. �e lineage b consists of strains isolated from europe and asia (gáper et al., 2016). �e presence of the two variable groups in europe was further con�rmed by molecular methods based on both efa region and lsu gene sequences comparison, so the f. fomentarius actually includes two sympatric cryptic species in europe (pristaš et al., 2013). �e present paper reviews the morphological variability of the f. fomentarius basidiomata with implications for reliable separation of f. fomentarius lineages/sublineages, basing on descriptions published in the literature. in contrast to the strain data (dresch et al., 2015; gáper et al., 2016), the morphological variability of f. fomentarius basidiomata is well documented. methods all reliable information on f. fomentarius basidiomata was collected and corroborated through the evaluation of literature in libraries and searches in online databases using google scholar, scifinder and web of knowledge. in this article, we present only a small fraction of the literature on the species within north america, namely mccormick et al. (2013a, b) because basidiomata morphology of f. fomentarius examined in their studies are consistent with formerly published descriptions (overholts, 1953; lowe, 1957; gilbertson, ryvarden, 1986). �ey examined macroand micromorphological characters of basidiomata sampled from multiple woody plant hosts and geographic regions in the united states (mccormick et al., 2013a, b). results and discussion general description of basidiomata macrocharacters: basidiomata (fig. 1) perennial, sessile, ungulate (occasionally applanate or triquetrous), tough, woody with a blunt margin; upper surface of pileus crustose, glabrous, prominently zoned (occasionally slightly zonate or no zonation) and shallowly furrowed, marginal part �nely tomentose or smooth; pore surface concave (occasionally �at to slightly concave or convex); pores round, small (2–5 per mm); context pale brown, so�-corky, tough and �brous, azonate or concentrically zoned, up to 1–3 cm thick; granular core of varying size developing at upper part of the context close to the substratum consisting of very thick-walled and irregularly shaped hyphae (sclerids); tube layers not distinctly (exceptionally distinctly) strati�ed, brown and becoming �lled with white mycelium. microcharacters: hyphal system trimitic; contextual generative hyphae thin-walled, hyaline with clamps, 2–5 µm in diam., inconspicuous; contextual sketetal hyphae thick-walled, aseptate, pale brown in koh, 3–8 µm in diam.; contextual m orphological variability of fom es fom entarius basidiom ata based on literature data s ve tla na g áp er ov á, j án g áp er , t er éz ia g aš pa rc ov á, k at eř in a n áp la vo vá , p et er p ris ta š 44 binding hyphae thick-walled, strongly branched, aseptate, pale brown, 1.5–4.5 µm in diam. cystidia none, cystidioles o�en present in the hymenium, thin-walled, fusoid, 24–40×3.5–7.5 µm, with a  basal clamp. basidia clavate, 4-sterigmate, usually with a  basal clamp, developed only early in the spring. basidiospores cylindric, hyaline, smooth, negative in melzer’s reagent. geographic variability of basidiomata above mentioned characters from basidioma macroand micromorphology basically follow the description given by ryvarden and gilbertson (gilbertson, ryvarden, 1986; ryvarden, gilbertson, 1993). �ey generally correspond well with those reported in other comprehensive polyporological monographs and recent studies, but we detected some di�erences. on the other hand, some european and asian data is identical (ryvarden, gilbertson, 1993; nuñez, ryvarden, 2001) (tab. 1). as described in tab. 1, basidiomata pileus and pore surface colours, basidioma size, depth of tube layer and basidiospore size are the most variable characters within species. it seems that pileus surface colour in europe is generally of lighter tones in comparison with basidiomata found in north america and asia. �e same pattern appears in pore surface colours, basidiomata from europe are generally of lighter tones. �e basidiomata width is 15 cm in north america and up to 40 cm in asia, on the other hand, it may reach up to 50 cm in europe. maximum depth of tube layer is 8 mm in europe and up to 10 mm in north america, in asia it varies from 2 to 16 mm and in fig. 1. fomes fomentarius basidiomata typically ungulate in shape on beech (fagus sylvatica) trunk. from m. šebesta (unpub., 2015) 45 europe it varies from 2 to 10 mm (jülich, 1984; breitenbach, kränzlin, 1986; gilbertson, ryvarden, 1986; zhao, zhang, 1992; ryvarden, gilbertson, 1993; bondartseva, 1998; nuñez, ryvarden, 2001; bernicchia, 2005; mccormick, 2013a; prasher, 2015). tab. 1. geographic variation in macroand micromorphological traits of the fomes fomentarius basidioma pileus surface colour europe: older party gray, marginal part light brown1,2; older party gray, blackish, yellowish or yellowish brown; marginal part light reddish brown3, silvery white, grayish, gray-brown to nearly black4; ocher to red brown when young, later light to dark gray5; brown when young, later grayish brown and gray6 asia: older party gray, marginal part light brown7; older party gray, blackish, yellowish or yellowish brown; marginal part light reddish brown3, gray, gray-brown to black8; gray, grayish brown, grayish black9 north america: brownish gray to nearly black10; gray, brown, blackish gray, cinnamon, bu�11 pore surface colour europe: pale brown1,3 to pale gray3; cream coloured when young, then light ocher to brownish5,6; ochraceous to gray2 asia: pale brown1,3 to pale gray3; brownish to brown or black brown to black8; light brown to grayish brown9 north america: brown to grayish brown10; bu�, brown, cinnamon11 basidioma size: width × height [cm] europe: up to 15 wide1; up to 20–40×5–15 (-20)3; 10–25 (-30)×15–205; up to 30–40 wide2; 5–50×3–256 asia: up to 15 wide7; up to 20–40×5–15 (-20)3; 3–20 (-40)×2–15 (-27)8; up to 12×189 north america: 2–15 wide10 depth of tube layer [mm] europe: 2–63; 2–5 (-8)5; 5–72; ca. 106 asia: 2–63,8 up to 168; 79; north america: up to 1010 basidia size [µm] europe: 23–25×7–91; 25–30×8–113; 21–25×7–92; 20–30×7–105; 25–30×8–116 asia: 23–25×7–97; 25–30×8–113 basidiospore size [µm] europe: 12–18 (-20)×4–71; 15–20×5–73; 18.5–19×5.5–65; 14–20×4.5–6.52; 6–24×5.5–6.56 asia: 12–18 (-20)×4–77; 15–20×5–73; 12–19×5–6.5 (-8.5)8; 15.6–18.6×5.1–6.19 north america: 12–18×4–710; 10–21.25×2.5–7.511 1 ryvarden, gilbertson, 1993; 2 bernicchia, 2005; 3 bondartseva, 1998; 4 schwarze, 1992; 5 breitenbach, kränzlin, 1986; 6 jülich, 1984; 7 nuñez, ryvarden, 2001; 8 zhao, zhang, 1992; 9 prasher, 2015; 10 gilbertson, ryvarden, 1986; 11 mccormick, 2013a concerning microscopic features, there are no signi�cant di�erences in the size of basidia in europe and asia. length of basidiospores from asia is not less than 12 µm. maximum width of basidiospores is up to 7 µm in europe, whereas basidiospores found in north america and asia may have up to 7.5 µm and 8.5 µm respectively (jülich, 1984; breitenbach, kränzlin, 1986; gilbertson, ryvarden, 1986; zhao, zhang, 1992; ryvarden, gilbertson, 1993; bondartseva, 1998; nuñez, ryvarden, 2001; bernicchia, 2005; mccormick, 2013a; prasher, 2015). m orphological variability of fom es fom entarius basidiom ata based on literature data s ve tla na g áp er ov á, j án g áp er , t er éz ia g aš pa rc ov á, k at eř in a n áp la vo vá , p et er p ris ta š 46 according to schwarze (1992) within the geographical area of distribution in europe, the colouration, size and shape of the basidioma can vary based on where the specimen has grown. �e colour of the pileus surface ranges from silvery white to nearly black although when it is wet it can appear to be black and when it is old and dry it can be bleached to an o�-white colour. �e colour is lighter at lower latitudes, at low elevations and on the south side of stems. despite his studies covered basidiocarps collected in di�erent places of origin in great britain and in mainland europe, they revealed no discernible pattern (schwarze, 1994). �ese di�erences have caused some fungal taxonomists to propose infraspeci�c divisions. �us, the black basidiomata were previously classi�ed as a fomes nigrescens, ungulina fomentaria subsp. nigricans, f. fomentarius subsp. nigricans, or f. fomentarius var. nigrescens. �e lighter coloured basidiomata were classi�ed as a f. fomentarius subsp. fomentarius. according to zhao, zhang’s interpretation of the chinese specimens examined, the basidiospores are also di�erent; in f. fomentarius subsp. nigricans they are ovoid to subglobose (9–11×6.5– 6.8 µm), while in f. fomentarius subsp. fomentarius they are cylindrical to oblong ellipsoid (12–19×5–6.5 µm) (zhao, zhang, 1992). similarly, the small basidiomata with closely concentrically zoned pileus surfaces associated with birches were previously classi�ed as a  polyporus fomentarius var. lineatus (velenovský, 1922). in addition, there are a  number of taxonomic synonyms, in the past described as separate taxa. basidiomata of f. fomentarius can range in size from ca. 5 to ca. 50 cm (tab. 1). the species have been observed in northern japan (hokkaido island) and northeastern china (changbai shan nature reserve; xiao xing’anling mts.) to have two morphologically different types basing on basidioma size (cheng, 2000). while the small-sized basidiomata (up to 20 cm wide) seem to be linked preferably with betula spp., less frequently with populus sp. and acer sp., as the hosts, the large-sized basidiomata (over 20 cm wide) were found mostly on acer sp., tilia amurensis, quercus liaoi and less frequently fraxinus mandshurica, ulmus sp. and betula platyphylla (beeches are not distributed either in hokkaido, or in the changbai shan nature reserve) (cheng, 2000). two types of f. fomentarius have also been reported from an old-growth beech (fagus japonica, f. crenata) and oak (quercus serrata, q. mongolica) forest in a cool temperate area of japan in the southern part of the abukuma mts. along the pacific coast. both types occurred frequently on beeches (yamashita et al., 2010). the two types of f. fomentarius basidiomata were found growing together in the various localities (cheng, 2000; yamashita et al., 2010). these data correspond well with those reported in the japanese polyporological monographs (imazeki, hongo, 1989; igarashi, 1989, 1994; takahashi, 2003). apart from size, basidiomata may range also in shape. according to gaudreau and co-authors it is either bracketor boot-shaped (gaudreau et al., 2005). the boot shaped form, however, is probably related to the following phenomenon. if a tree that f. fomentarius is growing on falls 47 down, the fungus will re-orient its direction of growth to account for its changed position, so specimens are sometimes seen with two different patterns of growth in the basidioma, at right angles to each other. this is to ensure that the fertile underside is always aligned with gravity so the basidiospores will fall into the air currents. according to læssøe and del conte (1996) this species in europe exists in two forms, the beech and the birch form, depending on the host. beech form is semicircle-shaped and brown. birch form is hoof-shaped, more slender and darker gray (læssøe, del conte, 1996). the two forms have been already recognized by lloyd in the early 20th century (lloyd, 1915). according to this author, both forms are hoof-shaped. beech form is larger with soft context and occurs in france. birch form is smaller with harder context and occurs in the usa and northern europe (lloyd, 1915). however, morphological analyses of 1 165 basidiomata samples within the malenovický kotel nature reserve (beskydy mts., the czech republic) using the general linear model (glm) confirmed a direct relationship between the shape of basidioma and its age rather than the host. the approximate age of transition of the basidiomata from the birch form to the beech form was around 5–6 years (rybovičová, 2010). but some of the existing above mentioned phenotypic differences can be based on the interactions between the genotype and environment (schwarze, 1999, 2004) and/or presence of different lineages/sublineages (gáper et al., 2016). however, no clear differences between basidiomata, useful for reliable separation of the lineages/sublineages, have been observed so far. nevertheless, a recent f. fomentarius basidiomata description in north american research articles (mccormick et al., 2013a, b) represents north american a1 sublineage (gáper et al., 2016). basidiomata of the f. fomentarius sublineage a1 are ungulate, applanate or triquetros; pileus surfaces gray, brown, blackish gray, cinnamon, buff; pore surfaces buff, brown, cinnamon, concave, flat to slightly concave or convex; pores 2–5 per mm; basidiospores 17.54(±0.05)×5.27(±0.03) µm (mean ±se) and ranged from 10.0–21.25×2.5–7.5 µm. colorus of external portions of the basidiomata (pileus and pore surface colours) are more variable than internal portions (context and tube layers) (mccormick et al., 2013a). in both asian and european literature such data is absent. as regards then, european literature alone, it is most possible that these descriptions are based on basidiomata of two other lineages/ sublineages and thus we cannot discriminate between them. conclusions �e analysis of data present in literature shows that no clear di�erences can be observed between basidiomata of fomes fomentarius which would be suitable for reliable separation of lineages/sublineages. within geographical area of distribution in the world, these characteristics can vary considerably. �e existing phenotypic di�erences m orphological variability of fom es fom entarius basidiom ata based on literature data s ve tla na g áp er ov á, j án g áp er , t er éz ia g aš pa rc ov á, k at eř in a n áp la vo vá , p et er p ris ta š 48 in f. fomentarius basidiomata morphology can be attributed either to di�erent lineages/sublineages, or to interactions between the genotype and its environment. �e phenotypic basidioma traits of the north american a1 sublineage are known. however, it is most possible that the descriptions in european literature are based on basidiomata of both sublineage a2 and lineage b, and thus we cannot discriminate between them. moreover, some european and asian macroand microcharacters are identical. �erefore, the following recommendations which would be suitable for reliable separation of the lineages/sublineages can be made based on the review: 1. basidiomata di�er macromorphologically mainly in pileus and pore surface colours, basidioma size and depth of tube layer, 2. basidiomata di�er micromorphologically mainly in basidiospore size, 3. a detailed study of each of these three attributes, especially within europe and asia, would be suitable for separation of the above mentioned lineages/sublineages. acknowledgments �is work has been supported by grants from the scienti�c grant agency of the ministry of education, science, research and sport of the slovak republic (vega no. 1/0286/17 and kega no. 025umb-4/2017) and the project sgs 16/přf/2016 from the faculty of sciences of the university of ostrava (czech republic). we wish to acknowledge the anonymous reviewers for their detailed and helpful comments on the manuscript. references baum, s., sieber, t.n., schwarze, f.w.m.r., fink, s. 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(2010). host preference and species richness of wood-inhabiting aphyllophoraceous fungi in a cool temperate of japan. mycologia, 102(1), 11–19. doi: 10.3852/09-008 zhao, j.d., zhang, x.q. (1992). �e polypores of china. bibliotheca mycologica, 145, 1–524. abstract currently two morphological species of the genus fomes (polyporaceae, basidiomycota) are known: f. fomentarius (l.) fr. and f. fasciatus (sw.) cooke. both species are very important in the decomposition of wood and in the nutrient cycling in forest ecosystems. moreover, f. fomentarius is also known as a source of medicinal and nutraceutical products. recently the existence of three separate its lineages/sublineages among f. fomentarius strains has been clearly established – a1 (the strains isolated from north america), a2 (only from europe) and b (from europe and asia). in this review the current knowledge of the morphological variability of f. fomentarius basidiomata has been summarized in respect of the reliable separation of its lineages/sublineages. microand macrofeatures and geographic variability of the basidiomata have been described. morphological traits of the f. fomentarius basidiomata can vary due to geographical distribution. �ese phenotypic di�erences can be based on the presence of several groups or interactions between the genotype and environment. however, no clear di�erences between basidiomata, useful for reliable separation of the lineages/sublineages, have been observed so far. a recent description in north american fomes research articles is based on basidiomata of the a1 sublineage. it is most possible that the descriptions in european literature are based on basidiomata of two other lineages/sublineages and thus we cannot discriminate between them. in the future a  detailed study of the macroand microtraits – pileus and pore surface colors, basidioma size, depth of tube layer, and basidiospore size – is therefore proposed for reliable separation of the lineages/sublineages of f. fomentarius. key words: basidioma, cryptic species, morphological variability received: [2016.05.16] accepted: [2016.09.21] zmienność morfologiczna owocników fomes fomentarius na podstawie danych z literatury streszczenie obecnie znane są dwa morfologiczne gatunki z rodzaju hubiak fomes (fr.) fr. (polyporaceae, basidiomycota): f. fomentarius (l.) fr. i f. fasciatus (sw.) cooke. obydwa są bardzo ważne w procesie rozkładu drewna oraz w obiegu składników pokarmowych w ekosystemach leśnych. ponadto, 51 f. fomentarius jest również uznany jako źródło produktów leczniczych i nutraceutycznych. ostatnio zostało wyraźnie określone istnienie trzech odrębnych linii rozwojowych its/sublinii pomiędzy szczepami f. fomentarius – a1 (szczepy wyizolowane w ameryce północnej), a2 (tylko w europie) i b (w europie i azji). w niniejszym przeglądzie aktualna wiedza o zmienności morfologicznej owocników f. fomentarius została zebrana w odniesieniu do rzetelnego wydzielenia linii rozwojowych/sublinii. zostały tu opisane mikroi makrocechy oraz zmienność geogra�czna owocników. cechy morfologiczne owocników f. fomentarius mogą się różnić ze względu na rozmieszczenie geogra�czne. różnice fenotypowe mogą bazować na obecności różnych grup lub interakcji pomiędzy genotypem a  środowiskiem. jednakże nie ma wyraźnych różnic między owocnikami, przydatnych do niezawodnego oddzielenia linii rozwojowych/sublinii, które do tej pory obserwowano. nowsze opisy gatunków z rodzaju fomes w ameryce północnej w artykułach naukowych opierają się na owocnikach z sublinii a1. jest bardzo prawdopodobne, że opisy w literaturze europejskiej oparte są na owocnikach dwóch pozostałych linii rozwojowych/sublinii, a  zatem nie możemy odróżniać ich między sobą. w przyszłości proponuje się do niezawodnego oddzielenia linii rozwojowych/sublinii f. fomentarius szczegółowe badania makroi mikrocech: kolorów kapelusza i powierzchni porów, rozmiaru owocnika, głębokości warstwy rurek i rozmiaru bazydiospor. słowa kluczowe: zróżnicowanie morfologiczne, gatunki kryptyczne, owocnik information on the authors svetlana gáperová she is the member of department of biology and ecology of faculty of natural sciences of matej bel university in banská bystrica (slovakia). she is interested in the biology and ecology of both woody plants and wood-decaying fungi in urban areas and monitoring of urban air pollution. ján gáper he is the member of department of biology and general ecology of faculty of ecology and environmental sciences of technical university in zvolen (slovakia), as well as the member of department of biology and ecology in faculty of natural sciences at university of ostrava in ostrava (czech republic). he is interested in the identi�cation and ecology of wood-decaying polypores, regarding the importance for ecosystem functioning. terézia gašparcová she is an internal phd student at faculty of natural sciences of matej bel university in banská bystrica (slovakia), department of biology and ecology, in study program “evolution of ecosystems and their protection”. �e topic of her dissertation is “diversity and distribution of the genus ganoderma in slovakia”. kateřina náplavová she is an internal phd student in faculty of sciences at university of ostrava (czech republic), department of biology and ecology in study program biology. �e topic of her dissertation is “diversity and distribution of wood-decaying polypores”. peter pristaš he is the head of department of microbiology in institute of biology and ecology of faculty of natural sciences at pavol josef šafárik university in košice (slovakia). he is interested in the molecular phylogeny and identi�cation of wide spectrum of microorganisms including bacteria, protozoa and fungi. m orphological variability of fom es fom entarius basidiom ata based on literature data 71 annales universitatis paedagogicae cracoviensis studia naturae, 1: 71–84, 2016, issn 2543-8832 noel a. gonzález-valdivia1, william cetzal-ix1*, saikat kumar basu2, isidra pérez-ramírez1, jesús f. martínez-puc1, peiman zandi3 1 instituto tecnológico de chiná, calle 11 entre 22 y 28, colonia centro chiná 24050, campeche, méxico, *rolito22@hotmail.com 2 department of biological sciences, university of lethbridge, lethbridge, ab canada t1k 3m4 3 institute of crop science, chinese academy of agricultural sciences, beijing 100081, people’s republic of china conservation of the genetic diversity of local corn (zea mays l.) in the yucatan peninsula, mexico introduction maize (zea mays l.) is an important cereal member which is historically associated with the original human populations that once populated and till now continues inhabiting the vast and biodiverse continents of the americas (serratos-hernández, 2000). corn seeds have been reported from several archaeological sites across the americas dating back as old as 5000 years (arteaga et al., 2016). several scholars and researchers consider that the origin of human is inherently associated with the evolution of corn, for example the ch’ol ethnicity is recognized with the phrase “men of corn” (cross-cortés, 2000). �e genetic diversity of this grass family (poaceae) member o�ers a perspective of its relevance as food, and numerous ways in which it is used by the indigenous communities across the americas, where the crop is reported to have originated (wellhausen et al., 1951). mexico is one of the most important centers of origin for maize and the most diverse representing 64 landraces (59 natives); along with innumerable varieties and “criollo” or natives hybrids have been identi�ed. �is monumental genetic diversity is the result of the mass selection of the species since it was domesticated from teosintes or similar to those mesoamerican teosintes ancestors (ine-conabio-sagarpa, 2008; arteaga et al., 2016). in the yucatan peninsula (yp) of mexico, three maize races (nal tel narrower, dzit bacal and xnu’uk nal or tuxpeño and two hybrids (nal xoy and x’mejen nal) with restricted distribution have been recorded (arias et al., 2007). �ese races and hybrids are disappearing fast as a result of technological displacement imposed by improved varieties and commercial hybrids, produced and distributed by giant multi-national corporations. �e genetic erosion due to the subsequent decrease 72 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di in seed banks of the local producers and growers, along with incentive policies of replacing native seeds for higher pro�ts and commercial interests further lead to the loss of local genetic diversity of maize landraces and germplasms in the countryside by both rural youth, as well as aging small and medium range producers “milpero” (buenrostro, 2009). such anthropogenic interferences have been creating a  scenario of gradual extinction of the native maize genetic diversity in the yp. climate change, another signi�cant challenge for agriculture and agri-production, also needs rapid adaptation options to deal with the production constraints. an important line of work in this regard is to rescue the local maize genetic diversity under a  restorative scheme that helps conserving local diversity threatened by several anthropogenic factors and to study their responses to new environmental rigors, defig. 1. localities of cultivation and conservation of native germplasms of corn in the yucatan peninsula, mexico 73 termining their ability to continue productivity under the context of highly variable climatic uncertainties. �e main objective of the research has been collecting seeds still used by farmers in their traditional production systems, identifying races and varieties used in yp area and documenting their life cycle and other usage identi�ed by key informants and maize growers and producers. material and methods �e study was designed based on the review of specialized literature on native corn (wellhausen et al., 1951; serratos-hernández, 2009; perales-rivera, golicher, 2011). to identify the diversity of races and varieties, specialized literature was reviewed and the �elds were located for identifying the possible races and varieties that have been recorded in the yp (tab. 1 – appendix); priority areas of cultivation and conservation of native germplasms of corn were visited (fig. 1). subsequently, seeds of landraces were purchased in �ve rural seed fairs (trade shows) in di�erent areas of the yp (fig. 2). it is important to mention that such traditional rural fairs are significant resource centers where seeds of local landraces and germplasms of maize are sold and exchanged among local farmers and producers. also in these local events short interviews were successfully conducted with producers, who answered questions about documenting their experiences and sharing traditional knowledge regarding the life cycles of various local maize varieties, the best time of planting and possible partnering with other cultivated species and the source of seeds. in each locality visited (fig. 1), several corncobs were collected to characterize morphology (number of rows per cobs, number of grains per row and along the cob) (fig. 3–4) and their performance or yield. results races, varieties and hybrids �e principal �nding with respect to native corn varieties registered in the yp is that these are still in places considered “maya centers of resistance”. however, the quantity and quality of seed available has been reduced, mainly due to severe drought conditions and increased pest pressures. nevertheless, some native varieties were di�cult to �nd, such as nal tel blanco or sak nal tel (fig. 3a), which is characterized by being precocious corn, high quality, �our, and drought tolerant. �e same situation appears true for all varieties to a  lesser or greater degree. on the other hand, the presence of two additional races has been recorded in the yp: palomero amarillo (yellow palomero) (fig. 3b) and tabloncillo (fig. 4). with respect to the conservation c onservation of the genetic diversity of local corn (zea m ays l.) in the y ucatan p eninsula, m exico 74 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di fig. 2. localities of rural seed fairs or corn (trade show) in the yucatan peninsula, mexico of maize germplasm, �ve varieties: red-white-yellow nal tel (fig. 3c), white-yellow dzit bacal (fig. 3d), white-yellow-red (also known as chac chu’ub) xnu’uk naal or tuxpeño, blue xnu’uk naal (also known as eh hu), yellow palomeroand tabloncillo and several hybrids: xnu’uk nal (cross between nal tel with tuxpeño), yellow nal xoy (possible tri-hybrid between nal tel and xnu’uk nal) (fig. 3g), white nal xoy (cross between nal tel, tuxpeño and variety), x’mejen nal (cross between nal tel and dzit bacal), chac xmejen nal (tri-hybrid between chac chu’ub and x’mejen nal) (fig. 3h), san pableño, red nal tel (cross between nal tel with chac ch’ub), yellow nal tel (possible cross between nal tel and nal xoy) were recovered (appendix 1 – tab. 1). relevant agroecological characteristics �e varieties registered are characterized by lifecycles, presenting three main classes: precoces (lower lifecycle at 80 days), intermediate (80–110 days) and late (over 110 days). in the germplasm enhancement of maize, the maya people have made cross 75 fig. 3. races, varieties and hybrids of corn; a  – nal tel blanco or sak nal tel, b – palomero amarillo (yellow palomero), c – red nal tel, d – yellow nal tel, e – white dzit bacal, f – eh hu, g – yellow nal xoy, h – chac xmejen nal (photo. w. cetzal-ix) c onservation of the genetic diversity of local corn (zea m ays l.) in the y ucatan p eninsula, m exico 76 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di fig. 4. tabloncillo race of corn (photo. w. cetzal-ix) tab. 1. provenances collected between 2015 and 2016 of native varieties of corn in yucatan and campeche, mexico * number of collections variety campeche* yucatan* lifecycle can dzit bacal 2 0 intermediate can nal tel 1 1 precocious can nal xoy 0 2 intermediate can san pableño 1 0 late can xmejen naal 0 2 intermediate can xnu’uk naal 0 1 late chac chu’ub 1 2 late chac xmejen naal 0 3 intermediate chak nal tel 1 1 precocious eh hu 3 2 late mixto 1 0 late yellow palomero 1 2 precocious pix cristo 2 0 late sak dzit bacal 1 0 intermediate sak nal tel 3 0 precocious sak nal xoy 0 2 intermediate sak tux 2 0 late sax tux (long insertion) 1 0 late sak xmejen naal 0 1 intermediate sak xnu’uk naal 1 0 late sak san pableño 3 0 late santa rosa 1 0 late tabloncillo 0 1 late tuxpeño 1 0 late 77 that allows to combine the characteristics of the lifecycle with the organoleptic characteristics of the grains, which has allowed the emergence in the collections of hybrid double and even triple. for example, chac xmejen nal (fig. 3h) is a reddish corn of intermediate lifecycle (75–80 days), probably from the cross product of chac chu’ub of red color with xmejen nal (hybrid between nal tel and xnu’uk nal) or with precocious varieties of nal tel of red color (chu’ub nal tel) (fig. 3c). of the 47 samples collected at di�erent sites visited, 10 belong to precocious varieties (nal tel and palomero), 14 to intermediate (dzit bacal, xmejen naal, nal xoy) and 23 correspond to late (xnu’uk naal and tabloncillo) types. �e characteristics of cobs of di�erent races, varieties and provenances that facilitate distinguishing them and their values are shown in table 1. of the total of 25 varieties registered, 18 were reported in campeche and 13 in yucatan (tab. 2). in quintana roo, it was not possible to �nd sources of native maize, except for xmejen naal; for which so far no data has been generated. quintana roo producers indicate that seeds have been lost due to recurrent drought occurring in the state. �e set of late-type corn, the xnu’uk naal race, shows the greatest number of records and provenances. �e varieties that were found during expeditions between 2015 and 2016 allowed to determine that native corn germplasm preserved in campeche and yucatan belong to the three groups that are subdivided according to their lifecycle and harvest. materials with long cycles (late) are prevalent in the state of campeche compared to precocious and intermediate types. �e opposite is observed in yucatan, where corns’ intermediate cycles were more common among communities where native corn is 6 3 16 4 10 7 0 4 8 12 16 20 precocious intermediate late pr ov en an ce s of n at iv e co rn campeche yucatan fig. 5. number of provenances in which native varieties are reported based on the lifecycle of the varieties collected in the states of campeche and yucatan, mexico c onservation of the genetic diversity of local corn (zea m ays l.) in the y ucatan p eninsula, m exico 78 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di cultivated; although late corns also represent a signi�cant proportion of the diversity of this crop (fig. 5). discussion �e diversity of races and varieties of corn in the yucatan peninsula that have been identi�ed in the region include races mentioned by wellhausen et al. (1951) and arias et al. (2007) with addition of tabloncillo and reventador (yellow palomero) races. �ere is evidence to some gene �ow that might have occurred due to the migration of people from rural areas to other parts of mexico, which have brought seeds that then thrive in their communities. �e other alternative is that this evidence may not have been detected in the peninsula of yucatan in previous studies. for example, the tabloncillo (fig. 4) race has been mentioned to be present in the gulf of mexico side. gene �ow has been mentioned by perales-rivera and golicher (2011) as an active process between nine provinces and six biocultural centers of origin and diversity of native maize in mexico. �e olotillo race, that has been mentioned by lazos and cheauvet (2012) to be found in campeche, is a possible reference to eh hu variety, dark purple, o�en located in di�erent sub-regions of the state and generally in the peninsular region. however, other authors report this as the eh hu that is part of the xnu’uk naal (tuxpeño) race. �e late varieties seem to dominate the current scenario of diversity of native maize germplasm in the yucatan peninsula, followed by varieties and hybrids with intermediate lifecycles. �is has been mentioned as a preference by arias et al. (2007). against the context of possible climate change scenarios, that predicts an increase in the frequency and intensity of the phenomenon of drought; thereby reducing the presence of early varieties that may limit the adaptation options for corn producers in the region. based on the results obtained, it can be concluded that the genetic diversity of native maize in the yucatan peninsula still remains; based on records and collections of germplasm conducted in 2015 and 2016, represented by the same range of races and varieties that have historically been reported for the region (wellhausen et al., 1951; arias et al., 2007; serratos-hernández, 2009; arteaga et al., 2016). cazares-sánchez et al. (2015) mentioned that nal tel has an average of 22 grains per row, under the results (26 grains per row) in varieties of cobs collected between 2015–2016. also, for varieties like xnu’uk naal, these authors mention an average of 40 grains per row, coinciding with the average obtained in the varieties that are presented in this study. for the state of campeche, a predominance of maize varieties of late lifecycle is detected and in the yucatan peninsula of intermediate lifecycle coinciding with arias et al. (2007), who mentioned that the region has been demonstrating declining use of precocious varieties and increasing the number of varieties with 79 longer cycles, which may result in a change of production systems to those of temporary irrigation. varieties with longer periods in their growth and developments tend to produce higher grain yield per unit area. �en the xnu’uk naal (tuxpeño) races are likely to continue to prevail in the region, while precocious nal tel race, belonging to the ancient races in mexico (wellhausen et al., 1951), appears to decline. particularly gallito white (sak nal tel), which was obtained only in small quantities in the chenes region in campeche, is quite vulnerable. a new contribution to the region constitutes the collections of the yellow palomero (near reventador precocious) race and the appearance has strong similarity in their morphometry to tabloncillo race; which may indicate germplasm movement by exchange of seeds or due to the impact of migrant population in the region. both materials were documented in the area of peto, in yucatan; that also highlights to be the main source of several varieties of native corn for the yucatan peninsula in this study. acknowledgments �e �rst author would like to acknowledge prodep itchn-ptc support of the project rescate de germoplasma de maíz criollo y acriollado en la península de yucatán (number of o�ce dsa/103.5/15/6793). references arias, l.m., latournerie, l., montiel, s., sauri, e. 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[in spanish] wellhausen, e., roberts, l., hernández, e., mangeldorf, p.c. (1951). razas de maíz en méxico. su origen, características y distribución. programa de agricultura cooperativo de la secretaría de agricultura y ganadería, fundación rockefeller, méxico. [in spanish] c onservation of the genetic diversity of local corn (zea m ays l.) in the y ucatan p eninsula, m exico 80 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di appendix 1 tab. 1. races, varieties and characteristics of native corn cob collected between 2015 and 2016 in the yucatan peninsula, mexico. yucatan peninsula states: cam – campeche, qroo – quintana roo, yuc – yucatan, c – colonical, cl – cylindrical, co – cónico, nd – no data collection cobs group race variety (hybrid*) provenances grain color cob long [cm] row per cob grains per row cob shape pr ec os io us nal tel sak nal tel suc-tuc [cam] white 11 14 30 c sak nal tel hopelchén [cam] white 11 15 18 c sak nal tel sahcabchén [cam] white 14 14 23 c chak nal tel xoy, peto [yuc] red 11 14 25 c chak nal tel tikinmul [cam] red 12 14 27 c k’an nal tel peto [yuc] yellow 11 14 28 c k’an nal tel suc-tuc [cam] yellow 14 16 28 c k’an nal tel xbox [yuc] yellow 10 12 24 c k’an nal tel chacsinkín, peto [yuc] yellow 13 15 28 c palomero yellow palomero peto [yuc] yellow 12 12 33 cl yellow palomero timul [yuc] yellow 9 13 24 cl yellow palomero tikinmul [cam] yellow 10 12 33 cl in te rm ed ia te dzit bacal chac dzit bacal hopelchén [cam] reddish nd nd nd nd k’an dzit bacal suc-tuc [cam] yellow nd nd nd cl k’an dzit bacal dzutoh, timul [yuc] yellow 14 14 33 cl sak dzit bacal dos lagunas norte, calakmul [cam] white 19 12 43 cl sak dzit bacal dzutoh, timul [yuc] white 13 12 35 cl sak dzit bacal peto [yuc] white 19 12 47 cl dzit bacal por nal tel sak xmejen naal* timul [yuc] white 12-16 12-14 29-35 cl sak xmejen naal* blanca flor, bacalar (qroo] white nd nd nd nd sak xmejen naal* sabacché [yuc] white 17 14 39 cl sak xmejen naal* dzutoh [yuc] white 15 14 32 cl k’an xmejen naal* peto [yuc] yellow nd nd nd nd 81 in te rm ed ia te k’an xmejen naal* dzutoh, timul [yuc] yellow 14 14 32 cl chac chu’ub por x’mejen nal chac xmejen naal* peto [yuc] red 16 15 32 c chac xmejen naal* box [yuc] dark red 14 13 32 co chac xmejen naal* dzutoh, timul [yuc] red 19 13 39 co xnu’uk naal por nal tel sak nal xoy* xoy, peto [yuc] white 19 12 46 co sak nal xoy* dzutoh, timul [yuc] white 16 16 27 co k’an nal xoy* xoy, peto [yuc] yellow 19 16 36 co xnu’uk naal x nal tel k’an nal xoy dzutoh, timul [yuc] yellow 17 12 38 co la te xn’´uk naal chac chu’ub dos lagunas norte, calakmul, [cam] red 17 13 27 cl chac chu’ub timul [yuc] red 16 13 36 cl chac chu’ub xoy, peto [yuc] red 17 16 42 cl pix cristo dos lagunas norte, calakmul [cam] white with red 20 12 33 cl pix cristo suc-tuc [cam] white with red 21 8 50 cl k’an xnu’uk naal peto [yuc] yellow nd nd nd nd sak xnu’uk naal sahcabchén [cam] white 18 14 42 cl sak xnu’uk naal sabacché [yuc] white 18 10 45 cl sak xnu’uk naal suc-tuc [cam] white 15 12 32 cl sak xnu’uk naal peto [yuc] white 18 14 26 cl eh hu xoy, peto [yuc] purple 18 12 44 cl eh hu dzutoh, timul [yuc] purple 16 12 39 cl eh hu xmabén, calak-mul [cam] purple nd nd nd nd eh hu suc-tuc [cam] purple 18 12 44 cl eh hu los laureles [cam] purple nd nd nd nd sak tux suc-tuc [cam] white nd nd nd cl sak tux xmabén, calak-mul [cam] white nd nd nd nd c onservation of the genetic diversity of local corn (zea m ays l.) in the y ucatan p eninsula, m exico 82 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di la te sax tux (long insertion) dos lagunas norte, calakmul [cam] white 16 14 34 cl santa rosa xmabén, calak-mul [cam] yellow nd nd nd nd mixed uxpeño nohacal [cam] white 17 16 4.3 cl los laureles [cam] yellow y rojizo nd nd nd nd sak san pableño sahcabchén [cam] white 17 14 41 cl sak san pableño sahcabchén [cam] white 15 13 22 cl can san pableño sahcabchén [cam] yellow 16 16 27 cl sak xnu’uk naal hopelchén [cam] white nd 12 37 cl tabloncillo tabloncillo peto [yuc] purple and white nd 8-12 47 cl 83 abstract �e production of native corn at regional level is greatly limited by the seasonality of rainfall, availability of adequate lands, poor fertility status of the soil, high input costs and constraints of resources of the local corn growers and/or producers. �e challenges of reduced cultivable area give very little opportunity for increasing production area in a  sustainable manner; it is important to note that the soil recover its fertility status through crop rotation and prolonged rest period (>25 years) known as sequential agroforestry system or “milpa”. during 2015, corn collections were performed in the yucatan peninsula, mexico that included �ve races from the yucatan (in localities of nohacal and peto) and campeche (calakmul, suc-tuc, sakabchen, i chek) states. �e races identi�ed were: 1) nal-tel (gallito), 2) dzit bacal, 3) xnu’uk naal (tuxpeño), 4) palomero and 5) tabloncillo. �e local varieties, pix cristo (knees of christ), eh hu (purple maize) and chac chu’ub (chac’s blood or red maize), are included within the tuxpeño (xnu’uk naal) race. �e land race of corn that is in imminent danger of extinction is nal-tel, characterized by its precocity and ability to escape periods of low rainfall; it is important to rescue it for adoption to the practices of local and regional production. �e adaptation of this race as a germplasm is important due to its resilience to climate change itself. palomero, tabloncillo, pix cristo, chac chu’ub and eh hu can thus be used in traditional food industry, to preserve the traditional knowledge and to provide opportunities for additional income for the local, rural communities. yellow palomero and tabloncillo races are new records of germplasm for the region; and hence it is essential to exchange their seeds among local producers and growers. key words: campeche, conservation, corn, cultivation, genetic diversity, mexico, yucatán received: [2016.09.07] accepted: [2016.10.20] zachowanie różnorodności genetycznej rodzimej kukurydzy (zea mays l.) na jukatanie w meksyku streszczenie produkcja rodzimej kukurydzy na poziomie regionalnym jest znacznie ograniczona przez sezonowość opadów, dostępność odpowiednich gruntów, zły stan żyzności gleby, wysokie koszty produkcji i ograniczenia zasobów lokalnych plantatorów lub producentów kukurydzy. apele o zmniejszenie powierzchni uprawnej dają bardzo małą szansę na zwiększenie powierzchni produkcyjnej w sposób zrównoważony; ważne jest, aby pamiętać, że gleba może odzyskać swój status płodności dzięki płodozmianowi oraz dłuższym okresom spoczynku (>25 lat), zwanym sekwencyjnym systemem agroleśnym lub „milpa”. w 2015 roku na jukatanie w meksyku przeprowadzono zbiory kukurydzy, które obejmowały 5 lokalnych odmian z regionu jukatan (w miejscowościach nohacal i peto) i campeche (calakmul, suc-tuc, sakabchen, i chek). zidenty�kowano następujące odmiany: 1) nal-tel (gallito), 2) dzit bacal 3) xnu’uk naal (tuxpeño), 4) palomero i 5) tabloncillo. lokalne odmiany pix cristo (kolana chrystusa), eh hu (kukurydza purpurowa) i chac chu’ub (kukurydza czerwona) obejmowały również odmianę tuxpeño (xnu’uk naal). nal-tel jest odmianą kukurydzy, która jest narażona bezpośrednio na niebezpieczeństwo wymarcia. charakteryzuje się wczesnym rozwojem i zdolnością przetrwania w okresach niskich opadów, co jest ważne dla jej ratowania w celu praktycznego zastosowania w lokalnej i regionalnej produkcji. przystosowania tej odmiany, jako plazmy zarodkowej, są ważne ze względu na jej odporność na zmiany klimatyczne. palomero, tabloncillo, pix cristo, chac chu’ub i eh hu mogą być stosowane w tradycyjnej produkcji spożywczej, w celu zachowania wiedzy oraz zapewnienia możliwości dodatkowego dochodu dla lokalnych społeczności wiejskich. żółte odmiany palomero i tabloncillo są nowymi archiwami plazmy zarodkowej dla regionu i dlatego niezbędna jest wymiana ich nasion wśród lokalnych producentów oraz plantatorów. słowa kluczowe: campeche, ochrona, kukurydza, uprawa, różnorodność genetyczna, meksyk, jukatan c onservation of the genetic diversity of local corn (zea m ays l.) in the y ucatan p eninsula, m exico 84 n oe l a . g on zá le zv al di vi a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, is id ra p ér ez -r am íre z, j es ús f . m ar tín ez -p uc , p ei m an z an di information on the authors noel a. gonzález-valdivia professor and researcher at the instituto tecnológico nacional, sni level 1; an agronomist specializing in the rescue, assessment and management of native germplasms adapted to the tropics. also interested in ethnobiology studies, ecology and sustainable development, which have recently resulted in innovative applications of traditional knowledge in obtaining human products such as biopesticides, medicinal and food products based on �torrecursos utility. he participated in the training of human resources and in the development of initiatives and projects with positive impacts on the rural environment. william cetzal-ix interested in systematics, taxonomy and conservation of neotropical orchids and ferns, as well as in �oristic studies of indicator species (epiphytes) conservation of forests of southeastern mexico, and knowledge and conservation of plants with potential uses (melliferous, medicinal and ornamental). saikat kumar basu traditionally trained in botany (plant sciences) and specializing in microbiology, works actively in various areas of plant sciences and environmental conservation. �e author works extensively on forage crops with particular reference to annual forage legume and medicinal herb and spice, fenugreek. currently he is working in biomolecular sciences �eld dealing with plant biotechnology and genetic engineering application in small grain cereals like wheat. eliana noguera-savelli interested mainly in systematics, taxonomy, �oristic and anatomy of neotropical vascular plants, developing research to generate knowledge on biodiversity, exploration of timber and non-timber forest resources and training of human resources to support knowledge, conservation and sustainable use of natural resources. isidra pérez-ramírez agronomist with a master of science in natural resources and rural development specializing in agroecosystem management. he is interested in the characterization and geographical distribution of plants in home gardens. jesús f. martínez-puc extensionist, researcher, and professor interested in the tropical apiculture. currently developing projects on pest control bees, nutritional quality of nectar and mellifera �ora. peiman zandi he was deeply trained in agronomy (crop science) and specializing in stress physiology, biotic/abiotic stresses and agroecology. he is also interested in working in di�erent areas of plant developmental biology, agroecology, plant nutrition, botany, plant breeding and genetics. 69 annales universitatis paedagogicae cracoviensis studia naturae, 2: 69–80, 2017, issn 2543-8832 doi: 10.24917/25438832.2.5 katarzyna możdżeń*, patrycja zagata leśnicka, mateusz ślęczka, magdalena greczek-stachura institute of biology, pedagogical university of kraków, podchorążych 2 st., 30-084 kraków, poland, *katarzyna.mozdzen@up.krakow.pl the photosynthetic activity of paramecium bursaria endosymbiotic algae in varying temperature conditions introduction microbial organisms are ideal to study adaptation to a variable environment. �ey are characterised by large population sizes, short generation time, and the ability to manipulate their environment in controlled conditions (jessup et al., 2004). paramecium bursaria ehrenberg 1831 is cosmopolitan organism, inhabiting standing or slowly �owing water with relatively high purity. p. bursaria forms the endosymbiotic relationship with algae of chlorella species (reisser, 1980). �is relationship is an unusual example of optional and mutualist interaction between the two species. �ere are up to several hundred symbiotic algae inside the cell of p. bursaria (karakashian et al., 1968) (fig. 1). �e symbiotic algae are enclosed in a perialgal vacuole membrane (derived from the host digestive vacuole), and this membrane protects the algae from the hosts lysosomal digestion (kodama, fujishima, 2005). �e host cell protects endosymbionts from infections by chlorella virus. paramecium supply algal cells with nitrogen components and the co2 necessary for photosynthesis (reisser, 1980; albers, wiessner, 1985; kodama, fujishima, 2005). green endosymbionts carry out photosynthesis and thus provide the host with maltose and oxygen (brown, nielsen, 1974); therefore, p. bursaria becomes completely or partially independent of the external source of food (sommaruga, sonntag, 2009). endosymbionts inside p. bursaria are sensitive to di�erent environmental factors, e.g., temperature. long-term exposure ciliates to low temperatures may cause changes in the early stages of development of their metabolism and consequently lead to the microorganisms extinction (unpublished). global warming leads to changes in the process of photosynthesis and respiration in autotrophic organisms. psii seems to be one of the most thermo-sensitive protein complex pigments, which regulates photosynthetic activity in algae, cyanobacteria, as well as in higher plants (strasser et al., s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 70 1995; morgan-kiss et al., 2006). chlorella vulgaris beijer. cells regulate photosynthetic processes at the level of lhcii polypeptides, chlorophyll molecules, as well as through the xanthophyll cycle, in response to di�erent temperatures and light intensities (wilson, huner, 2000). �e aim of this study was to investigate the e�ects of temperatures (21°c, 24°c, 27°c, 30°c, and 33°c) on the photosynthesis carried out by endosymbiotic green algae of two p. bursaria strains from warm climate (ardmore, usa) with an average annual temperature of +23.7°c (ard7) and from cold climate (kamchatka, russia) with an average air temperature of -6°c (kd64). fig. 1. paramecium bursaria with green endosymbionts – a; endosymbiotic algae isolated from the paramecium bursaria cell – b (photo. m. ślęczka) 71 material and methods �e experiments were conducted at the institute of biology of pedagogical university of kraków. �e study material was paramecium bursaria strains from (1) kamchatka (kd64) located in the asian part of russia (58°36ʹ40ʹʹn; 38°54ʹ44ʹʹe) and (2) ardmore (ard7) located in the south-eastern carter county, oklahoma, united states (34°10ʹ52ʹʹn; 97°07ʹ46ʹʹw). paramecium bursaria culture techniques green paramecium bursaria strains were grown on a lettuce medium with klebsiella pneumoniae (smc strain) (sonneborn, 1970). �e cultures were maintained under constant light/dark cycle (12l:12d) at 18°c, at light intensity 200 µmol m-2 s-1 for 7 days at the following temperatures: 21°c, 24°c, 27°c, 30°c and 33°c. chlorophyll a �uorescence chlorophyll a �uorescence was measured using a handy plant e�ciency analyser �uorimeter (hansatech instruments, united kingdom). a 1 ml sample, with green p. bursaria, was taken into glass cell, then the sample was darkened for 5 minutes the photosynthetic activity of param ecium bursaria endosym biotic algae in varying tem perature conditions fig. 2. chlorophyll a �uorescence parameters of paramecium bursaria strains: kd64 and ard7, incubated at di�erent temperatures; di�erent letters di�er signi�cantly according to the duncan test at p ≤ 0.05; n = 5 s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 72 to make the conditions needed to expire the light phase of photosynthesis. �e obtained results were analysed as follows: f0 – chlorophyll �uorescence intensity measured when all photosystem ii reaction centres are open, fm – maximal chlorophyll �uorescence intensity measured when all photosystem ii reaction centres are closed, fv – variable chlorophyll �uorescence (fm/f0), fv/f0 – e�ciency of the water-splitting complex on the donor side of psii and fv/fm – maximum quantum yield of psii. in addition, tfm – time needed for reaching fm (ms), rc/abs – index expression as the density of reaction centres (rc), pi – indicator of the functioning of psii, tro/rc – trapped energy �ux per cross section (rc) at t = 0, eto/rc – electron transport �ux per cross section (rc) at t = 0, and vj – relative change in chlorophyll �uorescence during the light phase of photosynthesis. emission �uorescence – spectro�uorimetry method measurement of blue-green and red �uorescence emission spectra were performed according to lichtenthaler et al. (2004) with a spectro�uorimeter (perkin-elmer ls55b, united kingdom) equipped with a liquid measuring device. measurements of �uorescence intensity in the range of blue-green light (430–650 nm) were performed at 390 nm and near and far red (650–800 nm) with blue 430 nm. �e slot for the excitation radius was 15 nm, and for the emitted 20 nm. results were analysed using fl winlab version no. 3.00. statistic analysis a parametric multi-factorial anova / manova test was used to compare the variables tested, based on multiple duncan homogeneous tests at p ≤ 0.05; n = 5. calculations were made using statso�, inc. (2014). statistica®12. program. results �e minimum (f0) and maximum (fm) �uorescence values for both strains increased with temperature. �e highest f0 values were observed at 30°c. parameters of maximum psii (fv/fm) photochemical e�ciency and maximum splash water yield a�er psii donor side (fv/f0) were highest at 18°c and lowest at 30°c for two paramecium bursaria strains (fig. 2). �e chlorophyll a �uorescence values were considerably di�erent at tested temperatures compared to the control group. �e chlorophyll �uorescence parameters (f0 and fm) were signi�cantly higher for the kd64 strain compared to the ard7 strain (fig. 2–3). �e blue-green and red �uorescence emission spectra in kd64 and ard7 strains were similar in the shape (fig. 4). �e increase of blue-green emission �uorescence 73 the photosynthetic activity of param ecium bursaria endosym biotic algae in varying tem perature conditions fi g. 3 . c hl or op hy ll a �u or es ce nc e pa ra m et er s [ sh ow n as % o f c on tr ol ] o f p ar am ec iu m b ur sa ri a st ra in s i nc ub at ed a t d i� er en t t em pe ra tu re s s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 74 fi g. 4 . � e bl ue -g re en (a ) a nd r ed (b ) � uo re sc en ce e m is si on s pe ct ra k d 64 a nd a rd 7 st ra in s of p ar am ec iu m b ur sa ri a in cu ba te d at d i� er en t t em pe ra tu re s 75 was clearly shown at two wavelengths. �e �rst peak was observed at a wavelength 450–460 nm, and the second peak was at 485–490 nm. �e blue-green �uorescence emissions at 21°c were similar to the control group. �e �uorescence emission increased with temperature. red �uorescence emission spectra for both strains of p. bursaria were characterised by a distinct peak at a wavelength 675–685 nm with an arm at 750 nm. fluorescence intensity increased at temperatures 24°c to 30°c. �e values of f450/f685, f450/f735, f485/f685, and f485/f735 for p. bursaria strains decreased with increasing temperature. �e parameter f450/f530 increased with temperature, and the highest value was observed at 30°c (tab. 1). tab. 1. fluorescence emission factors values of ard7 – (a) and kd64 – (b) strains incubated at di�erent temperatures. values shown as di�erent letters within the line di�er signi�cantly according to the duncan test at p ≤ 0.05; n = 5 ratio temperature [°c] 18 21 24 27 30 a b a b a b a b a b f450/f685 65.5a 49.1b 48.5b 45.9bc 40.9bcd 42.7bcd 36.90cd 44.2bcd 26.9e 35.2d f450/f735 652.7a 341.9ab 392.0ab 339.5ab 373.3ab 299.2ab 298.5ab 292.9ab 278.1b 262.6b f450/f530 1.43c 1.10g 1.46bc 1.27f 1.47b 1.27f 1.47b 1.30e 1.72a 1.40d f485/f685 72.3a 52.9bc 52.6bc 56.1b 44.6cd 49.3bcd 44.0cd 48.3bcd 29.2e 40.2d f485/f735 724.2a 394.0ab 466.3ab 391.6ab 407.6ab 341.1ab 323.5ab 320.6ab 301.0b 300.2b f685/f735 6.1a 5.4a 9.2a 6.6a 9.8a 8.0a 10.5a 7.4a 10.6a 8.5a discussion temperature has a major structuring e�ect at all levels of biological organisation. at the cell level, the temperature a�ects both energetic requirements and division rates, growth rate, and the decomposition and exchange of carbon dioxide and oxygen. �e functioning of the whole ecosystems depends on temperature (brown et al., 2004; savage et al., 2004). green algae tolerance is not the same for constant and variable temperature treatments (feder, hofmann, 1999). ciliates respond promptly and di�erently to environmental change (jiang, morin, 2004; esteban, finlay, 2007). in the present study, there was revealed a signi�cant e�ect of temperature on the photosynthetic activity of endosymbionts inside the paramecium bursaria cells, which originated from a warm and cold climate. one-week incubation of ciliates at di�erent temperatures caused changes in chlorophyll a �uorescence parameters (fig. 2–3), and an increase of �uorescence the photosynthetic activity of param ecium bursaria endosym biotic algae in varying tem perature conditions s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 76 emission intensity (fig. 4). �e values of emission �uorescence ratios (f450/f530 and f685/f730) were increased with temperature (tab. 1). �ese changes may indicate a decrease in the e�ciency of primary reactions occurring in psii and the activation of defence mechanisms of endosymbiont photosynthetic apparatus (lichtenthaler, rinderle, 1988; chemeris et al., 2004). at high temperatures, cell membrane permeability and damage of psii subunits increase (kota et al., 2002). �e changes are observed (i) in the structure of proteins and lipids, (ii) in the functioning of ion channels, (iii) disturbances in electron transport and in the reduction of electron acceptors, (iv) in the e�ciency of oxygen extraction, and (v) the dissipation of heat (weng, lai, 2005). high temperatures cause a blockade of energy transfer from the reaction centre to plastochinone (reigosa, weiss, 2001). changes in f450/f530 values indicate an increase in phenolic compounds, and changes in f685/f730 values indicate a decrease in chlorophyll content (lang et al., 1991; lichtenthaler et al., 2004). �e rate of p. bursaria metabolism depends on the number of endosymbiotic chlorella cells and their photosynthetic activity (weis, 1969). �e photosynthetic products of symbiotic green algae increase the tolerance to high temperatures of the host cell (iwatsuki et al., 1998). p. bursaria cells with chlorella algae are more tolerant to high temperatures than algae-free ciliates (miwa, 2009). �e studies on the e�ect of temperature on the morphology and physiology of algae show that the optimal growth temperatures for chlorella vulgaris range from 26°c to 34°c (mayo, 1997; ma et al., 2014). duncan et al. (2011) showed that paramecium from variable environments grow well at both 23°c and 35°c. at temperatures from 29°c to 39°c, chlorella sp. strain r-06/2 originating from geothermal source in rupite (bulgaria) is highly photosynthetically e�cient (gacheva, pilarski, 2008). under natural lighting conditions, the highest increase in chlorophyll content, carotenoids and proteins in c. vulgaris are observed at temperatures from 25°c to 30°c. under continuous light conditions and at temperatures from 30°c to 35°c, algae growth is minimal (sharma et al., 2012). changes in physiological properties are due to the endosymbiotic close relationship between paramecium and algae (reisser, 1986). according to mcauley et al. (1996), the host regulates the growth of symbiotic algae. in the present study, the higher di�erences in photosynthetic activity were observed in the kd64 strain from kamchatka (fig. 2–4; tab. 1). �e environmental stressors may cause many adverse changes in aquatic ecosystems, as well as for the economy and human health. �at is why it attaches great importance to ensuring continuous monitoring of waters, so that changes can be noted and appropriate corrective or preventive measures taken in the natural environment. given the signi�cant ecological role played by ciliates, it is important to understand how temperature a�ects the adaptation of organisms in their local environment. 77 conclusion �e study showed a signi�cant e�ect of temperature on the activity of the photosynthetic apparatus of the paramecium bursaria green endosymbionts. with an increase of temperature, changes in psii were observed. high temperature caused an increase of blue-green and red �uorescence emission of endosymbiotic algae. �e strain of p. bursaria from kamchatka (kd64) was more sensitive than the strain from admore (ard7). references albers, d., wiessner, w. 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(2004). recording chlorophyll �uorescence emission spectra with the perkin elmer �uorescence spectrometer ls 50. in: m. filek, j. biesaga-kościelniak, i. marcińska (eds.), analytical methods in plant stress biology. kraków: drukrol, 112–124. lichtenthaler, h.k., rinderle, u. (1988). �e role of chlorophyll �uorescence in the detection of stress conditions in plants. critical reviews in analytical chemistry, 19, s29–s85. ma, x., zheng, h., huang, h., liu, y., ruan, r. (2014). e�ects of temperature and substrate concentration on lipid production by chlorella vulgaris from enzymatic hydrolysates of lipid-extracted microalgal biomass residues (lmbrs). applied biochemistry and biotechnology, 174(4), 1631–1650. doi: 10.1007/s12010-014-1134-5 mayo, a.w. (1997). e�ects of temperature and ph on the kinetic growth of unialga chlorella vulgaris cultures containing bacteria. water environment research, 69(1), 64–72. doi: 10.2175/106143097x125191 mcauley, j.p., dorling, m., hodge, h. (1996). e�ect of maltose release on uptake and assimilation of ammonium by symbiotic chlorella (chlorophyta). journal of phycology, 32, 839–846. doi: 10.1111/j.0022-3646.1996.00839.x miwa, i. (2009). regulation of circadian rhythms of paramecium bursaria by symbiotic chlorella species. in: m. fujishima (ed.) endosymbionts in paramecium. microbiology monographs, 12, 83–110. morgan-kiss, r., ivanov, a.g., williams, j., khan, m., huner, n.p.a. (2002). di�erential thermal e�ects on the energy distribution between photosystem ii and photosystem i in thylakoid membranes of a psychrophilic and a mesophilic alga. biochimica et biophysica acta (bba)-biomembranes, 1561(2), 251–265. doi: 10.1016/s0005-2736(02)00352-8 reigosa, r.m.j., weiss, o. (2001). fluorescence techniiques. in: r.m. reigosa (ed.) handbook of plant ecophysiology techniques. dordrecht: �e netherlands kluwer academic publishers, 155–171. reisser, w. (1980). �e metabolic interactions between paramecium bursaria and chlorella spec. in the paramecium bursaria-symbiosis. archives of microbiology, 125, 291–293. doi: 10.1007/bf00446890 reisser, w. (1986). endosymbiotic associations of freshwater protozoa and algae. program protistolgy, 1, 195–214. savage, v.m., gillooly, j.f., brown, j.h., west, g.b., charnov, e.l. (2004). e�ects of body size and temperature on population growth. american naturalist, 163, 429–441. doi: 10.1086/381872 sharma, r., singh, g.p., sharma, v.k. (2012). e�ects of culture conditions on growth and biochemical pro�le of chlorella vulgaris. journal of plant pathology and microbiology, 3(5), 1–6. doi: 10.4172/21577471.1000131 sommaruga, r., sonntag, b. (2009). photobiological aspects of the mutualistic association between paramecium bursaria and chlorella. in: m. fujishima (ed.), endosymbionts in paramecium. microbiology monographs, 12, 111–130. doi: 10.1007/978-3-540-92677-1_5 sonneborn, t.m. (1970). methods in paramecium research. in: e.d.m prescott (ed.), methods in cell biology. new york: academy press, 241–339. strasser, r.j., strvastava, a., govindjee (1995). polyphasic chlorophyll a �uorescence transient in plants and cyanobacteria. photochemistry and photobiology, 61, 32–34. doi: 10.1111/j.1751-1097.1995. tb09240.x weis, d.s. (1969). regulation of host and symbiont population size in paramecium bursaria. experientia, 25(6), 664–6. 79 weng, j.h., lai, m.f. (2005). estimating heat tolerance among plant species by two chlorophyll �uorescence parameters. photosynthetica, 43, 439–444. doi: 10.1007/s11099-005-0070-6 wilson, k.e., huner, n.p. (2000).�e role of growth rate, redox-state of the plastoquinone pool and the trans-thylakoid deltaph in photoacclimation of chlorella vulgaris to growth irradiance and temperature. planta, 212(1), 93–102. doi: 10.1007/s004250000368 abstract �e aim of this study was to investigate the e�ect of higher temperatures on the photosynthesis of endosymbiotic chlorella sp. of two paramecium bursaria ehrenberg 1831 strains originating from regions with a warmer and colder climate (ardmore – usa and kamchatka – russia, respectively). a�er seven days of protozoa incubation at 18°c (control), 21°c, 24°c, 27°c, 30°c and 33°c, the chlorophyll a �uorescence measurements were carried out and �uorescence spectra were measured in blue-green and red light. as a result of the studies, a signi�cant e�ect of higher temperature on the photosynthesis process of p. bursaria endosymbionts was observed. weekly incubation at 33°c was lethal for both protozoan strains in comparison to the control temperature (18°c). �e blue-green �uorescence spectra were characterised by marked peaks at 450 nm and 490 nm. within the red light range, the peak was observed at about 690 nm with a lesser arm at 730 nm. endosymbionts from kamchatka were more sensitive to the temperature increase than algae from areas with relatively warm climates. key words: emission �uorescence, high temperatures, psii activity, spectro�uorimetry, chlorella vulgaris received: [2017.08.13] accepted: [2017.11.14] aktywność fotosyntetyczna endosymbiotycznych glonów paramecium bursaria w zróżnicowanych warunkach temperatury streszczenie celem niniejszej pracy było zbadanie wpływu podwyższonej temperatury na przebieg procesu fotosyntezy endosymbiontów z gatunku chlorella sp. dwóch szczepów paramecium bursaria ehrenberg 1831, pochodzących z terenów o niskich i wysokich temperaturach powietrza (ardmore – usa i kamczatka – rosja). po 7 dniach inkubacji pierwotniaków w każdej z temperatur 18°c (kontola), 21°c, 24°c, 27°c, 30°c i 33°c przeprowadzono pomiary �uorescencji chloro�lu a  i  wyznaczono widma emisji �uorescencji w  zakresie niebiesko-zielonym i czerwonym.w wyniku przeprowadzonych badań zaobserwowano istotny wpływ podwyższonej temperatury na proces fotosyntezy endosymbiontów p. bursaria. tygodniowa inkubacja w temperaturze 33°c była letalna dla obu szczepów pierwotniaka, w porównaniu z temperaturą kontrolną (18°c). widma emisji �uorescencji niebiesko-zielonej charakteryzowały wyraźnymi pikami przy 450nm i 490 nm. w  zakresie czerwonym pik zaobserwowano przy około 690 nm z  mało wyraźnym ramieniem przy 730 nm. endosymbionty szczepu pochodzącego z kamczatki były bardziej wrażliwe na wzrost temperatury od glonów pochodzących z terenów o stosunkowo ciepłym klimacie. słowa kluczowe: wysoka temperatura, aktywność psii, spektro�uorymetria, szczepy chlorella vulgaris information on the authors katarzyna możdżeń her scienti�c interests concentrate on the e�ects of di�erent environmental factors (light, ozone, heavy metals, allelopathic extracts) on the morphology and physiology plants cultivated, protected, and invasive species. patrycja zagata leśnicka she is graduate of doctoral studies at the institute of biology at the pedagogical university in kraków. her scienti�c interests concern microbiology and algology, and especially the endosymbionts of paramecium bursaria. the photosynthetic activity of param ecium bursaria endosym biotic algae in varying tem perature conditions s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 80 mateusz ślęczka he graduated from the environmental protection at the pedagogical university in kraków. his scienti�c interests concern environmental microbiology. magdalena greczek-stachura she is associate professor at the pedagogical university in cracow. her interest is in the broadly understood microbiology and algology. in recent years, her research has focused on endosymbiosis in a group of ciliata (animal protista). 196 on 24–27 may 2017, the xvii edition of the festival of science and art took place in cracow. the pedagogical university of the national education commission took custody of the organisation of this event for the third time. just like last year, the vice-rector for development, associate professor robert stawarz was the host of the festival. the main honorary patrons were the president of the city of cracow, the province governor and marshal of the lesser poland region, the minister of science and higher education, the minister of sport and tourism, the minister of national education, the european commission representation in poland, the president of the college of rectors of cracow and the national centre for research and development. traditionally, the centre of events was located on the main market square in cracow, where a scientific tent town was established (fig. 1). this year’s edition of the festival ran under the slogan “in harmony with nature”. on this occasion, the representatives of scientific centres prepared interesting expositions and demonstrations. at the main market square in cracow, thousands of people xvii festival of science and art in kraków “in harmony with nature”, kraków, 24–27th may 2017, poland xvii festiwal nauki i sztuki w krakowie „w zgodzie z naturą”, kraków, 24–27 maj 2017, polska w dniach 24–27 maja 2017 r. odbyła się xvii edycja festiwalu nauki i sztuki w krakowie. opiekę nad organizacją tego wydarzenia już po raz trzeci sprawował uniwersytet pedagogiczny im. komisji edukacji narodowej. podobnie jak w ubiegłym roku, rolę gospodarza tego festiwalu pełnił prorektor ds. rozwoju dr hab. robert stawarz. głównymi patronami honorowymi byli prezydent miasta krakowa, wojewoda i marszałek małopolski, minister nauki i szkolnictwa wyższego, minister sportu i  turystyki, minister edukacji narodowej, przedstawicielstwo komisji europejskiej w  polsce, przewodniczący kolegium rektorów szkół wyższych krakowa oraz narodowe centrum badań i rozwoju. tradycyjnie centrum wydarzeń zlokalizowane było na rynku głównym w  krakowie, gdzie powstało namiotowe miasteczko naukowe (ryc. 1). tegoroczna edycja festiwalu przebiegała pod hasłem „w  zgodzie z  naturą”. z  tej okazji przedstawiciele krakowskich ośrodków naukowych przygotowali interesujące ekspozycje i pokazy. na rynku głównym w  krakowie tysiące osób miało możliwość wzięcia udziału w  prezentacjach i  eksperymentach naukowych. dodatkowe annales universitatis paedagogicae cracoviensis studia naturae, 2: 196–203, 2017, issn 2543-8832 197 r eports were able to take part in presentations and scientific experiments. additional attractions awaited visitors at the premises of individual universities and institutions involved in the organisation of this year science festival. during the festival, one could visit museums, scientific and cultural institutes, scientific laboratories, astronomy observatory, etc. the official inauguration of the festival began with a concert performed by lora szarfan and henryk miśkiewicz and the performance of the big band of the academy of music in cracow. during the festival, the organizers encouraged visiting, among others, the railway control room of the cracow main station, and watching a play entitled “to dna” performed by students of the cracow state higher school of theatre of ludwik solski. the program, in addition atrakcje czekały na zainteresowanych w  siedzibach poszczególnych uczelni oraz instytucji zaangażowanych w  organizację tegorocznego święta nauki. w  trakcie festiwalu można było zwiedzać, m.in.: muzea, instytuty naukowe i  kulturalne, pracownie naukowe, obserwatorium astronomiczne itp. uroczysta inauguracja festiwalu rozpoczęła się koncertem w  wykonaniu lory szarfan i  henryka miśkiewicza oraz występem zespołu big band akademii muzycznej w krakowie. w trakcie festiwalu organizatorzy zaprosili do zwiedzania, m.in. nastawni kolejowej kraków główny oraz do obejrzenia spektaklu pt. „do dna”, w  wykonaniu studentów krakowskiej państwowej wyższej szkoły teatralnej im. ludwika solskiego. w  programie obok prelekcji i  doświadczeń laboratoryjnych znalazły się także sportowe fig. 1. view on a scientific town on cracow main square (photo. k. strzeżoń) ryc. 1. widok na naukowe miasteczko na krakowskim rynku głównym (fot. k. strzeżoń) 198 r ep or ts to the lectures and laboratory experiments, also included sports competitions, “naturally futsal!” art contest “nature is a priceless gift,” photographic contest “nature and human” and prof. walery goetl’s memory seminar devoted to contemporary environmental issues. during the cracow scientific festival, the collegium medicum of the jagiellonian university organised interesting lectures for primary and pre-secondary schools, as well as for the inhabitants of cracow. the staff of the haematology department and clinic prepared a lecture “what can be found in blood? blood – its ingredients and functions”. stories about the treatment of obesity “one day from the surgeon’s life – that is, what the surgeon actually does” could be heard at the ist department of general surgery. the answers to questions: “are there any useful bacteria that affect human health and where to look for them?” were provided during the workshops at the department of microbiology. new technical advances used in the treatment of the central nervous system were presented by scientists from the department of neurosurgery and neurotraumatology during the lecture “technology a precision in neurosurgery operations”. the department of pathomorphology presented the problem of smog and its effect on human lungs, “smog – is there anything to be afraid of ?” in the zodiac room of the jagiellonian university. researchers from the haematology department and clinic, during the lecture “how to share life…, the secrets of transplantation and everything about being a bone marrow donor”, talked about bone marrow transplant, “what does it mean to be a conscious donor? can blood rozgrywki „naturalnie futsal!”, konkurs plastyczny „natura bezcennym darem”, fotograficzny „natura a  człowiek” oraz seminarium pamięci prof. walerego goetla, poświęcone współczesnym problemom ochrony środowiska. podczas krakowskiego święta nauki collegium medicum uniwersytetu jagiellońskiego zorganizowało ciekawe wykłady dla szkół podstawowych, gimnazjalnych oraz dla mieszkańców krakowa. pracownicy katedry i kliniki hematologii przygotowali prelekcję „co tkwi w  kropli krwi? krew – jej składniki i  funkcje”. opowieści o  leczeniu otyłości „dzień z  życia chirurga – czyli czym właściwie zajmuje się lekarz chirurg” można było posłuchać w  i  katedrze chirurgii ogólnej. odpowiedzi na pytania: „czy istnieją pożyteczne bakterie, jaki wpływ mają na zdrowie człowieka i  gdzie ich szukać?” zostały udzielone podczas warsztatów w katedrze mikrobiologii. o nowych zdobyczach technicznych wykorzystywanych w  leczeniu ośrodkowego układu nerwowego opowiadali naukowcy z kliniki neurochirurgii i neurotraumatologii podczas wykładu „technologia a precyzja w  operacjach neurochirurgicznych”. katedra patomorfologii przedstawiła problem smogu i jego wpływu na płuca człowieka „smog – czy jest się czego bać?” w sali zodiakalnej uj. naukowcy z katedry i kliniki hematologii podczas wykładu „jak podzielić się życiem…, czyli tajniki transplantacji i wszystko o byciu dawcą szpiku kostnego” mówili o transplantacji szpiku kostnego, z  czym wiąże się bycie świadomym dawcą? czy krew może zmienić płeć? badacze z  kliniki gastroenterologii i  hepatologii przekonywali słuchaczy do zdrowego trybu życia podczas wykładu „zasady zdro199 r eports change sex?” researchers from the clinic of gastroenterology and hepatology persuaded their listeners to a healthy lifestyle during the lecture “principles of healthy nutrition”. on the other hand, the department of forensic medicine presented the toxicological and analytical aspects of using power-ups during the lecture “power-ups – current trends and selected cases of poisoning from the practice of the department of forensic medicine in cracow”. the scientists presented the selected cases of poisoning with power-ups as a warning, which were recorded in the department of forensic medicine in cracow. all faculties of the agricultural university of cracow also joined the festival (fig. 2–3). employees, postgraduates, and students from this university were encouraged to admire ecological thermodynamic engines, vegetation of roofs and green terraces, wego odżywiania”. natomiast katedra medycyny sądowej zaprezentowała aspekty toksykologiczne i  analityczne stosowania dopalaczy podczas prelekcji „dopalacze – obecne trendy i wybrane przypadki zatruć z praktyki zakładu medycyny sądowej w krakowie”. ku przestrodze naukowcy przedstawili wybrane przypadki zatruć dopalaczami, które odnotowano w  krakowskim zakładzie medycyny sądowej. w  organizację festiwalu włączyły się także wszystkie wydziały uniwersytetu rolniczego w krakowie (ryc. 2–3). pracownicy, doktoranci i  studenci z  tej uczelni zachęcali do podziwiania ekologicznych silników termodynamicznych, roślinności dachów i  tarasów zielonych, grzybków kefirowych, roślin in vitro, roślin energetycznych, żywności liofilizowanej itd. przygotowali również warsztaty z  układania bukietów, malowania fig. 2. tent of university of agriculture faculty of forestry (photo. k. strzeżoń) ryc. 2. namiot uniwersytetu rolniczego wydziału leśnego (fot. k. strzeżoń) 200 r ep or ts na mleku, czy ręcznego wyrobu drucianych zwierzątek. działalność kulturalną uniwersytetu rolniczego przedstawiły na scenie: studencki zespół góralski „skalni”, chór uniwersytetu rolniczego oraz zespół sygnalistów myśliwskich „hagard”. pracownicy z  zakładu fizjologii roślin instytutu biologii uniwersytetu pedagogicznego przygotowali doświadczenia z związane z procesem fotosyntezy i wpływem światła na rośliny. zakład zoologii bezkręgowców i  parazytologii prezentował szkodniki pasożytujące na produktach spożywczych, a  pracownia chemii postawiła w  tym roku na pokaz wpływu kwasów i  zasad na materiały organiczne. elementy biofizyki w życiu codziennym, czyli eksperymenty z  napięcia powierzchniowego, metod wykrywania biakefir fungi, in vitro plants, energy plants, lyophilized food, etc. they also prepared workshops in arranging bouquets, painting on milk, or making wire animals by hand. the cultural activities of the agricultural university were presented on stage by the student highlander band “skalni”, the choir of the agricultural university and the hunting signal team “hagard”. employees from the department of plant physiology at the institute of biology of the pedagogical university prepared experiments related to the process of photosynthesis and the effect of light on the plants. the department of invertebrate zoology and parasitology presented parasitic worms parasitizing on foodstuffs, and the chemistry laboratory this year decided to present the effect of acids fig. 3. a scheme for the management of natural resources presented by the faculty of forestry of the university of agriculture (photo. k. strzeżoń) ryc. 3. schemat dotyczący zarządzania zasobami przyrody prezentowany przez wydział leśny uniwersytetu rolniczego (fot. k. strzeżoń) 201 r eportsand alkali on organic materials. elements of biophysics in daily life, i.e. surface tension experiments, methods of detecting proteins in milk, water, and juices were presented by the department of biochemistry, biophysics, and biotechnology. the construction of wood, the secondary growth of trees, and the factors influencing the production of their biomass were discussed by the employees, postgraduates, and students of the department of ecology and environmental protection. the most interested were also able to carry out an independent dendrochronological analysis, allowing them to determine the age of a tree, the conditions of growth, and the dating of disturbances that took place in the past in its habitat. apart from the lectures at the main square, the university of economics organised, among others, a lecture and debate on organic food in the university buildings. the participants were also able to discuss the air quality in cracow and determine the directions of improving the cleanliness of the environment. the visitors could also participate in the workshops “design thinking” and admire the exhibition of eco-implementation prototypes. the debate “full-time studies and gainful employment – un harmony with the students nature”. in the open air, the cracow university of technology prepared exhibitions of student paintings and drawings as well as architectural models. experiments related to mechanics and electricity were presented by representatives of the faculty of physics, mathematics, and computer science. visitors could learn about iot solutions in a smart home system equipped with motion, light, temperature and humidity sensors, and about a mobile łek w  mleku, wodzie, sokach prezentował zakład biochemii, biofizyki i  biotechnologii. o budowie drewna, przyroście wtórnym drzew i  czynnikach wpływających na produkcję ich biomasy opowiadali pracownicy, doktoranci i  studenci zakładu ekologii i  ochrony środowiska. najbardziej zainteresowani mieli również możliwość przeprowadzenia samodzielnej analizy dendrochronologicznej, pozwalającej na określenie wieku drzewa, warunków wzrostu i  datowaniu zaburzeń, które miały miejsce w przeszłości w jego siedlisku. uniwersytet ekonomiczny oprócz pokazów, prelekcji na rynku głównym, zorganizował w  budynkach uczelni, m.in. wykład i  debatę o  żywności ekologicznej. uczestniczy mogli także podyskutować o  jakości powietrza w  krakowie i  wyznaczeniu kierunków udoskonalania czystości środowiska. odwiedzający mogli wziąć również udział w  warsztatach „desing thinking” i  podziwiać wystawę prototypów eko-wdrożeń. dla studentów zorganizowano także debatę „studia dzienne i praca zarobkowa – w zgodzie z naturą studenta”. w  plenerze politechnika krakowska przygotowała wystawy studenckich prac malarskich i  rysunkowych oraz modeli architektonicznych. doświadczenia związane z  mechaniką oraz elektrycznością prezentowali przedstawiciele wydziału fizyki, matematyki i  informatyki. odwiedzający mogli dowiedzieć się o  rozwiązaniach iot w systemie inteligentnego domu wyposażonego w czujniki ruchu, światła, temperatury i wilgotności oraz o mobilnym systemie monitoringu i sterowania. interaktywne pokazy budowy konstrukcji inżynierskich „zbuduj 202 r ep or ts monitoring and control system. interactive demonstrations of engineering structures “build your own bridge”, of individual protection measures for high-altitude works and medical rescue in construction and communication cases were organized by the faculty of civil engineering. the festival program also included the following: interactive transport games, vehicle motion simulations, prize contests, wind farm demonstrations, simulations of the effects of lack of ventilation in the cities and smog prevention measures, a heron fountain, a model of a well with a deep water pump, and a model of a biological wastewater treatment, a presentation of rocks from different regions of the world, the rinsing of gold and expensive stones, the operation of thermal imaging cameras, and others. it is also worth mentioning the 3d mapping exhibition on the building of the interdepartmental education and research centre “działownia”, organised by the students of the compute graphics scientific group and the computer graphics and multimedia specialists of the cracow university of technology. the department of foundry engineering of the mining and metallurgy academy organised three-dimensional scanning shows, and observations using the endoscopic camera “look inside the engine”. the faculty of physics and applied computer science paraphrased this year festival slogan “in harmony with physics!” and conducted experiments illustrating the formation of clouds, water and fire tornadoes, and lightning. representatives of the department of mining geodesy and environmental engineering performed chemical analyses of air, water, and soil. they also presented home-made water treatment swój most”, pokazy środków ochrony indywidualnej przy robotach wysokościowych i  działania ratownictwa medycznego w  wypadkach budowlanych oraz komunikacyjnych zorganizował wydział inżynierii lądowej. w  programie festiwalu znalazły się również: interaktywne gry transportowe, pokazy symulacji ruchu pojazdów, konkursy z  nagrodami, prezentacje laboratorium wiatrowego, symulacje skutków braku przewietrzania miast oraz sposobów walki ze smogiem, fotanna herona, model studni z pompą głębinową i biologiczne oczyszczanie ścieków, prezentacje skał z różnych rejonów świata, płukanie złota i drogich kamieni, działanie kamery termowizyjnej i  inne. warto też wspomnieć o  pokazie 3d mappingu na budynku międzywydziałowego centrum edukacyjno-badawczego „działownia”, zorganizowanym przez studentów koła naukowego grafiki komputerowej oraz specjalistów grafiki komputerowej i multimediów politechniki krakowskiej. wydział odlewnictwa akademii górniczo-hutniczej zorganizował pokazy skanowania trójwymiarowego, obser wacje z  wykorzystaniem kamery endoskopowej „zajrzyj do wnętrza silnika”. wydział fizyki i  informatyki stosowanej parafrazując tegoroczne hasło festiwalu na „w  zgodzie z  fizyką!”, przeprowadził doświadczenia ilustrujące powstawanie chmur, tornad wodnych i  ogniowych oraz piorunów. przedstawiciele wydziału geodezji górniczej i  inżynierii środowiska wykonywali analizy chemiczne powietrza, wody i  gleby. zaprezentowali także domowe metody uzdatniania wody oraz zapraszali do obserwacji mikroorganizmów występujących 203 r eportsmethods and invited people to observe microorganisms found in the rivers. additional attractions included thermal images in anti-smog masks, demonstrations of how the cyclone works, and the possibility of shaping the surface of sandboxes scanned by a kinect motion sensor. during the festival, one could meet robots imitating emotions, robots and vehicles powered by energy from renewable sources, and one could participate in workshops, “creative design of the future”. in addition to screenings of films, exhibitions and performances, a special guest took part in the finale of the xvii science festival, gheorghe zamfir, a world famous virtuoso of the pan flute. his concert under the title of nature: mater et magistra took place on may 26 in the st. catherine of alexandria church in cracow. he was accompanied by, among others, the orchestra of the pedagogical university and the academic choir headed by prof. adam korzeniowski. there were many attractions during this year meeting with the world of science. it is impossible to mention all the events here, which were prepared by the representatives of cracow universities and other scientific institutions. one can hope that next year we will meet again in the same place, in a group even bigger than before. the organisers will probably make every effort to surprise their guests once again. w rzekach. dodatkową atrakcją były zdjęcia termalne w maskach antysmogowych, pokazy jak działa cyklon oraz możliwość kształtowania powierzchni piaskownicy skanowanej za pomocą sensora ruchu kinect. podczas festiwalu można było spotkać roboty naśladujące emocje, roboty i  pojazdy zasilane energią ze źródeł odnawialnych, a także wziąć udział w warsztatach „twórcze projektowanie przyszłości”. oprócz pokazów filmów, wystaw i  spektakli, w  finale xvii festiwalu nauki wystąpił gość specjalny gheorgha zamfira, światowej sławy wirtuoz fletni pana. jego koncert pod hasłem natura: mater et magistra odbył się 26 maja w  kościele św. katarzyny aleksandryjskiej w krakowie. towarzyszyli mu, m.in. orkiestra uniwersytetu pedagogicznego oraz chór akademickim pod dyrekcją prof. adama korzeniowskiego. atrakcji podczas tegorocznego spotkania ze światem nauki było wiele. nie sposób wspomnieć tutaj o wszystkich wydarzeniach, które przygotowali przedstawiciele krakowskich uczelni i innych instytucji naukowych. można mieć nadzieję, że za rok znów się spotkamy w  tym samym miejscu, w  jeszcze większym gronie niż dotychczas. zapewne organizatorzy dołożą wszelkich starań, by po raz kolejny zaskoczyć swoich gości. anna kocoń1, katarzyna możdżeń2 1department of invertebrate zoology and parasitology, institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland, a_kocon@up.krakow.pl 2department of plant physiology, institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland 159 annales universitatis paedagogicae cracoviensis studia naturae, 2: 159–169, 2017, issn 2543-8832 doi: 10.24917/25438832.2.12 samuel robinson1, william cetzal-ix2, saikat kumar basu3* 1department of biological sciences, university of calgary, alberta, canada 2instituto tecnológico de chiná, tecnológico nacional de méxico, chiná, campeche, mexico 3department of biological sciences, university of lethbridge, alberta, canada, *saikat.basu@alumni.uleth.ca wild bee decline and conservation in north america pollination by wild bees one of the major challenges of the 21st century will be feeding an increasing number of people on smaller and smaller areas of fertile land. while much of our diet comes from crops which do not require any animal pollination (e.g., wheat, corn, rice), about 35% of global food production depends on pollinating animals, such as bees (klein et al., 2007). over 90% of the vitamin c in our diet – as well as a majority of other vitamins, minerals, antioxidants, and micronutrients – come from animal-pollinated crops (eilers et al., 2011). �is makes animal pollinators an important part of the human food supply and global economy, and conservation of pollinators will be a key factor in managing global food security. historically, most pollination worldwide has been done by wild bees, and in most undeveloped nations, a majority of pollination is still done by wild pollinators (�ies, moths, butter�ies, bats, and hummingbirds also provide some services). however, developed nations typically rely on large-scale pollination services provided by the european honey bee, apis mellifera l. honey bees were brought to north america by european settlers, and they have expanded into a  multi-billion dollar industry over the last 50 years. �e total value of pollination services worldwide is close to $178 billion usd (gallai et al., 2009). in the united states in 2009, the total value of crop pollination services from honey bees was $11.6 billion, while services from non-honey bees were about $3.4 billion, with much of this value coming from almond, apple, and sun�ower crops, which are all highly pollinator-dependent and grown in large amounts (calderone, 2012). while much of the general public are aware of honey bees and bumble bees in north america, they are o�en surprised to �nd out that this is a tiny fraction of the total number of bees! out of the 20.000 species of bees worldwide, there are about 4600 s am ue l r ob in so n, w ill ia m c et za l-i x, s ai ka t k um ar b as u 160 tab. 1. common families of bees in the united states, mexico, and canada family number of genera number of species examples andrenidae 12 1472 mining bee (andrena sp., panurgus sp.) fairy bee (perdita sp.) squash bees (peponapis sp.) apidae 83 1305 honey bee (apis mellifera) bumble bee (bombus sp.) long-horned bees (eucera sp., mellisodes sp.) carpenter bees (xylocopa sp.) stingless bees (melipona sp., trigona sp.) colletidae 9 288 cellophane bee (colletes sp., hylaeus sp.) halictidae 26 654 sweat bee (halictus sp., lasioglossum sp.)green bee (agapostemon sp., augochlora sp.) megachilidae 26 791 leafcutter bee (megachile sp.) mason bee (osmia sp.) carder bee (anthidium sp.) mellitidae 3 28 mellita sp., macropis sp. species of bees living in north america (ascher, pickering, 2017) (tab. 1, appendix – fig. 1–2), and most of them are in the desert southwest of the united states (wilson, carril, 2016). �is is mainly due to the local diversity of local �owering plants, as high �owering plant diversity generally translates into higher bee diversity (potts et al., 2003). �ere are 46 species of bumblebee living in north america (williams et al., 2014), but most wild bees are solitary. solitary bees do not have a strict caste system with workers, queens, and drones, and can live their lives independently from each other. however, many ‘solitary’ bees will start acting socially (guarding nests, feeding other bees larvae, acting as queens and workers) if enough of them are in the same area (williams et al., 2014). pollinator decline are wild bee populations declining? because of their large diversity, di�culties in identifying them, and the fact that they are not as well-known as honey bees, they are less well-sampled in north america, and even less so in mexico (freitas et al., 2009). historical insect collections have proved to be valuable sources of information, as we can compare ‘snapshots’ of bee populations through time at a given location. unfortunately, this has revealed that many wild bee populations are in decline. cameron et al. (2011) used over 73.000 museum specimens to study 8 common species of bumblebees across the continental us, and found that 4 out of 8 were in serious decline. �e international union for conservation of nature (iucn) lists 12 out of 37 north american bumble bees (that we have good data for) as being endangered or vulneraw ild bee decline and conservation in n orth a m erica 161 ble (iucn, 2017). solitary bees do not fare much better in this regard. biesmeijer et al. (2006) found that there have been declines in wild bee populations across britain and the netherlands, and more worryingly, there have also been declines in the plants that are pollinated by these bees! causes of decline �e factors that contribute to the decline of wild bees can also be complex, but there are some general drivers of pollinator decline: 1. land use changes globally, about 40% of land has been converted to agricultural land (foley et al., 2005). in mexico, large-scale deforestation for charcoal, cattle, or agricultural expansion has reduced native rainforests to almost 10% of their original extent (freitas et al., 2009). more than 50% of north american grasslands have been converted to agriculture (hoekstra et al., 2005), and in some parts of western north america, this number approaches almost 100%, especially for tall-grass prairie (samson et al., 1998). habitat destruction has 2 main e�ects on wild bee populations: the reduction of food sources (�owers) and the reduction of nest sites. wind-pollinated crops, such as corn, provide little nutrition for bees, and even crops like canola or alfalfa (which bees can bene�t from) can be unhelpful in the long run, as the �owers are only available during a few weeks of the summer. some bee species are only active for a few weeks during the summer, so if their foraging time does not overlap with the �owering of the crop, they have no food! crop �elds can be dangerous for bees, as tillage and irrigation can destroy nest sites in the ground, and pesticides can injure or confuse bees. agricultural areas typically have few species of �owering plants present, meaning that generalist bees can persist (kleijn et al., 2015), but that �oral specialists will be without the �owers that they need. finally, crop �elds in the us and canada are o�en very large; small bees o�en only have �ight ranges of a few hundred yards, meaning that �nding areas with both nest habitat and food becomes more and more di�cult as �elds get bigger and bigger. 2. honey bees honey bees, while being incredibly useful and pro�table for humans, can put pressure on wild bee populations. �ey do this in two ways: competition for �owering plants, and disease spillover. honey bee colonies can raise 150–200.000 bees per year (seeley, 1985), and the sheer number of foraging workers can drain nectar and pollen resources around their hives. �ey can also �y up to 5 km while foraging, while many solitary s am ue l r ob in so n, w ill ia m c et za l-i x, s ai ka t k um ar b as u 162 bees only �y a few hundred meters during their entire lifetime, meaning that honey bees have the potential to compete with many types of wild bees. in mexico, stingless bees (melipona beecheii bennett) can be directly attacked by africanised honey bees, and declines in native mayan beekeeping over the last 20 years have been associated with increases in numbers of honey bees (cairns et al., 2005). many bumblebee diseases are similar to those of honey bees, meaning that disease spillover can occur between them. for example, fürst et al. (2014) found that both nosema ceranae i. fr., f. feng, j.a. da silva, s.b. slemenda & n.j. pieniazek and deformed wing virus (2 diseases of honey bees) could easily reproduce in bumblebee colonies, and more worryingly, both diseases were found in wild populations of bumblebees across the united kingdom. diseases of solitary bees are not well-studied, but chalkbrood fungus from honey bees (ascosphaera aggregata skou) can also infect leafcutter bees (goulson, 2003). finally, beekeepers provide help to honey bees by controlling their pests and diseases, and by feeding them during �ower-free periods. �is make sense, given that their livelihoods are based on having a large worker force, but this can put wild bees at a distinct disadvantage when it comes to surviving alongside honey bees. 3. insecticides large amounts of attention have been devoted to the e�ect of pesticides on honey bees. in particular, neonicotinoid insecticides have been implicated in honey bee deaths. �ere is far less data examining the e�ects of pesticides on wild bees, but research does show that insecticides can change foraging behaviour and can cause increased rates of mortality in bumblebees. for example, laycock et al. (2014) found that bumblebees fed small amounts of pesticides in their diet tended to live shorter lives. sandrock et al. (2014) showed that mason bees (osmia bicornis l.) can also be a�ected, as they tended to produce less o�spring when fed small doses of pesticides throughout their lives. however, it is far less clear whether these small-scale e�ects of pesticides are causing reductions in wild bee populations, and it is more likely that a combination of land use changes, diet changes, and pesticide usage are driving declines in wild bee populations (goulson et al., 2015). 4. mitigation clearly, there are many things that threaten the existence of wild bee populations. but how can we help encourage robust wild bee populations, and lessen risks of extinction? even more importantly, how do we balance our desire for agriculture and urban development while still conserving wild bees? to do this, we need to move beyond the “only honey bees” mentality. we suggest that both individuals and local governments should work to establish wild bee habitats in order to preserve local species diversity and abundance. w ild bee decline and conservation in n orth a m erica 163 while we recognise that honey bees are extremely practical in the modern agricultural setting, we encourage people to begin thinking beyond just honey bees, and to consider wild pollinators as valuable parts of our ecosystems. worldwide, the number of pollinator-dependent crops have grown more than 300% in the last 50 years, while the number of honey bee colonies has only grown by 45% (aizen, harder, 2009). �is means that the number of honey bees has not kept pace with our growing demand for pollination, and this ‘pollination gap’ will continue to increase into the future. wild pollinators also provide insurance against honey bee die-o�s, which have increased in severity in many parts of the united states. additionally, honey bees are not necessarily the best pollinators for all crops! garibaldi et al. (2013) found that in over 41 worldwide crop types, wild pollinators increased yield across all fruit, nut, and seed crop types, but that honey bees only increased yield in 14% of the crop types. in other words, honey bees can supplement, but not replace, pollination by wild pollinators. greenleaf and kremen (2006) found that the yield of hybrids sun�owers was increased in �elds located close to natural areas, and that honey bees switched between �owers more frequently if there were wild bees present, meaning that wild bees can actually increase the e�ciency of honey bee pollination! in this way, wild pollinators have the potential to work alongside introduced pollinators if they establish stable populations. bee habitats should provide a  diverse set of food resources and stable year-toyear nesting sites for wild pollinators. �ese areas can extend the bee foraging period beyond the �owering crop season (basu, cetzal-ix, 2017) and provide undisturbed nesting sites so that populations can more easily persist from year-to-year. plant seed mixtures could include native wild�owers, grasses, and legumes, as well as �owering trees and shrubs. priority should be given to native plants, as they generally bene�t wild pollinators more than introduced plant species. �e plant species included in the mixture should be selected so that they do not all �ower simultaneously, but are spread across the �owering season so that they bene�t the widest range of pollinators (basu, cetzal-ix, 2017). hedgerows or other pollinator habitats will need to be tailored speci�cally for each region in which they are planted, and the xerces society (http:// xerces.org/guidelines/) provides recommendations for planting pollinator habitats, including lists of plant species across the continental united states and canada. but will creating new pollinator habitats take away farmland or urban areas and create ‘o�-limit’ zones for human development or activity? not necessarily, because many unused areas already exist in urban and rural environments where these pollinator habitats could be established. habitats could be established along fence lines, wind breaks, irrigation canals, highways, sections of golf courses, and municipal parks and gardens (basu, 2017; basu, cetzal-ix, 2017). pollinator habitats can have positive impacts on the richness and abundance of wild bee species, and they can provide s am ue l r ob in so n, w ill ia m c et za l-i x, s ai ka t k um ar b as u 164 bene�ts to farmers growing pollinator-dependent crops. for instance, kremen and m’gonigle (2015) found that planting hedgerows (lines of �owering shrubs and bushes) in the highly cultivated napa valley in california increased the abundance of both common and rare pollinators. not only did these hedgerows increase abundance and diversity locally, but they acted as sources of wild pollinators for the surrounding adjacent �elds, as �elds planted next to hedgerows tended to have higher numbers of pollinators present in them (morandin, kremen, 2013). even within highly disturbed agricultural or urban areas, there is still hope for wild pollinators! acknowledgements wci would like to acknowledge the support from tecnm project (chn-lgac-03-14) “caracterización de la �ora nativa de importancia apícola en campeche: estrategia para incrementar la productividad en las colonias de abejas melíferas en periodos críticos de �oración”. research assistance and funding provided to skb by performance seed, lethbridge, ab, canada for developing di�erent performa pollinator mixes (comprising of annual/perennial legumes, wild�owers, and grasses) is gratefully acknowledged. references aizen, m.a., harder, l.d. 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(2016). �e bees in your backyard: a guide to north america’s bees. new jersey: princeton university press. a ppendix 1 167 appendix 1 fig. 1. diversity of �ora of mexico pollinated by di�erent species of bees. bees: a–e – apis mellifera l., f – scaptotrigona sp., g – bombus sp., h – euglossa viridissima friese. plants: a – capraria mexicana moric. ex benth. (scrophulariaceae)., b – species from the family of grasses (poaceae), c – ximenia americana l. (olacaceae), d – ruellia nudi�ora (engelm. & a. gray) urb. (acanthaceae), e – okenia hypogaea schltdl. & cham. (nyctaginaceae), f – lu�a aegyptiaca mill. (cucurbitaceae), g – catasetum integerrimum hook. (orchidaceae), h – notylia barkeri lindl. (orchidaceae) (photo. w. cetzal-ix) s am ue l r ob in so n, w ill ia m c et za l-i x, s ai ka t k um ar b as u 168 fig. 2. some common western north american bee species. bees: a – apis mellifera l., b – osmia lignaria say provisioning their nests in a bee block, c – bombus ternarius say, d – halictus sp., e – agopostemon virescens abrams & eickwort, f – andrena lupinorum cockerell, g – bombus borealis kirby, h – halictus spp. plants: a – medicago sativa l. (fabaceae), c – syringa vulgaris l. (oleaceae), d, h – taraxicum o cinale f. h. wigg. (asteraceae), e – gaillardia aristata pursh (asteraceae), f – brassica napus l. (brassicaceae) (photo. s. robinson and p. birch) a ppendix 1 169 zanikanie i ochrona dzikich pszczół w ameryce północnej streszczenie pszczoły – to niezwykle różnorodna i  ważna grupa owadów; około 4600 gatunków pszczół występuje w ameryce północnej. dzikie pszczoły „świadczą usługi” w zakresie zapylania roślin, a ich działalność jest prawdopodobnie nawet więcej warta niż działalność pszczół miodnych na całym świecie. są one słabo zbadane na większości obszarów, ale na terenach, dla których mamy wiarygodne dane, liczebność ich populacji obniża się. wynika to głównie z  przekształcania obszarów półnaturalnych w  grunty rolne, a  nowsze dane wskazują również, że pszczoły miodne mogą także wpływać na ich populacje. aby zachować istniejące populacje pszczół, należy ustalić siedliska zapylaczy, w celu zachowania różnorodności dzikich zapylaczy w  krajobrazach wiejskich i  miejskich. w  miarę jak populacja ludzi rośnie wraz z  zapotrzebowaniem na produkcję rolną, musimy znaleźć sposoby „współpracy” z dzikimi zapylaczami i sposoby zachowania dla przyszłych pokoleń tysięcy gatunków pszczół, które żyją w ameryce północnej i w innych częściach świata. key words: causes of decline, conservation, diversity, wild bee received: [2017.08.25] accepted: [2017.11.10] 41 annales universitatis paedagogicae cracoviensis studia naturae, 3: 41–51, 2018, issn 2543-8832 doi: 10.24917/25438832.3.3 monika grandtke1*, mariusz kasprzak1, mateusz ciepliński1, ewa burda1, ariel durajski1, joachim siekiera2, leszek jerzak1 1faculty of biological sciences, university of zielona gora, prof. z. szafrana st.1, 65-516 zielona gora, poland, *m.grandtke@wnb. uz.zgora.pl 2rzeczna 17 st., 47-300 żywocice, poland analysis of peripheral blood white blood cell parameters in european white storks (ciconia ciconia l.) chicks that varies by sex introduction biology and veterinary sciences have established health indicators for haematological parameters. the analysis of white blood cells provides some insight into the health status of the animal at the moment when it was sampled, and it can reflect habitat quality, nutrition, and other environmental stressors. because of the ease in obtaining them, they are generally some of the most important data available for wildlife health monitoring (bounous et al., 2000; campbell, 2015). avian leukocytes include granulocytes (heterophils, eosinophils and basophils) and agranulocyte cells (lymphocytes and monocytes) (lashev et al., 2005; szabó et al., 2010). the quantity and quality of leukocyte cells are generally used to determine immune reactions and diseases (mitchell, johns, 2008; shen et al., 2009). furthermore, changes in leucocytes also occur when birds are subjected to stress and when environmental quality is degraded (fiorello et al., 2009; kamiński et al., 2014). some authors used varied haematological values as parameters to determine the effect of stress caused by soil pollution in white storks (kamiński et al., 2015). the relation between heterophils and lymphocytes (h/l ratio) has been used as a reliable stress indicator in birds. when a bird is stressed, an increase in the number of heterophils and a decrease in the number of lymphocytes is usually observed (krams et al., 2012). because of the increasing interest of zoos and public parks in wild species, some studies about wild and captive animals can be found in the literature (aengwanich et al., 2002; narkkong et al., 2011). the white stork (ciconia ciconia l.) is classified in the genus ciconia, family ciconiidae, which includes 19 species around the world (norton, whiteside, 2015). in m on ik a g ra nd tk e, m ar iu sz k as pr za k, m at eu sz c ie pl iń sk i, ew a b ur da , a rie l d ur aj sk i, jo ac hi m s ie ki er a, l es ze k je rz ak 42 recent years, a noticeable change in the white stork population has been observed in europe, together with a rapid decrease in reproductive success and an increase in mortality rates (bocheński, jerzak, 2006; gilbert et al., 2016). limited data exists regarding blood cells characteristics, blood cells size, and the haematological value of white storks (jerzak et al., 2010; kamiński et al., 2014). the aim our study was the qualitative and quantitative analysis of white blood morphometric elements of peripheral blood (determining the quantity, blood cells dimension and several haematological values) in white stork chicks. one of the aims was to indicate whether the sex relevantly influences the variety of the examined white blood indicators. material and methods object samples were collected during the 2016 breeding season (june/july) from 53 juvenile white storks (25 males and 28 females) in krapkowice (50°28ʹ30ʹʹn, 17°57ʹ55ʹʹe). the age of the nestlings varied from 18 to 53 days. age (with accuracy of 1–2 days) was determined by measurement of the bill length following the kania method (1988); therefore, the hatch date was found by back calculation. blood collection blood was collected from the cutaneous ulnar vein by 20g peripheral venous catheter and 10 ml syringe (campbell, 2015). material from each specimen was transferred to tubes with anticoagulant immediately after collection. standard tubes containing k2edta for 2 ml of blood (2 mg of anhydrous k2edta per 1 ml of blood) (medlab-products) have been used. laboratory analysis haematological parameters were assessed using routine manual methods. total wbc were counted using the natt and herrick method (campbell, 2015). blood smears were prepared immediately after collection, using the push slide technique. smears of blood were air-dried, fixed in methanol and stained with the may–grünwald stain method (robertson, maxwell, 1990). slides were observed under a nikon eclipse ni microscope with a digital camera (nikon ds-fi2). cells were observed under a 1000× magnification and classified as heterophils, eosinophils, basophils, lymphocytes, and monocytes, according to criteria specified by others authors (clark et al., 2009; jamroz, lucas, 1961). the h/l ratio was then calculated by dividing the number of heterophils by the number of lymphocytes (lentfer et al., 2015). the stress factor was eliminated during 43 a nalysis of peripheral blood w hite blood cell param eters in european w hite s torks (ciconia ciconia l.) chicks that varies by sex the process of obtaining blood samples from the birds by covering the head of the chicks. photos of the blood cells were taken using a nikon eclipse ni microscope with digital camera (nikon ds-fi2) and processed using the image software nis-elements basic research. to determine the genetic sex of young white storks, the multiplex pcr method was applied. two sets of primers were used. first were the primers specific for w chromosome for amplification of the female-specific sequence on w chromosome. the second set of primers was used to replicate the 18s ribosome gene, which serves as a positive control of the pcr reaction (kamiński et al., 2014; 2015). statistics analysis statistical analyses were performed using statistica 13.1 software (dell inc., 2016). the length and area of each type of leukocytes were measured on the blood smear. the mean, standard deviation, and maximum and minimum values for each variable were calculated. the results were given as mean ± sd. the shape ratio was marked, where 1/1 means round. haematological values between males and females were compared by kruskal-wallis tests, and the level of significance set at p < 0.05. results heterophils (fig. 1a) were single, round, or irregular cells (shape ratio 0.95/1 – tab. 1) with spindle or oval shaped cytoplasmic granules, representing 41.2 ± 7.36% of leukocytes for males and 44.4 ± 7.16% for females. the nucleus had 2–3 lobules with contained dense and dark-staining chromatin for both sexes. they were round and measured 11.14 ± 0.65 μm in diameter for males and 11.01 ± 0.48 μm for females (tab. 1). basophils were the smallest granulocytes, averaging 9.56 ± 0.78 μm in diameter for males and 9.13 ± 0.84 μm for females (tab. 1). with may–grünwald stain, their granules were vacuolated or clear, due to the bleaching effect of methanol fixation (fig. 1b). eosinophils (fig. 1c) had the diameter of 10.72 ± 0.49 μm for males and 10.97 ± 0.53 μm for females (tab. 1). the granules were brighter in colour when compared to the heterophil granules. the nucleus was lobed and mostly stained clear blue and contained red-orange, round or rod shape granules. the shape ratio was 0.90/1, which indicates that the cells varied in shape. monocytes were not frequently observed, and their diameter was 8.8 ± 4.87% and 7.6 ± 3.93%, respectively, for males and females (tab. 2). monocytes were the largest type of leukocytes found in the blood film. they were round or irregular in shape with a violet, kidney shaped nucleus and a light, pale blue cytoplasm (fig. 1d), measuring 13.40 ± 0.97 μm in diameter for males and 13.09 ± 1.05 μm for females (tab. 1). m on ik a g ra nd tk e, m ar iu sz k as pr za k, m at eu sz c ie pl iń sk i, ew a b ur da , a rie l d ur aj sk i, jo ac hi m s ie ki er a, l es ze k je rz ak 44 fig. 1. may – grünwald stained leukocytes in the blood film of a white stork (ciconia ciconia l.); a – heterophil, b – basophil, c – eosinophil, d – monocyte, e – large lymphocyte, f – small lymphocyte lymphocytes are second only to the heterophils in frequency in most species (tab. 2). lymphocytes (fig. 1e, f) had compact, dark nuclei and thin cytoplasm fringes of a blue colour. they were small and well differentiated with an average of 8.10 ± 0.66 μm and 8.28 ± 0.74 μm in diameter for large lymphocytes, respectively, for males and females, and 5.31 ± 0.65 μm and 5.57 ± 0.59 μm, respectively, for males and females for small lymphocytes (tab. 1). small lymphocytes had a shape ratio of 0.99/1, which indicated that the cells are round, while large lymphocytes had a shape ratio of 0.87/1, which proves that they are very diverse in shape. the h/l ratio was 4/4 for both sexes. 45 tab. 1. blood cell diameters in μm (mean ± sd) in white stork (ciconia ciconia l.) chicks parameter male female shape ratio number of cellsmean ± sd mean ± sd heterophils 11.14 ± 0.65 11.01± 0.48 0.95/1 100 eosinophils 10.72 ± 0.49 10.97 ± 0.53 0.90/1 100 basophils 9.56 ± 0.78 9.13 ± 0.84 0.98/1 75 monocytes 13.40 ± 0.97 13.09 ± 1.05 0.93/1 100 lymphocytes (small) 5.31 ± 0.65 5.57 ± 0.59 0.99/1 100 lymphocytes (large) 8.10 ± 0.66 8.28 ± 0.74 0.87/1 100 there were no statistically significant differences in male and female white stork tab. 2. white blood cells (wbc) in male and female white stork (ciconia ciconia l.) chicks parameter male n = 25 female n = 28 reference range from our study mean ± sd mean ± sd wbc [×109/l] 18.44 ± 3.69 18.52 ± 3.33 13.56 – 30.22 heterophils number [×103/μl] relative [%] 7.51± 1.53 41.20 ± 7.36 8.05 ± 1.56 44.00 ± 7.16 4.97 – 12.60 26.00 – 57.00 eosinophils number [×103/μl] relative [%] 1.73 ± 0.86 9.40 ± 4.41 1.97 ± 0.83 10.50 ± 3.68 0.45 – 3.47 3.00 – 17.00 basophils number [×103/μl] relative [%] 0.34 ± 0.30 1.80 ± 1.42 0.38 ± 0.37 1.90 ± 1.86 0.00 – 1.19 0.00 – 6.00 monocytes number [×103/μl] relative [%] 1.75 ± 1.38 8.80 ± 4.87 1.43 ± 0.85 7.60 ± 3.93 0.00 – 6.65 0.00 – 22.00 lymphocytes number [×103/μl] relative [%] 7.03 ± 1.89 37.20 ± 7.61 6.70 ± 1.80 36.00 ± 6.48 3.36 – 12.01 22.00 – 52.00 there were no statistically significant differences in male and female white stork discussion the wbc counts of white storks (tab. 2) were similar to those living at large, as reported by others authors studying storks (alonso et al., 1991; kamiński et al., 2014; kasprzak et al., 2006; lashev et al., 2005; montesinos et al., 1997; puerta et al., 1989; szabó et al., 2010). the white stork is a monomorphic species, and morphological differences between chicks of different sex do not exist. female white storks had a higher estimated wbc number compared to males examined in this study, but the difference was not significant (tab. 2). no significant differences were also observed between both of the sexes in white blood cell parameters (wbc frequencies). in our study, the heterophils were a nalysis of peripheral blood w hite blood cell param eters in european w hite s torks (ciconia ciconia l.) chicks that varies by sex m on ik a g ra nd tk e, m ar iu sz k as pr za k, m at eu sz c ie pl iń sk i, ew a b ur da , a rie l d ur aj sk i, jo ac hi m s ie ki er a, l es ze k je rz ak 46 the most frequent cellular type, representing 42.7 ± 7.32% of leukocytes for both sexes, as found by other authors (tab. 3) in storks (alonso et al., 1991; montesinos et al., 1997; santos, serra, 2006; szabó et al., 2010). nevertheless, some authors marked lymphocytes as the predominant cells type in storks, as presented in table 3 (aengwanich et al., 2002; lanzarot et al., 2005; puerta et al., 1989; salakij et al., 2003). as a result, juvenile white storks are another exception to the rule that lymphocytes are the most abundant white cell type in birds, together with the rheas rheidae (gallo et al., 2017), cinereous vulture aegypius monachus l. (seok et al., 2017), white-naped cranes antigone vipio pallas (rayhel et al., 2015), red-tailed amazon parrot amazona brasiliensis l. (vaz et al., 2015), barn owls tyto alba scopoli (szabó et al., 2014), cormorants phalacrocorax (gallo et al., 2013), black cockatoos calyptorhynchus banksi latham (le souëf et al., 2013), swans cygnus (milani et al., 2012), hyacinth macaws anodorhynchus hyacinthinus latham (kolesnikovas et al., 2012), bearded vultures gypaetus barbatus l. (hernández, margalida, 2010). eosinophils, basophils and monocytes occurred in smaller proportions and were in similar numbers to the white storks reported by other authors (alonso et al., 1991; lashev et al., 2005; puerta et al., 1989; szabó et al., 2010). tab. 3. comparative differential frequency of white blood cells in family ciconiidae j.e. gray (average ± sd); a – juvenile, b – adult; n – sample size; h – heterophils, e – eosinophils, b – basophils, m – monocytes, l – lymphocytesw a ut ho rs of st ud ie s o ur st ud y pu er ta e t a l. 19 89 m on te si no s et a l. 19 97 * la sh ev e t al . 2 00 5 a lo ns o et al . 1 99 1 sz ab ó et a l. 20 10 sa nt os a nd se rr a 20 06 la nz ar ot e t al . 2 00 5 a en gw an ic h et a l. 20 02 sa la ki j e t a l. 20 03 sp ec ie s white stork ciconia ciconia l. black stork ciconia nigra l. painted stork mycteria leucocephala pennant age a b a b a b n 53 24 23 6 7 80 48 36 10 12 h 42.70± 7.00 15.70 ± 2.10 67.90 41.80 ± 1.05 72.30 ± 5.20 61.00 ± 9.80 45.40 ± 15.90 41.60 ± 13.90 10.60 ± 9.49 40.30 ± 5.80 e 9.94± 4.03 23.70 ± 1.90 4.70 3.17 ± 0.48 4.80 ± 1.20 0.75 ± 0.91 5.60 ± 4.50 6.19 ± 3.91 12.00 ± 8.54 10.30 ± 1.10 b 1.84± 1.66 1.30 ± 0.20 0.30 0.67 ± 0.21 0.50 ± 4.60 0.38 ± 0.56 1.00 ± 2.50 – 1.00 ±1.33 – m 8.17± 4.40 3.10 ± 0.40 1.30 0.50 ± 0.22 1.90 ± 0.40 3.44 ± 2.30 3.50 ± 3.30 1.15 ± 0.95 – 1.50 ± 0.50 l 36.60± 6.50 55.30 ± 2.40 23.00 53.80 ± 1.05 20.40 ± 4.60 34.30 ± 9.10 44.40 ± 20.20 50.30 ± 13.10 76.10 ± 10.28 48.40 ± 4.50 h/l 4/4 2/6 7/2 4/5 7/2 6/3 4/4 4/5 1/8 4/5 *authors did not give ± sd 47 heterophils to lymphocyte ratio (h/l) (tab. 3) varies depending on the level of the stress reaction (krams et al., 2012). in this research, the ratio was 4/4, and it was the same as reported by other authors in juvenile black stork ciconia nigra l. (puerta et al., 1989; santos, serra, 2006) and adult painted stork mycteria leucocephala pennant (salakij et al., 2003). a similar ratio (4/5) was also noticed in an adult white stork (lashev et al., 2005) and in a juvenile black stork (lanzarot et al., 2005). the high ratio indicated a stress reaction; whereas, the ratio reported by alonso et al. (1991) was 7/2 for adult white storks. such a large variation in the h/l ratio indicates that it should only be used in comparisons of similar bird life conditions. our results provide comparative morphological characteristics and a guide for the identification of blood cells in white stork chicks. these may be useful for health management in the white storks, which are endangered species and it is beneficial for further study and related research. references aengwanich, w., mahasarakham u., tanomtong, a., pattanarungson, r. 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(2009). blood cell morphology, some hematological and serum biochemistry values of common kestrel (falco tinnunculus). journal of sustainable development, 1(2), 123. doi: 10.5539/jsd.v1n2p123 szabó, z., beregi, a., vajdovich, p., abonyi-tóth, z., mátrai, e., pazár, p., gaál, t. (2010). hematologic and plasma biochemistry values in white storks (ciconia ciconia). journal of zoo and wildlife medicine: official publication of the american association of zoo veterinarians, 41(1), 17–21. doi: 10.1638/2008-0164.1 szabó, z., klein, a., jakab, c. (2014). hematologic and plasma biochemistry reference intervals of healthy adult barn owls (tyto alba). avian diseases, 58(2), 228–231. doi: 10.1637/10715-111013-reg.1 vaz, f.f., locatelli-dittrich, r., sipinski, e.a.b., abbud, m.c., sezerban, r.m., schmidt, e.m.s., cavalheiro, m.l. (2015). hematologic and total plasma protein values in free-living red-tailed amazon parrot nestlings (amazona brasiliensis) in paraná state, brazil. journal of avian medicine and surgery, 29(3), 187–191. doi: 10.1647/2014-050 abstract the aim our study was qualitative and quantitative analysis of white blood morphometric elements of peripheral blood (determining the quantity, blood cells dimension and several haematological values) in white stork chicks. one of the aims was to indicate whether the sex relevantly influences the variety of the examined white blood indicators. white blood cells parameters of 53 white stork chicks, with molecularly marked sex, were examined. blood samples were collected in southern poland (around krapkowice town, near opole city). lymphocytes of white storks (mean 37% for females and males) were identified as round a nalysis of peripheral blood w hite blood cell param eters in european w hite s torks (ciconia ciconia l.) chicks that varies by sex m on ik a g ra nd tk e, m ar iu sz k as pr za k, m at eu sz c ie pl iń sk i, ew a b ur da , a rie l d ur aj sk i, jo ac hi m s ie ki er a, l es ze k je rz ak 50 cells with dark purple non-lobed, eccentrically positioned nucleus. among the whole population we differentiated small lymphocytes with diameters of 5.31 ± 0.65 μm in males and 5.57 ± 0.59 μm in females, and large lymphocytes with a diameter of 8.10 ± 0.66 μm and 8.28 ± 0.74 μm, respectively in females and males. monocytes (mean 8% for female and males) were the largest leukocytes found in the blood film of white storks, measuring 13.40 ± 0.97 μm for males and 13.09 ± 1.05 μm for females in diameter. the cytoplasm was abundant, and it stained blue-grey and very often contained vacuoles. heterophils (mean 42.7%) were the largest in granular leukocytes group. they were round and 11.14 ± 0.65 μm for males and 11.01 ± 0.48 μm for females in diameter. the nucleus of heterophils was lobed, usually with two or three lobes. the cytoplasm contained brick-red, elongated granules. eosinophils (mean 9.44%) were round cells, with a diameter of 10.72 ± 0.49 μm and 10.97 ± 0.53 μm, respectively, in males and females. the nucleus was lobed and mostly stained clear blue and contained red-orange, round or rod-shaped granules. basophils (mean 1.84%) were round and contained dark blue granules, with an average of 9.56 ± 0.78 and 9.13 ± 0.84 μm in diameter, for males and females. the nucleus was usually non-lobed. the h/l ratio was 4/4 for both sexes. no significant differences in levels and types of leukocytes between male and female juvenile storks have been observed. key words: white blood cells size, white stork, hematology, morphometric received: [2018.06.20] accepted: [2018.10.16] analiza parametrów białych krwinek krwi obwodowej piskląt bociana białego (ciconia ciconia l.) w zależności od płci streszczenie celem naszego badania była analiza jakościowa i  ilościowa morfometrycznych elementów krwi obwodowej (określenie ilości, wymiarów krwinek i kilku wartości hematologicznych) u piskląt bociana białego. jednym z celów było wskazanie, czy płeć ma istotny wpływ na różnorodność badanych wskaźników krwi białej. zbadano parametry białokrwinkowe 53 piskląt bociana białego, z molekularnie oznaczoną płcią. próbki krwi pobierano w południowej polsce (okolice miasta krapkowice, w pobliżu miasta opola. limfocyty bocianów białych (średnio 37% dla samic i  samców) zostały zidentyfikowane, jako okrągłe komórki z  ciemnofioletowym, ekscentrycznie usytuowanym jądrem. spośród całej populacji wyodrębniliśmy małe limfocyty: o  średnicy 5,31 ± 0,65 μm u  samców i  5,57± 0,59 μm u  samic oraz limfocyty duże o średnicy 8,10 ± 0,66 μm i 8,28 ± 0,74 μm, odpowiednio u samic i samców. monocyty (średnio 8% dla samic i samców) były największymi leukocytami znalezionymi we krwi obwodowej bociana białego, o średnicy 13,40 ± 0,97 μm u samców i 13,09 ± 1,05 μm u samic. cytoplazma była obfita i zabarwiona na niebiesko-szaro, często zawierała wakuole. heterofile (średnio 42,7%) były największymi krwinkami wśród granulocytów. były okrągłe i miały średnicę 11,14 ± 0,65 μm u samców i 11,01 ± 0,48 μm u samic. jądro heterofilów było płatowate, zwykle z dwoma lub trzema płatami. cytoplazma zawierała ceglasto-czerwone, wydłużone ziarnistości. eozynofile ( średnio 9,44%) były okrągłymi komórkami o średnicy 10,72 ± 0,49 μm i 10,97 ± 0,53 μm, odpowiednio u samców i samic. jądro było płatowate i przeważnie wybarwione na jasnoniebiesko, cytoplazma zawierała czerwono-pomarańczowe, okrągłe ziarnistości. bazofile (średnio 1,84%) były okrągłe i zawierały ciemnoniebieskie ziarnistości o średniej średnicy 9,56 ± 0,78 μm i 9,13 ± 0,84 μm u samców i samic. współczynnik h/l wynosił 4/4 dla obu płci. nie zaobserwowano istotnych różnic w liczbie jak i frekwencji poszczególnych rodzajów krwinek białych między młodymi bocianami płci męskiej i żeńskiej. słowa kluczowe: krwinki białe, bocian biały, hematologia, morfometria information about authors monika grandtke https://orcid.org/0000-0001-9472-252x phd student at the faculty of biological sciences at the university of zielona góra. she works at the university of zielona góra as a scientific and technical assistant. she is interested in the influence of the environment with varying degrees of pollution on the white and red blood cells parameters of storks. 51 mariusz kasprzak https://orcid.org/0000-0001-9088-8098 phd, focused on the relationship between the quality of the environment and the condition of the animals. assistant professor in the department of zoology at the faculty of biological sciences, university of zielona góra. mateusz ciepliński https://orcid.org/0000-0002-3386-9744 he is a phd student at the faculty of biological sciences at the university of zielona góra. he works at the university of zielona góra as a scientific and technical assistant. interested in fish diseases, hematology and biochemistry of fish. ewa burda she is a phd student at the faculty of biological sciences at the university of zielona góra. she works in the rehabilitation center for wild animals in zielona góra. her research work includes topics related to bioacoustics and monitoring in urban environment. she is interesting the avifauna synurbisation, veterinary and canine behaviour. ariel durajski he works in an animal clinic as a veterinary doctor assistant in zielona góra. currently writing a doctoral thesis on the obesity of companion animals. he is interested in veterinary, biology, science, and the nutrition of animals. joachim siekiera msc, active member of the silesian ornithological society. his research interests include birds’ ecology. for many years, he monitored the white storks’ population in opole province, s poland. leszek jerzak https://orcid.org/0000-0001-5332-279x he is a full professor in nature protection at the university of zielona gora, poland (faculty of biological sciences). he studies the biology and ecology of animals (especially the white stork and the magpie) and co-operates with research centres in germany, usa, ireland, and russia. a nalysis of peripheral blood w hite blood cell param eters in european w hite s torks (ciconia ciconia l.) chicks that varies by sex 105 annales universitatis paedagogicae cracoviensis studia naturae, 1: 105–114, 2016, issn 2543-8832 agnieszka chrustek, magdalena twarużek*, ewa zastempowska, jan grajewski department of physiology and toxicology, institute of experimental biology, faculty of natural sciences, kazimierz wielki university, bydgoszcz, chodkiewicza 30, 85-064 bydgoszcz, poland, *twarmag@ukw.edu.pl mycotoxin – induced apoptosis in swine kidney epithelial cells introduction apoptosis is a  programmed cell death that requires energy input and the activation of many genes controlled by a variety of proteins primarily of bcl-2 family. promoting proteins include, among others: bax, bad, bid, and bcl-xs, while the inhibiting ones are bcl-2, bcl-x1, bcl-w (haake, polakowska, 1993; ockner, 2001; elmore, 2007; stępień et al., 2007). programmed cell death can be caused by a number of factors such as uv and gamma radiation, cytostatics, bacteria, toxins, reactive oxygen species, oxidative stress or dna damage (stępień et al., 2007). process of apoptosis occurs in three stages. at �rst, the process induction signal is generated. in the second stage, caspases activation occurs. �en, during the third stage, phagocytosis of apoptotic bodies by macrophages can be observed (kopaczewska, kopaczewski, 2004). a given cell death can be distinguished on a basis of molecular, biochemical and morphological changes. �e latter include: condensation of cytoplasm and chromatin, shrinkage and nucleus fragmentation, as well as formation of blisters and apoptotic bodies. �ere is also the release of cytochrome c from mitochondria, decrease in mitochondrial potential, cell dehydration, increase in calcium ion concentration, activation of serine proteases and caspases, loss of asymmetry in cell membrane phospholipids distribution, as well as degradation of actin �laments and dna. apoptosis can occur in di�erent ways, depending on the pathway. �ere are external, internal, pseudo-receptor, sphingomyelin-ceramide and stress-induced types distinguished. in the �rst type, the pathway is induced by combining the ligand with a  membrane receptor, resulting in death signal transmission and caspase cascade activation. contradictory to that, the internal pathway, also known as mitochondrial, is invoked by, among others, the in�uence of reactive oxygen species, oxidative stress or dna damage. �is leads to the opening of mitochondrial channels and 106 a gn ie sz ka c hr us te k, m ag da le na t w ar uż ek , e w a za st em po w sk a, j an g ra je w sk i cytochrome c release. presence of pseudo-receptor pathway was observed in the natural killer (nk) cells and cytotoxic t lymphocytes, while sphingomyelin-ceramide pathway may be activated by viral infections or ionizing radiation. �e last apoptosis pathway (stress-induced) was discovered in 2000 (nakagawa et al., 2000). it arises from the accumulation of incorrectly folded proteins in the endoplasmic reticulum (kerr et al., 1972; haake, polakowska, 1993; ockner, 2001; fadeel, orrenius, 2005; elmore, 2007). apoptosis is an important physiological process occurring in the postnatal period and during embryogenesis. disturbance of this process can lead to the occurrence of such diseases as cancer, parkinson’s, huntington’s or alzheimer’s diseases, heart attack or brain stroke (elmore, 2007). process of apoptosis can be tested in many various ways (darzynkiewicz et al., 1997; smolewski, darzynkiewicz, 2003; elmore, 2007). �ere are cytometric and non-cytometric methods. �e latter include the cell morphology analysis using dyes such as hematoxylin and eosin or �uorochromes, e.g. dapi. changes are assessed by means of light, �uorescence and electron microscope. one of the apoptosis symptoms is dna fragmentation that can be analyzed using agarose gel electrophoresis and by comet assay. �e cytometric methods include: analysis of laser light scattering by apoptotic cells, study of cell membrane permeability disturbances using dyes such as: trypan blue, ethidium bromide, 7-aad (7-aminoactinomycin), as well as study of conformational changes in cell membrane using annexin v. �e study frequently analyses the phenomena associated with the participation of mitochondria in programmed cell death, e.g. mitochondrial potential drop using dyes (rhodamine 123), translocation of bax to the mitochondria or the release of cytochrome c from mitochondria into cytoplasm by means of western blotting using antibodies conjugated with a �uorochrome (darzynkiewicz et al., 1997; smolewski, darzynkiewicz, 2003). mycotoxins are secondary metabolites produced by a  wide range of di�erent molds including aspergillus, penicillium, fusarium and stachybotrys spp. biosynthesis of these compounds can take place during storage and processing of raw materials and during plant growth as well. mycotoxins are a  potential threat to the health of animals and humans. �ey enter the organism not only by ingestion, but also through inhalation of contaminated air. secondary metabolites of molds can cause mycotoxicosis, which causes damage to internal organs and skin tissues (grajewski, 2006). �e number of adverse e�ects of mycotoxins on an animal organism have been shown, e.g. dermotoxic, estrogenic, hepatotoxic, immunotoxic, carcinogenic, mutagenic, teratogenic and toxic actions to the hematopoietic system (wróbel, 2014). �e most common mycotoxins include: a�atoxins, fumonisins, ochratoxin a, trichothecenes (t2, ht2, deoxynivalenol) and zearalenone. trichothecenes are secondary metabolites of fusarium that occur in cereals and cereal products. �ey have 107 m ycotoxin – induced apoptosis in sw ine kidney epithelial cells dermatoxic and hemorrhagic action (chen et al., 2008; liu et al., 2014). a�atoxins are primarily synthesized by aspergillus and are being found in peanuts, spices, dried fruits, cotton pulp and maize grain (twarużek et al., 2013). a�atoxin b1 is the most toxic metabolite. �e milk can contain a�atoxin m1 (zastempowska et al., 2016). a�atoxins are potent poisons having teratogenic, mutagenic and carcinogenic properties (wróbel, 2014; liu et al., 2015a; peng et al., 2016). �ere is also ochratoxin a synthesized by penicillium and aspergillus, and occurs in cereal grain, dried fruits, co�ee, spices, animal-origin food and wine (kosicki et al., 2016). it causes irreversible damage to the nephrons and reveals carcinogenic and immunotoxic action (grajewski et al., 2007; zhang et al., 2009; chopra et al., 2010; liu et al., 2015b). �e aim of this study was to analyze the impact of mycotoxins: a�atoxin b1, ochratoxin a  and toxin t2 on the cells of swine kidney (sk) epithelial cell line and their apoptosis. materials and methods cell culture and treatment conditions �e sk (swine kidney epithelial) cell line derived from the department of physiology and toxicology of the kazimierz university of bydgoszcz were the material for research. �e cells were cultured in mem (minimum essential medium eagle) supplemented with antibiotics (penicillin and streptomycin) and 5% fetal calf serum. cultures (adjusted to 4×105 cells/ml) were grown for 24 h at 37°c, at the access for 5% co2 and at 95% humidity. �en cells were incubated for 24 h with mycotoxins in concentrations chosen experimentally and based on literature: t2 toxin at 2.5 µm and 25 µm dose, a�atoxin b1 at concentrations of 10 µm and 30 µm and ochratoxin a of 50 µm and 80 µm. cell line sk cultured in mem supplemented with antibiotics and fetal calf serum constituted the control. analysis of apoptosis flow cytometer muse cell analyzer, as well as muse annexin v & dead cell kit (merck) were used for evaluation of apoptosis. �e cytometer uses laser-based �uorescent detection and microcapillary technology to deliver quantitative cell analysis. cells used in the culture were separated from the base using 0.25% trypsin and then incubated for 20 min with muse annexin v & dead cell kit in accordance with the attached protocol. muse annexin v & dead cell kit contains �uorescently labelled annexin v, which, as the coagulant, binds with negatively charged phospholipids such as phosphatidylserine moving outside the plasma membrane upon the onset of apoptosis. �is procedure allows to detect cells in various stages of apoptosis: live cells, dead cells, early and late apoptotic cells. 108 a gn ie sz ka c hr us te k, m ag da le na t w ar uż ek , e w a za st em po w sk a, j an g ra je w sk i results and discussion studies have shown sensitivity of sk cell line on mycotoxins, namely t2 toxin, a�atoxin b1 and ochratoxin a (tab. 1, fig. 1–2). �e data indicates a relationship between the dose of mycotoxins and the occurrence of apoptosis. �e highest percentage of apoptotic cells was observed in those treated with a�atoxin b1 at a dose of 30 µm, and ochratoxin a at both concentrations. �ere were no clear di�erences in the e�ect of ochratoxin a at both concentrations, i.e. 50 µm and 80 µm (tab. 1, fig. 1–2). mycotoxin the least interacting on sk cell line turned out to be t2 toxin. tab. 1. in�uence of mycotoxins on the presence of apoptosis in the swine kidney epithelial cell line toxin (dose) live [%] early apoptotic [%] late apoptotic [%] total apoptotic [%] dead [%] control 93.90 5.35 0.15 5.50 0.60 t2 (2,5 µm) 93.04 3.36 3.53 6.90 0.06 t2 (25 µm) 73.55 15.10 11.25 26.35 0.10 a�atoxin b1 (10 µm) 85.90 12.80 0.70 13.50 0.60 a�atoxin b1 (30 µm) 55.05 26.05 14.55 40.60 4.35 ochratoxin a (50 µm) 31.49 64.77 3.74 68.51 0 ochratoxin a (80 µm) 39.59 52.47 7.94 60.41 0 mycotoxins such as trichothecenes (t2 toxin, ht2, deoxynivalenol), a�atoxins, ochratoxin a, fumonisins and zearalenone can lead to the cell death. most o�en, apoptosis and necrosis can be observed. apoptosis is a programmed cell death and, in contrast to necrosis, is the active process that needs some energy input and gene activation (majno, joris, 1995; trump et al., 1997; stępień et al., 2007). during this process, chromatin condensation, dna fragmentation, formation of apoptotic bodies and phagocytosis take place. programmed cell death is a physiological process occurring in the postnatal period and during embryogenesis. disorders during its course can lead to the occurrence of many diseases (elmore, 2007). in contrast to apoptosis, necrosis is a pathological process a�ecting the cell groups. it brings cells to swell, causes disintegration of organelles and induces in�ammation (majno, joris, 1995; trump et al., 1997). �ese studies have con�rmed the ability of selected mycotoxins to induce apoptosis. our study showed that t2 toxin proved to be the least toxic mycotoxin (at least at the concentrations used in the experiment). �e t2 toxin is derived from trichothecenes group. �e exact mechanism of this substance action is not known. it is believed that t2 toxin is an inhibitor of protein synthesis, induces lipid peroxidation and 109 m ycotoxin – induced apoptosis in sw ine kidney epithelial cells inhibits synthesis of dna and rna (doi et al., 2008). it is well known that t2 toxin induces apoptosis (shinozuka et al., 1997; li et al., 1997; wang et al., 2012), but the research of molecular mechanism of this process is still unknown. scientists suspect that apoptosis can probably occur through the extrinsic pathway with the use of membrane receptors and their ligand (chen et al., 2008). �e death receptors include tnf (tumor necrosis factor), while tnf-alpha or fasl as its ligand (haake, polakowska, 1993; ockner, 2001; fadeel, orrenius, 2005; elmore, 2007). combination of the relevant molecules activates the caspase cascade, which in consequence leads to death. chen et al. (2008) studied the e�ects of t2 on human chondrocytes. �eir results showed the increment of fas, p53, bax, caspase-3 and a decrease of bcl2 and, therefore, indicated the presence of apoptosis. it is believed that it might have occurred with the use of an external pathway. �e researchers suggest that the t2 toxin can cause kashin-beck disease, which causes damage to the joints (chen et al., 2008). when administered orally, parenterally and in contact with the skin, t2 induces apoptosis in thymocytes, in the thymus lymphocytes, hematopoietic cells, epithelial cells of the intestinal crypts, hepatocytes and keratinocytes in animals. �is process was also noted in the red and white pulp of the spleen, and in peyer’s patches as well (chen et al., 2008; doi et al., 2008; liu et al., 2014). it is worth mentioning that the toxin penetrates the placenta and a�ects the fetus, among others, by damaging its brain (doi et al., 2008). in addition to t2, ochratoxin a at concentrations of 50 µm and 80 µm was also tested in the present study. it has been shown that both doses caused apoptosis in about 60% of the cells. ochratoxin a is classi�ed as a human carcinogen. liu et al. (2015b) studied the e�ect of the substance on het-1a cells (cancer of the esophagus). fig. 1. in�uence of mycotoxins on the presence of apoptosis in the swine kidney epithelial cell line 0 20 40 60 80 100 control t2 2,5µm t2 25µm a�atoxin b1 10µm a�atoxin b1 30µm ochratoxin a 50µm ochratoxin a 80µm [%] toxin live total apoptotic 110 a gn ie sz ka c hr us te k, m ag da le na t w ar uż ek , e w a za st em po w sk a, j an g ra je w sk i fig. 2. �e occurrence of apoptosis in the swine kidney (sk) epithelial cells a�er treatment with mycotoxins at di�erent concentrations: a – control, b – 2.5 µm t2, c – 25 µm t2, d – 10 µm a�atoxin b1, e – 30 µm a�atoxin b1, f – 50 µm ochratoxin a, g – 80 µm ochratoxin a. �e cells have been divided into four distinct populations: live (the lower le� square), early apoptotic (right lower square), late apoptotic (right upper square) and dead (le� upper square). results have been expressed as percentages 111 m ycotoxin – induced apoptosis in sw ine kidney epithelial cells �e dna strand breaks were observed along with chromosomal aberrations, and arrest of the cell cycle in the g2 phase. moreover, there was an increase of caspase-3, which suggests the occurrence of apoptosis in het-1a cells. elevated activation of caspase-3 was also observed in a study upon the immunotoxicity of ochratoxin a in cells of h9 line (human t lymphocytes). in addition, changes were seen at the cell morphology level (condensed cytoplasm, chromatin condensation, apoptotic bodies, swollen mitochondria), the increase in tnf-alpha, and a decrease of il-2. it is suggested that necrosis has the priority over apoptosis at higher concentrations of ochratoxin a. �e researchers turned their attention to the possibility of the apoptosis occurrence through the mitochondrial pathway, otherwise known as internal pathway (darif et al., 2016). mitochondrial channels are opened under the in�uence of stress-inducing factors and cytochrome c is released into the cytoplasm. �is compound binds to cytoplasmic factor apaf 1 and inactive caspase-9. �en, activation of caspase-9 and executive caspases takes place, which leads to cell death (elmore, 2007; stępień et al., 2007; darif et al., 2016). ochratoxin a has nephrotoxic, hepatotoxic and immunotoxic action. other studies have shown the dose-dependent manner increase in cytotoxicity on neural/nerve cells (zhang et al., 2009). in the present experiment, we demonstrated a  dose-dependent manner e�ect of a�atoxin b1 on the sk line cells. a dose of 10 µm resulted in an increase of apoptotic cells by 8%, while 30μm dose by 35% in comparison to the control version. a�atoxin b1 interferes with the porcine oocyte maturation by inducing the oxidative stress, and also causes the occurrence of excessive autophagy and apoptosis (liu et al., 2015a). both apoptosis and autophagy are a  type of programmed cell death (stępień et al., 2007). autophagy is present in all eukaryotic cells and is activated in a  response to a shortage of development-inducing nutrients, as well as damage due to toxins. it is characterized by a lack of the nucleus fragmentation, degradation of the golgi apparatus or the endoplasmic reticulum, and partial chromatin condensation. peng et al. (2016) studied the role of mitochondria, death receptors and endoplasmic reticulum in the toxin-invoked pathways of thymocytes apoptosis induced by a�atoxin b1. �e study revealed the importance of external and internal pathways in the occurrence of apoptosis. it was also noted that the mycotoxin induces apoptosis, but also necrosis in human lymphocytes (al-hammadi et al., 2014). conclusions �e studies presented in this article have shown the impact of mycotoxins such as t2 toxin, a�atoxin b1 and ochratoxin a on the sk line cells. �e scienti�c literature con�rms the toxic e�ects of mycotoxins on a variety of cell lines of animal, as well as human organisms. unfortunately, we still can not explain the molecular mechanisms 112 a gn ie sz ka c hr us te k, m ag da le na t w ar uż ek , e w a za st em po w sk a, j an g ra je w sk i of apoptosis occurrence in cells. it is also possible the existence of other types of cell death, e.g. necrosis, mitotic catastrophe, or autophagy as the reaction to fungal toxins. references al-hammadi, s., marzouqi, f., al-mansouri, a., shahin, a., al-shamsi, m., mensah-brown, e., souid, a. 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(2009). ochratoxin a  induces apoptosis in neuronal cells. genes & nutrition, 4(1), 41–48. doi: 10.1007/s12263-0080109-y abstract apoptosis, as the programmed cell death, plays a signi�cant role in proper functioning of an organism, both in the postnatal period and during embryogenesis. disturbances in this process can lead to the occurrence of several dysfunctions, e.g. cancer, stroke, alzheimer’s disease and others. apoptosis can be triggered by factors such as oxidative stress, free radicals, uv radiation and cytotoxic drugs, but also mycotoxins, e.g. a�atoxins, ochratoxin a  and trichothecenes. �ese toxins are produced primarily by fungi of aspergillus, penicillium, fusarium and stachybotrys genera. �e aim of the study was to investigate the e�ect of mycotoxins on the occurrence of apoptosis in the swine kidney (sk) epithelial cells. for this purpose, trichothecene t2 toxin was used at a concentration of 2.5 µm and 25 µm, a�atoxin b1 at a dose of 10 µm and 30 µm, and ochratoxin a concentrations of 50 µm and 80 µm. �e results were assessed using �ow cytometer muse cell analyzer (merck). studies have shown high sensitivity of the cell line sk on mycotoxins. apoptosis was caused by all kinds of toxins and depended on the dose of ex114 a gn ie sz ka c hr us te k, m ag da le na t w ar uż ek , e w a za st em po w sk a, j an g ra je w sk i amined substance. t2 toxin at a concentration of 2.5 µm caused apoptosis in 6.9% of the cells, whereas at a concentration of 25 µm in 26.35% of the cells. a�atoxin b1 used at concentrations of 10 µm and 30 µm caused apoptosis in 13.5% and 40.6% of the cells, respectively. �e use of ochratoxin a in concentrations of 50 µm and 80 µm caused the occurrence of apoptosis respectively in 68.51% and 60.41% of the cells. key words: a�atoxins, apoptosis, mycotoxins, ochratoxin a, swine kidney cell line, t2 toxin received: [2016.08.16] accepted: [2016.10.26] apoptoza w komórkach nabłonkowych nerki świni indukowana mykotoksynami streszczenie istotną rolę w prawidłowym funkcjonowaniu organizmu odgrywa apoptoza jako programowa śmierć komórki, zarówno w  okresie poporodowym, jak i  podczas embriogenezy. zaburzenia tego procesu mogą prowadzić do wystąpienia różnych schorzeń, np. raka, udaru mózgu, choroby alzheimera i innych. apoptoza może być efektem takich czynników jak: stres oksydacyjny, wolne rodniki, promieniowanie uv, leki cytotoksyczne, w tym również mykotoksyny. do najczęstszych mykotoksyn należą: a�atoksyny, ochratoksyna a  i  trichoteceny. są one produkowane głównie przez grzyby aspergillus, penicillium, fusarium oraz stachybotrys. celem pracy było zbadanie wpływu mykotoksyn na występowanie apoptozy w  komórkach nabłonkowych nerki świni (sk). w  eksperymencie zastosowano toksynę t2 w  stężeniu 2,5 μm i  25 μm, a�atoksynę b1 w dawce 10 μm i 30 μm oraz ochratoksynę a w stężeniu 50 μm i 80 μm. wyniki oceniano za pomocą cytometru przepływowego muse cell analyzer (�rmy merck). badania wykazały wysoką wrażliwość linii komórkowej sk na mykotoksyny. apoptoza była wywoływana przez wszystkie rodzaje toksyn i była zależna od dawki badanej substancji. toksyna t2 w stężeniu 2,5 μm powodowały apoptozę u 6,9% komórek, podczas gdy w stężeniu 25 μm stwierdzono jej obecność u 26,35% komórek. a�atoksyna b1 stosowane w stężeniach 10 μm i 30 μm powodowały apoptozę komórek odpowiednio u 13,5% i 40,6% komórek. zastosowanie ochratoksyny a w stężeniu 50 μm i 80 μm powodowało wystąpienie apoptozy u 68,51% oraz 60,41% komórek. słowa kluczowe: a�atoksyny, apoptoza, mykotoksyny, ochratoksyna a, linia komórkowa sk, t2 toksyna information on the authors agnieszka chrustek she is interested in the identi�cation and analysis of pathways leading to the occurrence of cell death (apoptosis, necrosis, mitotic catastrophe). �e research are carried out using light, �uorescence and electron microscopy, �ow cytometry and western blot method. magdalena twarużek her main research interests revolve around the following thematic groups: the occurrence of mold and mycotoxins in food, feed and environment of humans and animals, the e�ect of molds and mycotoxins on human and animal health and the use of in vitro culture in the evaluation of contamination. ewa zastempowska �e subject of her scienti�c interests are microorganisms that are causative agents of mammary gland in�ammation (mastitis) in cows. she is also interested in problems related to virulence, drug resistance and cytotoxicity of the pathogenic microorganisms isolated from the animal milk and the presence of mycotoxins in milk produced by molds as well. jan grajewski professor of agricultural sciences. his research interests include food and feed safety in aspects of molds and their secondary metabolites – mycotoxins. 70 annales universitatis paedagogicae cracoviensis studia naturae, 3: 70–79, 2018, issn 2543-8832 doi: 10.24917/25438832.3.5 iwona konieczna1*, grzegorz rut1, angelika kliszcz2 1institute of biology, pedagogical university of cracow, podchorążych 2 st. 30-084 kraków, poland, *i.konieczna08@gmail.com 2department of agrotechnology and agricultural ecology, university of agriculture in kraków, mickiewicza st. 21, 31-120 krakow, poland photosynthetic activity of daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens and triticum aestivum l. in the presence of copper and vanadium salts introduction heavy metals are elements that naturally occur in the form of oxides in ores and rocks. they are released through the processing and incineration of various types of waste, transportation vehicles, the mining industry, and metallurgy, including burning energy materials in power plants, cogeneration plants, boiler rooms, and industrial plants (kabata-pendias, pendias, 1999). these include micronutrients, elements necessary for the proper growth and development of organisms (including copper, zinc, chromium, iron) and the trace elements that are dispensable (e.g., cadmium, lead, mercury, vanadium). although, both types of micronutrients are necessary in many biochemical processes of living organisms, trace elements in excess produce toxic effects disrupting metabolic processes (kabata-pendias, pendias, 1999). due to the ability of intensive migration and accumulation, heavy metals are a factor causing toxic contamination of soils, water, and air (karczewska et al., 2008). the toxicity of heavy metals depends on the degree of their concentration and the path of penetration into the environment, the chemical form in which they occur, the type of their interaction with other metals, and the species and age of the organisms (maciejewska, 2003). copper (cu) is an element strongly bound in surface layers of soil and does not move into its profile. in moderate climate, it remains in soils between 1 and 3 thousand years (bowen, 1979). this element occurs in all types of waters. its content is very different and depends on the surrounding geological formation and impurities (formicki, 2010). due to the strong sorption of other substances, copper usually accumulates in bottom sediments. sometimes, it is transported with water in the form of a colloidal suspension of various salts or minerals (kabata-pendias, pendias, 1979; 1999). 71 p hotosynthetic activity of d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens and triticum aestivum l. in the presence of copper and vanadium salts most often, such situations occur in mining and metallurgical areas, where the permissible content of this metal can be exceeded several thousand times. besides the industrial emissions, the sources of copper in the soil are mineral and organic fertilizers, plant protection chemicals, and municipal waste. tolerance of plants to excess copper is relatively high, and they can grow in environments contaminated with this element (kabata-pendias, pendias, 1999). plants are relatively resistant to copper, but its excess causes disturbances in metabolism and, as a consequence, the copper contamination limits growth and development. excess copper mainly damages the dna of cells, manifesting in changes in the permeability of cell membranes by increasing the secretion of some cations and anions (e.g., k+, po43-). it also causes disturbances in electron flow in the process of photosynthesis (maksymiec, 1997). the other toxic element found in the environment is vanadium (v). the biological role of vanadium is involved in the metabolism of lipids, phosphotransferases, sugars, and monoamine oxidases. it is also involved in the sodium-potassium and calcium-magnesium systems. vanadium is needed mainly in the metabolism of algae cells in which it stimulates photosynthesis. additionally, it is engaged in the reduction of hydrogen peroxide through the vanadium-bromoperoxidase enzyme, thus acting as an electron transporter (v3+-v4+-v5+). for higher plants, vanadium is not necessary for proper functioning. however, in small concentrations, it positively influences the synthesis of chlorophyll. it also works catalytically in the process of nitrogen fixation by bacteria of the genus rhizobium and free-living bacteria. in plants, only the biological reduction of vanadium from thevo3form to the vo2+ form is protective, and the vanadium is harmful at a higher degree of oxidation (morell et al., 1986). the aim of this study was to determine the effect of copper and vanadium salts with different molar concentrations on the photosynthetic activity of carrots (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) and winter wheat (triticum aestivum l.). the assessment of the chlorophyll content and chlorophyll fluorescence kinetics parameter in young specimens were measured. material and methods carrot seeds and wheat grains were put on sterilised petri dishes wetted with distilled water, and stored in the dark, at room temperature. after 9 days, similar morphologically seedlings in pots with sand were planted and cultivated for 5 weeks. in the first week, sand cultures with seedlings were watered regularly with both distilled water (10 ml each) and steiner’s medium (10 ml each). in the next four weeks, the plants were treated once a week with copper (cuso4, copper (ii) sulphate) and vanadium salts (h4no3v, azanium; oxido(dioxo)vanadium, according to iupac) with both mentioned salts at concentrations of 0.6 mm and 3 mm (10 ml each) and with steiner’s iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 72 medium (10 ml each). the control was only watered with both distilled water (10 ml each) and steiner’s medium (10 ml each) (fig. 1). the determination of chlorophyll content and measurement of chlorophyll a fluorescence after five weeks of carrot and wheat cultivation, the total chlorophyll (chl) content was measured using a spad chlorophyll meter (geomor-technik) at room temperature according to the procedure provided by the manufacturer. at room temperature, chlorophyll a fluorescence measurements using a fms-1 fluorometer (fluorescence monitoring system) of hansatech instruments england were made, according to the method used by strasser et al. (2000). in order to quench the phase of bright photosynthesis, carrot and wheat leaves were adapted to the darkness for 20 minutes using clips. after that time, by treating the plants with a light intensity of 1500 μmol m–2 s–1 for 1 s, measurements of the parameters f0 (zero fluorescence), fm (maximum fluorescence), fv = (fm – f0) (variable fluorescence), and fv/fm (maximum photochemical efficiency psii) were measured (kalaji, łoboda, 2010). statistical analysis the results were analysed using anova/manova parametric statistical tests, single and multifactor, based on the duncan test, with p ≤ 0.05. average values from 10 infig. 1. sand cultures of cultivated carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) and wheat (triticum aestivum l.) treated with copper and vanadium salts (photo. i. konieczna) 73 dependent repetitions were tested, along with the standard deviation (± sd). the calculations were made using statsoft, inc. (2017) – statistica (data analysis software system), version 13. www.statsoft.com. results and discussion heavy metals accumulated in the soil disturb the basic physiological functions of microorganisms. they lead to an imbalance in the processes of the decomposition and metabolism of organic matter, inhibit the growth and development of plants, and reduce their resistance to pathogens (becker et al., 2006; yanai et al., 2006). plants collect heavy metals from the soil through the root system and the air through leaf blades. the most easily collected metals are from the soil in the form of free ions, and metals in the form of complexes, which are activated by substances secreted from the roots of plants (inal et al., 2007). the amount of heavy metals taken in by plants depends on the type of metal, the form of its occurrence, its content in the soil, and the species of the plant that assimilates it (jin et al., 2005). the excessive accumulation of heavy metals in plants adversely affects their physiological and biochemical processes (możdżeń et al., 2016; 2017). heavy metals alter photosynthesis, respiration, and mineral uptake. they contribute to the degradation of cell organelles and, in extreme cases, lead to the death of the entity (jing et al., 2007). the cause of any changes in the metabolic activity of plants can be an uncontrolled increase in harmful substances that cause cell-toxic oxidative stress (bartosz, 2003). in the presence of heavy metals, various life activities of organisms are inhibited, including protein activity through the formation of covalent bonds (mcgrath et al., 2001), the reduction of the stomatal conductance by increasing the synthesis of abscisic acid (parys et al., 1998), water decomposition reactions caused by damage to the oxygen-releasing complex (gonzalez-mendoza et al., 2007), lipid synthesis in thylakoids and protein antenna system (joshi, mohanty, 2004), dark reactions (burzyński, żurek, 2007), and the reduction of psii activity caused by slowing down the reduction and oxidation of plastochinone molecules (strasser et al., 1995). the photosynthetic response of plants to stress is partly the result of changes in the chlorophyll fluorescence reaction; whereas, the chlorophyll fluorescence efficiency is related to the chlorophyll content (kalaji, łoboda, 2010). chlorophyll a, is an important indicator of photosynthesis that converts solar energy absorbed by other molecules of chlorophyll into energy, which is used by plants in biological processes. therefore, the chlorophyll content inevitably affects the fluorescence, ultimately affecting plant photosynthesis. the key organelles of chlorophyll synthesis are chloroplasts. they participate in the process of photosynthesis, in the assimilation of nitrogen and sulphur, and also in the metabolism of proteins and nucleic acids. the presence of heavy metals p hotosynthetic activity of d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens and triticum aestivum l. in the presence of copper and vanadium salts iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 74 in chloroplasts contributes to their ultrastructural changes, including the increase in the number and size of plastoglobules, the disorganization of grana and thylakoids (solymosi, bertrand, 2012), the disturbance of electron transport in the process of photosynthesis, and the induction of chlorosis (lewis et al., 2001; yruela, 2005). in this experiment, the negative effect of copper and vanadium salts on the chlorophyll content and the photosystem ii (psii) activity in carrot and wheat plants were observed (fig. 2). measurements of the total chlorophyll (chl) content in d. carota showed clear changes in the pigment content at the highest copper salt concentration (3 mm cuso4) and in each of the vanadium solutions, in relation to the control plants (fig. 2a). the chl content in t. aestivum generally was decreased in plants treated with 3 mm h4no3v (fig. 2b). similarly, chlorophyll reduction has been reported, e.g., in the leaves of common bean (phaseolus vulgaris l.), tomato (solanum lycopersicon l.), radish (raphanus sativus l.), and broad bean (vicia faba l.) (perez-espinosa et al., 2002; gad, 2005; kandil, 2007; yavakumar et al., 2007). limitation of chl production is connected with direct inhibition by heavy metals of enzymes involved in chlorophyll a and b synthesis, which play an important role in chloroplast membrane formation and participate in photosynthesis, and thus affect chlorophyll fluorescence (gruca-królikowska, wacławek, 2006). tab. 1. the chlorophyll a fluorescence parameters of carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – a and wheat (triticum aestivum l.) – b plants watered in the growth phase with copper (cuso4) and vanadium (h4no3v) salt solutions, at various concentrations (0.6 mm and 3 mm) pa ra m et er control concentration of the solution [mm] cuso4 h4no3v 0.6 3 0.6 3 a b a b a b a b a b f0 249.6 200.2 228.6 225.3 160.8 224.0 210.6 206.0 121.1 228.3 fm 1541.6 1251.8 1514.4 1490.3 104.9* 1496.3 1308.1 1342.3 751.3* 1412.0 fv 1292.0 1068.5 1285.8 1265.0 88.1* 1272.3 1097.3 1136.3 630.9* 1183.8 fv/fm 0.84 0.85 0.85 0.85 0.08* 0.85 0.75 0.85 0.42* 0.84 f0 – zero fluorescence, fm – maximum fluorescence, fv = (fm – f0) – variable fluorescence, fv/fm – maximum photochemical efficiency psii; mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 the kinetics of chlorophyll a fluorescence illustrates both short-term and longterm effects of heavy metal ions on photosynthesis, and it is expressed using chlorophyll fluorescence parameters (kalaji, łoboda, 2010). the parameters of chlorophyll a fluorescence allow non-invasive determination of the efficiency of primary photosynthesis reactions (krupa, baszyński, 1995). in experiments, a statistically significant effect was indicated with 3 mm solutions of copper ions (cuso4) and vanadium (h4no3v) on photosynthetic activity of young plants, expressed as measurements of 75 kinetics of chlorophyll a fluorescence. for wheat plants, the copper and vanadium salts did not cause significant changes in the chlorophyll a fluorescence (tab. 1). the increase in the value of zero fluorescence (f0) was most probably caused by the limitation of the ability to transmit excitation energy in antenna complexes psii or reducing the energy absorption efficiency to psii as a result of protein dissociation (murkowski, 2002). changes in the maximum fluorescence index (fm) indicated a reduced ability to reduce electron acceptors in psii. the reduction in variable fluorescence efficiency (fv) by thylakoid damage was caused, in which a large part of the excitation energy dissipated in the form of heat, and consequently decreased the activity psii. the maximum photochemical efficiency (fv/fm) for healthy plants under control conditions in the full development phase is assumed to be 0.83 (björkmann, demming, 1987). lowering the value of this parameter indicates the reduced efficiency of electron transport, the degradation of proteins, and the inactivation of psii reaction fig. 2. the chlorophyll content in carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – (a) and wheat (triticum aestivum l.) – (b) leaves, grown with copper (cuso4) and vanadium (h4no3v) salts solutions, at various concentrations (0.6 mm and 3 mm); mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 p hotosynthetic activity of d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens and triticum aestivum l. in the presence of copper and vanadium salts iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 76 centres. this parameter is considered a basic indicator of photosynthesis (kalaji, łoboda, 2010; puła et al., 2019). the main method of tolerating or neutralising the toxicity of metals is the biosynthesis by organisms of various cellular biomolecules. this includes the induction of proteins, organic acids, glutathione, phytochelatins, or cellular secretions, including flavonoids and phenolic compounds, specific amino acids (proline and histidine) and hormones (salicylic acid, jasmonic acid and ethylene). when the above-mentioned strategies are not able to stop heavy metal poisoning, the balance of cellular systems in plants is disturbed and leads to the increased induction of reactive oxygen species (ros). in order to alleviate the harmful effects of free radicals, plant cells have developed an antioxidant defence mechanism, which consists of the activation of enzymatic antioxidants acting as “scavengers” of free radicals. these include superoxide dismutase (sod), catalase (cat), ascorbic peroxidase (apx), glutathione peroxidase (gpx) and glutathione reductase (gr), and non-enzymatic antioxidants, such as ascorbate (asa), glutathione (gsh), carotenoids, alkaloids, tocopherols, proline, and phenolic compounds (flavonoids, tannins and lignin) (emamverdian et al., 2015). although plants constantly create new defence strategies against stress, and some of the biological molecules involved in metal detoxification have antiradical, chelating and antioxidant effects, heavy metals still cause a disturbance of the metabolic balance of plants. exploitation and regulation of defence mechanisms depend on many factors, including on the plant species, the stage of growth, and the type of heavy metal (ociepa-kubicka, ociepa, 2012; emamverdian et al., 2015). conclusion in this experiment, the copper and vanadium salts effect on photosynthetic activity of carrot and wheat plants depend on salts concentrations. significant effects of 3 mm solutions of copper ions and vanadium on photosynthetic activity of carrot plants were observed. for wheat plants, the copper and vanadium salts did not cause significant changes in the chlorophyll a fluorescence. chlorophyll content decreased with increasing of salt concentrations. references becker, j.m., parkin, t., nakatsu c.h., wilbur, j.d., konopka, a. 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(2019). activity of the photosynthetic apparatus in phaseolus vulgaris under the cd stress. notulae botanicae horti agrobotanici cluj-napoca, 47(2), 405–411. doi: 10.15835/nbha47111328 solymosi, k., bertrand, m. (2012). soil metals, chloroplasts, and secure crop production: a review. agronomy for sustainable development, 32, 245–272. doi: 10.1007/s13593-011-0019-z strasser, r.j., srivastava, a., govindjee (1995). polyphasic chlorophyll a fluorescence transient in plants and cyanobacteria. photochemistry and photobiology, 61, 32–42. strasser, r.j., srivastava, a., tsimilli-michael, m. (2000). the fluorescence transient as a tool to characterize and screen photosynthetic samples, probing photosynthesis: mechanism, regulation and adaptation. in: m. yunus, u. pathre, p. mohanty (eds.), probing photosynthesis. london: taylor and francis, 443–480. yanai, j., fang-jie, z., mcgrath, s.p., kosaki, t. (2006). effect of soil characteristics on cd uptake by the hyperaccumulator thlaspi caerulescens. environmental pollution, 139, 167–174. doi: 10.1016/j. envpol.2005.03.013 yavakumar, k., jaleel, ch.a., vijarengan, p. (2007). changes growth, biochemical constituents, and antioxidant potentials in radish (raphanus sativus l.) under cobalt stress. turkish journal of biology, 31, 127–136. yruela, i. (2005). copper in plants. brazilian journal of plant physiology, 17, 145–156. doi: 10.1590/ s1677-04202005000100012 79 abstract the aim of this study was to investigate the influence of copper and vanadium ions on the photosynthetic activity of carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) and winter wheat (triticum aestivum l.). measurements of the total chlorophyll content were performed using a spad chlorophyll meter, and the basic chlorophyll a fluorescence parameters were determined using a fms-1 fluorometer – hansatech. aqueous solutions of copper (cuso4) and vanadium (h4no3v) salt with molar concentrations of 0.6 mm and 3 mm were used. the control group consisted of plants watered with distilled water. with an increase in the concentration of heavy metal ions, a decrease in the content of chlorophyll both in carrots and in wheat was observed, and significant changes in the activity of the photosystem ii were demonstrated. statistically significant effects of 3 mm solutions of copper and vanadium ions on photosynthetic activity were only shown in d. carota l. subsp. sativus. in t. aestivum l., none of the heavy metal ions induced significant changes in the values of chlorophyll a fluorescence. key words: photosynthesis, carrot, wheat, heavy metals, copper sulphate, ammonium metavanadate received: [2018.04.09] accepted: [2018.11.05] aktywność fotosyntetyczna daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens i triticum aestivum l. w obecności jonów miedzi oraz wanadu streszczenie celem niniejszej pracy było zbadanie wpływu jonów miedzi i wanadu na aktywność fotosyntetyczną marchwi uprawnej (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) i pszenicy ozimej (triticum aestivum l.). przeprowadzono pomiary całkowitej zawartości chlorofilu – chlorofilomierzem spad oraz wyznaczono podstawowe parametry fluorescencji chlorofilu a  przy użyciu fluorymetru fms-1 – hansatech. w badaniach wykorzystano wodne roztwory soli miedzi (cuso4) i wanadu (h4no3v), o stężeniach molowych 0,6 mm, i  3 mm. grupę kontrolną stanowiły rośliny podlewane wodą destylowaną. zarówno u marchwi, jak i u pszenicy, wraz ze wzrostem koncentracji jonów metali ciężkich, zaobserwowano zmniejszenie zawartości chlorofilu oraz wykazano istotne zmiany w aktywności fotosystemu ii. pomiary kinetyki fluorescencji chlorofilu a jedynie u roślin d. carota wykazały istotny statystycznie wpływ 3 mm roztworów jonów miedzi i wanadu na aktywność fotosyntetyczną badanych roślin. u t. aestivum l. żaden z jonów metali ciężkich nie wywoływał znaczących zmian w wartościach parametrów fluorescencji chlorofilu a. key words: fotosynteza, marchew, pszenica, metale ciężkie, siarczan miedzi (ii), metawanadan amonu information about authors iwona konieczna https://orcid.org/0000-0002-9538-8516 she is a student of biology. she is interested in allelopathic interaction between plants and effect of heavy metals on plant morphology and physiology. grzegorz rut https://orcid.org/0000-0001-6719-3060 his research topic concern the course of basic physiological processes in plants (photosynthesis, respiration, chlorophyll fluorescence, transpiration), concentration chlorophyll, antocyanins, and reactions of plants, mainly mosses (polytrichum piliferum, mnium undulatum), ferns (platycerium bifurcatum), and vascular plants (oxalis acetosella, o. corniculata) on abiotic stress factors (excess and deficiency of solar radiation, hypoxia, anoxia, excess heavy metals, herbicides, salinity, changes in carbon dioxide concentration, drought stress, and low temperature). angelika kliszcz she is focusing on enhancing the understanding of the influence of different factors on soil structure and fertility. particularly, she is investigating the interaction of plants with the physical, chemical, and biological properties of the soil. she is also interested in organic methods of plant production and soil-enriching substances. p hotosynthetic activity of d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens and triticum aestivum l. in the presence of copper and vanadium salts 124 annales universitatis paedagogicae cracoviensis studia naturae, 2: 124–134, 2017, issn 2543-8832 doi: 10.24917/25438832.2.10 sylwia śliwińska-wilczewska*, arkadiusz knitter, daria cisło, adam latała institute of oceanography, university of gdansk, gdynia, poland; *ocessl@edu.pl allelopathic activity of the baltic picocyanobacterium synechocystis sp. introduction �e term “allelopathy” as �rst de�ned by molisch (1937), who indicated that these are positive or negative biochemical interactions between plants. rice (1979) included into this de�nition the interaction between micro-organisms, and inderjit and dakshini (1994) indicated that allelopathic activity is also present in the aquatic environment, especially between phytoplankton species. allelopathic interaction is widespread and can occur in all aquatic ecosystems (gross, 2003). moreover, cyanobacterial allelopathy can be found in all aquatic environments where these primary producers are able to release active allelopathic compounds which can a�ect the functioning of the whole ecosystem (żak, kosakowska, 2015). production of these allelopathic compounds is an important adaptation by which some cyanobacteria can achieve a competitive advantage over other primary producers (poniedziałek et al., 2015). allelopathy may be also one of the factors affecting the formation of massive and harmful cyanobacterial blooms in aquatic environments. over the past few decades, the world’s coastal waters have experienced an increase in the number of harmful bloom events (anderson et al., 2012). recent studies indicate that blooms of picocyanobacteria have grown signi�cantly in the last decades (sorokin, zakuskina, 2010), so it is important to determine the allelopathic interactions between the dominant species of picocyanobacteria. �e production and release of allelopathic compounds is an intriguing concept in which substances are secreted by the interacting species. cyanobacteria are e�ective producers of many bioactive metabolites, including both acute toxins and allelopathic compounds (burja et al., 2001; berry et al., 2008; leão et al., 2012). mazur-marzec et al. (2015) described baltic cyanobacteria as a rich source of novel metabolites, e.g., curacin a, apratoxin d, dolastatin 10 and largazole, and di�erent peptides, e.g., anabaenopeptins, cyanopeptolins, aeruginosins, spumigins, and nostocyclopeptides. however, 125 a llelopathic activity of the b altic picocyanobacterium synechocystis sp. many of cyanobacteria remain unknown. moreover, production of allelopathic substances is common, but not identical for all species forming massive blooms, and there are no clear reasons for the synthesis of these compounds (gross, 2003; legrand et al., 2003). it is possible that their function enables the deterrence of predators and the inhibition of co-occurring species of phytoplankton (liu et al., 2010). in addition, it is suggested that in the baltic sea cyanobacterial allelopathy may cause their dominance a�er the maximum concentration of cells is formed by environmental factors (suikkanen et al., 2004). moreover, available studies indicate that the baltic cyanobacteria may be allelopathic to other species, and this e�ect is dependent not only on the selected species, but even a strain of cyanobacteria (śliwińska et al., 2011). �e use of biological tests is the �rst step in identifying which group of allelopathic compounds is responsible for causing harmful e�ects in the aquatic environment (fistarol et al., 2004). �e main aim of this study was to estimate the allelopathic interaction between di�erent baltic strains of picocyanobacteria of the genus synechocystis. in this study, the in�uence of allelopathic compounds on the growth of analysed species was investigated in monocultures and in mixed cultures. �ese results indicate that the allelopathic e�ect of synechocystis sp. may be connected with the formation of a massive cyanobacterial bloom of these organisms in many aquatic ecosystems. material and methods �e experiments were conducted on the three strains of the picocyanobacteria of the genus synechocystis (ba-121, ba-122 and ba-153) (fig. 1). �e strains were isolated from the coastal zone of the gulf of gdańsk (southern baltic sea) and are maintained as uni-algal cultures in the culture collection of baltic algae (ccba) at the institute of oceanography, university of gdańsk, poland (latała et al., 2006). �e tests on the ‘batch cultures’ were carried out in 25 ml glass erlenmeyer �asks containing sterilised f/2 medium (guillard, 1975). �e media were prepared from baltic water with a salinity of 8 psu, which was �ltered through whatman gf/c glass �bre �lters, and autoclaved. analysed picocyanobacteria were grown 7 days in constant conditions of 20°c and 8 psu, under a 16:8 h light:dark cycle at 10 μmol photons m-2s-1 and these were the control treatment conditions. fluorescent lamps (cool white 40w, sylvania, usa) were used as a source of irradiance. �e intensity of par was measured using a li-cor quantum-meter with a cosine collector. �e donor and target picocyanobacteria were acclimated to these culture conditions for 7 days; a�erwards, actively growing cultures were used for the establishment of the allelopathic experiment. allelopathic interactions in monocultures were determined by using the modi�ed method proposed by suikkanen et al. (2004). allelopathic interaction was studied by adding the �ltrate obtained from picocyanobacterial culture of s yl w ia ś liw iń sk aw ilc ze w sk a, a rk ad iu sz k ni tte r, d ar ia c is ło , a da m l at ał a 126 synechocystis sp. ba-153 to tested picocyanobacteria synechocystis sp. ba-121 and ba-122. �e culture of ba-153 was �ltered through 0.45-µm pore size macherey-nagel mn gf-5 �lters. �e cell-free �ltrate (v = 2 ml) was added to 25 ml erlenmeyer �asks containing the tested cyanobacteria (v = 20 ml). in all experiments, the ratio of picocyanobacterium to target species in erlenmeyer �asks was adjusted to 1:1 based on the chlorophyll a (chl a) content (�nal chl a concentration in the experimental cultures was 0.4 µg chl a ml-1). control samples were prepared by adding mineral medium f/2 with a volume equal to the added cell-free �ltrate. �e number of cells in the experimental cultures was determined a�er the 1st and 7th day of the cyanobacteria exposure to the picocyanobacterial �ltrate. tests were conducted in triplicate and all analysed species were obtained from the early exponential growth phase. allelopathic interactions in mixed cultures were determined by using the modi�ed method proposed by ji et al. (2011). allelopathic activity was studied by adding the culture of donor picocyanobacteria synechocystis sp. ba-153 to the tested organisms: synechocystis sp. ba-121 or ba-122. in all experiments, the �nal chl a concentration in the experimental cultures was 0.4 µg chl a ml-1. �e culture of donor picocyanobacteria (v = 2 ml) was added to 25 ml erlenmeyer �asks containing the tested cyanobacteria (v = 20 ml). as controls, both synechocystis sp. ba-121 and ba-122 were cultured individually. controls consisted in the addition of 2 ml of �ltrated f/2 medium to 25 ml erlenmeyer �asks containing 20 ml of cell suspensions of the same cyanobacteria species. we measured the change in the density of target picocyanobacteria by directly counting the number of cells a�er the 1st and 7th day. �ree replicate �asks were used for each treatment, and all analysed species were obtained from the early exponential growth phase. �e experiments lasted 7 days. �e number of cells was counted using �ow cytometer bd accuri™ c6 plus (fig. 2, 3). events are recorded in list mode. to avoid generating large �les, samples can be run for 40s at a delivery rate of 14 µl min-1, and the number of events is kept at less than 1.000 per second. events are recorded in standard �lters: 670 lp (detector fl3) and 675/25 (detector fl4) (marie et al., 2005). fig. 1. picocyanobacterial strains of the genus synechocystis used in this study: a) ba-121, b) ba-122 and c) ba-153. scale bars = 10 µm 127 analysis of variance (anova) was used to test for di�erences in analysed parameters between the target cultures treated with picocyanobacterial cell-free �ltrates or living cells and the control over the experimental period. a post hoc test (tukeys hsd) was used to show which treatments for growth signi�cantly di�ered from the control and from each other. data are reported as mean ± standard deviation (sd). levels of signi�cance were * p < 0.05. �e statistical analyses were performed using the statistica® 13.1 so�ware. results �e e�ect of the cell-free �ltrate addition obtained from strain of synechocystis ba-153 on the growth of synechocystis ba-121 and ba-122 a�er 1 and 7 days of exposition to the �ltrates are shown in �gure 4. �e results demonstrated that the addition of a llelopathic activity of the b altic picocyanobacterium synechocystis sp. fig. 3. cytograms obtained with mixed cultures of picocyanobacterial strains of the genus synechocystis analysed using a bd accuri™ c6 plus �ow cytometer fig. 2. cytograms obtained with monocultures of three picocyanobacterial strains of the genus synechocystis: a) ba-121, b) ba-122 and c) ba-153 analysed using a bd accuri™ c6 plus �ow cytometer s yl w ia ś liw iń sk aw ilc ze w sk a, a rk ad iu sz k ni tte r, d ar ia c is ło , a da m l at ał a 128 cell-free �ltrate from ba-153 decreased the number of cells of ba-121 compared to their control. a�er the �rst day of the experiment, for a �ltrate addition obtained from synechocystis sp. ba-153 growth inhibition of synechocystis sp. ba-121 expressed as a percent of culture density (% of control) constituted 45% (p < 0.05). on the other hand, it was found that the addition of �ltrate obtained from synechocystis sp. ba-153 did not a�ect the number of cells of synechocystis sp. ba-121 a�er 7 day of exposition (p > 0.05; fig. 4a). in addition, it was observed that the cell-free �ltrate obtained from synechocystis sp. ba-153 did not a�ect the number of cells of synechocystis sp. ba-122 (p > 0.05, fig. 4b). fig. 5. �e e�ect of the addition of living cells from synechocystis sp. ba-153 cultures on the growth of a) synechocystis sp. ba-121 and b) synechocystis sp. ba-122 a�er 1st and 7th day of exposition, expressed as a number of cells (n). �e values refer to means (n = 3, mean ± sd). asterisk indicates signi�cant di�erence compared with control (p < 0.05) fig. 4. �e e�ect of the addition of cell-free �ltrate from synechocystis sp. ba-153 cultures on the growth of a) synechocystis sp. ba-121 and b) synechocystis sp. ba-122 a�er 1st and 7th day of exposition, expressed as a number of cells (n). �e values refer to means (n = 3, mean ± sd). asterisk indicates significant di�erence compared with control (p < 0.05) n um be r o f c el ls (n ∙ 10 5 ) time (d) time (d) n um be r o f c el ls (n ∙ 10 5 ) time (d) a a b b 129 a�er the addition of live cells of the strain synechocystis ba-153 to synechocystis ba-121 culture, the decline in growth was observed (fig. 5a). on the 1st day of the experiment, the minimum cell response of synechocystis sp. ba-121 constituted 40% (p < 0.05) in comparison to the control treatment. on the other hand, the addition of live cells of strain synechocystis ba-153 culture stimulated the growth of synechocystis sp. ba-122, and at 7th day of experiment growth increased by 94% relative to control (p < 0.05, fig. 5b) discussion �e study of the allelopathy phenomenon focuses mainly on understanding the e�ects of allelopathic compounds on a variety of target organisms. �e modes of the action of allelochemicals depend on the nature of the interaction between donor and target organisms and the activity of these compounds. �ere are some reports of allelopathic e�ects, such as growth stimulation or inhibition caused by di�erent cyanobacteria (suikkanen et al., 2006), but no information about the allelopathic potential of baltic picocyanobacterium of genus synechocystis on co-existing picocyanobacterial strains has been found. �erefore, the main goal of this study was to investigate the in�uence of metabolites obtained from picocyanobacterium synechocystis sp. in monocultures and in mixed cultures. in most cases, allelopathic compounds cause the death of target organisms or reduce their growth rate and biomass (rzymski et al., 2014). moreover, inhibition of the growth of the target organism by production allelopathic compounds is relatively widespread and the most frequently reported mode of action of cyanobacteria (issa, 1999; schagerl et al., 2002). in this study, we have demonstrated that the picocyanobacterium strain of the genus synechocystis ba-153 caused allelopathic e�ects against other strains of picocyanobacteria. it was found that strain ba-121 was strongly inhibited by strain ba-153 in both the mixed culture and cell-free �ltrates. picocyanobacteria plays an important role in aquatic ecosystems but not much is known about their allelopathic activity. information about the ability to allelopathic interactions of other picocyanobacterium synechococcus sp. was described by śliwińska-wilczewska et al. (2016a). in this study, the authors showed that the addition of the cell-free �ltrate obtained from the baltic picocyanobacterium synechococcus sp. had a signi�cant inhibitory e�ect on nodularia spumigena. additionally, śliwińska-wilczewska et al. (2016b) showed the sensitivity of the diatom navicula perminuta to allelopathic compounds produced by this picocyanobacterium. in this study, it was demonstrated that analysed picocyanobacterium reveals allelopathic activity that was regulated by the intensity of light, temperature, and salinity. śliwińska et al. (2011) also showed that all analysed strains of synechococcus sp. demonstrated the allelopa llelopathic activity of the b altic picocyanobacterium synechocystis sp. s yl w ia ś liw iń sk aw ilc ze w sk a, a rk ad iu sz k ni tte r, d ar ia c is ło , a da m l at ał a 130 athic activity and signi�cantly decreased the number of cells of chlorella vulgaris. �e authors concluded that all three strains of baltic picocyanobacterium from the genus synechococcus (ba-120, ba-124, and ba-132) showed allelopathic activity; however, the ba-124 strain had the greatest negative impact on the growth of analysed green algae c. vulgaris. moreover, martins et al. (2008) describe the biological activities of marine cyanobacteria strains belonging to the genus synechocystis and synechococcus isolated from atlantic coast of portugal. �e authors screened marine picocyanobacterial secondary metabolites for cytotoxic activity using primary rat hepatocytes and human hl-60 cells. in this study, the authors showed that marine picocyanobacterial strains of the genus synechocystis and synechococcus produce substances with inhibitory effects on prokaryotic cells and with apoptotic activity in eukaryote cell lines. moreover, hamilton et al., (2014) demonstrated a signi�cant e�ect of synechococcus strain on dark preference of black perch embiotoca jacksoni. costa et al. (2015) demonstrated that marine picocyanobacterium cyanobium sp. inhibited nannochloropsis sp. growth. moreover, paz-yepes et al. (2013) have found a clear growth impairment of two strains of synechococcus sp. (cc9311 and wh8102) when they are cultured in the presence of synechococcus sp. cc9605. it is believed that selective inhibition of the growth of the target organism may a�ect the succession of selected cyanobacteria in an aquatic ecosystem (legrand et al., 2003). our results showed that synechocystis allelopathy should be considered when estimating the potential interactions between picocyanobacteria in aquatic ecosystems. moreover, the results may in part explain the reasons for achieving a competitive advantage of picocyanobacterium synechocystis in many aquatic ecosystems, including the baltic sea. it was also found that the addition of live cells of picocyanobacterium strain ba-153 stimulated the growth of synechocystis sp. ba-122. suikkanen et al. (2005) also indicated that cyanobacteria may stimulate natural plankton assemblages. �e authors showed that cyanobacterial �ltrates stimulated other cyanobacteria in the community. �e numbers of snowella sp. and pseudanabaena sp. were signi�cantly higher in all cyanobacterial treatments than in the control. moreover, the addition of n. spumigena �ltrate signi�cantly increased the numbers of both n. spumigena and anabaena sp. and aphanizomenon sp. �ltrate and caused an over 50-fold increase in the cell numbers of aphanizomenon sp. in the community. in the present study, picocyanobacterial exudates had the stimulatory e�ects on the growth of other picocyanobacterium from the same genus. �is suggests that the cyanobacteria released some autostimulatory, which accelerated the growth of the same and related species. our results con�rm that the picocyanobacterium synechocystis sp. ba-153 is able to produced allelopathic compounds. unfortunately, information on which metabolites mediate this and many other known allelopathic interactions are still scarce (leão et 131 al., 2012). �e identi�cation and isolation of the compounds responsible for the allelopathic e�ect detected in synechocystis sp. in our studies will require further research. �ese results showed the allelopathic activity of baltic synechocystis, which either causes the inhibition or stimulation of the growth of selected picoplanktonic cyanobacteria. many studies indicated that cyanobacteria produced a wide spectrum of secondary metabolites (e.g., berry et al., 2008). it is believed that allelopathy may be one of the important factors a�ecting the formation of massive cyanobacterial blooms in aquatic environments (gross, 2003; legrand et al., 2003). despite the seriousness of the problem, relatively little is known about the allelopathic e�ect between picocyanobacteria in the baltic sea. to evaluate the signi�cance of the allelopathy phenomenon, more detailed studies are needed about the production and secretion of active allelopathic compounds in order to better understand the mechanisms of their e�ects on the surrounding ecosystem. acknowledgements �e authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. �is study was supported by bmn grants, poland, no. 538-g245-b568-17. references anderson, d.m., cembella, a.d., hallegrae�, g.m. 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(2006). allelopathy of baltic sea cyanobacteria: no evidence for the role of nodularin. journal of plankton research, 28, 543–550. doi: 10.1093/plankt/�i139 śliwińska, s., jodłowska, s., latała, a. 2011. eko�zjologiczne i allelopatyczne właściwości pikoplanktonowej sinicy synechococcus sp. acta geographica silesiana, 1, 63–66. śliwińska-wilczewska, s., pniewski, f., latała, a. (2016a). allelopathic interactions between synechococcus sp. and nodularia spumigena under di�erent light conditions. allelopathy journal, 37(2), 241–252. śliwińska-wilczewska, s., pniewski, f., latała, a. (2016b). allelopathic activity of the picocyanobacterium synechococcus sp. under varied light, temperature and salinity conditions. international review of hydrobiology, 101, 1–9. doi: 10.1002/iroh.201501819 żak, a., kosakowska, a. (2015). �e in�uence of extracellular compounds produced by selected baltic cyanobacteria, diatoms and dino�agellates on growth of green algae chlorella vulgaris. estuarine, coastal and shelf science, 167, 113–118. doi: 10.1016/j.ecss.2015.07.038 abstract allelopathic compounds produced by picocyanobacteria could a�ect the growth and development of biological systems. �e main aim of this study was to investigate the in�uence of unknown allelochemicals obtained from picocyanobacterium synechocystis sp. ba-153 in monocultures and in mixed cultures. in this study, we demonstrated that synechocystis sp. ba-153 caused allelopathic e�ects against other strains of picocyanobacteria. it was found that synechocystis sp. ba-121 was strongly inhibited by synechocystis sp. ba-153 in both the mixed culture and cell-free �ltrates. on the other hand, the addition of live picocyanobacterial culture of synechocystis sp. ba-153 stimulated the growth of synechocystis sp. ba-122. �ese results showed the allelopathic activity of synechocystis sp. ba-153, which can cause either the inhibition or stimulation of growth of selected picoplanktonic cyanobacteria. key words: allelopathy, picocyanobacteria, synechocystis sp., growth, mixed culture, monoculture received: [2017.05.27] accepted: [2017.10.19] zjawisko oddziaływania allelopatycznego bałtyckiej pikoplanktonowe j sinicy synechocystis sp. streszczenie związki allelopatyczne produkowane przez pikoplanktonowe sinice mogą wpływać na wzrost i funkcjonowanie gatunków �toplanktonu w ekosystemach wodnych. głównym celem niniejszej pracy było wykazanie wpływu wciąż nierozpoznanych związków allelopatycznych produkowanych przez pikoplanktonową sinicę synechocystis sp. ba-153 w monokulturach i hodowlach mieszanych. na podstawie uzyskanych danych wykazano, że pikoplanktonowa sinica synechocystis sp. ba-153 wykazywała oddziaływanie allelopatyczne na inne szczepy pikoplanktonowych sinic z tego samego rodzaju. stwierdzono, że szczep synechocystis sp. ba-153 silnie hamował wzrost synechocystis sp. ba-121, zarówno w hodowlach mieszanych, jak i w eksperymentach z dodaniem przesączu. z drugiej strony, dodanie żywej kultury synechocystis sp. ba-153 stymulowało wzrost synechocystis sp. ba-122. uzyskane wyniki potwierdzają aktywność allelopatyczną synechocystis sp. ba-153 i wskazują, że może ona zarówno hamować, jak i stymulować wzrost wybranych szczepów pikoplanktonowych sinic. słowa kluczowe: allelopatia, pikoplanktonowe sinice, synechocystis sp., wzrost, hodowle mieszane, monokultury a llelopathic activity of the b altic picocyanobacterium synechocystis sp. s yl w ia ś liw iń sk aw ilc ze w sk a, a rk ad iu sz k ni tte r, d ar ia c is ło , a da m l at ał a 134 information on the authors sylwia śliwińska-wilczewska she is interested in the allelopathy of cyanobacteria and microalgae, in particular of picocyanobacteria synechococcus sp. in her study, the in�uence of allelochemicals on the growth, chlorophyll �uorescence, and photosynthesis irradiance curves of di�erent phytoplankton species was investigated. she is also investigating what in�uences environmental factors have on produced allelopathic compounds on algae and cyanobacteria. arkadiusz knitter �e �eld of his interest is allelopathic interactions of picocyanobacterium synechocystis sp. in monocultures and mixed cultures. he uses a �ow cytometer to determine allelopathic interactions between picocyanobacteria. daria cisło she is interested in allelopathy of picocyanobacteria, in particular of picocyanobacterium synechocystis sp. she is investigating what in�uence allelopathic compounds have on those organisms in monocultures and mixed cultures. adam latała he has wide experience in ecophysiology and ecotoxicology of marine benthic and planktonic algae. his particular interest is on the in�uence of the main environmental factors, such as salinity, temperature, and light on the photosynthesis, photoacclimation, �uorescence, respiration, and the growth of algae from natural communities and cultured under laboratory conditions, including the use of �uorescence techniques to determine algal and cyanobacterial ecophysiology and ecotoxicology. he is the curator of culture collection of baltic algae (ccba) at the institute of oceanography, university of gdańsk. actually, ccba maintains more than 100 baltic strains from three taxonomic groups: blue-green algae, green algae, and diatoms. 114 annales universitatis paedagogicae cracoviensis studia naturae, 2: 114–123, issn 2543-8832 doi: 10.24917/25438832.2.9 zofia konarzewska, sylwia śliwińska-wilczewska*, adam latała institute of oceanography, university of gdańsk, gdynia, poland, *ocessl@edu.pl allelopathic effect of the baltic picocyanobacterium synechococcus sp. on selected diatoms introduction �e allelopathy phenomenon was originally de�ned as a harmful e�ect between plants. �is de�nition was used for the �rst time by professor hans molisch (1937). as the result of the development of science and a better understanding of this phenomenon, the de�nition of allelopathy has evolved. �e international allelopathy society (ias, 1996) established the de�nition of allelopathy as the process in which the compounds secreted by di�erent organism (vascular plant, algae, bacteria and fungi) in�uenced the growth and development of the both plant and animal organisms (legrand et al., 2003). �ese chemical substances were called allelopathic compounds or allelochemicals (le�aive, ten-hage, 2007), and organisms used them to protection against predators, and avoid competitors (granéli et al., 2008). in aquatic ecosystems, allelopathic interactions are still not very well recognised, because the distance between organisms which are able to secreting secondary metabolites is crucial to observe the allelopathic e�ect (wolfe, 2000). allelopathy in freshwater, brackish, and marine ecosystems depends on the production and secretion of active allelopathic compounds and their e�ective spread to other organisms (lewis, 1986). �erefore, to demonstrate allelopathy between photoautotrophs, scientists need to perform a number of laboratory experiments. it has been proven that several groups of phytoplankton, such as cyanobacteria, dino�agellates, prymnesiophytes, green algae, and diatoms product allelopathic compounds (subba rao, smith, 1995; chiang et al., 2004; żak, kosakowska, 2015). moreover, published data indicated that allelopathic e�ects have the competitive advantage of some phytoplankton species by eliminating physiologically more sensitive species (sarkar et al., 2006). moreover, the structure of phytoplankton (legrand et al., 2003) and the formation of cyanobacterial and algal blooms may be determined by allelopathic 115 interactions (smayda, 1997; weissbach et al., 2010). massive and harmful cyanobacterial and algal blooms are a serious problem a�ecting not only the ecology but also the economy of the aquatic ecosystem (anderson, 1989; anderson et al., 2002). massive blooms are formed by cyanobacteria (also picocyanobacteria from the genus synechococcus), green algae, dino�agellates, and diatoms (beardall, 2008; błaszczyk et al., 2010). little is known about the in�uence of cyanobacteria on diatoms (sivonen, jones 1999), and the speci�c mechanism of action of the allelopathic interactions is unexamined. it is impossible to demonstrate allelopathy in natural conditions, which is why it is important to contribute controlled experiments in laboratory environment. �erefore, in this study, we investigated the allelopathic e�ect of picocyanobacterium synechococcus sp. on diatoms nitzschia fonticola (grunow) grunow, fistulifera saprophila (lange-bertalot & bonik) lange-bertalot, navicula perminuta grunow and amphora co�eaeformis (c.agardh) kützing. in this study, the in�uence of picocyanobacteria was examined by adding the cell-free �ltrate of synechococcus sp. to studied diatoms. �e results of this experiment may provide further information about allelopathic interactions between cyanobacteria and diatoms, which could be important to the understanding of cyanobacterial and algal blooms in aquatic ecosystems, including the baltic sea. material and methods �e experiments were conducted on the strain of picocyanobacterium synechococcus sp. (ba-124) and four diatoms: nitzschia fonticola (ba-34), fistulifera saprophila (ba56), navicula perminuta (ba-30), and amphora co�eaeformis (ba-16) (fig. 1). �ese species are typical representatives of baltic diatoms; therefore, they were selected for the allelopathic experiment. �e strains were isolated from the coastal zone of the gulf of gdańsk (southern baltic sea) and are maintained as uni-algal cultures in the culture collection of baltic algae (ccba) at the institute of oceanography, university of gdańsk, poland (latała et al., 2006). �e tests on the ‘batch cultures’ were carried out in 25 ml glass erlenmeyer �asks containing sterilised f/2 medium (guillard, 1975). �e media were prepared from baltic water, which was �ltered through whatman gf/c glass �bre �lters, and autoclaved. analysed organisms were grown 7 days in constant conditions of 18°c and 8 psu, under a 16:8 h light : dark cycle at 10 μmol photons·m-2 s-1 and this were the control treatment conditions. fluorescent lamps (cool white 40w, sylvania, usa) were used as source of irradiance. �e intensity of par was measured using a li-cor quantum-meter with a cosine collector. �e donor and target organisms were acclimated to these culture conditions for 7 days; a�erwards, actively growing cultures were used for the establishment of the allelopathic experiment. allelopathic interactions in monocultures were determined by using the modi�ed method proposed by suikkanen et al. (2004). allelopathic interaction a llelopathic effect of the b altic picocyanobacterium synechococcus sp. on selected diatom s zo fia k on ar ze w sk a, s yl w ia ś liw iń sk aw ilc ze w sk a, a da m l at ał a 116 was studied by adding the �ltrate obtained from the picocyanobacterial culture of synechococcus sp. ba-124 to tested diatoms. �e culture of picocyanobacteria was �ltered through 0.45-µm pore size macherey-nagel mn gf-5 �lters. �e cellfree �ltrate (v = 10 ml) was added to 25 ml erlenmeyer �asks containing the tested diatoms (v = 10 ml). in all experiments, the ratio of picocyanobacterium to target species in erlenmeyer �asks was adjusted to 1:1, based on the chlorophyll a content (�nal chlorophyll a concentration in the experimental cultures was 0.4 µg chl a ml-1). control samples were prepared by adding mineral medium f/2 with a volume equal to the added cell-free �ltrate. tests were conducted in triplicate, and all analysed species were obtained from early exponential growth phase. �e number of cells was counted using �ow cytometer bd accuri™ c6 plus. events are recorded in list mode. to avoid generating large �les, samples can be run for 40s at a delivery rate of 14 µl·min-1, and the number of events is kept at less than 1000 per second. events are recorded in standard �lters: 670 lp (detector fl3) and 675/25 (detector fl4) (marie et al., 2005). �e number of cells in the experimental cultures was determined a�er the 1st and 7th day of the diatoms exposure to the picocyanobacterial �ltrate. fig. 1. diatoms strains used in this study: nitzschia fonticola (a), fistulifera saprophila (b), navicula perminuta (c) and amphora co�eaeformis (d); scale bars = 10 µm a b c d 117 chlorophyll a �uorescence was measured with a pulse amplitude modulation (pam) �uorometer (fms1, hansatech), using a 594 nm amber modulating beam with 4 step frequency control as a measuring light. analysed species were taken for chlorophyll �uorescence analysis a�er the 7th day of exposure to the �ltrate. before measurements, each sample taken from the culture was �ltered through 13 mm glass �bre �lters (whatman gf/c). before starting the experiment, the �lter sample was adapted in the dark for about 10 minutes. �e maximum quantum yield of psii photochemistry (fv/fm) and e�ective quantum yield of psii photochemistry (φpsii) was calculated (campbell et al., 1998). analysis of variance (one-way anova) was used to test for di�erences in analysed parameters between the target cultures treated with picocyanobacterial cellfree �ltrates and the control over the experimental period. data are reported as mean ± standard deviation (sd). levels of signi�cance were * p < 0.05. �e statistical analyses were performed using the statistica® 13.1 so�ware. results �e e�ect of the cell-free �ltrate addition obtained from synechococcus sp. cultures on the growth of analysed diatoms nitzschia fonticola, fistulifera saprophila, navicula perminuta and amphora co�eaeformis a�er 1 and 7 days of exposition to the �ltrates fig. 2. �e e�ect of the addition of cell-free �ltrate from synechococcus sp. ba-124 cultures on the growth of nitzschia fonticola (a), fistulifera saprophila (b), navicula perminuta (c) and amphora co�eaeformis (d) a�er the 1st and 7th days of exposition, expressed as a number of cells (n). �e values refer to means (n = 3, mean ± sd); asterisk indicates signi�cant di�erence compared with control (p < 0.05) a llelopathic effect of the b altic picocyanobacterium synechococcus sp. on selected diatom s zo fia k on ar ze w sk a, s yl w ia ś liw iń sk aw ilc ze w sk a, a da m l at ał a 118 are shown in �gure 2. �e results showed that addition of cell-free �ltrate from synechococcus sp. increased the number of cells of n. fonticola and f. saprophila compared to their control. a�er the 7th day of the experiment for a �ltrate addition, growth stimulation of n. fonticola and f. saprophila constituted 105% and 107%, respectively (anova, p < 0.05). in addition, it was observed that the cell-free �ltrate obtained from picocyanobacterium did not a�ect the number of cells of n. perminuta and a. co�eaeformis (anova, p > 0.05). �e e�ect of the addition of cell-free �ltrate from synechococcus sp. cultures on the �uorescence parameters fv/fm and фpsii of analysed diatoms a�er one week of exposition is showed in �gure 3. �e study indicated that the addition of cell-free �ltrate obtained from synechococcus sp. signi�cantly a�ected the fv/fm and фpsii of n. fonticola and n. perminuta. a�er the 7th day of exposition, it was noted that the addition of the �ltrate resulted in increase of �uorescence parameters of analysed diatoms, which was higher by 29% and 4%, respectively, for fv/fm and 56% and 14%, respectively for фpsii, compared to a control (anova, p < 0.05). based on the results, it was found that the �ltrate obtained from synechococcus sp. also caused signi�cant changes of the фpsii value of f. saprophila. a�er one week of the experiment, the �uorescence parameter of this diatom was higher by 6%, compared to control (anova, p < 0.05). on the other hand, it was noted that �ltrate obtained from picocyanobacterium caused the inhibition of the fv/fm parameter of a. co�eaeformis, which constituted 96% (anova, p < 0.05). fig. 3. �e e�ect of the addition of cell-free �ltrate from synechococcus sp. ba-124 cultures on the �uorescence parameters fv/fm and фpsii of nitzschia fonticola (a), fistulifera saprophila (b), navicula perminuta (c) and amphora co�eaeformis (d) a�er the 7th day of exposition; the values refer to means (n = 3, mean ± sd); asterisk indicates signi�cant di�erence compared with control (p < 0.05) 119 discussion it is believed that cyanobacterial allelopathy may be one of the important factors affecting the formation of massive algal blooms in the aquatic ecosystems (e.g., antunes et al., 2012; rzymski et al., 2014). picocyanobacteria plays an important role in aquatic ecosystems but not much is known about their allelopathic activity. �erefore, the main aim of this study was to investigate the allelopathic e�ect of picocyanobacterium synechococcus sp. on growth and �uorescence parameters of nitzschia fonticola, fistulifera saprophila, navicula perminuta, and amphora co�eaeformis. allelopathic e�ects of the growth of the target organism by the production allelopathic compounds are the most frequently reported mode of the action of cyanobacteria and microalgae (rzymski et al., 2014). in this study, we have also demonstrated that picocyanobacterium caused allelopathic e�ects against baltic diatoms. �e results showed that addition of cell-free �ltrate from synechococcus sp. increased the number of cells of n. fonticola and f. saprophila. in addition, it was observed that the cell-free �ltrate obtained from picocyanobacterium did not a�ect the number of cells of n. perminuta and a. co�eaeformis. only a few studies have documented the in�uence of the allelopathic e�ect of cyanobacteria on diatoms species. information about the allelopathic interactions of synechococcus sp. on baltic diatom n. preminuta was described by śliwińska-wilczewska et al. (2016). in this study, the authors demonstrated that the addition of the cell-free �ltrate obtained from the picocyanobacterium had an inhibitory e�ect on analysed diatom. moreover, the study reveals that allelopathic activity was regulated by the intensity of light, temperature, and salinity. keating (1977, 1978) also demonstrated the growth inhibition of selected diatom by the addition of �ltrate obtained from a lake in which cyanobacteria was dominated. similar results were reported by la�orgue et al. (1995), who showed that the low biomass of fragilaria crotonensis kitton in lake aydat was the result of the allelochemicals obtained from anabaena sp. more detailed data on the allelopathic e�ects of baltic cyanobacteria on diatoms were provided by suikkanen et al. (2004). in this study, the allelopathic e�ects of baltic nodularia spumigena mertens ex bornet & flahault, anabaena lemmermannii p.g.richter and aphanizomenon �osaquae ralfs ex bornet & flahault on the diatom �alassiosira weiss�ogii (grunow) g.fryxell & hasle were demonstrated. �e authors showed that, a�er a single addition of the �ltrate, all three analysed cyanobacteria had a generally negative e�ect on the tested diatom. moreover, on the basis of the research, it was noted that the diatom of t. weiss�ogii showed some tolerance for a single addition of the �ltrate, but its growth a�er repeated additions of the �ltrate was signi�cantly inhibited. di�erent characteristics, such as membrane permeability, may contribute to the sensitivity of some phytoplankton species to allelopathic compounds (suikkanen et al., 2004). �e sensitivity of organisms may also depend on the nature of a llelopathic effect of the b altic picocyanobacterium synechococcus sp. on selected diatom s zo fia k on ar ze w sk a, s yl w ia ś liw iń sk aw ilc ze w sk a, a da m l at ał a 120 allelopathic compounds to which they are exposed, because the same target organisms may react di�erently to the �ltrate derived from di�erent donor organisms. �ere are also only a few reports on the growth stimulation of target organisms caused by allelochemicals produced by cyanobacteria. suikkanen et al. (2005) showed that some baltic cyanobacteria may stimulate natural plankton communities. �e authors found that the addition of n. spumigena �ltrate signi�cantly increased the numbers of cells of n. spumigena and anabaena sp. in our study, allelochemicals produced by synechococcus sp. had the stimulatory e�ects on the growth of diatoms n. fonticola and f. saprophila. �is indicated that the picocyanobacteria released some compounds that accelerated the growth of selected and co-existing diatoms. another possible e�ect of allelopathic compounds is their e�ect on the photosynthetic activity of target organisms. measurements of chlorophyll a �uorescence are non-invasive methods. moreover, �uorescence measurements are a useful tool in the study of the allelopathy and ecophysiology of cyanobacteria and microalgae (prince et al., 2008). in this study, it was found that the addition of �ltrate stimulated and inhibited the �uorescence parameters fv/fm and φpsii of analysed diatoms. published data indicated that the allelopathic compounds produced and released by some cyanobacteria can a�ect the photosynthesis, and detailed studies have shown that they act mainly on the photosystem ii (psii). issa (1999) presented the inhibitory e�ect of the donor oscillatoria sp. and calothrix sp. on photosynthesis of chlorella fusca shihira & r.w.krauss. more detailed studies were conducted prince et al. (2008). �e authors analysed the impact of karenia brevis (c.c.davis) gert hansen & ø.moestrup on �uorescence parameter fv/fm of target diatoms amphora sp., asterionellopsis glacialis (castracane) round and skeletonema costatum (greville) cleve. inhibition of photosynthetic yield by extract obtained from k. brevis coincided with a negative e�ect on the growth of target organisms. �e e�ect of allelopathic compounds may be due to the di�erent sensitivity of the target species (mulderij et al., 2003), and this may explained the dominance of some species over other photoautotrophs. it is believed that selective stimulation or inhibition of the target organism may a�ect the succession of selected cyanobacteria and microalgae in aquatic ecosystem (legrand et al., 2003). �e present research demonstrated the allelopathic e�ect of synechococcus sp. either inhibited or stimulated the growth and �uorescence parameters of selected baltic diatoms. to evaluate the signi�cance of the picocyanobacterial allelopathy, more detailed studies are needed. �ese results showed that synechococcus sp. allelopathy should be considered when estimating the potential interactions between picocyanobacteria and diatoms in aquatic ecosystems. acknowledgements �e authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. �is study was supported by bmn grants, poland, no. 538-g245-b568-17. 121 references anderson, d.m. 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(2016). allelopathic activity of the picocyanobacterium synechococcus sp. under varied light, temperature and salinity conditions. international review of hydrobiology, 101, 1–9. doi: 10.1002/iroh.201501819 subba rao, d.v., pan, y., smith, s.j. (1995). allelopathy between rhizosolenia alata (brightwell) and the toxigenic pseudo-nitzschia pungens f. multiseries (hasle). in: p. lassus, g. arzul, e.e. le denn, p. gentien, c. marcaillou (eds.), harmful marine algal blooms. paris: lavoiser intercept ltd, p. 681–686. weissbach, a., tillmann, u., legrand, c. (2010). allelopathic potential of the dino�agellate alexandrium tamarense on marine microbial communities. harmful algae, 10, 9–18. doi: 10.1016/j.hal.2010.05.007 wolfe, g.v. (2000). �e chemical defense ecology of marine unicellular plankton: constraints, mechanisms, and impacts. biological bulletin, 198, 225–244. żak, a., kosakowska, a. (2015). �e in�uence of extracellular compounds produced by selected baltic cyanobacteria, diatoms and dino�agellates on growth of green algae chlorella vulgaris. estuarine, coastal and shelf science, 167, 113–118. doi: 10.1016/j.ecss.2015.07.038 abstract it is commonly believed that the structure of phytoplankton and the formation of cyanobacterial and algal blooms may be explained by allelopathic interactions. �e main aim of this study was to investigate the allelopathic e�ect of picocyanobacterium synechococcus sp. on the following growth and �uorescence parameters: the maximum quantum yield of psii photochemistry (fv/fm), and the e�ective quantum yield of psii photochemistry (φpsii) of selected diatoms – nitzschia fonticola, fistulifera saprophila, navicula perminuta and amphora co�eaeformis. in this study, it was demonstrated that picocyanobacterium caused allelopathic 123 e�ects against baltic diatoms. �e results showed that the addition of cell-free �ltrate from synechococcus sp. increased the number of cells of n. fonticola and f. saprophila. moreover, it was found that picocyanobacterium was stimulated �uorescence parameters of n. fonticola, f. saprophila, and n. perminuta. on the other hand, it was noted that �ltrate obtained from picocyanobacterium caused the inhibition of fv/fm parameter of a. co�eaeformis. �e results of this experiment may provide further information about allelopathic interactions between baltic picocyanobacteria and diatoms that are crucial to the understanding of algal blooms in aquatic ecosystems. key words: allelopathy, baltic sea, diatom, growth, �uorescence, picocyanobacteria received: [2017.07.22] accepted: [2017.10.27] zjawisko oddziaływania allelopatycznego bałtyckiej pikoplanktonowej sinicy synechococcus sp. na wybrane gatunki okrzemek streszczenie powszechnie uważa się, że oddziaływanie allelopatyczne może mieć wpływ na strukturę �toplanktonu oraz tworzenie masowych zakwitów sinic oraz mikroglonów. głównym celem niniejszych badań było wykazanie oddziaływania allelopatycznego pikoplanktonowej sinicy synechococcus sp. na wzrost oraz parametry �uorescencji, charakteryzujące maksymalną wydajność fotosystemu ii (fv/fm) oraz rzeczywistą wydajność fotosystemu ii na świetle (φpsii) wybranych gatunków okrzemek: nitzschia fonticola, fistulifera saprophila, navicula perminuta i amphora co�eaeformis. zaobserwowano, że pikoplanktonowa sinica wykazywała allelopatyczne właściwości na badane bałtyckie okrzemki. na podstawie uzyskanych danych zanotowano, że dodanie przesączu z sinicy synechococcus sp. stymulowało wzrost n. fonticola i f. saprophila. ponadto zaobserwowano znaczący wzrost parametrów �uorescencji u n. fonticola, f. saprophila i n. perminuta. w pracy również wykazano, że przesącz powodował obniżenie wartości parametru fv/fm u a. co�eaeformis. wyniki uzyskane w niniejszej pracy dostarczają szerszych informacji odnoście oddziaływania allelopatycznego, występującego pomiędzy wybranymi gatunkami sinic i okrzemek, które w znaczący sposób mogą przyczynić się do zrozumienia pojawiających się w wielu ekosystemach wodnych masowych zakwitów sinic i mikroglonów. słowa kluczowe: allelopatia, morze bałtyckie, okrzemki, wzrost, �uorescencja, pikoplanktonowe sinice information on the authors zo�a konarzewska �e �eld of her interest is allelopathic interactions of picocyanobacterium synechococcus sp. and baltic diatoms in monocultures. in her studies, she uses �ow cytometer and pam �uorymetry to determine allelopathic interactions between picocyanobacteria and selected diatoms – nitzschia fonticola, fistulifera saprophila, navicula perminuta and amphora co�eaeformis. sylwia śliwińska-wilczewska in her research, she is interested in the allelopathy of cyanobacteria and microalgae, and particularly picocyanobacteria synechococcus sp. allelopathy plays an important role in interspeci�c competition and contributes to cyanobacterial bloom maintenance. in her study, the in�uences of allelochemicals on the growth, chlorophyll �uorescence, and photosynthesis irradiance curves of di�erent phytoplankton species were investigated. she is also investigating what in�uences environmental factors have on produced allelopathic compounds on algae and cyanobacteria. adam latała his subjects of research include wide experience in the ecophysiology and ecotoxicology of marine benthic and planktonic algae, the in�uence of the main environmental factors such as salinity, temperature, and light on the photosynthesis, photoacclimation, �uorescence, respiration, and growth of algae from natural communities and cultured under laboratory conditions, and the use of �uorescence techniques to study algal and cyanobacterial ecophysiology and ecotoxicology. a llelopathic effect of the b altic picocyanobacterium synechococcus sp. on selected diatom s 97 annales universitatis paedagogicae cracoviensis studia naturae, 2: 97–103, 2017, issn 2543-8832 doi: 10.24917/25438832.2.7 joanna puła*, angelika kliszcz department of agrotechnology and agricultural ecology, university of agriculture in kraków, mickiewicza ave. 21, 31-120 krakow, poland, *rrpula@cyf-kr.edu.pl the yield of jerusalem artichoke plant helianthus tuberosus l. grown in various combinations of fertilisation – preliminary research introduction jerusalem artichoke – helianthus tuberosus l. is commonly known in poland as ‘topinambur’. it is a plant whose history goes back to the pre-columbian times, when in the eastern and southern parts of north america, topinambur was cultivated by indigenous peoples. �e jerusalem artichoke was brought to europe in the early 17th century (jasińska, kotecki, 2003). despite the many advantages of tubers, both in nutritional aspects (content of inulin, high content of vitamins and minerals) and the ability to use it in the human diet (tubers can be fried, cooked, marinated, pickled), until now, this plant has not found a widespread use and the area of cultivation is still small (cieślik, 1998; cieślik, filipiak-florkiewicz, 2000). in poland, it is cultivated on a small scale, mostly for feed and consumer purposes and as an energy crop and decorative plant. �e plant can be used for energy purposes in a twofold way: harvesting green fodder for biogas production or harvesting tubers for bioethanol production. assuming a triple cut of green fodder and yield within 100 t∙ha-1, biogas production can reach a level of 53.500 m3. in the case of straw (50 t∙ha-1), it is possible to generate 900 gj during combustion, and as far as the use of tubers is concerned, it has been calculated that, from 25 t∙ha-1, 2.600 dm3 of ethanol can be obtained (piskier, 2004). �is plant is successfully planted around the forest as bait for forestry quarry (mainly wild boars) to minimize losses in root crops in adjacent crop �elds. �e latin name helianthus tuberosus was proposed by carl linnaeus in 1753; however, the popular name is topinambur and, in english literature, the jerusalem artichoke. �e origin of this �rst nomenclature is related to the indian tribe topinamba in south america, from where this native plant was taken over by the natives of the north american continent. one of the theories that explain the english name of this plant is derived from the organoleptic properties of this plant. namely, the cooked jo an na p uł a, a ng el ik a k lis zc z 98 tubers are similar in taste and texture to the receptacle of the globe artichoke (cynara scolymus l.). on the other hand, the reference to jerusalem artichoke may just be a phonetic simpli�cation of the italian word (girasole) in the 17th century, in response to the di�culty of pronunciation (kays, nottingham, 2008). in poland, there are mainly two cultivar varieties: cv. albik, elongated, white tubers and cv. rubik, oval tubers with purple and red skin. other foreign varieties such as ‘boston red’, ‘fuseau’, or ‘golden nugget’ are also available (makowski, 2014). �is species belongs to the compositae family (now asteraceae). �ese plants produce raised stems of about 3–4 m height and underground stolons, at the ends of which are formed tubers of various shapes and di�erent colours, depending on the variety (forkiewicz et al., 2007). �is species is extremely tolerant to the environment. it does not require intensive cultivation, and stands poor sites in nutrients and water quite well. it reproduces by seeds (but it is quite easy to create hybrids in a natural state and as a short-day plant, in our latitude, it will not be able to develop mature seeds before frosts) as well as vegetatively by tubers (jasińska, kotecki, 2003). tubers winter in the soil and start growing quite fast in spring. �e plant shading the ground dynamically is much better at dealing with weeds, which gives it an advantage over other root crops, especially in relation to the potato. �ere is no need to plant tubers every year, as the inulin content helps them resist the frosts up to -30°c. nowadays, the plant is also used to establish energy crops from which biomass is obtained for further processing, achieving an average energy of 96.2 mj per hectare of plantation in poland (average yield of dry matter obtained in experiment multiplied by accepted calori�c value for this species 15.93 mj∙kg-1) (podlaski et al., 2010). one of the many positive impacts on the environment is the low demand for phosphorus in the aboveground parts of plants (0.05–0.15% dm), which reduces the risk of water eutrophication in its environment (kays, nottingham, 2008). in addition, antonkiewicz and jasiewicz (2003) found that the plant exhibited heavy metal accumulation along with soil contamination level (respectively in decreasing order: cd, zn, ni, cu and pb), which could be used in phytoremediation of degraded areas. �is plant, thanks to its similarity to the sun�ower, acts as a reservoir of pollen and nectar for bene�cial insects in the wild, especially in the late summer when there is no other source of food for them. decorative, yellow �owers are used in bouquets and garden design. however, the cultivation of the jerusalem artichoke also exhibits negative aspects. as a result of plants density and susceptibility to the fungal disease caused by sclerotinia sclerotiorum (lib.) de bary, there is a risk of a high volatility of dry matter yields. as podlaski et al. (2010) pointed out, this variability can range from 2.7 to 9.7 t∙ha-1. it is a plant classi�ed according to tokarska-guzik et al. (2011, 2012) as an 99 the yield of jerusalem artichoke plant h elianthus tuberosus l. grow n in various com binations of fertilisation – prelim inary research alien species with the status of invasive taxa. rapidly spreading, and being stable in the environment by producing large amounts resistant to adverse environmental conditions with tubers that are deeply embedded in the soil. material and methods �e study was based on a �eld experiment in 2016 at experimental station of department of agrotechnology and agricultural ecology at the university of agriculture in kraków, 50°05ʹ08.5ʹʹn; 19°51ʹ08.3ʹʹe. �e experimental area was on sandy soil with a ph of 6.36, the contain of total c-org 1.14% of dm (biochar was derived from coniferous wood wastes, which contains: total c (77%), ash (5%) and volatile matter (18%); this fertiliser is attested by national institute of public health-national institute of hygiene, no pzh/ht-3146/2016), n 0.13%, p2o5 31.00 and k2o 7.67 and mg 8.10 mg 100g soil. total precipitation between april and november was 55.3 mm, according to the long-period total precipitation on a level of 68.2 mm (1961–1990). �e average daily temperature was 13.5°c compared to the long-period mean temperature of 12°c (1961–1990). �e experiment was established as a factorial design with two factors (varieties and fertilisation) in a randomised complete block design with three replications. �e cv. albik and cv. rubik were cultivated in 5 combinations of fertilisation in 5 replications: (1) control object – without mineral fertilisation and biochar, (2) mineral fertilisation npk: 100:80:100 kg·ha-1, (3) biochar 10 t·ha-1, (4) biochar 10 t·ha-1 and npk 100:80:100 kg·ha-1, and (5) biochar 5 t·ha-1 and npk 50:40:50 kg·ha-1. fertilisation was applied early in the spring prior to establishing the cultivation. tubers were planted at 1 m spacing and at 0.5 m spacing between plants in a row. no pesticides were used in the crop. tubers harvest was made at the end of november when the plant stems were already dried. for the measurement of morphological features, tubers of 2 randomly selected plants were taken from one row. �is is equivalent to 4 square meters per row. �e yield was obtained from 6 plants which were grown in the row. �e area of the plot was 18 square meters. data was subjected to the analysis of variance (anova). means (n = 5) were separated using hsd tukey range test at the 0.05 signi�cance level. �e results discussed in this paper are a part of the long-term �eld experiment. results and discussion �e applied various kinds of fertilisation in jerusalem artichokes cultivation di�erentiated tubers yield (tab. 1). signi�cant di�erences in tuber yield between varieties were found. higher yields were noted for the cv. rubik. signi�cant di�erences were also objo an na p uł a, a ng el ik a k lis zc z 100 served between the fertilisation objects, but only for the ‘albik’ (tab. 1). �e highest effect of increasing yield was obtained using mineral fertilisation (object 2) and biochar with a full dose of mineral fertilisers (object 4). �e di�erence between the smallest and the largest yield was over 9 t (cv. albik) and about 6 t (cv. rubik). in case of the ‘rubik’, there were no signi�cant di�erences in the yield of tubers between the kind of fertilisation. �e application of biochar (objects 3 and 4) also signi�cantly a�ected the yield of tubers. �e cv. albik also yielded worse under control conditions than the cv. rubik. �e average yield of tubers for the cv. albik was (regardless of fertilisation objects) 24.11 t·ha-1 and for cv. rubik 33.18 t·ha-1. according to prośba-białczyk (2007), the yield of tubers of these varieties of jerusalem artichoke in favourable weather conditions without fertilisation and chemical protection can be about 40 t·ha-1. in that paper, one can also notice that cv. albik yields better than cv. rubik. �is is in contrast to the results obtained in this study, because the control object (without fertilisation) was 19.35 t·ha-1 for ‘albik’ and 34.27 t·ha-1 for ‘rubik’ (tab. 1). according to applied fertilisation, cv. albik produced on average smaller and more diversi�ed tubers than cv. rubik (tab. 1). nevertheless, based on the statistical analysis, no signi�cant di�erences were found between fertilisation objects and varieties. �e results show that the most optimum combination of fertilisation, where the largest biomass of tubers ‘albik’ were recorded (objects 3 and 4), and for ‘rubik’ (objects 3 and 5). similar results according to the reduced doses of mineral fertilisation were obtained in the studies of kocsis et al. (2007). during the evaluation of the number of tubers from one plant, signi�cant di�erences between varieties in the size of this parameter and some combination of fertilistab. 1. average tubers yield and the biomass of a single tuber of helianthus tuberosus l. ‘albik’ and ‘rubik’ objects average tubers yield [t·ha-1] �e biomass of a single tuber [g] ‘albik’ ‘rubik’ ‘albik’ ‘rubik’ 1 19.35 a 17.45–20.30 34.27 b 30.35–36.35 37.31 a 32.00–52.00 49.14 a 33.66–58.46 2 28.45 b 24.35–34.45 32.07 b 23.35–40.15 43.68 a 34.67–56.00 53.22 a 20.40–65.60 3 22.32 a 19.05–25.70 34.13 b 33.95–34.25 44.31 a 13.16–62.11 54.74 a 30.24–83.48 4 27.85 b 19.20–33.60 35.68 b 27.85–40.45 44.61 a 17.46–65.93 51.49 a 33.71–72.50 5 22.57 a 15.95–26.95 29.75 b 25.45–32.40 43.94 a 19.46–56.36 55.48 a 34.50–75.50 s 24.11 a 33.18 b 42.77 a 52.81 b 1 – control object without mineral fertilisation and biochar, 2 – mineral fertilisation npk: 100-80-100 kg·ha-1, 3 – biochar 10 t·ha-1, 4 – biochar 10 t·ha-1 and npk, 5 – biochar (5 t·ha-1) and npk 50-40-50 kg·ha-1, s – mean values; a, b – di�erent letters relate to the signi�cant di�erences according to tukey test, n = 5, p = 0.05 101 ation were recorded (fig. 1). in the case of the ‘albik’ variety, the most favourable conditions for the development of tubers were the conditions where a full dose of mineral fertilisation (object 2) and full dose of npk with the addition of biochar (object 4) were applied. under these conditions, one plant produced, on average, from about 42 to more than 46 tubers. on the other objects these values ranged from 32 to about 36 tubers per plant. in turn, the ‘rubik’ variety, in this experience, was di�erent from the ‘albik’ variety. �e highest number of tubers (over 38) was from one plant, where only full biochar was used (object 3). conclusions based on the �eld experiment, it was found that the addition of biochar derived from coniferous wood biomass could be useful in the fertilisation of the jerusalem artichoke. �e yield of tubers and the morphological features of helianthus tuberosus l. cv. albik and cv. rubik were varied and dependent on the fertilisation variants applied. in the studied conditions of the habitat for yields of the jerusalem artichoke, the most e�ective were the fertilisation of the soil with a full dose of biochar and mineral fertilisation. acknowledgements �e research was �nanced by the ministry of science and higher education of the republic of poland. fig. 1. number of tubers per one plant helianthus tuberosus ‘albik’ and ‘rubik’: 1 – control object without mineral fertilisation and biochar, 2 – mineral fertilisation npk: 100-80-100 kg·ha-1, 3 – biochar 10 t·ha-1, 4 – biochar 10 t·ha-1 and npk, 5 – biochar (5 t·ha-1) and npk 50-40-50 kg·ha-1, 6 – mean values; a, b – di�erent letters relate to the signi�cant di�erences according to hsd tukey test, n = 5, p = 0.05 the yield of jerusalem artichoke plant h elianthus tuberosus l. grow n in various com binations of fertilisation – prelim inary research jo an na p uł a, a ng el ik a k lis zc z 102 references antonkiewicz, j., jasiewicz, cz. (2003). ocena przydatności topinamburu (helianthus tuberosus l.) do �toremediacji gleby zanieczyszczonej cd. pb. ni. cu i zn. archiwum ochrony środowiska, 29(4), 81–87. [in polish] cieślik, e. (1998). zawartość składników mineralnych w bulwach nowych odmian topinamburu (helianthus tuberosus l.). zeszyty naukowe ar im. h. kołłątaja w krakowie, 342, 23–30. [in polish] cieślik, e., filipiak-florkiewicz, a. (2000). topinambur (helianthus tuberosus l.) – możliwości wykorzystywania do produkcji żywności funkcjonalnej. żywność. nauka. technologia. jakość, 7(1), 73–81. [in polish] florkiewicz, a., cieślik, e., filipiak-florkiewicz, a. (2007). wpływ odmiany i terminu zbioru na skład chemiczny bulw topinamburu (helianthus tuberosus l.). żywność. nauka. technologia. jakość, 3(52), 71–81. [in polish] jasińska, z., kotecki, a. (2003). szczegółowa uprawa roślin. tom 1. wrocław: axa, 383–392. [in polish] kays, s.j., nottingham, s.f. (2008). biology and chemistry of jerusalem artichoke helianthus tuberosus l. crc press, 1–27. kocsis, l., kaul, h.-p., praznik, w., liebhard, p. (2007). ein�uss des erntetermins auf den krautund knollenertrag unterschiedlicher sorten von ‘topinambur’ (helianthus tuberosus l.) im semiariden produktionsgebiet österreichs. p�anzenbauwissenscha�en, 11(2), 67–76. [in german] makowski, d. (2014). topinambur – uniwersalna roślina. http://www.farmer.pl/produkcja-roslinna/ inne-uprawy/’topinambur’-8211-uniwersalna-roslina.51868.html [in polish] piskier, t. (2004). topinambur – alternatywne źródło energii. czysta energia, 12, 12–13. [in polish] podlaski, s., chołuj, d., wiśniewski, g. (2010). produkcja biomasy z roślin energetycznych. postępy nauk rolniczych, 2, 163–174. [in polish] prośba-białczyk, u. (2007). produkcyjność topinamburu (helianthus tuberosus l.) uprawianego bez nawożenia. fragmenta agronomica (xxiv), 4(96), 106–112. 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[in polish] abstract jerusalem artichoke is a perennial plant, which originates from north america. tubers are characterised by high nutritional and energy values and can therefore be a source of food for humans and animals. �e interest in tubers of helianthus tuberosus l. in the diet of man is primarily due to the content of inulin and fructooligosaccharides, minerals, vitamins, and amino acids in them. in addition, it is a plant that is not demanding in agrotechnical conditions. hence, the interest in cultivation this plant has increased. �e main objective of the experiment was to evaluate the e�ect of biochar on the average yield of tubers and some morphological characteristics of plants. �e study was conducted in conditions of a �eld experiment in 2016 at the experimental station of the university of agriculture in kraków. two varieties: ‘albik’ and ‘rubik’, were grown in the experiment with di�erent fertilisation variants. biochar from the coniferous wood industry and mineral fertilisers were used. �e ‘rubik’ variety yields better than the ‘albik’ variety under tested soil conditions, and the combined use of biochar and the basic dose of mineral fertilisation gives the best yields in the cultivation of jerusalem artichoke. key words: jerusalem artichoke, helianthus tuberosus l. cv. albik, cv. rubik, biochar, size and numbers of tubers received: [2017.09.01] accepted: [2017.11.13] 103 plon słonecznika bulwiastego helianthus tuberosus l. uprawianego w różnych kombinacjach nawozowych – badania wstępne streszczenie topinambur (słonecznik bulwiasty) helianthus tuberosus l. to wieloletnia roślina, która pochodzi z ameryki północnej. bulwy charakteryzują się wysokimi właściwościami odżywczymi, dlatego mogą być źródłem pożywienia dla człowieka i  paszą dla zwierząt. ponadto wykorzystywane są jako biomasa do produkcji energii i  etanolu. zastosowanie bulw w  diecie człowieka wynika przede wszystkim z  zawartości w  nich inuliny i fruktooligosacharydów, minerałów, witamin oraz aminokwasów. jest to roślina o małych wymaganiach agrotechnicznych, stąd też w  ostatnich latach wzrasta zainteresowanie jej uprawą, zwłaszcza na glebach słabych. głównym celem eksperymentu była ocena wpływu biowęgla na plon bulw i niektóre cechy morfologiczne topinamburu. doświadczenie polowe przeprowadzono w 2016 roku w stacji doświadczalnej katedry agrotechniki i ekologii rolniczej uniwersytetu rolniczego w krakowie. uprawiano dwie odmiany topinambura: ‘albik’ i ‘rubik’, przy zróżnicowanym nawożeniu: 1 – obiekt kontrolny bez nawożenia mineralnego i biowęgla, 2 – npk: 100-80-100 kg∙ha-1, 3 – biowęgiel 10 t∙ha-1, 4 – biowęgiel 10 t∙ha-1 i npk, 5 – biowęgiel (5 t∙ha-1) i npk 50-40-50 kg∙ha-1. na tle porównywanych obiektów nawozowych stwierdzono, że najlepszą kombinacją nawożenia w uprawie słonecznika bulwiastego było zastosowanie biowęgla wraz z nawozami mineralnymi. słowa kluczowe: słonecznik bulwiasty, helianthus tuberosus l. cv. albik i cv. rubik, biowęgiel, wielkość i liczba bulw information on the authors joanna puła her research is connected with agrotechnology in plant cultivation and plant ecology. presently, she is interest in the use of the biomass of plants and other organic fertiliser like biochar in agriculture. angelika kliszcz she is focusing on enhancing the understanding of the in�uence of di�erent factors on soil structure and fertility. particulary, she is investigating the interaction of plants with the physical, chemical, and biological properties of the soil. she is also interested in organic methods of plant production and soilenriching substances. the yield of jerusalem artichoke plant h elianthus tuberosus l. grow n in various com binations of fertilisation – prelim inary research 95 annales universitatis paedagogicae cracoviensis studia naturae, 1: nr 95–104, 2016, issn 2543-8832 roman lysiuk1*, yeva zaritska2, roman darmohray1 1 danylo halytsky lviv national medical university, lviv, ukraine, *pharmacognosy.org.ua@ukr.net 2 �e state scienti�c-research control institute of veterinary medicinal products and feed additives, lviv, ukraine investigation of microelements contents in aerial parts of agrimonia eupatoria l., collected in lviv region (ukraine) introduction medicinal plants have a long history of use in therapy throughout the world and still play an important role in traditional medicine. �e use of herbal products as the �rst choice in self-treatment of minor conditions continues to expand rapidly across the world. �is makes the safety of herbal products an important public health issue. �us, medicinal plants and herbal products must be safe for the patient (consumer) (kosalec et al., 2009). man-made environmental pollution can largely a�ect heavy metal contamination levels of herbal materials. it includes emissions from factories, leaded petrol, agrochemicals such as cadmium-containing fertilizers, organic mercury and arsenic-based pesticides that are still in use in some countries (who guidelines for…, 2007). herbal products can be contaminated at any stage of production, from growing conditions to open-air drying, preserving, and manufacturing (e.g. release from lead-containing utensils). harmful contaminants may also originate from the conditions in which the medicinal plants are cultivated, post-harvest treatment of herbal material (e.g. fumigants), and �nished product manufacturing stages (e.g. organic solvent residues) (chan, 2003; kosalec et al., 2009). �e genus agrimonia l. of the family rosaceae juss. is known to include valuable medicinal plants. in particular, agrimonia eupatoria l. is indigenous to middle and northern europe, temperate regions of asia and north america (gruenwald et al., 2000). �e material of commerce is usually imported from bulgaria, hungary and croatia (wichtl, 2004). polyphenols are major constituents of a. eupatoria and exempli�ed by phenolic acids, �avonoids, ellagitannins, and procyanidins. among �avonoids, two groups of compounds were characterised: the �rst group consists of �avanols described as 96 r om an l ys iu k, y ev a za rit sk a, r om an d ar m oh ra y quercetin and kaempferol derivatives, and the second one are �avones containing luteolin or apigenin as aglycone moiety. agrimoniin is reported as a major ellagitannin occurring in common agrimony in high quantities (granica et al., 2015). �e major compounds in its aqueous-alcoholic extract are �avonoids (0.33%), great amounts of tannins (10.08%), as well as phenolic acids (2.26%), including luteolin 7-o-glucoside and apigenin 7-o-glucoside, luteolin 7-o-sophoroside, luteolin 7-o (6”acetylglucoside), acacetin 7-o-glucoside; protocatechuic, vanillic, and p-hydroxybenzoic acids (shabana et al., 2003); apigenin, luteolin, kaempferol (wichtl, 2004; barnes et al., 2007), quercetin, quercitrin, and glycosides (barnes et al., 2007). �e monograph for agrimony herb is included into the british herbal pharmacopoeia, british and european pharmacopoeias, complete german commission e, martindale (35th edition) (barnes et al., 2007). a standardised medicinal plant material (european pharmacopoeia…, 2013), consisting of dried �owering tops of a. eupatoria l., contains minimum 2.0% of tannins, expressed as pyrogallol (dried drug); besides of macroscopic and microscopic authentication, for its quality control thin layer chromatography is used (identi�cation c), applying isoquercitroside and rutin as reference solutions, with further uv detection at 365 nm, spraying with solution of diphenylboric acid aminoethyl ester in methanol. kurkina et al. (2013) recommend to use cynaroside (luteolin-7-o-glucoside) as a standard sample for purposes of the standardisation of a. eupatoria herb. agrimony is well known for its bene�cial e�ects in various diseases such as liver complaints, gall-bladder stones; diarrhea, edemas and kidney problems. approved by commission e common applications comprise diarrhea, in�ammation of the skin, in�ammation of the mouth and pharynx. �is species is used internally for mild, nonspeci�c, acute diarrhea, in�ammation of kidney and bladder, diabetes and childhood bedwetting, cholestasis, in�ammation of oral and pharyngeal mucosa; externally for poorly healing wounds, chronic pharyngitis, psoriasis, seborrhoeic eczema, as well as in hip-baths for lower abdominal conditions. it is applied in chinese medicine as a hemostyptic. it is also used for certain forms of cancer and as an anthelmintic (gruenwald et al., 2000). agrimony is stated to possess mild astringent and diuretic properties. it has been used for mucous colitis, grumbling appendicitis, urinary incontinence, cystitis and as a gargle for acute sore throat and chronic nasopharyngeal catarrh (barnes et al., 2007). in experimental model of type 1 diabetes antihyperglycaemic, insulin-releasing and insulin-like activities of agrimony’s aqueous extract, given orally to mice, have been demonstrated (swanston-flatt et al., 1990; gray, flatt, 1998). �e herb is shown to be a perspective therapeutic, especially for treatment of social signi�cant diseases such as diabetes and obesity, accompanied by low-grade in�ammation. accumulated evidence suggests that antioxidant activity of a. eupatoria might be due to chemical 97 structure of polyphenols and/or herb ability to activate the endogenous antioxidant defence systems (ivanova et al., 2011). due to antioxidant properties of �avonoids the herb has shown to exert neuroprotective e�ects (lee et al., 2010). �e ethanolic-aqueous extracts of herba agrimoniae showed the immunostimulant activity in inbred mice resulting in a  rise of phagocytic activity and phagocytic index of peritoneal macrophages, in a  bactericide action of disintegrated peritoneal cells on escherichia coli cells and in an increased lysozyme activity of supernatants of disintegrated peritoneal cells. �e extract of herba agrimoniae stimulated peritoneal cells to the increased peroxidase activity (bukovsky, blanarik, 1994). signi�cant uricolytic activity has been documented for agrimony infusions and decoctions (15% w/v), following their oral administration to male rats at a  dose of 20 ml/kg body weight (equivalent to 3 g as a  dry drug). diuretic activity was stated to be minimal and elimination of urea unchanged (giachetti et al., 1986; barnes et al., 2007). finally, marked antibacterial activity against staphylococcus aureus and α-haemolytic streptococci has been reported for agrimony (petkov, 1986). �erefore, the aforementioned data concerning valuable pharmacological e�ects of a. eupatoria encourage all-side investigation of the medicinal plant since its application in the phytopharmaceutical practice is still underestimated. with that purpose in mind, we have summarized current scienti�c data concerning its biological activity, paying a special attention to those, which might be useful for renal impact, and also carried out analysis of its trace elements. materials and methods trace elements in the investigated plant material were quantitatively determined by atomic absorption spectrometry with electrothermal atomization (aas/ea) with a zeeman background correction a�er total microwave – assisted digestion (mineralization) of samples by means of their conversion into soluble forms and further determination for concentration in the solutions (a method of standard additions) in sealed analytical autoclaves. reagents all the reagents were of analytical grade; highly purified water was obtained in accordance with the requirements of the european pharmacopoeia (2013). as a matrix modifier for electrothermal atomisation palladium was used (10 g/dm3). the blank solutions of manganese, copper, selenium, lead and cadmium with the certified value of the mass ion concentration of 1.0 mg/cm3 were applied. reference solutions of the specified elements were prepared by a  step dilution of the blank with 1% nitric acid solution from 1 mg/cm3 concentration to 10 µg/cm3 (solution investigation of m icroelem ents contents in aerial parts of agrim onia eupatoria l., collected in lviv region (u kraine) 98 r om an l ys iu k, y ev a za rit sk a, r om an d ar m oh ra y a), followed the obtained solution a by treatment with the same reagent to a concentration of 0.1 µg/cm3. to construct a calibration graph reference solutions of di�erent concentrations of the trace elements were used (within the operating range for determination of the spectrophotometer), obtained from the blank solutions (0.1 g/cm3) by the standard addition method (european pharmacopoeia…, 2013). preparation of the samples �e procedure for a sample preparation was based on a total mineralization technique with a  mixture of nitric acid and highly puri�ed water in the reaction chamber of te�on analytical autoclave with microwave decomposition of the samples under conditions of high pressure. accurately weighted samples (0.6 g) of the subjected plant materials, placed in a  te�on vessel, were introduced directly into the reaction chamber of an autoclave, adding 3 ml of highly puri�ed water, followed by addition of 6 ml of 65% nitric acid. �e closed autoclave was heated in a microwave digestion apparatus “milestone start d”. programme of temperature changes in autoclaves during the period of mineralization is presented in table 1. tab. 1. parameters of temperature changes over time of mineralization in autoclaves stage time [min. sec.] temperature [°c] radiation power [w] 1 5.00 80 ≤350 2 3.30 160 ≤800 3 4.30 195 ≤1000 4 14.00 195 ≤800 �e resulting test solution was cooled a�er mineralization and quantitatively transferred from the reaction chamber of the autoclave into a 25 cm3 volumetric �ask, diluting its contents to volume with highly puri�ed water. �e solutions of the samples were analyzed on atomic absorption spectrometer (varian aa 240 zeeman atomic absorption spectrometer), electrothermal atomiser equipped with a graphite cuvette (gta 120 zeeman graphite tube atomizer) and autosampler (psd 120 programable sample dispenser). �e physical and technological parameters, followed in determining the quantitative content of the microelements were presented (tab. 2). �e plant material (herba agrimoniae) consisting of dried aboveground parts of a. eupatoria was collected at �owering period (june, 2016) in the suburbs of lviv, western part of ukraine and identi�ed by the authors. 99 tab. 2. parameters of atomic absorption spectrometry for analysis of trace elements in plant materials trace element wavelength of absorption (resonance) lines [nm] voltage of resonance radiation lamp [ma] flow rate of argon [l/min] ashing temperature [°c] atomization temperature [°c] slit width, [nm] manganese 279.50 10 0.30 700 2400 0.20 copper 327.40 10 0.30 800 2300 0.50 selenium 196.00 10 0.30 1000 2600 1.00 lead 283.30 5 0.30 400 2100 0.50 cadmium 228.80 5 0.30 250 1800 0.50 a composite sample weighing about 10 g consisted of about �ve subsamples taken within an area of about 10 m2. all samples were placed in polyethylene bags and transported to the laboratory on the day of sampling (gałuszka et al., 2015). results and discussion chronic kidney disease is associated with low concentration of serum selenium and lower platelet glutathione peroxidase (gpx) activity (kuo, tarng, 2010). ceballos -picot et al. (1996) demonstrated lower serum levels of glutathione and plasma gpx activity in renal failure patients. plasma selenium is reduced in dialysis patients; there is also signi�cant de�ciency and downregulation of glutathione peroxidase, copper-zinc superoxide dismutase, and manganese superoxide dismutase in renal dysfunction (kuo, tarng, 2010). in laboratory animals, parenteral administration of organic and inorganic selenium (210 to 12 000 μg/kg) has been shown to protect against cisplatin-induced nephrotoxicity. �e glutathione (gsh) peroxidases are the best-characterized selenoproteins, although other circulating selenoproteins also have antioxidant functions (ebadi, 2007). antal et al. (2010) consider agrimonia eupatoria herb as the plant material able to accumulate higher se amounts, when this element is present in adequate amounts in the soil. cynaroside (luteolin-7-o-β-d-glucoside), another signi�cant substance of agrimony herb, was comparable to the well-known medicinal preparation lespenephryl with respect to hypoazotemic action. a technology has been developed for producing a medicinal form of cynaroside in 0.05 g tablets (mamatkhanova et al., 2009). many contaminants occur naturally in the ground and the atmosphere, such as radionuclides and metals. some arise from past or present use of agents that pollute the environment and subsequently medicinal plants, such as factory emissions or persistent chemical residues. due to their excessive use and disposal, contaminants from environmental sources may even be present if a herb is organically grown. metals are investigation of m icroelem ents contents in aerial parts of agrim onia eupatoria l., collected in lviv region (u kraine) 100 r om an l ys iu k, y ev a za rit sk a, r om an d ar m oh ra y widely distributed throughout nature and occur freely in soil and water. as they are likely to be present in many food sources, it is important to reduce the total population exposure to toxic elements by minimizing contamination of herbal products (who guidelines for…, 2007; kosalec et al., 2009). agrimony is listed by the council of europe as a natural source of food �avouring (category n2). �is category indicates that agrimony can be added to foodstu�s in small quantities, with a possible limitation of an active principle (as yet unspeci�ed) in the �nal product (barnes et al., 2007). scienti�c data concerning contents of trace elements of agrimony herb are rather limited and are related to several publications (štro�eková et al., 2006, 2008; antal et al., 2010; gałuszka et al., 2015), where the samples of herba agrimoniae from slovakia, romania and poland were analysed. �erefore, it seems important to investigate contents of trace elements in the herbal drug, collected in wild in the western ukraine, for comparison of the found outcomes with the known ones related to the subjected medicinal plant material. �e content of the elements in above-ground portions of a. eupatoria, collected in two di�erent slovak regions, determined by the method of radionuclide x-ray �uorescence (source 238pu) was the following: mn – 36.81 µg/g, cu – 7.70 µg/g, pb – 2.26 µg/g (nové mesto nad váhom) and mn – 25.07 µg/g, cu – 7.53 µg/g, pb – 2.51 µg/g (ružomberok) (štro�eková et al., 2006). metals contents for agrimony herb samples, grown in non-chemical treated meadows in nitra (slovakia), performed using x-ray �uorescence analysis (xrf) (source 238pu for lead, and 241am for cadmium) and galvanostatic stripping chronopotentiometric analysis (scp) were determined as follows: pb – ≤ 1.95 µg/g, cd – ≤ 1.96 µg/g (xrf) and pb – 1.043 µg/g, cd – 0.279 µg/g, se – 0.175 µg/g (scp). xrf is an o�cial method in european pharmacopoeia (2013), it is described as x-ray spectrometry (štro�eková et al., 2008). for the plant samples, collected from limestone soils in two localities of the aninei mountains (banat region, romania), believed to be unpolluted (far away from roads, kilometers outside villages or towns), outstanding se contents (332 μg/kg) was measured for a. eupatoria herb from lisvar hill; the samples from ciopliaia hill yielded 61 μg/kg of selenium (antal et al., 2010). a semiquantitative multi-element analysis by inductively coupled plasma mass spectrometry (icp-ms) of aboveground parts of agrimony, collected in a residential area of the city of kielce, (góry świętokrzyskie), south-central poland, allowed determining the following contents of the trace elements in the subjected plant material: mn – 44 mg/kg, cu – 8.0 mg/kg, pb – 0.001 mg/kg, cd – 0.001 mg/kg (gałuszka et al., 2015). �e found quantities of trace elements in the analyzed by us samples of herba agrimoniae, collected from wild in the suburbs of lviv, are as follows: mn – 46.9 µg/g, cu – 7.9 µg/g, se – 0.16 µg/g, pb – 0.15 µg/g, cd – 0.05 µg/g. 101 comparing the received outcomes with the other known investigations, it might be concluded that the contents range of the trace elements in herba agrimoniae, collected from various locations in europe, comprise for mn – 25.07–46.9 µg/g; cu – 7.53–8.0 µg/g; se – 0.061–0.332 µg/g; pb – 0.15–2.51 µg/g; cd – 0.05–0.279 µg/g. �e who recommends the following limits for heavy metals in herbal drugs: 10 mg/kg for lead and 0.3 mg/kg for cadmium (quality control methods…, 1998; who guidelines for…, 2007). �erefore, the analysed by us herbal drug complies with the requirements of the who concerning the contents of lead and cadmium. conclusions and recommendations with an objective to determine quality characters, as well as contents of 5 trace elements (manganese, copper, selenium, lead, cadmium) in the aboveground parts of agrimony, o�cial herbal drug, grown wild in the western part of ukraine, there has been carried out an atomic absorption spectroscopy with electrothermal atomization a�er mineralization. �e received research outcomes were compared with results of other known scienti�c investigations of the subjected medicinal plant material. �e determined yield of microelements allows recommending the herbal drug for further biological investigations, as well as a promising nephroprotective medicine. references antal, d.s., canciu, c.m., dehelean, c.a., anke, m. (2010). how much selenium do medicinal plants contain? results of a research on wild-growing species from western romania. analele universităţii din oradea – fascicula biologie, 17(1), 23–28. barnes, j., anderson, l.a., phillipson, j.d. (2007). herbal medicines. 3rd ed. london: pharmaceutical press. bukovsky, m., blanarik, p. (1994). immunomodulative e�ects of ethanolic aqueous extracts of herba agrimoniae, flos chamomillae and flos calendulae cum calyce. farmaceutiky obzor, 63, 149–156. ceballos-picot, i., witko-sarsat, v., merad-boudia, m., nguyen, a.t., �évenin, m., jaudon, m.c., zingra�, j., verger, c., jungers, p., descamps-latscha, b. (1996). glutathione antioxidant system as a  marker of oxidative stress in chronic renal failure. free radical biology and medicine, 21(6), 845–853. doi: 10.1016/0891-5849(96)00233-x chan, k. (2003). some aspects of toxic contaminants in herbal medicines. chemosphere, 52, 1361–1371. doi: 10.1016/s0045-6535(03)00471-5 ebadi, m.s. 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(2006). determination of heavy metals in samples of di�erent states by radionuclide x-ray �uorescence. acta facultatis pharmaceuticae universitatis comenianae, 53, 268–274. štro�eková, o., planková, a., jánošová, v., sýkorová, m., havránek, e. (2008). determination of fe, zn, pb, cd and se content in medical plants by x-ray fluorescence analysis and galvanostatic stripping chronopotentiometric analysis. acta facultatis pharmaceuticae universitatis comeniae, 55, 219–229. who guidelines for assessing quality of herbal medicines with reference to contaminants and residues. (2007). geneva: who press. wichtl, m. (2004). herbal drugs and phytopharmaceuticals: a handbook for practice on a scienti�c basis. 3rd ed. stuttgart: medpharm gmbh scienti�c publishers. 103 abstract �e genus agrimonia l. of the family rosaceae juss. is known to include valuable medicinal plants. agrimony is well known for its bene�cial e�ects in various diseases such as liver complaints, ga\ll-bladder stones; diarrhea, edemas and kidney ailments. �e contents of microelements in aerial parts of agrimonia eupatoria, an o�cial herbal drug, collected from wild samples in the suburbs of lviv city, western ukraine, have been determined by atomic absorption spectroscopy a�er mineralisation in the microwave milestone start d. �e found quantities of trace elements in the analysed samples were as follows: manganese – 46.9 µg/g, copper – 7.9 µg/g, selenium – 0.16 µg/g, lead – 0.15 µg/g, cadmium – 0.05 µg/g. �e received research outcomes were compared with results of other known scienti�c investigations of the medicinal plant material. �e determined amounts of lead and cadmium in the investigated samples of the herbal drug complied with the world health organization (who) requirements. �e special attention is paid to pharmacological e�ects, related to its renal impact, contents of selenium and phenolic compounds that allow considering the herbal drug as a promising nephroprotective agent. key words: аас/еа (atomic absorption spectroscopy with electrothermal atomization), agrimonia eupatoria, microelements, renoprotective, selenium, trace elements received: [2016.06.13] accepted: [2016.09.27] badanie zawartości mikroelementów w nadziemnych częściach agrimonia eupatoria l., zebranych w obwodzie lwowskim (ukraina) streszczenie rzepik pospolity agrimonia eupatoria l. z rodziny rosaceae juss. należy do cennych roślin leczniczych. jest dobrze znany ze swoich korzystnych działań w różnych chorobach, takich jak: dolegliwości wątroby, kamienie pęcherzyka żółciowego, biegunka, obrzęki i dolegliwości nerek. próbki do badań z tego o�cjalnego leku ziołowego pozyskano ze stanu naturalnego w okolicach lwowa (zachodnia ukraina). zawartość mikroelementów w częściach nadziemnych a. eupatoria zbadano metodą atomowej spektroskopii absorpcyjnej, po mineralizacji mikrofalowej w milestone start d. w analizowanych próbkach znaleziono śladowe ilości pierwiastków, takich jak: mangan – 46,9 ug/g, miedź – 7,9 ug/g, selen – 0,16 ug/g, ołów – 0,15 ug/g, kadm – 0,05 ug/g. uzyskane wyniki badań porównano z innymi znanymi z literatury badaniami na materiale roślinnym z tego gatunku. stwierdzone w badanych próbkach ilości ołowiu i kadmu są zgodne z wymogami światowej organizacji zdrowia (who). szczególną uwagę zwrócono na efekty farmakologiczne tego substratu, związane z jego wpływem na nerki, zawartością selenu i związków fenolowych, które pozwalają uznać ten lek za obiecujący środek chroniący nerki. słowa kluczowe: аас/еа (spektroskopia absorpcji atomowej z atomizacją elektrotermiczną), agrimonia eupatoria, mikroelementy, nefroprotekcja, pierwiastki śladowe, selen information on the authors roman lysiuk he carries out a  focused analysis of plant species for selenium contents and the marker substances of �avonoid structure, that cause nephroprotective (hypoazotemic) activity, and investigates renal impact of their extracts. research interests also comprise principles and improving of teaching of pharmacognosy for english-medium students. author of over 55 scienti�c publications, including 3 manuals, 1 electronic textbook, 5 educational and methodical recommendations. investigation of m icroelem ents contents in aerial parts of agrim onia eupatoria l., collected in lviv region (u kraine) 104 r om an l ys iu k, y ev a za rit sk a, r om an d ar m oh ra y yeva zaritska scienti�c researcher at laboratory of instrumental methods of control of the state scienti�c-research control institute of veterinary medicinal products and feed additives, lviv, ukraine. her scienti�c publications are dedicated to atomic absorption spectrometry with electrothermal atomization of veterinary medicinal products, food supplements and plant materials. roman darmohray he carries out the pharmacognostic investigations of the asteraceae, fabaceae, apiaceae, polygonaceae as promising sources of bioactive substances, their chemotaxonomic studies. he is an author of above 115 scienti�c, educational and methodical works. 22 annales universitatis paedagogicae cracoviensis studia naturae, 3: 22–37, 2018, issn 2543-8832 doi: 10.24917/25438832.3.2 terézia beck1*, ján gáper2,3, martin šebesta2, svetlana gáperová1 1faculty of natural sciences, matej bel university, tajovského 40, 974 01 banská bystrica, slovak republic, *terezia.gasparcova@umb.sk 2faculty of ecology and environmental sciences, technical university in zvolen, t. g. masaryka 24, 960 63 zvolen, slovak republic 3faculty of sciences, university of ostrava, chittussiho 10, 710 00 ostrava, czechia host preferences of wood-decaying fungi of the genus ganoderma in the urban areas of slovakia introduction ganoderma p. karst. 1881 (basidiomycota, polyporales) is a cosmopolitan genus (richter et al., 2015) with the greatest diversity in the tropical regions (ryvarden, melo, 2014). fungi of this genus cause white rot of living or dead deciduous and coniferous trees of the wide range of species worldwide (schwarze, ferner, 2003; bernicchia, 2005; ryvarden, melo, 2014). the species of ganoderma, especially ganoderma lucidum complex, are also known due to their various medicinal properties (zhou et al., 2015). the basidiocarps of these polypores grow annually or can be perennial, sessile, or stipitate, with matte or a laccate pileus surface, cream to dark purplish-brown-coloured context and creamed-coloured pore surface, brownish after touching (ryvarden, melo, 2014). the genus ganoderma is characterised by ovoid, echinulate basidiospores with a truncate apex, two layered walls and interwall pillars between endosporium and exosporium (ryvarden, 1991; moncalvo, 2000; ryvarden, melo, 2014). the ganoderma species that occur in europe are g. adspersum (schulzer) donk, g. applanatum (pers.) pat., g. carnosum pat., g. lucidum (curtis) p. karst., g. pfeifferi bres., g. resinaceum boud., and g. valesiacum boud. (sokół, 2000; bernicchia, 2005; papp, szabó 2013; ryvarden, melo, 2014). the presence of all these species was also recorded in slovakia (gáper, 1998; gáperová, 2001; gašparcová et al., 2017a). the interspecific genetic diversity of ganoderma species occurring in slovakia was also studied by pcr-rflp analysis of its nrdna and with newly applied method maldi-tof ms (pristaš, gáperová, 2001; gašparcová et al., 2017b; pristaš et al., 2017). host preferences of european ganoderma species were analysed by several authors. g. adspersum has synanthropic character of distribution (kotlaba, 1984; 23 h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia gáper, 1998; sokół, 2000; gáperová, 2001; kotlaba, pouzar, 2009a; papp, szabó, 2013). in southern and warmer regions, it also occurs in forests (kotlaba, pouzar, 2009a). in europe, it was recorded on 43 genera of trees (breitenbach, kränzlin, 1986; ryvarden, gilbertson, 1993; bernicchia, 2005; ryvarden, melo, 2014). g. applanatum is the most common european species known as the “artists fungus” (sokół, 2000; ryvarden, melo, 2014). it grows mostly in forests (kotlaba, 1984; sokół, 2000; papp, szabó, 2013) and rarely in urban areas (gáperová, 2001). in europe, it was recorded on 35 genera of trees (breitenbach, kränzlin, 1986; bernicchia, 2005; ryvarden, melo, 2014). g. resinaceum often grows in urban areas such as parks and gardens, near roads, etc. (kotlaba, 1984; gáper, 1998; sokół, 2000; gáperová, 2001; kotlaba, pouzar, 2009b), but in warmer regions it also occurs in forest environment (kotlaba, pouzar, 2009a). it prefers hosts of the genus quercus, but it is known from many other deciduous trees (kotlaba, 1984; breitenbach, kränzlin, 1986; bernicchia, 2005; ryvarden, melo, 2014). g. pfeifferi is typical synanthropic species that prefers old beeches in parks, but in southern europe, in denmark (sokół, 2000) and hungary (papp, szabó, 2013) follows occurence of old beech forests. however, it was also noted on acer, aesculus, fraxinus, prunus, quercus and ulmus (bernicchia, 2005; ryvarden, melo, 2014). g. lucidum is the type species of the genus (richter et al., 2015) and it belongs to the g. lucidum complex (zhou et al., 2015). it often colonises old deciduous trees in parks and botanical gardens, but it occurs mainly in forests (sokół, 2000; bernicchia, 2005). basidiocarps of g. lucidum grow mostly on oaks, but also on other trees (kotlaba, 1984; tortić, 1985; sokół, 2000; karadelev et al., 2008; papp, szabó, 2013; ryvarden, melo, 2014). in europe, it grows on 24 woody plants genera (bernicchia, 2005; ryvarden, melo, 2014). g. carnosum belongs together with g. lucidum to the ‘g. lucidum complex’ (ryvarden, melo, 2014). similar to g. lucidum, it occurs mainly in forest environment, but it can also appear in parks or botanical gardens (sokół, 2000; gáperová, 2001). it grows usually on conifers (ryvarden, melo, 2014) mainly abies alba mill. (kotlaba, 1984; bernicchia, 2005). in europe, it colonises trees of 10 genera (breitenbach, kränzlin, 1986; bernicchia, 2005; ryvarden, melo, 2014). g. valesiacum is central european species that occurs at higher altitudes in natural coniferous forests (plank, wolkinger, 1981; kotlaba, 1984; sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014). it is very rare european species and there are no data on the synanthropic character of this species (sokół, 2000). it was recorded on larix (kotlaba, 1984; sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014) and probably on picea abies (l.) h.karst. (kotlaba, 1984; sokół, 2000). te ré zi a b ec k, j án g áp er , m ar tin š eb es ta , s ve tla na g áp er ov á 24 there are only few studies focusing on distribution of fungi of this genus in slovakia and on their host preferences. most of them confirm the presence of this genus in urban areas (kotlaba, 1984; gáper, 1998; gáperová, 2001; boleková, sliacka, 2015). a more comprehensive analysis of ganoderma’s host range in forests in slovakia was presented by gašparcová et al. (2017a). the aim of the present study was to identify the host preferences of the species of this genus occurring in the urban areas of slovakia, and the analysis of the location of basidiocarps on the colonised trees. material and methods data on the occurrence, host preferences, and topology of ganoderma basidiocarps on the colonised trees/wood were obtained by field research and from the information given for herbarium specimens located in slovak national museum – natural history museum in bratislava and national museum – natural history museum in prague (czechia). the field research of authors was carried out in the period of 1989–2017 in the urban areas of slovakia. the basidiocarps obtained during field research were deposited in the herbarium of the department of biology and ecology of the faculty of natural sciences, matej bel university, banská bystrica, slovakia. all basidiocarps used in the present study were collected in the districts bánovce nad bebravou, banská bystrica, banská štiavnica, bardejov, bratislava, čadca, detva, dunajská streda, galanta, hlohovec, kežmarok, komárno, košice, krupina, levice, levoča, liptovský mikuláš, lučenec, malacky, martin, michalovce, myjava, nitra, nové mesto nad váhom, nové zámky, pezinok, piešťany, poltár, poprad, považská bystrica, prešov, prievidza, rožňava, ružomberok, senica, spišská nová ves, stará ľubovňa, trebišov, trenčín, trnava, veľký krtíš, vranov nad topľou, zlaté moravce, zvolen, žiar nad hronom and žilina. all obtained data were recorded in a database (gáperová, krátka, 2002) that is currently based on a total of 34 input data divided into 5 categories: i – characteristics of tree, ii – characteristics of localities, iii – characteristics of fungus, iv – general data (in relation to a specific finding), v – vitality of tree. the database is connected with digital maps in the gis environment (gáperová, krátka, 2002; 2003; krátka et al., 2004). basidiocarps were determined using scientific mycological literature (breitenbach, kränzlin, 1984; ryvarden, 1991; sokół, 2000; bernicchia, 2005; antonín, 2006; holec et al., 2012; ryvarden, melo, 2014). microscopic studies were performed with the use of a motic microscope (motic company, germany), on preparations mounted in 5% koh. the topology of basidiocarps was evaluated for each recorded species separately. places of the occurrence of basidiocarps on trees were divided into the following categories: stump, root swelling, dead trunk base, dead standing/laying trunk, living trunk 25 base, living trunk, others. the category ‘others’ includes some parts of host woody plants that do not belong to the previous categories (e.g., tree branch, dead wood) or in the case of herbarium specimens for which places of basidiocarps occurrence were not recorded. for nomenclature of fungi, the index fungorum (cooper, kirk, 2018) database was followed. for nomenclature of woody plants, the checklist by marhold and hindák (1998) and the international plant names index database (ipni, 2012) were followed. results and discussion a total of 263 collections of basidiocarps were analysed to detect host preferences of fungi of the genus ganoderma in urban areas of slovakia (appendix 1 – tab. 1). we confirmed the presence in urban areas of slovakia all the species recorded in the country, namely ganoderma applanatum, g. adspersum, g. carnosum, g. lucidum, g. pfeifferi, g. resinaceum and g. valesiacum. we found out that ganoderma adspersum is the most common species of the genus ganoderma occurring in urban areas of slovakia (fig. 1). it was recorded on 24 species of host woody plants (appendix 1 – tab. 1). a wide range of hosts is associated with the synanthropic character of this species in our country. the basidiocarps most often occurred on aesculus hippocastanum l. (18% of all findings) and tilia cordata mill. (17% of all findings). similar to our results, gáperová (2001) recorded g. adspersum mainly on the introduced tree a. hippocastanum and the native t. cordata in urban environments of slovakia. moreover, fungi also grew on other planted non-native trees, mainly gleditsia triacanthos l. (8%), prunus serrulata lindl. (3%) and robinia pseufig. 1. representation of species of the genus ganoderma in urban areas of slovakia 39% 24% 4% 14% 2% 17% 0% ganoderma adspersum g. applanatum g. carnosum g. lucidum g. pfei�eri g. resinaceum g. valesiacum h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia te ré zi a b ec k, j án g áp er , m ar tin š eb es ta , s ve tla na g áp er ov á 26 doacacia l. (3%). our findings have not proved oaks as the most common substrate/ host that was recorded by some authors (kotlaba, 1984; tortić, 1985; gáper, 1998; papp, szabó, 2013). however, in the forest environments of slovakia, g. adspersum was found to grow mainly on quercus robur l. (gašparcová et al., 2017a). according to kotlaba and pouzar (2009a), g. adspersum is a parasitic species often found on planted trees in urban areas, because inner parts of the forest complexes of northern and northwest european countries are too cold. however, in the southern warmer climatic regions of europe, including moravia (czechia) and slovakia, g. adspersum also colonises native trees in forests (kotlaba, pouzar, 2009a). our research confirms its synanthropic character and frequent occurrence on non-native trees (39% of all findings) in urban areas, which was also realised by other authors (kotlaba, 1984; gáper, 1998; sokół, 2000; gáperová, 2001; kotlaba, pouzar, 2009a; papp, szabó, 2013). the basidiocarps of g. adspersum were found growing mainly on the base and along the living trunk, and also saprotrophically on stumps (fig. 2). it corresponds well to the published data (breitenbach, kränzlin, 1986; sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014). in the forests of slovakia, it was recorded mainly on the trunk base of host trees (gašparcová et al., 2017a). g. applanatum is often misidentified as g. adspersum (fig. 3), but g. applanatum has smaller basidiospores, paler context, tubes layers separated by thin layers of the context and often attacked by larvae of agathomya wankowici schnabl. (sokół, 2000; ryvarden, melo, 2014). in urban areas of slovakia, g. applanatum was recorded on 18 species of host trees, mostly deciduous (appendix 1 – tab. 1). over 45% of records come from decomposing wood of different hosts. in our study, we found 9% of all records on european beech. other studies confirm its frequent occurrence on fagus spp. (kotlaba, 1984; tortić, 1985; gáper, 1998; sokół, 2000; karadelev et al., 2008; papp, szabó, 2013). in the forests of slovakia, g. applanatum was also found growing predominantly on fagus sylvatica l. (38% off all findings). however, compared to urban areas, it was recorded only on 11 species of host trees in forests (gašparcová et al., 2017). although more records of this species are known from the forest environment (gašparcová et al., 2017), g. applanatum is the second most widespread species in urban areas of slovakia (fig. 1). we cannot confirm that it grows rarely in urban environments, as reported by gáperová (2001). however, according to sokół (2000), g. applanatum occurs in towns more often than typical synanthropic species. in urban areas of slovakia, g. applanatum was recorded mainly on stumps and the living trunks of host trees (fig. 2). some authors also reported that g. applanatum decomposes mainly stumps and dead trunks of deciduous trees, rarely of conifers. it can also colonise living trees, mostly the bases of the trunks (sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014). in the forests of slovakia, it was found to grow mainly on trunks and less often on stumps (gašparcová et al., 2017). 27 the laccate species g. carnosum was recorded only on 5 genera of host trees (appendix 1 – tab. 1). it is a rare species (4% of all records) in the urban areas of slovakia (fig. 1). we found out that g. carnosum grew on a decomposing wood (mostly of unknown trees – more than half of all findings). our results do not confirm published data that it colonises mainly abies alba mill. (kotlaba, 1984; tortić, 1985; breitenbach, kränzlin, 1986; sokół, 2000; bernicchia, 2005; karadelev et al., 2008; ryvarden, melo, 2014); however, in the forest ecosystems of slovakia, it was recorded predominantly on this tree species (gašparcová et al., 2017). we recorded one new deciduous host tree, tilia, that has not yet been mentioned in the literature. we found g. carnosum growing saprotrophically mainly on stumps (fig. 2), which confirms the information found in the literature (bernicchia, 2005). the species of g. lucidum and g. carnosum are very similar and difficult to distinguish (papp, szabó, 2013), but g. carnosum has larger basidiocarps with darker to blackish pilear surface, and wider spores, and it prefers coniferous trees (sokół, 2000; h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia fig. 2. topology of the basidiocarps of ganoderma spp. on host trees 4 5 21 33 16 4 19 0 1 21 5 10 1 26 0 1 2 1 1 1 5 1 0 2 3 26 2 3 2 0 9 16 12 1 4 0 0 2 0 0 2 0 0 0 1 0 0 0 0 0 5 10 15 20 25 30 35 dead trunk dead trunk base living trunk living trunk base others root swelling stump g. valesiacum g. pfeifferi g. resinaceum g. lucidum g. carnosum g. applanatum part of the woody plant number of records te ré zi a b ec k, j án g áp er , m ar tin š eb es ta , s ve tla na g áp er ov á 28 ryvarden, melo, 2014). g. lucidum was recorded on 4 species of host woody plants (appendix 1 – tab. 1), although most data do not specify trees species (65% of all findings). our results confirm the most common occurrence of this fungus on oaks (16% of all findings) as reported earlier by some authors (kotlaba, 1984; tortić, 1985; sokół, 2000; karadelev et al., 2008; papp, szabó, 2013; ryvarden, melo, 2014). we recorded 1 new species of the host tree – armeniaca vulgaris lam. the topology of g. lucidum basidiocarps was not recorded in most cases. in addition, it grew in approximately the same amount on stumps, living trunk bases, living trunks, and root swellings (fig. 2), which partially confirms data presented by sokół (2000), who stated that it grows on trunk and roots of hardwoods. however, our results do not confirm that g. lucidum grows predominantly saprotrophically, as some authors reported (sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014). more data on the topology of basidiocarps are needed to draw more precise conclusions. g. pfeifferi can be often confused with g. adspersum due to several similar features of basidiocarps (sokół, 2000), but g. pfeifferi has the wrinkled and cracked resinous pilear layer and wider basidiospores (sokół, 2000; ryvarden, melo, 2014). in the urban areas, we recorded g. pfeifferi only 4 times on 3 host species, mainly on fagus sylvatica (appendix 1 – tab. 1). other studies confirm the occurrence predominantly on beeches (kotlaba, 1984; tortić, 1985; sokół, 2000; papp, szabó, 2013; ryvarden, melo, 2014). our results do not show the occurrence of g. pfeifferi on conifers, although szczepkowski and konsenczjusz (2006) reported it was probably recorded on abies sp. we also found 1 new host species for this fungus – tilia platyphyllos scop. in the forests of slovakia, there are a few records of g. pfeifferi basidiocarps, mainly on fagus sylvatica (gašparcová et al., 2017a). g. pfeifferi was found to grow in the same number on stumps and paraswitically on the base of trunks (fig. 2). the occurrence on living trunk base confirms published data (bernicchia, 2005; ryvarden, melo, 2014). g. resinaceum can be confused with g. lucidum and g. pfeifferi, but g. lucidum has not resinous layer on the crust (ryvarden, melo, 2014), its basidiospores have a much coarser ornamentation, and it always forms a stem (kotlaba, pouzar, 2009b). g. pfeifferi has wider spores and a darker context (ryvarden, melo, 2014). g. resinaceum is the third most widespread ganoderma species in the urban areas of slovakia (fig. 1). it was found growing on 15 species of host trees (appendix 1 – tab. 1), mainly on quercus cerris l. (23% of all records), which confirms published data (kotlaba, 1984; tortić, 1985; sokół, 2000; bernicchia, 2005; papp, szabó, 2013; ryvarden, melo, 2014). our data prove the synanthropic character of the distribution of g. resinaceum and its rare occurrence in forests (kotlaba, 1984; gáper, 1998; sokół, 2000; gáperová, 2001; kotlaba, pouzar, 2009b). we found that g. resinaceum has various topologies of basidiocarps, but its basidiocarps are formed mainly on the living trunk base (fig. 2). 29 our results confirm that g. resinaceum grows rather parasitically on the base of trunks (breitenbach, kränzlin, 1986; bernicchia, 2005; ryvarden, melo, 2014) than saprotrophically on dead trunks or stumps (sokół, 2000). ganoderma valesiacum can be misidentified as g. lucidum or g. carnosum. it differs from them with very short or rudimentary stipe and often cracked laccate crust exposing white context (sokół, 2000; ryvarden, melo, 2014). however, the taxonomic status of g. valesiacum is uncertain (unclear relationships to g. lucidum and g. carnosum) and sequencing is needed to solve this problem (ryvarden, melo, 2014). in the urban areas of slovakia, we recorded this species on larix sp. (only 1 record – fig. 1); although, sokół (2000) reported that no data on the synanthropic character of this species exist. in the forests of slovakia, it was also recorded on larix (gašparcová et al., 2017a). it confirms published data that g. valesiacum is a very rare european species known from larix (plank, wolkinger, 1981; kotlaba, 1984; sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014). g. valesiacum basidiocarps were growing on the living trunk of host tree (fig. 2), but, as some authors reported, it to grow saprotrophically on the stumps and logs of larix (kotlaba, 1984; sokół, 2000; bernicchia, 2005; ryvarden, melo, 2014), since it was found in the forests of slovakia (gašparcová et al., 2017a). fig. 3. ganoderma adspersum (schulzer) donk on tilia cordata mill. in trenčianske teplice (northwestern slovakia) – a (photo. s. gáperová), and ganoderma applanatum (pers.) pat. on salix caprea l. in špania dolina (central slovakia) – b (photo. m. šebesta) h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia te ré zi a b ec k, j án g áp er , m ar tin š eb es ta , s ve tla na g áp er ov á 30 conclusion in the urban areas of slovakia, we recorded all 7 european ganoderma species. the most common species is g. adspersum. most often, it colonised non-native aesculus hippocastanum and native tilia cordata (but not quercus). it grows mostly at the base and along the living trunk and saprotrophically on stumps. g. applanatum is the second most widespread species in the urban areas of slovakia. it was found mainly on a decomposing wood of unidentified trees and also on beeches. it grows mostly on the stumps, and on the living trunks of host trees. g. resinaceum is the third most widespread in the urban areas of slovakia. it grows mainly on quercus. it was recorded mainly on the living trunk base. g. carnosum was recorded only once on abies alba. it grows saprotrophically, mainly on the stumps. g. lucidum was recorded on a total of 4 species of host woody plants and mostly on oaks. according to available data (in most cases have not been noticed), it grows on stumps, living trunk bases, living trunks, and root swellings. the rarest species of the genus ganoderma are g. pfeifferi and g. valesiacum. g. pfeifferi grows mainly on fagus sylvatica. it was recorded in the same numbers on the stumps and bases of living trunks. in the urban areas, surprisingly, we also recorded the typical forest species g. valesiacum on the living trunk of larix sp. acknowledgments this work has been supported by grants from the scientific grant agency of the ministry of education, science, research and sport of the slovak republic (vega no. 1/0286/17 and kega no. 025umb-4/2017). we wish to acknowledge the anonymous reviewers for his/her detailed and helpful comments of the manuscript. references antonín, v. 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(2015). global diversity of the ganoderma lucidum complex (ganodermataceae, polyporales) inferred from morphology and multilocus phylogeny. phytochemistry, 114, 7–15. doi: 10.1016/j.phytochem.2014.09.023 33 appendix 1 tab. 1. host preferences of species of the genus ganoderma species of the genus ganoderma total number of records woody plant taxa number of records ganoderma adspersum 102 abies sp. 1 acer platanoides l. 1 a. pseudoplatanus l. 3 aesculus hippocastanum l. 18 armeniaca vulgaris lam. 1 celtis occidentalis l. 1 cerasus avium (l.) moench 2 crataegus laevigata (poir.) dc 1 fagus sylvatica l. 2 fraxinus excelsior l. 7 gleditsia triacanthos l. 8 juglans regia l. 2 larix sp. 1 morus nigra l. 2 negundo aceroides moench 1 picea abies (l.) p. karst. 3 populus alba l. 2 p. nigra l. 3 prunus serrulata lindl. 3 quercus petraea (matt.) liebl. 2 quercus sp. 2 robinia pseudoacacia l. 3 salix caprea l. 1 sorbus aucuparia l. 2 spirea veitschii hemsl. 1 tilia cordata mill. 17 tilia sp. 1 t. platyphyllos scop. 2 unidentified 9 ganoderma applanatum 64 abies sp. 1 acer pseudoplatanus l. 1 alnus glutinosa (l.) gaertn. 1 carpinus betulus l. 2 cerasus avium (l.) moench 3 fagus sp. 4 fagus sylvatica l. 2 fraxinus excelsior l. 1 fraxinus sp. 1 negundo aceroides l. 1 h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia te ré zi a b ec k, j án g áp er , m ar tin š eb es ta , s ve tla na g áp er ov á 34 ganoderma applanatum picea abies (l.) karst. 1 populus alba l. 2 p. nigra l. 1 populus sp. 1 p. tremula l. 2 prunus domestica l. 1 prunus sp. 1 quercus robur l. 3 quercus sp. 2 robinia pseudoacacia l. 4 salix caprea l. 2 s. fragilis l. 1 salix sp. 2 tilia cordata mill. 1 t. platyphyllos scop. 1 tilia sp. 2 unidentified 20 ganoderma carnosum 11 abies alba mill. 1 alnus glutinosa (l.) gaertn. 1 larix sp. 1 quercus sp. 1 tilia sp. 1 unidentified 6 ganoderma lucidum 37 alnus sp. 1 a. glutinosa (l.) gaertn. 1 armeniaca vulgaris lam. 1 carpinus betulus l. 2 carpinus sp. 1 castaneae sativa mill. 1 quercus sp. 6 unidentified 24 ganoderma pfeifferi 4 acer platanoides l. 1 fagus sylvatica l. 2 tillia platyphyllos scop 1 35 ganoderma resinaceum 44 acer campestre l. 1 a. platanoides l. 1 a. pseudoplatanus l. 1 aesculus hippocastanum l. 4 celtis occidentalis l. 2 fagus sylvatica l. 1 fraxinus excelsior l. 2 gleditsia triacanthos l. 5 juglans regia l. 1 negundo aceroides moench 1 populus sp. 1 quercus cerris l. 10 q. robur l. 4 quercus sp. 1 robinia pseudoacacia l. 5 salix alba l. 1 sophora japonica l. 1 unidentified 2 ganoderma valesiacum 1 larix sp. 1 h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia te ré zi a b ec k, j án g áp er , m ar tin š eb es ta , s ve tla na g áp er ov á 36 abstract ganoderma (basidiomycota, polyporales) is a cosmopolitan genus with the greatest diversity in the tropics. it causes white rot of a wide range of woody plants all over the world. in europe, 7 species of the genus ganoderma grow: ganoderma adspersum, g. applanatum, g. carnosum, g. lucidum, g. pfeifferi, g. resinaceum and g. valesiacum. all of them also occur in slovakia. the aim of the present study was to identify the presence of ganoderma species in the urban areas of slovakia and to find out their host preferences and the topology of the basidiocarps on the colonised trees. a total 263 findings of ganoderma obtained by our own field research and the processing of records from herbarium items located in natural museums in bratislava (slovakia) and prague (czechia) were analysed to detect their ecological characteristics. the occurrence of all 7 ganoderma species was recorded in the urban areas of slovakia. the most common species is g. adspersum (39% of all records) with the widest range of host woody plants (24 species). g. applanatum is the second most widespread species in the urban areas of slovakia growing mostly on a decomposing wood of unknown trees (31% of all findings) and also on beeches (9% of all findings). g. resinaceum is the third most widespread in the urban areas, and it grew mainly on quercus. g. lucidum was most often found on unknown trees (65% of all findings) and also on oaks (16% of all findings). the rarest ganoderma species are g. valesiacum (only 1 record on larix sp.), g. pfeifferi (4 records, mainly on fagus sylvatica), and g. carnosum (4% of all records, mainly on a decomposing wood of unknown trees). key words: ganoderma, polyporoid fungi, basidiomata, host tree, wood-decaying fungi received: [2018.03.02] accepted: [2018.09.24] żywicielskie preferencje grzybów z rodzaju ganoderma rozkładających drewno w środowisku miejskim na słowacji streszczenie lakownica – ganoderma p. karst. (basidiomycota, polyporales) to kosmopolityczny rodzaj o największej różnorodności w  tropikach. lakownice powodują białe zgnilizny wielu różnych drzewostanów na całym świecie. w europie, w tym także na słowacji, występuje siedem gatunków lakownic: ganoderma adspersum (l. europejska), g. applanatum (l. spłaszczona), g. carnosum (l. brązowoczarna), g. lucidum (l. zółtawa), g. pfeifferi (l. czerwonawa), g. resinaceum (l. jasnomiąższowa) oraz g. valesiacum (l. walezyjska). celem niniejszych badań było zidentyfikowanie obecności poszczególnych gatunków lakownic w  zurbanizowanym środowisku słowacji oraz określenie ich preferencji żywicielskich, a także topologii bazydiokarpów na skolonizowanych drzewach. w badaniu wykorzystano 263 okazy lakownic zebranych w trakcie badań terenowych oraz oznaczonych rekordów, pochodzących ze zbiorów zielnikowych z muzeów przyrodniczych w bratysławie (słowacja) oraz w pradze (czechy). w zurbanizowanym środowisku słowacji odnotowano występowanie wszystkich 7 gatunków lakownic. najczęściej notowywanym gatunkiem jest g. adspersum (39% wszystkich rekordów), z  najszerszym spektrum drzew żywicielskich (24 gatunki). g. applanatum jest drugą najbardziej rozpowszechnioną lakownicą w zurbanizowanycm środowisku na słowacji. rośnie głównie na rozkładającym się drewnie nieidentyfikowalnych gatunków drzew (31% wszystkich rekordów), a także na bukach (9% wszystkich rekordów). g. resinaceum jest trzecim najbardziej rozpowszechnionym gatunkiem i rośnie głównie na dębach. natomiast g. lucidum zazwyczaj występowała na nieidentyfikowalnym drewnie (65% wszystkich rekordów) oraz na dębach (16% wszystkich rekordów). najrzadsze lakownice to: g. valesiacum (tylko jedno znalezisko na larix sp.), g. pfeifferi (4 rekordy, głównie na fagus sylvatica) oraz g. carnosum (4% wszystkich rekordów, zwłaszcza na nieidentyfikowalnym gatunkowo rozkładającym się drewnie). słowa kluczowe: ganoderma, grzyby polyporowe, basidiomaty, drewno żywicielskie, grzyby rozkładające drewno 37 information on the authors terézia beck https://orcid.org/0000-0002-3179-8380 she is an internal phd student at faculty of natural sciences of matej bel university in banská bystrica (slovakia), department of biology and ecology, in study program „evolution of ecosystems and their protection”. the topic of her dissertation is „diversity and distribution of the genus ganoderma in slovakia”. ján gáper he is the member of department of biology and general ecology of faculty of ecology and environmental sciences of technical university in zvolen as well as the member of department of biology and ecology of faculty of natural sciences of university of ostrava in ostrava. he is interested in the identification and ecology of wood-decaying polypores, regarding the importance for ecosystem functioning. martin šebesta he is an internal phd student at faculty of ecology and environmental sciences of technical university in zvolen (slovakia), department of biology and general ecology, in study program “ecology and biodiversity conservation”. the topic of his dissertation is “diversity and ecology of wood-decay polypores in the urban environment”. svetlana gáperová she is the member of department of biology and ecology of faculty of natural sciences of matej bel university in banská bystrica. she is interested in the biology and ecology of both woody plants and wood-decaying fungi in urban areas and monitoring of urban air pollution. h ost preferences of w ood-decaying fungi of the genus g anoderm a in the urban areas of s lovakia 115 annales universitatis paedagogicae cracoviensis studia naturae, 1: 115–126, 2016, issn 2543-8832 sylwia śliwińska-wilczewska*, kinga gergella, adam latała institute of oceanography, university of gdańsk, av. piłsudskiego 46, 81-378 gdynia, poland, *ocessl@ug.edu.pl allelopathic activity of the synechococcus sp. (cyanobacteria, chroococcales) on selected cyanobacteria species introduction allelopathy may be one of the factors contributing to the formation and maintenance of cyanobacterial blooms, which strongly a�ect coastal marine ecosystems and cause economic problems for commercial aquaculture (gross, 2003). furthermore, some species of cyanobacteria are able to produce and release secondary metabolites that may be harmful to microorganisms, phytoand zooplankton, crustaceans, �sh and even humans (stal et al., 2003; mazur-marzec et al., 2015). cyanobacteria are known to produce a wide range of secondary metabolites with various biological actions. some of them, termed allelopathic compounds, have been shown to play a role in allelopathy (le�aive, ten-hage, 2007). �e precise mode of action of allelopathic compounds remains relatively poorly known due to methodological di�culties. it is believed that the allelopathic compounds may be responsible for the natural selection of organisms, competition and ecological succession (legrand et al., 2003). moreover, it was indicated that some cyanobacteria are able to produce and release allelopathic compounds that a�ect the growth and development of other organisms (gross, 2003; żak, kosakowska, 2015). �is can be important for many areas of science and industry. secondary metabolites isolated from cyanobacteria can be used in medicine, agriculture, as herbicides or insecticides, and maybe even in the creation of drugs (berry et al., 2008; hernández-carlos, gamboa-angulo, 2011). generally, the blooms of cyanobacteria that develop each summer in the freshwater and brackish ecosystems are composed of two di�erent groups: the large, colony-forming, filamentous cyanobacteria and small-sized picocyanobacteria from the genus synechococcus and synechocystis. picocyanobacteria fraction may comprise as much as 80% of the total cyanobacterial biomass and contribute as much as 50% of the total primary production of a cyano116 s yl w ia ś liw iń sk aw ilc ze w sk a, k in ga g er ge lla , a da m l at ał a bacterial bloom (stal et al., 2003). picocyanobacteria strain of the genus synechococcus are very important organisms in the world’s oceans, however, the information about allelopathic interactions between picoand �lamentous cyanobacteria in aquatic ecosystems are scarce. �e main aim of this work was to estimate the allelopathic interaction of picocyanobacterium synechococcus sp. on selected cyanobacteria. in this study, the in�uence of allelopathic compounds on the growth, cell-morphology, photosynthetic pigments, chlorophyll �uorescence and performance of photosynthesis in the analysed species was investigated by single and multiple addition of cell-free �ltrate obtained from picocyanobacterium synechococcus sp. in this experiment we investigated the e�ect of single and multiple addition of cell-free �ltrate obtained from synechococcus sp. on selected picoand �lamentous cyanobacteria: synechocystis sp., geitlerinema amphibium, nodularia spumigena and nostoc sp. future studies should examine the allelopathic activity of di�erent strains of picocyanobacteria, including synechocystis and aphanocapsa. material and methods �e experiments were conducted on the picocyanobacterium synechococcus sp. (ba124) and the cyanobacteria geitlerinema amphibium (ba-13), nodularia spumigena (ba-15), nostoc sp. (ba-81) and synechocystis sp. (ba-153). �e strains were isolated from the coastal zone of the gulf of gdańsk (southern baltic sea) and are maintained as unialgal cultures in the culture collection of baltic algae (ccba) at the institute of oceanography, university of gdańsk, poland (latała et al., 2006). �e tests on batch cultures were carried out in 25 ml glass erlenmeyer �asks containing sterilized f/2 medium (guillard, 1975). �e media were prepared from baltic water with a  salinity of about 8 psu, which was �ltered through whatman gf/c glass �ber �lters, and autoclaved. analyzed cyanobacteria were grown 7 days in constant conditions of 20°c and 8 psu, under a 16:8h light : dark cycle at 10 μmol m-2·s-1 and this were the control treatment conditions. fluorescent lamps (cool white 40w, sylvania, usa) were used as source of irradiance. �e intensity of par was measured using a li-cor quantum-meter with a cosine collector. �e donor and target cyanobacteria were acclimated to these culture conditions for 7 days; a�erwards, actively growing cultures were used for the establishment of the allelopathic experiment. allelopathic interactions were determined by using the method proposed by śliwińska-wilczewska et al. (2016a). allelopathic interaction was studied by adding the single and multiple cell-free �ltrate obtained from picocyanobacterial culture to tested cyanobacteria. �e culture of synechococcus sp. was �ltered through 0.45 µm pore size macherey-nagel mn gf-5 �lters. �e cell-free �ltrate (v = 2 ml) was added to 25 ml erlenmeyer �asks containing the tested cyanobacteria (v = 20 117 ml). in all experiments, the ratio of picocyanobacterium to target species in erlenmeyer �asks was adjusted to 1:1 based on the chlorophyll a content (�nal chlorophyll a concentration in the experimental cultures was 0.8 µg chl a ml-1). control samples were prepared by adding mineral medium f/2 with a volume equal to the added cell-free �ltrate. to simulate the e�ects of continuously released picocyanobacterial allelochemicals on target species, picocyanobacterial �ltrates were added daily to the target cultures for one week. �e �rst addition was made as described above. subsequent additions were made by removing 2 ml of test volume, used for cell counts each day and replacing it with an equal volume of fresh �ltrate or control medium. tests were conducted in triplicate and all analysed species were obtained from early exponential growth phase. �e identi�cation of allelopathic compounds is a di�cult and time-consuming task and will need further investigation. culture density was determined by the number of cells and optical density (od). �e number of cells was counted using bürker chamber and od was measured spectrophotometrically at 750 nm with a multiskan go �ermo scienti�c uv-vis spectrophotometer. �e results of cell counts and respective od measurements were then used to determine the linear correlation between them for each species. determined relationships were then used to estimate the number of cells in the experimental cultures a�er 1st, 3rd and 7th day of the cyanobacteria exposure to the picocyanobacterial �ltrate. in addition, the morphological changes of target cyanobacteria were documented using a nikon eclipse 80i microscope with a camera nikon dsu2. in this work the concentration of photosynthetic pigments of target organisms was measured by spectrophotometric method a�er one week of exposure to the picocyanobacteria cell-free �ltrate. chlorophyll a, carotenoids and phycobilins were determined with a �ermo scienti�c spectrophotometer uv-vis multiscan go using 1 cm glass cuvette. �e concentration of pigments was calculated according to equations provided by je�rey and humphrey (1975), strickland and parsons (1972) and bennett and bogorad (1973). chlorophyll a  �uorescence was measured with a  pulse amplitude modulation (pam) �uorometer (fms1, hansatech), using 594 nm amber modulating beam with 4 step frequency control as a measuring light. analysed species were taken for chlorophyll �uorescence analysis a�er 7th day of exposure to the �ltrate. before measurements, each sample taken from the culture was �ltered through 13 mm glass �ber �lters (whatman gf/c). before starting the experiment, the �lter sample was adapted in the dark for about 15 minutes. fluorescence parameters such as maximum psii quantum e�ciency (fv/fm) and e�ective psii quantum e�ciency (φpsii) were calculated (campbell et al., 1998). a llelopathic activity of the synechococcus sp. (c yanobacteria, c hroococcales) on selected cyanobacteria species 118 s yl w ia ś liw iń sk aw ilc ze w sk a, k in ga g er ge lla , a da m l at ał a �e measurements of oxygen evolution were carried out on the 7th day of the experiment by using clark-type oxygen electrode (chlorolab 2, hansatech). temperature was controlled at 20°c with a cooling system lauda (e100, germany). illumination was provided by a high intensity probe-type light array with 11 red led’s centered on 650 nm. irradiance was measured with a quantum sensor (quantitherm, hansatech). dark respiration was estimated from o2 uptake by cells incubated in the dark. experimental data were �tted to the photosynthesis-irradiance curve using equation (jassby, platt, 1976). analysis of variance (anova) was used to test for di�erences in analysed parameters between the target cyanobacteria cultures treated with picocyanobacterial cell-free �ltrates and the control over the experimental period. a post hoc test (tukey’s hsd) was used to show which experiments of picocyanobacterial �ltrate a�ected the growth of target cyanobacteria di�erently. data is reported as mean ± standard deviation (sd). levels of signi�cance were: * p < 0.05. �e statistical analyses were performed using the statistica® 12 so�ware. results �e e�ect of the cell-free �ltrate addition obtained from synechococcus sp. cultures on the growth of geitlerinema amphibium, nodularia spumigena, nostoc sp. and synechocystis sp. a�er 1, 3 and 7 days of exposition to the �ltrates are shown in �gure 1. �e results showed that addition of cell-free �ltrate from synechococcus sp. decreased the number of cells of g. amphibium, n. spumigena and nostoc sp. compared to their control. on the basis of the results it was found that the �ltrate obtained from picocyanobacterium had the strongest e�ect on g. amphibium. a�er the 7th for a  single and multiple �ltrate addition obtained from synechococcus sp. growth inhibition of g. amphibium expressed as a  percent of culture density (% of control) constituted 48% and 56% respectively (p < 0.05). it was also observed, that the single addition of cell-free �ltrate obtained from synechococcus sp. signi�cantly decreased the number of cells of n. spumigena and nostoc sp. (p < 0.05). on the 1st, 3rd and the 7th day of the experiment, the minimum cell response of n. spumigena constituted 94%, 81% and 79% (p < 0.05) respectively, and for nostoc sp. the percent of culture density constituted 87%, 81% and 82% (p < 0.05) respectively, in comparison to the control treatment. moreover, on the 3rd and the 7th day of the experiment, the minimum cell response of n. spumigena a�er multiple addition of the cell-free �ltrate, constituted 55% and 63%, respectively (p < 0.05). in addition, it was observed that the cell-free �ltrate obtained from synechococcus sp. did not a�ect the number of cells of synechocystis sp. (p > 0.05). �e morphological changes of the target cyanobacteria a�er the picocyanobacterial cell-free �ltrate addition was shown in �gure 2. it was shown that the addition 119 fig. 1. �e e�ect of the a) single and b) multiple additions of cell-free �ltrate from synechococcus sp. cultures on the growth of geitlerinema amphibium (ba-13), nodularia spumigena (ba-15), nostoc sp. (ba-81) and synechocystis sp. (ba-153) a�er 1, 3 and 7 days of exposition to the �ltrates, expressed as a percent of culture density (% of control). �e values refer to means (n = 3, mean ± sd). asterisk indicates signi�cant di�erence compared with control fig. 2. �e cells morphology of a) geitlerinema amphibium, b) nodularia spumigena, c) nostoc sp. and d) synechocystis sp. for a) control sample and b) in the experiments with the addition of cyanobacterial cell-free �ltrate a�er 7 days of exposure of cell-free �ltrate caused a decline of pigmentation and cell lysis of g. amphibium, n. spumigena and nostoc sp. compared to the control culture. in contrast, it was observed that the cell-free �ltrate obtained from synechococcus sp. had no e�ect on the picocyanobacterium synechocystis sp. �e e�ect of the single and multiple additions of cell-free �ltrate from synechococcus sp. cultures on the pigment contents of g. amphibium a�er one week of exposition is shown in �gure 3. �e results showed that allelochemicals released by picocyanobacterium synechococcus sp. signi�cantly decreased the chlorophyll a and phycobilins of g. amphibium compared to their control (p < 0.05). a�er one week of exposition, it was noted that the single addition of the �ltrate resulted in decrease of chlorophyll a and phycobilins in the cells of analysed cyanobacterium, which was lower by 28% and 50%, respectively, compared to a control (p < 0.05). based on the results, it was found that the multiple addition of cell-free �ltrate obtained from synechococcus sp. also caused a llelopathic activity of the synechococcus sp. (c yanobacteria, c hroococcales) on selected cyanobacteria species 120 s yl w ia ś liw iń sk aw ilc ze w sk a, k in ga g er ge lla , a da m l at ał a a signi�cant change in the pigment contents of g. amphibium cells. a�er one week of the experiment, chlorophyll a of this cyanobacterium was lower by 44% compared to control (p < 0.05). in addition, in this study it was observed that the cell-free �ltrate obtained from synechococcus sp. did not a�ect the carotenoids content of analysed cyanobacterium (p > 0.05). �e e�ects of picocyanobacterial cell-free �ltrate on chlorophyll a �uorescence after 7 days of incubation are shown in �gure 4. it was observed, that the single addition of cell-free �ltrate from synechococcus sp. signi�cantly stimulated the values of φpsii (p < 0.05). moreover, the multiple additions of cell-free �ltrate from synechococcus sp. fig. 3. �e e�ect of the a) single and b) multiple additions of cell-free �ltrate from synechococcus sp. cultures on the pigment contents: chlorophyll a  (chl a), carotenoids (car) and phycobilins (phyco) for geitlerinema amphibium (ba-13) a�er 7 days of exposure. �e values refer to means (n = 3, mean ± sd). asterisk indicates signi�cant di�erence compared with control fig. 4. �e e�ect of the single and multiple additions of cell-free �ltrate from synechococcus sp. cultures on the �uorescence parameter fv/fm and φpsii for geitlerinema amphibium (ba-13) a�er 7 days of exposure. �e values refer to means (n = 3, mean ± sd). asterisk indicates signi�cant di�erence compared with control inhibited the values of the analysed parameters fv/fm and φpsii which amounted to 94% (p > 0.05) and 57% (p < 0.05) in comparison to the control, respectively. p-e curves for analysed g. amphibium treated with single and multiple cell-free �ltrate obtained from synechococcus sp. are presented in �gure 5. it was demonstrated that the investigated cyanobacterium was sensitive to the picocyanobacterial cell-free 121 �ltrate. in this study the in�uence of picocyanobacterial allelochemicals on the maximum photosynthesis (pm) of the tested cyanobacterium was noted and its value constituted 31% and 27% (p < 0.05), respectively, in comparison to the control treatment. discussion allelopathic interaction may play a  signi�cant role in an aquatic ecosystem (gross, 2003). allelopathy is considered one of the factors promoting and maintaining cyanobacterial blooms in freshwater, brackish and marine ecosystems around the world. although the allelopathy phenomenon is common in aquatic ecosystems, the mode of action of allelopathic compounds produced by picocyanobacteria on cyanobacteria remains poorly investigated. inhibition of growth of the target organism by production of allelopathic compounds is relatively widespread and the most frequently reported mode of action of cyanobacteria (issa, 1999; schagerl et al., 2002). in this study it was demonstrated that the addition of cell-free �ltrate from synechococcus sp. decreased the number of cells of �lamentous cyanobacteria geitlerinema amphibium, nodularia spumigena and nostoc sp. compared to their control. �e results also showed that the allelochemicals produced by the picocyanobacterium had the strongest e�ect on g. amphibium. it was also observed, that for g. amphibium, the e�ect of single cyanobacterial �ltrate additions was stronger than the e�ect of multiple additions. surprisingly, it was observed that the cell-free �ltrate obtained from synechococcus sp. did not a�ect the number of cells of picocyanobacterium synechocystis sp. picocyanobacteria of the genus synechococcus plays an important role in aquatic ecosystems but not much is known about its allelopathic activity. information about the ability of allelopathic interactions of picocyanobacterium synechococcus sp. was described by ślifig. 5. p-e curves for geitlerinema amphibium (ba-13) for control and a) single and b) multiple additions of cell-free �ltrates obtained from synechococcus sp. cultures a�er 7 days of exposure. �e values refer to mean ± sd (n = 3) a llelopathic activity of the synechococcus sp. (c yanobacteria, c hroococcales) on selected cyanobacteria species 122 s yl w ia ś liw iń sk aw ilc ze w sk a, k in ga g er ge lla , a da m l at ał a wińska-wilczewska et al. (2016a). in this work the authors showed that addition of the cell-free �ltrate obtained from the picocyanobacterium synechococcus sp. had a  signi�cant inhibitory e�ect on n. spumigena. authors described that the longer the exposure time, the slower the growth of the analysed �lamentous cyanobacterium, and on the 7th day of experiment the minimum cells response constituted about 60% in comparison to control treatment. in another work schagerl et al. (2002) also demonstrated growth in inhibition of cyanobacteria anabaena cylindrica and microcystis �os-aquae a�er adding the �ltrate from the cyanobacterium anabaena torulosa. also issa (1999) investigated the e�ect of allelopathic compounds produced by oscillatoria angustissima and calothrix parietina on microcystis aeruginosa, synechococcus sp., scytonema hofmonni, anabaena spiroides, phormidium mölle, nostoc muscorum, oscillatoria angustissima and calothrix parietina. author noted that cyanobacteria of the genus oscillatoria, calothrix, nostoc and anabena were resistant to allelopathic compounds released by analysed cyanobacteria. moreover, the recovery of g. amphibium growth a�er a multiple �ltrate additions may have been due to, e.g., its ability to metabolise allelochemicals, as suggested by suikkanen et al. (2004). it is believed that selective inhibition of growth of the target organism may a�ect the succession of selected cyanobacteria in aquatic ecosystem (legrand et al., 2003). �ere are only several reports indicating that the chemical compounds can cause structural and morphological changes in target cells (gantar et al., 2008). in the present study it was shown that the tested allelopathic compounds obtained from synechococcus sp. caused restriction of pigmentation and cell lysis of all analysed �lamentous cyanobacteria. gantar et al. (2008) also noted that a�er being exposed to the crude extract from fischerella sp., cells of chlamydomonas sp. showed distinctive morphological and structural changes. �e electron microscopy revealed degeneration of thylakoids and disappearance of other cell structures including the nucleus. �e results may in part explain the reasons for achieving competitive advantage of picocyanobacterium synechococcus sp. over other �lamentous cyanobacteria in many aquatic ecosystems. �e precise mode of action of allelopathic compounds remains relatively poorly known due to methodological di�culties. �erefore, there are only a few reports that allelopathic compounds may a�ect the pigments content, chlorophyll �uorescence and photosynthesis of target organisms (gross et al., 1991; śliwińska-wilczewska et al., 2016b). in the present study it was shown that the tested allelopathic compounds obtained from synechococcus sp. caused restriction of pigmentation and inhibition of photosynthetic activity of analysed g. amphibium compared to the control culture. literature data indicates that allelopathic compounds produced by cyanobacteria can a�ect the photosynthesis, and detailed studies have shown that they act mainly on the photosystem ii (psii). gross et al. (1991) showed that �scherellin a produced by cy123 anobacterium fischerella muscicola, and the compounds released by trichormus doliolum and oscillatoria late-virens inhibited activity of psii in selected cyanobacteria and microalgae. in addition, śliwińska-wilczewska et al. (2016b) showed that the addition of cell-free �ltrate from synechococcus sp. cultures grown under varied light, temperature and salinity signi�cantly inhibited the values of fv/fm of the diatom navicula perminuta. moreover, authors noted that the lowest values of pm in n. perminuta were observed a�er addition of cell-free �ltrate obtained from synechococcus sp. grown at 25°c, and was 49% lower than for the control group. inhibition of photosynthesis, the major physiological process of competing phytoplankton species, can be a defensive strategy of co-occurring cyanobacteria (gross, 2003). �erefore, allelopathic interaction may result in inhibition of pigments content, photosynthesis and growth of target organisms which in part explain the domination of picocyanobacteria in many aquatic ecosystems. many studies indicated that cyanobacteria produced a wide spectrum of secondary metabolites, a  source of a  new bioactive and natural compounds, which can be used in medicine and pharmaceutical industry as antibacterial, antiviral and antifungal compounds and even against tumor cells (berry et al., 2008; hernández-carlos, gamboa-angulo, 2011). �e compounds produced by cyanobacteria can also be used in agriculture as herbicides, or insecticides, and may also have potential applications in biotechnology (le�aive, ten-hage, 2007). it is also believed that allelopathy may be one of the important factors a�ecting the formation of massive cyanobacterial blooms in the aquatic environment (gross, 2003; legrand et al., 2003). species forming harmful blooms in many places in the world are a serious problem, both ecologically and economically. despite the seriousness of the problem, relatively little is known about the inhibitory e�ect of secondary metabolites of cyanobacteria on coexisting organisms in the baltic sea. �erefore, providing new information on the extent of the e�ect of allelopathic cyanobacteria, it may also be important for better understanding of the worldwide intensi�cation of massive phytoplankton blooms in various aquatic ecosystems. acknowledgements �e authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. �is study was supported by bmn grants, no. 538-g245-b211-16. references bennett, a., bogorad, l. 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(2011). metabolites from freshwater aquatic microalgae and fungi as potential natural pesticides. phytochemistry reviews, 10(2), 261–286. doi: 10.1007/ s11101-010-9192-y gantar, m., berry, j.p., �omas, s., wang, m., perez, r., rein, k.s. (2008). allelopathic activity among cyanobacteria and microalgae isolated from florida freshwater habitats. federation of european microbiological societies, 64, 55–64. doi: 10.1111/j.1574-6941.2008.00439.x gross, e.m. (2003). allelopathy of aquatic autotrophs. critical reviews in plant sciences, 22, 313–339. doi: 10.1080/713610859 gross, e.m., wolk, c.p., jüttner, f. (1991). fischerellin, a  new allelochemical from the freshwater cyanobacterium fischerella muscicola. journal of phycology, 27, 686–692. doi: 10.1111/j.00223646.1991.00686.x guillard, r.r.l. (1975). culture of phytoplankton for feeding marine invertebrates. in: w.l. smith, m.h. chanley (eds.), culture of marine invertebrate animals. new york: plenum press, 26–60. issa, a.a. (1999). antibiotic production by the cyanobacteria oscillatoria angustissima and calothrix parietina. environmental toxicology and pharmacology, 8, 33–37. doi: 10.1016/s1382-6689(99)00027-7 jakubowska, n., szeląg-wasielewska, e. (2015). toxic picoplanktonic cyanobacteria – review. marine drugs, 13(3), 1497–1518. doi: 10.3390/md13031497 jassby, a.d., platt, t. (1976). mathematical formulation of the relationship between photosynthesis and light for phytoplankton. limnology and oceanography, 21, 540–547. doi: 10.4319/lo.1976.21.4.0540 je�rey, s.w., humphrey, g.f. (1975). new spectrophotometric equations for determining chlorophylls a, b, c1 and c2 in higher plants, algae and natural phytoplankton. biochemie und physiologie der p�anzen, 167, 191–194. latała, a., jodłowska, s., pniewski, f. 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(2015). baltic cyanobacteria – a source of biologically active compounds. european journal of phycology, 50, 343–360. doi: 10.1080/09670262.2015.1062563 schagerl, m., unterrieder, i., angeler, d.g. (2002). allelopathy among cyanoprokaryota and other algae originating from lake neusiedlersee (austria). international review of hydrobiology, 87, 365–374. doi: 10.1002/1522-2632(200207)87:4<365::aid-iroh365>3.0.co;2-b stal, l.j., albertano, p., bergman, b., bröckel, k., gallon, j.r., hayes, p.k., sivonen, k., walsby, a.e. (2003). basic: baltic sea cyanobacteria. an investigation of the structure and dynamics of water blooms of cyanobacteria in the baltic sea – responses to a changing environment. continental shelf research, 23, 1695–1714. doi: 10.1016/j.csr.2003.06.001 strickland, i.d.h., parsons, t.r. (1972). a practical handbook of seawater analysis. journal of the fisheries research board of canada, 167, 1–310. 125 suikkanen, s., fistarol, g.o., granéli, e. (2004). allelopathic e�ects of the baltic cyanobacteria nodularia spumigena, aphanizomenon �os-aquae and anabaena lemmermannii on algal monocultures. journal of experimental marine biology and ecology, 308, 85–101. doi: 10.1016/j.jembe.2004.02.012 śliwińska-wilczewska, s., pniewski, f., latała, a. (2016a). allelopathic interactions between synechococcus sp. and nodularia spumigena under di�erent light conditions. allelopathy journal, 37(2), 241–252. śliwińska-wilczewska, s., pniewski, f., latała, a. (2016b). allelopathic activity of the picocyanobacterium synechococcus sp. under varied light, temperature and salinity conditions. international review of hydrobiology, 101, 1–9. doi: 10.1002/iroh.201501819 żak, a., kosakowska, a. (2015). �e in�uence of extracellular compounds produced by selected baltic cyanobacteria, diatoms and dino�agellates on growth of green algae chlorella vulgaris. estuarine, coastal and shelf science, 167, 113–118. doi: 10.1016/j.ecss.2015.07.038 abstract picocyanobacterium synechococcus sp. is very important but still poorly understood component of marine and freshwater ecosystems. in this study, the e�ect of single and multiple addition of cell-free �ltrate obtained from synechococcus sp. on selected cyanobacteria synechocystis sp., geitlerinema amphibium, nodularia spumigena and nostoc sp. was investigated. �e species present in this work are groups of aquatic phototrophs known to co-occur in the baltic sea. �e study showed that the picocyanobacterial cell-free �ltrate inhibits the growth and changes the cell morphology of �lamentous cyanobacteria g. amphibium, n. spumigena and nostoc sp. it was shown that the addition of cell-free �ltrate caused a decline of pigmentation and cell lysis of g. amphibium, n. spumigena and nostoc sp. compared to the control culture. in addition, it was observed that the �ltrate obtained from synechococcus sp. did not a�ect the synechocystis sp. it was found that the �ltrate obtained from picocyanobacterium had the strongest e�ect on growth of g. amphibium, therefore for this cyanobacteria additional experiments were performed to show whether the �ltrate a�ected also photosynthetic pigments, chlorophyll �uorescence and photosynthesis. �e study proved that the picocyanobacterial allelopathic compounds reduce the e�ciency of photosynthesis, which results in the inhibition of growth of target organisms. �is way of interaction may explain the formation of almost monospeci�c cyanobacterial blooms in many aquatic ecosystems, including the baltic sea. key words: allelopathy, cyanobacteria, �uorescence, growth, photosynthesis, photosynthetic pigments received: [2016.06.08] accepted: [2016.09.06] zjawisko oddziaływania allelopatycznego sinicy synechococcus sp. (cyanobacteria, chroococcales) na wybrane gatunki sinic streszczenie pikoplanktonowa sinica synechococcus sp. jest bardzo ważnym, lecz nadal słabo poznanym składnikiem wodnych ekosystemów. w  przeprowadzonych badaniach określono wpływ pojedynczo i  wielokrotnie dodawanego przesączu uzyskanego z  synechococcus sp. na wybrane gatunki pikoplanktonowych i  nitkowatych sinic: synechocystis sp., geitlerinema amphibium, nodularia spumigena oraz nostoc sp. badane gatunki sinic występują w tych samych ekosystemach i znane są z odgrywania istotnej roli w morzu bałtyckim. w pracy wykazano, że przesącz wpływał hamująco na wzrost nitkowatych sinic g. amphibium, n. spumigena oraz nostoc sp. nie zanotowano natomiast istotnych zmian liczebności u pikoplanktonowej sinicy synechocystis sp. dla wszystkich gatunków badanych sinic zostały wykonane analizy zmian morfologicznych, które zaszły w ich komórkach pod wpływem dodania przesączu uzyskanego z kultur synechococcus sp. na podstawie uzyskanych wyników wykazano, że przesącz powodował utratę pigmentacji i lizę komórek nitkowatych sinic. ponadto dla sinicy g. amphibium zostały wykonane dodatkowe eksperymenty, na podstawie których stwierdzono, że dodany przesącz hamował �uorescencję chloro�lu a, tempo a llelopathic activity of the synechococcus sp. (c yanobacteria, c hroococcales) on selected cyanobacteria species 126 s yl w ia ś liw iń sk aw ilc ze w sk a, k in ga g er ge lla , a da m l at ał a fotosyntezy, a także wpływał znacząco na zawartość barwników fotosyntetycznych. w pracy wykazano, że związki allelopatyczne produkowane i uwalniane przez synechococcus sp. ograniczały sprawność fotosyntezy, co skutkowało zahamowaniem wzrostu organizmów targetowych. tego rodzaju oddziaływanie może wyjaśniać tworzenie się prawie monogatunkowych zakwitów sinic w wielu zbiornikach wodnych, w tym również w morzu bałtyckim. słowa kluczowe: allelopatia, sinice, �uorescencja, wzrost, fotosynteza, barwniki fotosyntetyczne information on the authors sylwia śliwińska-wilczewska currently she is researching the allelopathic activity of picocyanobacteria. recently she has examined that the synechococcus sp. reveals allelopathic activity on the photosynthesis and chlorophyll �uorescence, which results in the inhibition of growth of analyzed target species. she has also discovered that picocyanobacterium synechococcus sp. produces and releases allelopathic compounds which have negative in�uence on di�erent green algae, diatoms and �lamentous cyanobacteria. kinga gergella �e �eld of her interest is allelopathic interactions of phytoplankton, in particular of baltic microalgae and cyanobacteria. she is investigating what in�uence allelopathic compounds have on those organisms. in her studies she uses innovative methods: analyzing chlorophyll a �uorescence and measuring rate of photosynthesis to determine what impact allelochemicals have on algae. adam latała he has wide experience in ecophysiology and ecotoxicology of marine benthic and planktonic algae. he is interested in in�uence of the main environmental factors, such as salinity, temperature and light, on the photosynthesis, photoacclimation, �uorescence, respiration and growth of algae from natural communities and cultured under laboratory conditions. he uses �uorescence techniques to determine algal and cyanobacterial ecophysiology and ecotoxicology. 59 annales universitatis paedagogicae cracoviensis studia naturae, 2: 59–68, 2017, issn 2543-8832 doi: 10.24917/25438832.2.4 sylwia śliwińska-wilczewska1*, agata cieszyńska2, adam latała1 1institute of oceanography, university of gdańsk, gdynia, poland, *ocessl@edu.pl 2institute of oceanology of the polish academy of science, department of marine physics, marine biophysics laboratory, sopot, poland the impact of temperature and photosynthetically active radiation on the growth and pigments concentration in baltic picocyanobacterium synechococcus sp. introduction chrococcoid cyanobacteria strain of the genus synechococcus appears in marine, brackish, and freshwater ecosystems. picoplanktonic organisms show a lot of adaptations, which enable them to spread in aquatic environments and to dominate and occupy the niches inaccessible for other photoautotrophs. owing to the fact that picocyanobacteria exhibit the small size of cells and possess an advantageous surface area to volume ratio, they can assimilate trace amounts of nutrients. �erefore, in oligotrophic regions of seas and oceans, picoplankton can compete successfully with larger algae and determine the primary production of the whole water ecosystem (six et al., 2007; richardson, jackson, 2007). moreover, thanks to their small size and despite the absence of the gas vesicles, picocyanobacteria can �oat e�ectively in the ocean water. �e distribution and growth intensity of picocyanobacteria are determined by their optimal ecological requirements, such as light and temperature. �ese factors in�uence metabolic processes, photosynthetic pigments, photosynthetic activity, and consequently, the rate of cell division and growth. cyanobacterial strains show outstanding acclimation capability, which enable them to survive in environments with a great variability in conditions. some cyanobacteria are tolerant to high temperatures up to 70°c and can survive at low intensities of about 5 μmol m-2 s-1 (stal et al., 2003). picocyanobacteria are a major component of oceanic ecosystems. moreover, picocyanobacteria are important contributors to primary production in the ocean, particularly in warm nutrient-poor waters (stockner, 1988). �is suggests that the contribution of picocyanobacteria in temperate seas should be most important in the summer, when the water temperature is the highest and the seasonal thermocline limits the s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 60 supply of nutrients to the upper layer (agawin et al., 1998). unfortunately, coherent investigations of physiological adaptations of picocyanobacterium synechococcus sp. in aforementioned basins are scarce. �erefore, the objective of this work was to characterise the ecophysiological features of three di�erent baltic picocyanobacterial strains of the genus synechococcus. �e study was focused on estimating the e�ect of par and temperature on the wide range of changes in cell concentration and photosynthetic pigment content (chl a and car). �e autecological studies of the cyanobacterial species and the recognition of their reactions to the main environmental factor, such as light intensity and temperature, could be of great importance to recognise the phenomenon of picocyanobacterial blooms in aquatic ecosystems and an important step in the way to improved understanding of bio-geo-physical couplings in the water mediums. material and methods �e experiments were conducted on three di�erent picocyanobacterial strains from the genus synechococcus: red ba-120, green ba-124 and brown ba-132. depending on pigment content, strains from the genus synechococcus are classi�ed as red strains with phycoerythrin, green strains rich in phycocyanin, and brown strains with phycourobilin and phycoerythrin (mazur-marzec et al., 2013; jodłowska, śliwińska, 2014). �e strains were isolated from the coastal zone of the gulf of gdańsk (southern baltic sea) in late spring of 2002 and are maintained as unialgal cultures in the culture collection of baltic algae at the institute of oceanography, university of gdańsk, poland (latała et al., 2006). tests on the ‘batch cultures’ were carried out in 25 ml glass erlenmeyer �asks containing sterilised f2 medium (guillard, 1975). �e media were prepared from arti�cial sea water with a salinity of about 8. �e strains were incubated under a 16:8 h light : dark cycle at four par intensities (10, 100, 190, and 280 μmol m-2 s-1) and at four temperatures (10, 15, 20, and 25°c). as a lighting source �uorescent lamps, sylvania cool-white 40 w and sylvania 100 w halogen lamps for more intense light were used. �e intensity of par was measured by a licor li-189 quantum-meter with a scalar collector. �e cultures were acclimated to every culture condition for 2 days, and then they served as inoculum for experimental cultures where the initial number was 106 cells per ml. �e test cultures were grown in three replicates and were incubated for one week. a�er that time in the exponential growth phase, the concentration of cells and contents of pigments were measured. �e number of synechococcus sp. cells was determined using bd accuri™ c6 �ow cytometer (becton dickinson, new jersey, usa). �e amount of picocyanobacteria was counted at delivery rate of 14 µl min-1 and identi�ed using the standard optical filter 670 nm (fl3) and 675/25 nm (fl4) (marie et al., 2005). 61 chlorophyll a and carotenoids were extracted with cold 90% acetone in the dark for 4 hours at -60°c. to remove cell debris and �lter particles, the pigment extract was centrifuged at 13000 rpm for 2 minutes (sigma 2-16p, osterode am harz, germany). �e extinction was determined at 750, 665, and 480 nm with a uv-vis spectrophotometer – du 530 beckman using 1 cm glass cuvette. �e concentration of carotenoids was calculated according to strickland and parsons (1972) with the formula: car (µg ml-1) = 4(e480-e750)va/vb, while the concentration of chlorophyll a was estimated with the formula: chl a (µg ml-1) = 11.236(a665-a750)va/vb, derived from the factor by strickland and parsons (1972), where va – extract volume (ml) and vb – sample volume (ml). �e e�ects of par and temperature on the cell concentration and pigment content were examined by two-way analysis of variance method (anova) at a signi�cance level of p < 0.05, and statistical calculations were performed using the statistica® 13.1 program. results �e concentration of cells of investigated synechococcus strains were signi�cantly (p < 0.05) a�ected by intensity and temperature (fig. 1). �e highest cell concentrations for strains ba-124 and ba-132 occurred in the highest light intensity (280 µmol m-2 s-1) and the highest temperature (25°c), and they were about 15.8-fold and 5.1fold, respectively higher than the concentrations in the lowest light intensity of 10 µmol m-2 s-1 and 10°c. for ba-120, it was found that the highest cell concentrations occurred in the scenario of 190 µmol m-2 s-1 and the 25°c. comparing the results derived within all experiments, the highest cell concentration (36·106 cell ml-1) was noted in for ba-124 in the scenario mentioned above (280 µmol m-2 s-1, 25°c), and the lowest cell concentration was observed for ba-120 (8·106 cell ml-1) in 280 µmol m-2 s-1, 10°c (fig. 1). �e other striking observation is that the abundance of ba-120 in the highest par is lower than the abundance in 190 µmol m-2 s-1. �is tendency repeats for each temperature level (fig. 1a). in general, for the three strains of synechococcus, the cell-speci�c pigmentation was negatively a�ected by high par (fig. 2). �e highest concentration of pigments were observed at the lowest light intensity (10 µmol m-2 s-1) and high temperature (25°c), except for chl a and car in ba-124 cells, where the highest pigments content was noted at 10 µmol m-2 s-1 and 10°c. �e cell-speci�c composition of pigments content was di�erent for di�erent strains. �e highest values of chl a and car were observed in the red strain ba-120 (0.23 pg cell-1 and 0.09 pg cell-1, respectively); whereas, in the green strain ba-124, the maximum value of this pigments was about 0.10 pg cell-1 and 0.07 pg cell-1, respectively. the im pact of tem perature and photosynthetically active radiation on the grow th and pigm ents concentration in b altic picocyanobacterium synechococcus sp. s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 62 discussion according to the present results, par and temperature are important factors controlling the growth of picocyanobacteria. �e signi�cance of light and temperature was also emphasised by jasser and arvola (2003), who pointed to the impact of these factors on the abundance and also the distribution of picocyanobacteria in the water medium. moreover, following some already published research, it can be claimed that temperature and par may determine the abundance of the marine synechococcus community (jasser, arvola, 2003; jasser, 2006; flombaum et al., 2013; jodłowska, śliwińska, 2014). fig. 1. cell number of three strains of synechococcus: ba-120 (a), ba-124 (b) and ba-132 (c) on day 7 of culturing at four par levels and at four temperatures. values are means (± sd); n = 3 of independent replicates. �e scales in the sub-�gures were prepared di�erently for di�erent strains. �is was done intentionally in order to facilitate reading the values in sub-�gs a and c 63 in this study, it was found that elevated par and temperature have generally a positive e�ect on cell concentration for synechococcus sp. however, the adaptation to very low-light and low-temperature conditions are also known for picoplanktonic organisms. for instance, ibelings (1996) noted that picocyanobacteria can bene�t from low light intensity reaching high growth rates. moreover, this statement is consistent with the observations of picocyanobacterial high abundance at (or near) the end of the euphotic zone in coastal and o�shore marine waters (callieri et al., 2005; callieri, fig. 2. �e photosynthetic pigment content (pg cell-1): a – chlorophyll a, b – carotenoids for three stains of synechococcus: ba-120 (a), ba-124 (b) and ba-132 (c) on day 7 of culturing at four par levels and at four temperatures. values are means (± sd); n = 3 of independent replicates the im pact of tem perature and photosynthetically active radiation on the grow th and pigm ents concentration in b altic picocyanobacterium synechococcus sp. s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 64 2010). additionally, under culture conditions, some major picoplanktons demonstrated the ability to survive and resume growth a�er periods of total darkness. such a pronounced capacity for survival in the dark would enable these picoplanktonic organisms to survive the seasonal rhythm of winter darkness and sinking into the aphotic zone (antia, 1976). on the other hand, kana and glibert (1987a,b) showed that synechococcus wh7803 could also grow at an intensity as high as 2000 μmol m-2 s-1, but only if the cultivation of the strain was preceded by the acclimation in several intermediate intensities. despite showing similar general growth responses to changeable environmental conditions, each strain also presents its speci�city. for instance, a kind of growth saturation was measured for ba-120 between 190 and 280 μmol m-2 s-1 at each temperature level, where the abundance decreased along with par increase (fig. 1a). moreover, the abundance functions dependent on par and temperature are di�erent for di�erent strains. for example, for both ba-124 and ba-132 par and temperature have a positive impact on the abundance in the whole domain but the plot lines di�er (fig. 1b–c). surface and near-surface populations experience extremely variable light and temperature conditions (millie et al., 1990), and intensity is an important factor that a�ects the composition of photosynthetic pigments (prézelin, 1981). �e results obtained for the two strains of synechococcus ba-120 and ba-132 indicated that high intensity and low temperature had a generally negative e�ect on chl a cell concentration. for ba-124, the maximum value of chl a content was noted at the lowest light and temperature. �e green strain ba-124 is rich in pc; whereas, the other two strains, red ba-120 and brown ba-132, have pe as the dominating photosynthetic pigment. picocyanobacteria with a high concentration of pc are chromatically better adapted to harvest longer wavelengths of par than those with pe as a dominating pigment. �erefore, such picocyanobacteria (with pc) usually dominate in surface euphotic waters. on the other hand, the strains rich in pe usually occur deeper (callieri, 2010). moreover, picocyanobacteria, thanks to their high concentration of photosynthetic pigments, may occur in low light intensity waters (stal et al., 2003). �e above is consistent with derived results, which showed that ba-120 and ba-132 are characterised by higher a pigment composition than ba-124. ba-120 and ba-132 are these strains that are more adapted to live in deeper parts of the water column than ba-124. note that changes in the pigment content and in the ratio of di�erent pigments results in the optimisation of photosynthetic e�ciency (defew et al., 2004). in the two strains of synechococcus (ba-120 and ba-132), the highest car content were observed at the lowest par and the highest temperature, whilst the highest car content in ba-124 was measured at the lowest par and temperature. carotenoids have a dual role in the cell, which is to maintain a high capacity for photosynthetic light absorption and to provide protection against photooxidation. �is feature ad65 ditionally explains why synechococcus sp. is able to grow successfully both in the surface layer and in deeper waters (stal, walsby, 2000; stal et al., 2003; jodłowska, śliwińska, 2014). �e experiments on synechococcus strains demonstrated their tolerance to elevated light levels and temperatures. �ese strains were able to change the composition of photosynthetic pigments to use light quanta better and to protect themselves from unfavourable environmental factors. �e ability of synechococcus to sustain their growth in low light and temperature conditions and their lack (green and brown strain) or low photoinhibition (red strain) in exposure to high light intensities and high temperatures could give picocyanobacteria an advantage in changeable aquatic ecosystems. �is study indicates the need to conduct further research on synechococcus sp. as a whole and separately for each strain of this genus. only dedicated studies will bring detailed information about these microorganisms living not only in the baltic sea. acknowledgements �e authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. �is study was supported by bmn grants, poland, no. 538-g245-b568-17. references agawin, n.s., duarte, c.m., agusti, s. 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(1988). phototrophic picoplankton: an overview from marine and freshwater ecosystems. limnology and oceanography, 33, 765–775. doi: 10.4319/lo.1988.33.4part2.0765 strickland, i.d.h., parsons, t.r. (1972). a practical handbook of seawater analysis. journal of the fisheries research board of canada, 167, 1–310. abstract �e experiments on three baltic picocyanobacterial strains of synechococcus (ba-120 – red strain, ba-124 – green strain and ba-132 – brown strain) were conducted at four scalar irradiances in photosynthetically active radiation (par) and four temperature levels. �e main aim of this work was to estimate the e�ect of environmental conditions (temperature and par) on cell concentration and photosynthetic pigments (chlorophyll a – chl a and carotenoids – car) contents. �e ranges 67 of par and temperatures were 10, 100, 190, 280 μmol m-2 s-1 and 10, 15, 20, 25°c, respectively. �e experiment was carried in a medium of salinity of 8. �e number of synechococcus sp. cells was determined using a bd accuri™ c6 �ow cytometer. �e pigments contents were determined by a spectrophotometric method. in this work, it was found that elevated intensity and temperature have, on average, a positive e�ect on cell concentration for synechococcus sp. �e highest cells concentrations were noted at the highest par (280 µmol m-2 s-1) and the highest t (25°c) for green and brown strains (ba-124 and ba-132, respectively) and at 190 µmol m-2 s-1 and 25°c for red strain (ba-120). comparing the strains at each par level and temperature, the highest cell concentration was noted in green strain (36·106 cell ml-1), while the lowest was observed in red strain (8·106 cell ml-1). in general, in the two strains of synechococcus (ba-120 and ba-132), the highest car and chl a contents were observed at the lowest light intensity and the highest temperature. on the other hand, car and chl a maximum content in ba-124 were noted at the lowest light and temperature. �e experiments on synechococcus strains demonstrated their high capacity to acclimate to a wide range of par and temperature levels. �e three strains of synechococcus showed adaptation capabilities, since they were able to change the composition of their photosynthetic pigments to use light quantity better and to protect the cells from the unfavourable e�ect of elevated light and temperature. key words: picocyanobacteria, growth, par, photosynthetic pigments, temperature received: [2017.07.14] accepted: [2017.09.22] wpływ temperatury i natężenia światła na wzrost i zawartość barwników fotosyntetycznych u bałtyckiej pikoplanktonowej sinicy synechococcus sp. streszczenie badania przeprowadzono na trzech szczepach bałtyckiej pikoplanktonowej sinicy z rodzaju synechococcus (ba-120 – szczep czerwony, ba-124 – szczep zielony oraz ba-132 – szczep brązowy), które hodowano w czterech różnych warunkach oświetlenia fotosyntetycznie aktywnego (par): 10, 100, 190, 280 μmol m-2 s-1 oraz w czterech temperaturach: 10, 15, 20 i 25°c. głównym celem pracy było określenie wpływu wybranych zakresów par i temperatury na wzrost komórek oraz zawartość barwników fotosyntetycznych u analizowanych szczepów sinicy. eksperyment przeprowadzono w medium o stałym zasoleniu równym 8. liczebność komórek określana była za pomocą cytometru przepływowego bd accuri™ c6. pomiar oraz estymację zawartości barwników fotosyntetycznych określono metodą spektrofotometryczną. na podstawie uzyskanych danych wykazano, że wysoka intensywność światła oraz wysoka temperatura mają na ogół pozytywny wpływ na liczebność komórek synechococcus sp. stwierdzono, że szczep zielony (ba-124) oraz szczep brązowy (ba-132) osiągnęły największą liczebność komórek w najwyższym oświetleniu (280 µmol m-2 s-1) i najwyższej temperaturze (25°c). szczep czerwony ba-120 wykazał najwyższy wzrost w 190 µmol m-2 s-1 i 25°c. ponadto zaobserwowano, że szczep zielony posiadał najwyższą liczebność komórek, która po tygodniu eksperymentu wyniosła 36·106 kom. ml-1, natomiast szczep czerwony był najmniej liczny (8·106 kom. ml-1). ogólnie, szczepy ba-120 oraz ba-132 najwyższą zawartość chloro�lu a oraz barwników karotenoidowych wykazały w najniższym oświetleniu i najwyższej temperaturze. maksymalną zawartość tych barwników szczep ba-124 osiągnął w najniższej temperaturze oraz natężeniu światła. na podstawie przeprowadzonych doświadczeń wykazano wysoką aklimatyzację szczepów synechococcus sp. do szerokiego zakresu natężenia par oraz temperatury. wszystkie badane szczepy pikoplanktonowej sinicy były zdolne do zmiany proporcji swoich barwników fotosyntetycznych. tego rodzaju umiejętność pozwala im lepiej wykorzystywać dostępne promieniowanie w procesie fotosyntezy oraz daje ochronę w przypadku ekspozycji na niekorzystne warunki środowiskowe. słowa kluczowe: pikoplanktonowe sinice, wzrost, par, barwniki fotosyntetyczne, temperatura the im pact of tem perature and photosynthetically active radiation on the grow th and pigm ents concentration in b altic picocyanobacterium synechococcus sp. s yl w ia ś liw iń sk aw ilc ze w sk a, a ga ta c ie sz yń sk a, a da m l at ał a 68 information on the authors sylwia śliwińska-wilczewska she is interested in allelopathy of cyanobacteria and microalgae, in particular, of picocyanobacteria synechococcus sp. her study includes the in�uence of allelochemicals on the growth, chlorophyll �uorescence, and photosynthesis irradiance curves of di�erent phytoplankton species. she is investigating what in�uences environmental factors have on produced allelopathic compounds on algae and cyanobacteria. agata cieszyńska she is focusing on improving the understanding of the in�uence of di�erent environmental conditions on phytoplankton blooms in the baltic sea. she is working on available data-bases and numerical models. presently, she is working on the determination of the impact of ambient environment on picocyanobacteria growth on the basis of previously arranged laboratory experiments. additionally, she is using the laboratory results to develop a numerical algorithm for the baltic picocyanobacteria life cycle. adam latała wide experience in ecophysiology and ecotoxicology of marine benthic and planktonic algae. in�uence of the main environmental factors such as salinity, temperature and light on the photosynthesis, photoacclimation, �uorescence, respiration and growth of algae from natural communities and cultured under laboratory conditions. use of �uorescence techniques to determine algal and cyanobacterial ecophysiology and ecotoxicology. 129 annales universitatis paedagogicae cracoviensis studia naturae, 1: 129–143, 2016, issn 2543-8832 anatoliy a. khapugin1,2*, tatyana b. silaeva2, yulia n. utorova2 1 mordovia state nature reserve, republic of mordovia, temnikov district, pushta, russia 2 mordovia state university, republic of mordovia, saransk, russia, *hapugin88@yandex.ru three maples (acer l., aceraceae juss.) in the republic of mordovia, russian federation introduction genus acer l. (maple) includes a large number of species distributed worldwide, but especially in the northern hemisphere. maples are deciduous trees from 12 m to 35 m in height, rarely evergreen or sometimes shrubs (morselli, 1989). classically this genus is considered belonging to the aceraceae juss. family (dicotyledoneae) (takhtajan, 1987; cronquist, 1988; cherepanov, 1995). however, in recent time many studies using pollen morphology (müller, leenhouts, 1976), biochemistry of plants (umadevi, daniel, 1991) and molecular sequence data (gadek et al., 1996; savolainen et al., 2000; apg iii, 2009; buerki et al., 2009) con�rm inclusion of this genus into the sapindaceae s. lato. �ere are up to 148 wild or cultivated acer species widely distributed throughout north america, eurasia and north africa (olson et al., 1974). republic of mordovia is located in central russia. its territory lies on the border of the forest and forest-steppe zones in central russia. eastern part of the republic of mordovia covers the north-west of the volga upland, and its western part is located on the west of the oka-don lowland. �erefore, high habitats diversity is observed within this area. coniferous and mixed forests are distributed in the west and northwest mordovia. broad-leaved forests are located in the central and eastern parts of the region. forest-steppe landscapes dominate in the east and south-east of mordovia (yamashkin, 1998, 2012). within mordovia four maple species are known. a. platanoides l. is widely distributed species which inhabits plant communities of broad-leaved and mixed forests (silaeva et al., 2010). a. campestre l. (field maple) is vulnerable maple species which is on the north-eastern border of its range in the republic of mordovia (utorova et al., 2014). it is known in six districts (insar, kadoshkino, kovylkino, kochkurovo, 130 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va and ruzaevka district, as well as in neighborhoods of saransk). a. campestre is included in the regional red data book (resolution of the government…, 2015). a. tataricum l. (tatarian maple) is species distributed in mordovia sparsely, as well as in adjacent regions. it is known predominantly within �oodplain broad-leaved forests from 13 districts of the republic of mordovia. �ere is a lack of data on the favorable environmental conditions for a. tataricum existing. a. negundo l. (ash-leaved maple) is alien species known in all districts of mordovia (silaeva et al., 2010), as well as in all adjacent regions. �is tree species is aggressive invasive plant included in the black data book of central russia (vinogradova et al., 2010). better knowledge of ecology and biology of a. negundo is an important task in limiting its penetration into natural ecosystems within secondary range. we aimed to investigate three maple species (acer campestre, a. tataricum, a. negundo) in conditions of central russia on example of the republic of mordovia due to the lack of data on the ecology of these species in european part of russia. material and methods �e �eld investigations were carried out following aleksandrova (1964) and khapugin et al. (2014). for a. campestre and a. tataricum, ecological conditions of habitats have been evaluated using the data on the ecological preferences of vascular plant species which grow with maples together. distribution of plant species of the accompanying �ora on the ecological groups in relation to water has been carried out according to the classi�cation of shennikov (1950) with separation of main groups (hygrophytes, mesophytes, xerophytes) and intermediate groups (hygromesophytes, mesohygrophytes, xeromesophytes, mesoxerophytes). ecological scale proposed by tsyganov (1983) has been used in distribution of plant species of the accompanying �ora on the ecological groups in relation to the lighting/shading. �e �eld investigations of a. campestre populations were carried out in three locations of kadoshkino district of mordovia: adashevo (53.925 n, 44.377 e), latyshovka (53.961 n, 44.384 e) and insar station (54.060 n, 44.310 e) in years 2013–2015 (fig. 1). within all locations, we established square plots (10×10 m) to investigate the accompanying �ora and determine the forest’s stand formula. forest’s stand formula has been determined as percent of individuals of forest stand in the canopy layer per 100 m2 (area of one square plot). in adashevo location and insar station the total number of a. campestre individuals was counted. in latyshovka we established close to each other 2 square plots (10×10 m) (latyshovka 1 and latyshovka 2) to investigate the number of 1-year-old a. campestre seedlings (seed reproduction). for this purpose, we established 10 study plots (1×1 m) within each of large (10×10 m) established plots. in insar station we have counted number of age-bearing a. campestre individuals per 100 m2. 131 �e �eld investigations of a. tataricum populations were carried out in bolshie berezniki district of mordovia (54.176 n, 46.181 e) in years 2014–2015 (fig. 1). we established four square plots (10×10 m) in di�erent habitats: �oodplain meadow (plot  1), lime-pine forest (plot 2), �oodplain broad-leaved forest (plot 3), and plant community with a. negundo dominance (plot 4). we investigated the number of a. tataricum individuals per established plot, composition of accompanying �ora; forest stand formula was determined for each of studied plant communities. on the basis of the vascular �ora of herb and shrub layers of studied plant communities, we calculated weighted mean (diekmann, 2003) of environmental indicator values for several ecological factors (light, temperature, continentality, moisture, reaction ph, nutrient) according to ellenberg et al. (2001). in this calculation, rij is the response of species i in sample plot j, and xi is the indicator value of species i. �en, weighted mean of all values of those plant species presented in the plot was calculated to estimate the environmental indicator values for factors: weighted mean = �e �eld investigation of a. negundo populations were carried out in bolshie berezniki district of mordovia (54.176 n, 46.181 e) and on wasteland in saransk (54.164 n, 45.150 e) in years 2014–2015 (fig. 1). within each of these localities, we fig. 1. allocation of studied localities with acer species in the republic of mordovia. symbols: rhombs – localities with acer campestre in kadoshkino district: 1 – insar station, 2 – latyshovka, 3 – adashevo; 4 (circle) – locality with acer tataricum and acer negundo populations in bolshie berezniki district; 5 (square) – locality with acer negundo in saransk three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 132 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va established 2 large square plots (5×5 m) to investigate the number of 1-year-old a. negundo seedlings (seed reproduction). for this purpose, we established 10 study plots (1×1 m) within each of large (25 m2) established plots. �e �eld investigations of a. negundo seed reproduction were carried out in established plot no. 4 for a. tataricum study (see above). �e �eld investigations of a. negundo seed reproduction were carried out in the abandoned wasteland with ruderal vegetation in saransk. statistical analysis was performed in ms excel and past (hammer et al., 2001). results and discussion acer campestre l. – field maple as the result of investigations, we found that number of a. campestre individuals per 1 plot in adashevo widely varied from 8 to 115 di�erent-age plants (240 in total). in insar locality, this parameter was less variable – from 12 to 53 di�erent-age individuals (163 in total). mean values varied from 32.2 to 40 individuals per 1 established plot (tab. 1). tab. 1. number of acer campestre individuals per 1 established plot parameter locality adashevo insar station m 40.00 32.20 m 15.80 7.30 min 8.00 12.00 max 115.00 53.00 note: m – mean value, m – error of the mean, min – minimal value, max – maximal value investigations of the number of a. campestre 1-year-old individuals in latyshovka showed that this parameter varied from 8 (latyshovka 1) to 31 (latyshovka 2) individuals per 1 established plot (tab. 2). in average the number of a. campestre 1-year-old individuals varied from 12 to 19.4. tab. 2. number of 1-year-old individuals of acer campestre per 1 established plot parameter locality latyshovka 1 latyshovka 2 m 13.00 19.40 m 2.60 3.30 min 8.00 12.00 max 22.00 31.00 note: m – mean value, m – error of the mean, min – minimal value, max – maximal value 133 however, seed reproduction is of little importance in maintaining of a. campestre populations in mordovia since very few individuals of �eld maple reach the generative age (utorova et al., 2014). �is suggest with our results: only 11.7% (19 of 163) bearing-age individuals of a. campestre were found within established plots in the insar station locality (tab. 3). tab. 3. characteristics of bearing-age individuals in acer campestre population in the insar locality number of plots number of bearing-age individuals (percentage of total number of individuals [%]) average tree girth [cm] average tree diameter on the breast height [cm] 1 2 (5.10) 21.00 5.90 2 4 (10.50) 10.00 4.20 3 4 (33.30) 8.40 3.10 4 7 (13.20) 15.60 8.20 5 2 (9.50) 6.40 2.20 �e lack of generative individuals in a. campestre populations is completely o�set by an active vegetative propagation by root o�springs. as a result, the group arrangement of young individuals of a. campestre is observed around the parent individual (fig. 2). ecological conditions of certain habitats with a. campestre could be assessed on the basis of ecological preferences of plant species of �ora accompanying to this species. table 4 includes data on the distribution of plant species of accompanying �ora on ecological groups in relation of plants to water. fig. 2. spatial arrangement of acer campestre individuals amongst woody plant species at established plots 1 and 5. symbols: – acer campestre,  – acer platanoides,  – fraxinus excelsior l.,  – tilia cordata mill., + – euonymus verrucosa scop., m – quercus robur l., t– lonicera xylosteum l., × – sorbus aucuparia l., – – corylus avellana l. three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 134 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va table 4 shows that plant community with a. campestre has been formed in conditions of su�cient moisture. such conditions are typical to the deciduous forests in the river valleys. as seen from table 4, mesophytes group includes the highest percentage of species (53.7%). among them, there are a. platanoides, lathyrus vernus (l.) bernh., stellaria graminea l. and others. other ecological groups include less number of species. tab. 4. ecological groups on the basis of relation of plants to water in plant community with acer campestre ecological group number of species total number of species [%] mesophytes 36 53.70 hygrophytes 9 13.40 xeromesophytes 8 11.90 mesohygrophytes 7 10.40 mesoxerophytes 6 8.90 hygrophytes 1 1.40 total 67 100.00 table 5 includes data on the distribution of plant species of the accompanying �ora on the ecological groups in relation of plants to the lighting/shading. as seen in table 5, the group of shade-tolerant species dominates (65.6%). it includes such species as vicia sylvatica l., milium e�usum l., lonicera xylosteum l. and others. it indicates the signi�cant shading in plant communities with a. campestre. however, group of light-demanding species includes 29.8% plants of the accompanying �ora. such situation is typical for deciduous forest exposed to anthropogenic pressure (burova, feklistov, 2007). tab. 5. ecological groups of plants in relation to the lighting/shading in plant community with acer campestre ecological group number of species total number of species [%] shade-tolerant 44 65.60 light-demanding 20 29.80 shade-demanding 3 4.40 total 67 100.00 an ecological-coenotical characteristic of the �ora, which is accompanying a. campestre, is presented in table 6 with distributing of plant species on several ecological-coenotical groups. 135 tab. 6. ecological-coenotical characteristics of the �ora which is accompanying acer campestre in the investigated plant communities ecological-coenotical group number of species total number of species [%] forest 43 63.80 meadow 16 23.70 weed 6 8.90 forest swamp 2 2.80 total 67 100.00 species of forest group are predominant (63.8% of total species number) in the accompanying �ora of investigated plant community with a. campestre. among them, there are asarum europaeum l., galium odoratum (l.) scop., platanthera bifolia (l.) rich. and others. group of meadow species is located on the second place with 16 species (23.7%). among them, there are campanula patula l., stellaria graminea, ranunculus acris l. and others. presence of meadow species indicates penetration of these plants through forest roads, cuttings, forest edges and clearings. �ey also contribute to the penetration weed plants (6 species; 8.9%) into the plant community with a. campestre. as a whole, the plant community with a. campestre is a typical broadleaved forest, being under anthropogenic in�uence. acer tataricum l. – tatarian maple we investigated accompanying �ora of each of 4 established plots in bolshie berezniki district. established plot 1 was �oodplain meadow plant community with high abundance of fragaria viridis l. (19%), alopecurus pratensis l. (8%), and carex praecox schreb. (6%). established plot was located in lime-pine forest where urtica dioica l. (5%), pteridium aquilinum (l.) kuhn. (4%), dryopteris carthusiana (vill.) h.p. fuchs (3%), stellaria media l. (3%) are dominant in herb layer; shrub layer was represented by rosa cinnamomea l., euonymus verrucosa, rubus idaeus l., ribes nigrum l. established plot 3 was located in �oodplain lime-oak forest with dominance of aegopodium podagraria l. (7%), glechoma hederacea l. (5%) in herb layer. established plot 4 was located in �oodplain plant community formed by invasive a. negundo representing about 80% of forest stand; urtica dioica (15%) and glechoma hederacea (7%) are dominant in herb layer; shrub layer is represented by padus avium mill. and a. negundo. vegetation projective cover varied from 35% (lime–oak) to 80% (�oodplain meadow) (tab. 7). however, the number of a. tataricum individuals was not determined by this parameter. �e highest number of tatarian maple individuals was observed in �oodplain lime–oak forest. perhaps this can be explained by the fact that �oodplain deciduous forest is a typical habitat for a. tataricum. �e lowest number of tatarian maple individuals was found in �oodplain forest with dominance of a. negundo (tab. 7). three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 136 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va �is invasive tree species is capable of forming multilevel plant communities with high density of the forest canopy oppressing native species (kostina et al., 2016). investigated plant community was probably formed in place of well-moistened forest edge or meadow. �is is consistent with presence of meadow and forest edge species (filipendula vulgaris moench, rumex confertus willd.) and plants of deciduous forests (glechoma hederacea, angelica sylvestris l.) in this seminatural plant community. tab. 7. characteristics of established plots with acer tataricum parameter plots 1 2 3 4 projective cover [%] 80.00 35.00 65.00 65.00 number of acer tataricum individuals per plot 32.00 28.00 38.00 26.00 light 7.00 5.10 5.50 6.10 temperature 5.70 5.30 5.20 5.50 continentality 4.50 4.40 4.30 3.70 moisture 5.00 5.70 5.50 6.40 reaction 6.80 6.10 6.70 7.00 nutrient 5.20 5.30 5.80 6.60 forest stand composition [%] – 70ps20tc10qr+ap 50qr40tc10pt+ul 80an20at+ul+qr note: an – acer negundo, ap – acer platanoides, at – acer tataricum, ps – pinus sylvestris l., tc – tilia cordata, qr – quercus robur, pt – populus tremula l., ul – ulmus laevis pall �e analysis of the vascular �ora of herb and shrub layers of studied plant communities was carried out according to ellenberg et al. (2001). detrended correspondfig. 3. detrended correspondence analysis (dca) ordination diagram of plots established in plant communities with acer tataricum 137 ence analysis of total set of environmental indicator values (fig. 3) shows that the lowest number of a. tataricum individuals were found in the most moist and shaded plant communities (established plots 2 and 4). �us, the excessive moisture of habitat adversely a�ects the seed reproduction and development of a. tataricum seedlings, despite the fact that tatarian maple is a species of �oodplain broadleaved forests. �e highest number of a. tataricum individuals was found under conditions of su�cient light and moderate moisture (established plot 1). �us, light and moisture are the most signi�cant environmental factors for a. tataricum populations in the republic of mordovia. we selected 25 a. tataricum samaras per each established plot to reveal its morphological characteristics. as it is seen from table 8, samara of a. tataricum reached 3.7 cm in length and 0.9 cm in width as a mean. tab. 8. characteristics of acer tataricum samaras parameter length of samara [cm] width of samara [cm] м 3.70 0.90 m 0.30 0.30 min 2.80 0.40 max 4.20 2.00 note: m – mean value, m – error of the mean, min – minimal value, max – maximal value we analysed ecological preferences of plant species of �ora accompanying to a. tataricum to estimate ecological conditions of habitats with tatarian maple. table 9 includes data on the distribution of plant species of accompanying �ora of ecological groups in relation of plants to water. �e mesophytes group dominates with 52 species (51.4%) which are typical for habitats with moderate moisture level. among them, there are fragaria vesca, alopecurus pratensis, quercus robur and others. xeromesophytes (18.8% of total species number) are located on the second place. most of them were found on the established plot 1 located in meadow habitat. among them, there are festuca rubra l., elymus (=elytrigia) repens (l.) gould, bromus arvensis l. and others. plants con�ned to habitats with excessive moisture (hygrophytes, hygromesophytes) were presented by 14 species in sum. �ey include urtica dioica, athyrium �lix-femina (l.) roth, lysimachia nummularia l., l. vulgaris l., filipendula ulmaria (l.) maxim. and others. table 10 includes data on the distribution of plant species of the accompanying �ora of the ecological groups in relation of plants to the lighting/shading. as it is seen from table 10, both light-demanding and shade-tolerant groups contain approximately equal number of species. �is is explained by the fact, that a. tataricum is capable to grow both in open, as well as in woodland habitats. three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 138 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va tab. 9. ecological groups on the basis of relation of plants to water in plant community with acer tataricum ecological group number of species total number of species [%] mesophytes 52 51.40 xeromesophytes 19 18.80 mesohygrophytes 10 9.90 hygrophytes 9 8.90 hygromesophytes 5 4.90 mesoxerophytes 5 4.90 xerophytes 1 0.90 total 101 100.00 tab. 10. ecological groups of plants in relation to the lighting/shading in plant community with acer tataricum ecological group number of species total number of species [%] shade-tolerant 46 45.50 light-demanding 45 44.50 shade-demanding 10 9.90 total 101 100.00 tab. 11. ecological-coenotical characteristics of the �ora which is accompanying acer tataricum in the investigated plant communities ecological-coenotical group number of species total number of species [%] forest 51 50.10 meadow 24 23.40 weed 15 14.70 forest swamp 10 9.70 steppe 1 0.90 total 101 100.00 an ecological-coenotical characteristic of the �ora accompanying a. tataricum is presented in table 11 with distribution of plant species on several ecological-coenotical groups. forest group contains the highest number of species (50.1%). among them, there are asarum europaeum, galium odoratum, platanthera bifolia, rubus saxatilis l., scrophularia nodosa l. and others. second place is occupied by group of meadow plants (24 species) that indicates ability of a. tataricum to grow in �oodplain meadows. among them, there are betonica o�cinalis l., carex vulpina l., agrostis capillaris l. and others. �e presence of weed species (14.7%) in accompanying �ora indicates disturbance of habitats with tatarian maple. among them, there are arctium lappa l., taraxacum o�cinale wigg. s.l., polygonum convolvulus l. and others. �us, 139 a. tataricum populations are capable to grow in open, as well as in woodland habitats of �oodplain including under anthropogenic pressure. acer negundo l. – ash-leaved maple seed reproduction of a. negundo was determined within the established plot 4 to study a. tataricum populations (study plots 1, 2), as well as in saransk at wasteland overgrown by ash-leaved maple (study plots 3, 4). brief description of established plot in bolshie berezniki district of mordovia was listed above in section about a. tataricum. vegetation cover of wasteland in saransk is presented by ruderal plant species. among them, conyza (=erigeron) canadensis (l.) cronquist, arctium lappa, lactuca serriola l., artemisia vulgaris l., taraxacum o�cinale are dominant. as a result of investigations, we found that seed reproduction of a. negundo was signi�cantly higher in seminatural plant community with dominance of its invasive species. although there is no shading that inhibits a. negundo juveniles (kostina et al., 2016), number of 1-year-old individuals of ash-leaved maple in saransk was less in 1.7–2.6 time in comparison with habitat in bolshie berezniki district of mordovia (tab. 12). probably, more dry conditions of wasteland in saransk decrease the seed germination of a. negundo that is consistent with results of other researchers (erfmeier et al., 2011; lamarque et al., 2013). tab. 12. number of 1-year-old individuals of acer negundo per study plot in saransk and in bolshie berezniki district of mordovia parameter bolshie berezniki district saransk 1 2 3 4 м 66.00 71.00 27.00 39.00 m 8.60 12.50 4.80 9.10 min 43.00 36.00 14.00 18.00 max 95.00 108.00 42.00 67.00 note: m – mean value, m – error of the mean, min – minimal value, max – maximal value conclusions acer campestre populations have relatively low level of seed reproduction (13.0–19.4 of 1-year-old individuals per 100 m2). investigation of the number of generative individuals shows that only 11.7% of plants reach generative age. active vegetative propagation of a. campestre suggests maintaining the �eld maple population on the north-eastern border of its range. as a result, group arrangement of a. campestre individuals is observed. analysis of the accompanying �ora indicates that a. campestre is con�ned to �oodplain broadleaved forests and it capable to grow under moderate disturbance of habitats. three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 140 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va a. tataricum is capable to grow both in open (meadows, forest edges, banks of water bodies) and in woodland habitats in �oodplains. maintenance and development of a. tataricum populations are provided primarily by a su�cient amount of light and moisture. �e shading, appearing under the canopy of some tree plants (e.g. a. negundo forest stand), has depressing e�ect on a. tataricum seedlings development. �e analysis of the accompanying �ora re�ects the ecological-coenotical con�nement of a. tataricum to plant communities of both forests and meadows in �oodplains. participation of weed species (14.7%) in accompanying �ora indicates the ability of tatarian maple to develop in seminatural habitats. signi�cantly smaller number of a. negundo seedlings (in 1.7–2.6 times) in the urban environment can indicate initial stage of ash-leaved maple invading in this habitat. in opposition, in seminatural plant community with a. negundo dominance, its seeds reproduction has a large level, probably due to indirect facilitation of the adult trees for the growth of its own seedlings by suppressing of native plants in this habitat. �is is consistent with results of investigations of this phenomenon in relation to exactly a. negundo (saccone et al., 2010), as well as other plant species (e.g. siemann, rogers, 2003). obtained results about interactions of several maple species (a. tataricum and a. negundo) show the relevance of these studies in relation to study of closely related tree species in conditions of their joint existence. references aleksandrova, v.d. 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(2010). acer negundo invasion along a successional gradient: early direct faciliation by native pioneers and late indirect facilitation by conspeci�cs. new phytologist, 187, 831–842. doi: 10.1111/j.1469-8137.2010.03289.x savolainen, v., fay, m.f., albach, d.c., backlund, a., van der bank, m., cameron, k.m., johnson, s.a., lledó, m.d., pintaud, j.-c., powell, m., sheahan, m.c., soltis, d.e., soltis, p.s., weston, p., whitten, w.m., wurdack, k.j., chase, m.w. (2000). phylogeny of the eudicots: a newly complete familial analysis based on rbcl gene sequences. kew bulletin, 55, 257–309. doi: 10.2307/4115644 shennikov, a.p. (1950). ecology of plants. moscow: sovetskaya nauka. [in russian] siemann, e., rogers, w.e. (2003). changes in light and nitrogen availability under pioneer trees may indirectly faciliate tree invasions of grasslands. journal of ecology, 91, 923–931. doi: 10.1046/j.13652745.2003.00822.x silaeva, t.b. kiryukhin, i.v., chugunov, g.g., levin, v.k., mayorov, s.r., pismarkina, e.v., ageeva, a.m., vargot, e.v. (2010). vascular plants of the republic of mordovia (synopsis of �ora). saransk: publisher of the mordovia state university. [in russian] takhtajan, a.l. (1987). system of magnoliophyta. leningrad: academy of sciences. [in russian] tsyganov, d.n. (1983). phytoindication of ecological regimes in the mixed coniferous-broad-leaved forest subzone. moscow: nauka. [in russian] three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 142 a na to liy a . k ha pu gi n, t at ya na b . s ila ev a, y ul ia n . u to ro va umadevi, i., daniel, m. (1991). chemosystematics of the sapindaceae. feddes repertorium, 102, 607– 612. doi: 10.1002/fedr.19911020711 utorova, yu.n., khapugin, a.a., silaeva, t.b. (2014). about ecology of acer campestre l. (aceraceae) on north-eastern limit of the range. environment and ecology research, 2(1), 8–13. doi: 10.13189/ eer.2014.020102 vinogradova, yu.k., mayorov, s.r., khoroon, l.v. (2010). �e black book of flora of central russia. alien plant species in ecosystems of central russia. moscow: geos. [in russian] yamashkin, a.a. (1998). physical and geographical conditions and landscapes of the republic mordovia. saransk: publisher of the mordovia state university. [in russian] yamashkin, a.a. (2012). geographical atlas of republic of mordovia. saransk: publisher of the mordovia state university. [in russian] abstract genus acer in the republic of mordovia is presented by four plant species: acer campestre l., a. negundo l., a. platanoides l. and a. tataricum l. for a. campestre and a. tataricum we investigated accompanying �ora composition and conducted its analysis; characteristics of populations were carried out. seed reproduction of a. negundo was investigated in seminatural and anthropogenically disturbed habitats. seed reproduction is of little importance for the maintenance of a. campestre populations on the north-eastern border of its range. �is is o�set by an active vegetative propagation by root o�springs. all in all, very few a. campestre individuals reach the generative age. a. tataricum is capable to grow in open, as well as in woodland habitats in �oodplains. su�cient light and moderate moisture are the most signi�cant environmental factors for a. tataricum populations. depressing of tatarian maple is observed under conditions of shading in plant community with a. negundo dominance. analysis of the accompanying �oras of a. campestre and a. tataricum show the coenotical con�nement of these maple species. seed reproduction of a. negundo was signi�cantly higher in seminatural habitats with dominance of ash-leaved maple than it was in urban environment. probably, this is a manifestation of indirect facilitation of the adult a. negundo tree canopy for the growth of its own seedlings by oppressing other plants (e.g. a. tataricum seedlings). �us, we showed relevance for investigations of interactions between closely related tree species. key words: acer campestre, acer negundo, acer platanoides, acer tataricum, invasive species, population, rare species, republic of mordovia received: [2016.06.23] accepted: [2016.08.22] trzy klony (acer l., aceraceae juss.) w republice mordowii (federacja rosyjska) streszczenie rodzaj klon w republice mordowii reprezentowany jest przez cztery gatunki: acer campestre l., a. negundo l., a. platanoides l. i a. tataricum l. dla gatunków a. campestre i a. tataricum zbadano skład �ory towarzyszącej oraz przeprowadzono charakterystykę populacji. zbadano również reprodukcję nasion a. negundo w siedliskach półnaturalnych i antropogenicznie zaburzonych. na północno-wschodniej granicy zasięgu występowania a. campestre reprodukcja nasion ma niewielkie znaczenie dla utrzymania jego populacji. jest to równoważone przez aktywne rozmnażanie wegetatywne poprzez odrosty korzeniowe. niewiele osobników a. campestre osiąga wiek generatywny. a. tataricum rozwija się, zarówno w  miejscach otwartych, jak i w siedliskach leśnych, na obszarach zalewowych. odpowiednie warunki świetlne i umiarkowana wilgotność są najbardziej istotnymi czynnikami środowiskowymi dla populacji a. tataricum. nieprawidłowo rozwinięte osobniki klonu tatarskiego obserwowano w  warunkach zacienienia, w zbiorowisku roślinnym z dominacją a. negundo. analiza �ory towarzyszącej a. campestre oraz a. tataricum pokazała cenotyczne uzależnienie tych gatunków. reprodukcja nasion a. negundo była znacząco wyższa w półnaturalnych siedliskach z dominacją jesieniolistnych klonów, niż w środowisku miejskim. 143 prawdopodobnie jest to przejaw niebezpośredniego ułatwiania wzrostu siewek a. negundo w starszych drzewostanach poprzez zdominowanie innych roślin (np. siewek a. tataricum). w ten sposób wykazano znaczenie interakcji pomiędzy blisko spokrewnionymi gatunkami drzew. słowa kluczowe: acer campestre, acer negundo, acer platanoides, acer tataricum, gatunki inwazyjne, populacja, gatunki rzadkie, republika mordowii information on the authors anatoliy a. khapugin at present he studies population ecology and biology of rare and endangered plant species in central russia. also, recently he has carried out investigations of pyrogenic successions in forest ecosystems under conditions of southern boundary of the taiga zone. �e results of his investigations are dedicated to biology and ecology of several rare and endangered plants. additionally, he has investigated the biology and ecology of several alien invasive plants species. tatyana b. silaeva her scienti�c studies are dedicated to research of distribution and ecology of rare plant species in central russia and adjacent areas. she works in the �eld of nature conservation. she has also investigated invasion and penetrating of aggressive alien plants species in the republic of mordovia. yulia n. utorova her scienti�c interests are devoted to investigation of acer species in the republic of mordovia. she is interested in their ecology, biology and natural conditions of habitats. three m aples (acer l., a ceraceae juss.) in the r epublic f m ordovia, r ussian federation 175 annales universitatis paedagogicae cracoviensis studia naturae, 3: 175–178, 2018, issn 2543-8832 ii nationwide microbiological scientific conference microbs, organised by foundation for science promotion and development tygiel, was held from the 17th to 18th of may. dwór dwikozy, a manor house near the city of sandomierz, was chosen as a venue for this year’s conference iteration. media patronage was assumed by, among others, biotechnologia.pl and labportal.pl web portals. ms. assoc. prof. anna sikora from institute of biochemistry and biophysics of the polish academy of sciences in warsaw was this year’s guest of honour. the opening ceremony, held by the tygiel foundation representatives along with the guest of honour – phd anna sikora, took place in the forenoon of the 17th of may. after the ceremony, ms. assoc. prof. anna sikora gave the first of two invited lectures entitled “cross-feeding of lactate among bacteria of gastrointestinal tract and hydrogen-yielding microbial communities”. next, a lecture entitled “non-conventional yeast yarrowia lipolytica – valuable source of nutritions” was presented by ms. phd monika jach from department of biotechnology and environmental sciences of the john paul ii catholic university of lublin. w  dniach 17–18 maja 2018 roku, odbyła się iii ogólnopolska mikrobiologiczna konferencja naukowa microbs. na miejsce tegorocznej edycji wybrany został dwór dwikozy, położony w miejscowości o tej samej nazwie, koło sandomierza. organizatorem konferencji była fundacja na rzecz promocji nauki i  rozwoju tygiel. patronat medialny nad tym wydarzeniem objęły, m.in. portale biotechnologia.pl i labportal.pl. gościem honorowym tegorocznej konferencji była pani dr hab. anna sikora z instytutu biochemii i biofizyki polskiej akademii nauk w warszawie. w godzinach przedpołudniowych 17 maja, uroczystego otwarcia konferencji dokonali przedstawiciele fundacji tygiel wraz z  zaproszonym gościem honorowym panią dr hab. anną sikorą, która zaraz po otwarciu wygłosiła jeden z  dwóch proszonych referatów zatytułowany „transformacja mleczanu do maślanu (ang. cross-feeding of lactate) w  przewodzie pokarmowym i  bioreaktorach fermentacji wodorowych”). kolejny referat pt. „niekonwencjonalne drożdże yarrowia lipolytica – cenne źródło składników odżywczych” zaprezentowała pani dr monika jach z  wydziału biotechnologii i  nauk o  środowisku iii nationwide microbiological scientific conference microbs, dwikozy, may 17th–18th, 2018 iii ogólnopolska mikrobiologiczna konferencja naukowa microbs, dwikozy 17–18 maja, 2018 176 r ep or ts after a short coffee break, organisers invited all participants to take part in the scientific session, during which the following lecturers presented their papers: jakub gębalski from the department of pharmacy of the ludwik rydygier collegium medicum in bydgoszcz (“effects of fluoroquinolones, ascorbic acid and rutoside on proteus mirabilis biofilm formation”), arkadiusz gruca from the department of geography and biology of the pedagogical university of cracow (“an effect of bound to surface, fotoactive, zno nanoparticles on chosen bacteria strains”), aleksandra kurowska from the department of biology of the warsaw university (“the influence of heavy metals on the growth of sulfate reducing bacteria”), jana przekwas from the department of pharmacy of collegium medicum of the nicolas copernicus university in toruń (“impact of natural bee-derived products on biofilm formation by proteus mirabilis rods isolated from chronic wounds infections”), zuzanna orwat from the department of environmental and technical sciences of the universitas opoliensis (“influence of plant extract from dionaea muscipula on pathogenic microorganisms”), from the same department, adriana pacia (“antimicrobial activity of pelargonium essential oil on patogenic microorganisms”), and katarzyna kosiorek from the institute of biochemistry and biophysics of the polish academy of sciences in warsaw (“adaptive and industrial potential of lactococcus lactis strains determined by the presence of plasmidic genes”). after a dinner break, organisers invited all participants to take part in a tour of the vineyard nearby. the tour included sightseeing of the “winnica sandomierska (sandkatolickiego uniwersytetu lubelskiego jana pawła ii. tego samego dnia po krótkiej przerwie kawowej, organizatorzy zaprosili uczestników do udziału w sesji naukowej, w której wystąpili m.in. następujący prelegenci: jakub gębalski z  wydziału farmaceutycznego collegium medicum im. ludwika rydygiera w bydgoszczy („wpływ fluorochinolonów, kwasu askorbinowego oraz rutozydu na tworzenie biofilmu przez szczepy proteus mirabilit”), arkadiusz gruca z  wydziału geograficzno-biologicznego uniwersytetu pedagogicznego im. ken w krakowie („wpływ fotoaktywnych, związanych z powierzchnią, nanocząstek zno na wybrane szczepy bakterii”), aleksandra kurowska z  wydziału biologii uniwersytetu warszawskiego („wpływ metali ciężkich na wzrost bakterii redukujących siarczany”), jana przekwas z  wydziału farmaceutycznego collegium medicum uniwersytetu mikołaja kopernika w  toruniu („wpływ naturalnych produktów pochodzenia pszczelego na zdolność tworzenia biofilmu przez pałeczki proteus mirabilis izolowane z  zakażeń ran przewlekłych”), zuzanna orwat z wydziału przyrodniczo-technicznego uniwersytetu opolskiego („oddziaływanie ekstraktu roślinnego z  dionaea muscipula na mikroorganizmy patogenne”), adriana pacia z  tej samej jednostki („działanie pelargoniowego olejku eterycznego na mikroorganizmy patogenne”) oraz katarzyna kosiorek z instytutu biochemii i  biofizyki polskiej akademii nauk w  warszawie („adaptacyjny i  przemysłowy potencjał szczepów lactococcus lactis warunkowany obecnością genów plazmidowych”). po przerwie obiadowej, organizatorzy konferencji zaprosili wszystkich uczestników konferencji na wycieczkę do pobliskiej 177 r eportsomierska vineyard)”, a short vineyard history lecture, and local wine degustation. after their return to dwór dwikozy, participants took part in an integration evening. in the forenoon of the next day, the 18th of may 2018, a poster session took place. the following lecturers presented the results of their work in a form of scientific posters: mariusz cycoń from the department of pharmacy and laboratorial medicine of the medical university of katowice (“microbial degradation of vancomycin in the soil environment”), magdalena zaborowska from the department of environment shaping and agriculture of the university of warmia and mazury in olsztyn (“response of oxidoreductases to contamination of soil with bisphenol a”), paulina cholewińska from the department of biology and animal husbandry of the wrocław university of environmental and life sciences (“an effect of various compositions of the preparations based on nanosilver and mineral sorbents on mesophilic bacteria number in animal origin material”), andrzej jurkowski from the department of biological sciences of the university of zielona góra (“influence of the fe3o4 nanoparticles size on selected culture parameters of probiotic bacteria l. plantarum 299v”), karolina dobrosz from the department of biotechnology and environmental sciences of the john paul ii catholic university of lublin (“map kinases: from yeast to human”), marta michalak from the department of biology and animal husbandry of the of the wrocław university of environmental and life sciences (“influence of nutrition on methanogenesis in cattle”), paulina onopiuk from the department of biotechnology and environmental sciences winnicy. w programie wycieczki znalazły się zwiedzanie „winnicy sandomierskiej”, degustacja wina, a  także krótki wykład o  historii tej winnicy. po zakończeniu wycieczki i  powrocie do dworu dwikozy, uczestnicy wzięli udział w wieczorze integracyjnym. następnego dnia, tj. 18 maja 2018 roku, w  godzinach przedpołudniowych odbyła się sesja posterowa. problematykę swoich badań zaprezentowali tu następujący prelegenci: mariusz cycoń z  wydziału farmaceutycznego z  oddziałem medycyny laboratoryjnej w sosnowcu śląskiego uniwersytetu medycznego w  katowicach („mikrobiologiczny rozkład wankomycyny w środowisku glebowym”), magdalena zaborowska z  wydział kształtowania środowiska i  rolnictwa uniwersytetu warmińsko-mazurskiego w olsztynie („reakcja oksydoreduktaz na zanieczyszczenie gleby bisfenolem a”), paulina cholewińska z  wydziału biologii i hodowli zwierząt uniwersytetu przyrodniczego we wrocławiu („wpływ różnych kompozycji preparatów na bazie nanosrebra i sorbentów mineralnych na kształtowanie się liczby bakterii mezofilnych w  materiale pochodzenia zwierzęcego”), andrzej jurkowski z wydziału nauk biologicznych uniwersytetu zielonogórskiego („wpływ rozmiaru nanocząstek fe3o4 na wybrane parametry hodowli bakterii pro biotycznych l. plantarum 299v”), karolina dobrosz z  wydziału biotechnologii i  nauk o  środowisku katolickiego uniwersytetu lubelskiego jana pawła ii („kinazy map: od drożdży do człowieka”), marta michalak z  wydziału biologii i  hodowli zwierząt uniwersytetu przyrodniczego we wrocławiu („wpływ żywienia na metanogenezę u bydła”), paulina onopiuk z  wydziału biotechnologii i  nauk o  środowisku katolickiego uniwersy178 r ep or ts of the john paul ii catholic university of lublin (“influence of salinity on growth and metabolic activity of photorhabdus temperate”), and beata madras-majewska from the department of animal sciences of the warsaw university of life sciences – sggw (“selected methods for the assessment of the microbiological quality of lime and acacia honey”). after the poster session was concluded, a thematic summary of the conference took place. like in the years before, the main goals of the microbs conference were the following: the creation of a discussion forum, the exchange of knowledge regarding application, cultivation, environmental conditions, construction, and metabolism of the microorganisms, as well as encouragement for the cooperation of specialists from different scientific fields which could result in innovation and the practical application of microbiological research. these main goals were achieved for the third time, and the conference has attracted a large group of young scientists interested in microbiology. tetu lubelskiego jana pawła ii („wpływ zasolenia na wzrost i aktywność metaboliczną photorhabdus temperata”) oraz beata madras-majewska z wydziału nauk o zwierzętach szkoły głównej gospodarstwa wiejskiego w warszawie („wybrane metody oceny jakości mikrobiologicznej miodów lipowych i  akacjowych”). po zakończeniu sesji posterowej nastąpiło podsumowanie tematyczne całej tegorocznej konferencji microbs. jak co roku, głównymi jej celami były: stworzenie forum dyskusyjnego oraz wymiana wiedzy na temat zastosowania, poznania warunków rozwoju, cech środowiska bytowania, budowy oraz przejawów życiowych mikroorganizmów, a  także zachęcenie specjalistów z  różnych dziedzin do nawiązywania współpracy naukowej, która mogłaby zaowocować zwiększeniem innowacyjności i  aplikacyjności prowadzonych badań. już po raz trzeci cele te zostały zrealizowane, a konferencja z roku na rok zrzesza coraz to większą grupę młodych naukowców, zaineresowanych tematyką mikrobiologiczną. all abstracts are available under the following web address/ wszystkie tegoroczne abstrakty pokonferencyjne są dostępne w formie elektronicznej: http://www.bc.wydawnictwo-tygiel.pl/public/assets/232/iii%20og%c3%b3lnopolska%20 mikrobiologiczna%20konferencja%20naukowa%20microbs.%20abstrakty.pdf arkadiusz gruca institute of biology, pedagogical university of cracow, podchorążych 2 st., 30-084 kraków, arkadiusz.gruca@up.krakow.pl 55 annales universitatis paedagogicae cracoviensis studia naturae, 1: 55–67, 2016, issn 2543-8832 anna kocoń, magdalena nowak-chmura* department of invertebrate zoology and parasitology, institute of biology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, *mnowak@up.krakow.pl the treatment of attacks by the ticks ixodes ricinus in the selected areas in żywiec and silesian beskids (silesian province – southern poland) introduction �e city of bielsko-biała, which performs functions of the main city of the bielsko agglomeration, and which is the centre of culture, commerce, tourism and industry, together with the towns of: ujsoły (49°28´56˝ n, 19°08´20˝ e) and złatna (49°29´42˝ n, 19°10´26˝ e) are located in the southern poland, near the border with slovakia and are areas attractive to tourists. both cities are located in the żywiec landscape park, in the municipality of ujsoły. numerous mountain ranges and hiking trails, rivers, bicycle paths, forests and green areas create ideal places for resting, walking and trips, not only during the spring and summer periods. ixodes ricinus l. 1758 – a  tick, occurs throughout poland, mainly in deciduous forests, mixed, in wet habitats. less frequently or not at all it isn’t recorded in dry pine forests and in sunny areas, peat bogs or swamps (siuda, 1993). due to warmer winters, a large number of rodents and forest animals, the occurrence of i. ricinus increases in the spring and summer periods, what results in a higher risk of attacks of the parasite, especially for the forest workers, farmers, tourists and residents, as well as forest and household animals. in natural conditions in poland i. ricinus is characterised by the presence of two peaks of seasonal activity: the largest, spring – at the turn of spring and summer, and the smaller one, autumn – between summer and autumn (lachmajer et al., 1958; kolpy, 1963; siuda et al., 2001; nowak et al., 2009). �e constantly changing climatic conditions, e.g. lighting, humidity, temperature, a�ect the abundance of i. ricinus (estrada-peña, 2001). �is parasite is reservoir and a vector of pathogens, such as, e.g. the thick-borne encephalitis virus (tbe), borrelia burgdorferi s.l. in summer causing the lyme disease, babesia sp., and other disease-causing microorganisms, e.g. rickettsia slovaca, coxiella 56 a nn a k oc oń , m ag da le na n ow ak -c hm ur a burnetti, anaplasma phagocytophilum (siuda, 1998; nowak-chmura, siuda, 2012). �e areas of the silesian province, including the mountain areas of the żywiec, silesian and small beskids, with diverse �ora and fauna, a  large number of forests, characterised by harsh mountain climate, are little explored in terms of risk of the tick attacks, therefore, studies were undertaken both in the urban and rural areas, which were then compared to the results of studies from the other parts of poland. materials and methods for the harvesting of ticks the so-called �agging method was used (siuda, 1993). it is a  commonly used method for studies on the occurrence and number of ticks in the given area. it is based on a collection, using a �annel �ag with the dimensions of 100×70 cm, in places where we can �nd hungry and active ticks, occurring most o�en along the paths, trails, roads, forest passageways, everywhere where there is an easy access to the moving host. during each collection, the vegetation was swept to the height of about 1m, a�er several cuts the presence of ticks was checked on the fabric and they were removed using a pair of tweezers, and then placed into labelled tubes �lled with 96% ethanol. �e collected specimens were examined and diagnosed in morphological and taxonomic terms using a stereoscopic magnifying glass olympus zx and a light microscope olympus. keys were used for marking the ticks from the polish fauna of siuda (1993) and nowak-chmura (2013), their species a�liation and stage of development were marked. �e laboratory tests were conducted in the department of invertebrate zoology and parasitology of the institute of biology of the pedagogical university in kraków. for the studies of the seasonal activity of i. ricinus the method of research areas was used (daniel et al., 1986), which consists of the collection with the method of �agging the ticks from the strictly determined areas with the area of about 100 m2. each separate research �eld has been previously tested for the presence of ticks. �e live and hungry ticks were placed in dry tubes of �uid maintenance, then the ticks were counted, the species was determined, as well as the stage of development, and they were released back to the test area. observations of the occurrence and spread of the parasite in bielsko-biała and in ujsoły and złatna were performed on six selected stations in 2013 three times, and four times in two areas. in total 20 tests were performed, from april to october in the a�ernoon hours (4–7 pm) in dry, hot weather. �e list of research positions is: (1) park in wapienica in bielsko-biała – located near a  busy cieszyńska street, the wapieniczanka river runs through the park, the silence there invites the residents to rest; (2) włókniarz park in bielsko-biała – the area located near the city centre, numerous playgrounds are visited by parents with children. it is also an attractive place 57 fig. 1. �e location of research to study the seasonal activity of ixodes ricinus in ujsoły. field research: a) – forest meadow near the river of biała soła, b) – forest clearing in danielka, c) – forest path in ujsoły (photo. a. kocoń) a) b) c) the treatm ent of attacks by the ticks ixodes ricinus in the selected areas in żyw iec and s ilesian b eskids s ilesian province – southern p oland) 58 a nn a k oc oń , m ag da le na n ow ak -c hm ur a for cyclists and walkers; (3) straceńskie boulevards in bielsko-biała – the area near the creek, it is characterised by the presence of mixed forests, deciduous, it is o�en visited by the residents of the city, cyclists, tourists, as well as it is a place for people walking their dogs; (4) błonia of bielsko – a rest place for the parents with children, athletes, active people, numerous hiking trails are nearby; (5) the campsite in złatna – a place o�en frequented by tourists, residents of the village, a pond is located nearby; (6) the forest path in ujsoły – a forest habitat, a living habitat of many rodents, forest animals, the bystra brook runs nearby. studies on the seasonal activity of i. ricinus were conducted in 2015 in the area of the town of ujsoły, from april 15th to october 7th. collections were recorded once a week from 3 pm to 7.30 pm in the hot and dry weather, conducive to the presence of the parasite. �ree areas were selected for the observation: (1) the forest meadow near the river of biała soła – a position lying near the deciduous and coniferous complex, nearby there is the river of biała soła (ujsoła), a place of living of the forest animals: wild boars, roe deer, deer, and many reptiles and rodents; (2) the forest glad in danielka – there is a hiking trail nearby, the grassy area, and a mixed forest; (3) the forest path in ujsoły – the area is seldom visited by the locals, but willingly by the collectors of the undergrowth, it is mainly the habitat for the forest animals and rodents, there is the bystra stream nearby (fig. 1). results in the course of the study it has been demonstrated that all collected ticks belonged to the species of ixodes ricinus. in studies of fauna of ticks in 2013, 60 specimens of i. ricinus were collected and in studies on the seasonal activity of i. ricinus the collected number was 875 specimens. �e occurrence of this species was found in all research areas, with the exception of one position: włókniarz park in bielsko-biała. during the research on the occurrence of i. ricinus in 2013, 60 specimens of ticks were collected, including 27 males, 23 females and 10 nymphs. no larvae was collected (tab. 1, fig. 2). i. ricinus was numerous in spring and summer periods, less frequent in summer and autumn periods (fig. 3). in the wapienica park the largest number of specimens of the parasite was collected: 19 females and 17 males; no nymphs and larvae were collected. no ticks were collected in autumn months. i. ricinus occurred among the shaded vegetation, close to the path in the park, near the river, playground. �ese were places frequented by people. no occurrence of i. ricinus was stated in the area of the włókniarz park. �is may be due to the regularly mowed grass, as well as the presence of a large number of birds and dogs in this area, which are attacked by ticks, who carry them out from the park. however, the absence of the parasites in this area can not be excluded. not many ticks 59 tab. 1. �e number of active specimens of ixodes ricinus in studies on recreational areas of bielsko-biała and beskid żywiecki research positions number of ixodes ricinus larva nymph male female wapienica park 0 0 19 17 włókniarz park 0 0 0 0 straceńskie boulevards 0 1 2 2 błonia of bielsko 0 1 3 0 tent area in złatna 0 0 1 0 forest path in ujsoły 0 8 4 2 total 0 10 29 19 fig. 2. developmental stages of ixodes ricinus: a – larva, b – nymph, c – female, d – male (photo. from the collections of the department of invertebrate zoology and parasitology ib pu in kraków) were collected in the straceńskie boulevards: there were only 2 males, 2 females and 1 nymph. �e reason for the small numbers can be the large sunlight of the terrain. in the period of the greatest activity of ticks in may, in the research position in błonie in bielsko 3 males and 1 nymph were collected. in later studies no ticks were found; this area is also frequently mowed. in the campsite of złatna during four studies the occurrence of only 1 male was stated. in order to determine the actual risk of tick attacks longer research should be conducted. �is place is not too frequented by people and the treatm ent of attacks by the ticks ixodes ricinus in the selected areas in żyw iec and s ilesian b eskids s ilesian province – southern p oland) 60 a nn a k oc oń , m ag da le na n ow ak -c hm ur a animals; it is characterised by a higher than average humidity of the area. �e forest path in ujsoły is the second region in terms of the number of collections of i. ricinus: 8 nymphs, 4 males and 2 females were collected. numerous small rodents occur in the studied area, which are usually the hosts of i. ricinus nymphs and larvae. in 2015, in the period from april 15th to october 7th, the weekly studies of the seasonal activity of i. ricinus were conducted on three selected positions in the town of ujsoły (fig. 1). based on 26 test collections and observations of the course of the activity rhythm of the ticks, an increased activity was stated in the spring period and a slight increase of the autumn activity. a total of 875 hungry specimens of active ticks i. ricinus were collected (tab. 2). di�erences were registered in the occurrence and abundance of the development stages of the tick. females accounted for the largest number of 328 specimens, then nymphs – 320, males – 226, larvae were the least numerous – only 1 specimen was noted (tab. 2). �e average temperature in the course of the studies was 22–21°c, the average value of atmospheric pressure was 1016 hpa, while the average air humidity was 40.5% (tab. 3). �e rhythm of the seasonal activity of i. ricinus in individual research positions was as follows: (1) forest meadow near the river of biała soła (fig. 1a) – a total of 258 ticks was noted, including 102 nymphs, 91 females, 64 males and 1 larva (tab. 4). in this research �eld the smallest number of hungry and active parasites was collected; the reason may be that the area is sunny and there is a large availability of forest animals, such as: roe deer, deer, wild boars and a  large number of reptiles, including lizards, as the main hosts, what creates a smaller probability of �nding a tick with the fig. 3. �e number of ticks common in individual months set 21 7 24 5 2 1 0 0 5 10 15 20 25 30 april may june july august september october n um be r of s pe ci m en ts months 61 lagging method. �e increase of tick activity was noted in the period from may 8th to june 3rd. in the period from june 10th to july 22nd there was a decline in the parasite numbers. a slight increase of tick activity was observed in the period from july 29th to september 9th. �e last collections, from september 16th to october 7th, show the drop in the activity of i. ricinus (tab. 2); (2) forest clearing in danielka (fig. 1b) – the largest number of specimens was collected, a total of 326 specimens, including 122 nymphs, 120 females, 84 males and 0 larvae (tab. 4). �e proximity of the forest, hiking trails and shady places creates good conditions for the occurrence of the parasite in the whole area, waiting for its host. �e increase in activity coincided with the spring and summer period – may 8th to june 24th. from july 1st to july 22nd the decline of the parasite numbers was observed. a slight increase of the parasite activity was noted in the period from july 29th to september 9th. from september 16th to october 7th the drop in the tick activity was observed (tab. 2); (3) forest path in ujsoły (fig. 1c) – in total 291 tick specimens were collected: 117 females, 96 nymphs, 78 males, 0 larvae (tab. 4). �is area is covered with dense vegetation, there is a lot of low bushes, among which the parasite can wait for his host, mainly for rodents and small reptiles (lizards). �e greatest increase of tick activity was observed in the period from may 8th to june 24th 2015. in july, from 1st to 22nd, a drop in the parasite numbers was noted. from july 29th to september 9th a slight increase in the tick activity was observed and the collections from september 16th to october 7th show a decrease in the activity of i. ricinus (tab. 2). tab. 2. �e number of hungry collected specimens of ixodes ricinus in ujsoły in 2015 research �eld research time �e number of observed specimens of ticks male female nymph larva total forest meadow near the river of biała soła 15.04–24.06 31 43 68 0 142 1.07–26.08 21 33 23 1 78 2.09–7.10 12 15 11 0 38 forest clearing in danielka 15.04–24.06 39 59 80 0 178 1.07–26.08 26 36 23 0 85 2.09–7.10 19 25 19 0 63 forest path in ujsoły 15.04–24.06 40 59 64 0 163 1.07–26.08 25 41 21 0 87 2.09–7.10 13 17 11 0 41 tab. 3. average climatic conditions for research positions in the course of research; area 1 – a  forest meadow near the river of biała soła, area 2 – in the forest clearing in danielka, area 3 – a forest path in ujsoły environmental factor area 1 area 2 area 3 air temperature [°c] 22.00 22.00 21.00 humidity [%] 40.50 40.50 40.50 atmospheric pressure [hpa] 1016 1016 1016 the treatm ent of attacks by the ticks ixodes ricinus in the selected areas in żyw iec and s ilesian b eskids s ilesian province – southern p oland) 62 a nn a k oc oń , m ag da le na n ow ak -c hm ur a tab. 4. �e number of active specimens of ixodes ricinus in studies of seasonal rhythm of activity on area of the village ujsoły research area number of ixodes ricinus larva nymph male female forest meadow near the river of biała soła 1 102 64 91 forest clearing in danielka 0 122 84 120 forest path in ujsoły 0 96 78 117 total 1 320 226 328 discussion ixodes ricinus, due to the medical and veterinary importance, is one of the most dangerous external parasites among the arthropods and the most frequently registered non-nest tick among 19 tick species that constantly occur in poland (siuda et al., 2000; biaduń, krasnodębski, 2007; nowak-chmura, siuda, 2012), including the areas of southern poland. �e main places that favour the occurrence of this species include the shady and moist habitats, as well as deciduous and mixed forests; it is less o�en met in the mountains, it also avoids sunny and dry places, e.g. dry pine forests (siuda, 1993). areas attractive to tourists and recreational areas are particularly prone to the occurrence of this common tick, this is mainly due to the easy access to the hosts, who willingly spend time outdoor with their children, pets, they walk on the trails and forest paths. given the particular risk of tick attacks and the risk of the transmission of the tick-borne diseases, such recreational areas were selected for the research in bielsko-biała and two smaller towns – ujsoły and złatna. in bielsko-biała, ujsoły and złatna, located in beskid żywiecki, the su�cient research has not been conducted so far on the occurrence of i. ricinus. it results from the conducted own collections and observations, that i. ricinus occurs commonly both in the rural areas of ujsoły and złatna, and in the urban ones, in the city of bielsko-biała (kocoń, 2014). �e researches prove, that ticks are more o�en present in the polish cities; it is slowly becoming a common phenomenon, moreover, they are potentially infected with disease-causing pathogens (pet’ko et al., 1997; karbowiak, siuda, 2001; siuda et al., 2005; kiewra, sobczyński, 2007). due to the fact that i. ricinus is a very good vector of pathogens of the transmission diseases, its medical and veterinary importance is very signi�cant. �e increasing spread of this species to new areas, as well as a large number of diseases transmitted by these parasites, is the result of the occupation of new habitats, and this is due to, among others, climatic changes (wójcik-fatla et al., 2009). �e increase in temperature and air humidity may lead to the increased risk of contact with ticks, which play an important role in the transmission of pathogenic microorganisms and transferring of pathogens to humans an animals (derdáková et 63 al., 2003; cuber et al., 2010). pathogens in the body of this parasite can stay for a long time, as well as they may also be transferred to its hosts. in poland, the most dangerous diseases transmitted by i. ricinus include, among others: lyme disease, tick-borne encephalitis, human granulocytic anaplasmosis and babesiosis (nowak-chmura, 2013). �e rhythm of the seasonal activity of i. ricinus depends largely on the geographical location. �e one peak, autumn-winter-spring rhythm, occurs in the mediterranean area (senevet, 1937). heading north, the seasonal activity of the parasite changes, the spring-summer period of activity increases, and the winter one can disappear (siuda, 2002). in poland there are two peaks of activity: the �rst one, the most prominent one – the spring peak (between spring and summer) and a less pronounced one – autumn (between summer and autumn) (kolpy, 1963; siuda et al., 2001; nowak et al., 2009). most studies on the course of the seasonal rhythm of activity of i. ricinus were conducted in the province of lesser poland (urbanowicz, 2000; siuda et al., 2001; nowak, 2001; góra, 2006; solarz et al., 2007; szubryt, 2014). �e researchers found the presence of the clear spring peak in the months of april/may, and the lack or a small increase of the autumn activity of ticks. in the north part of the country, also two peaks of activity were observed: the largest one – spring, and the smaller one – autumn (wegner et al., 1997; humiczewska, 2007; stańczak et al., 2012). own studies performed once a week in the a�ernoon in the area of ujsoły from april to october con�rm the occurrence of a clear increased tick activity in the spring and summer months (april, may, june), and a lower autumn activity (september, october). �e research of the seasonal activity rhythm of i. ricinus is very important due to the growing epidemiological importance of this parasite in poland, and can facilitate the identi�cation of the areas, where the threat of tick attacks occurs in the country, and a better understanding of the mechanism of the circulation of dangerous pathogens in nature. residents of ujsoły and the surrounding area, as well as numerous tourists, workers of the forestry and agriculture, are exposed to the attack by the parasite during its highest spring activity and to a lesser degree in the period of the autumn activity. on the basis of such studies it is possible to determine when, at what time, there is a danger of meeting i. ricinus waiting for its host, what can increase the attention of people. �ere is a possibility of protection against the attacks of the common tick, the following protections should be used for this: (1) use clothing suitable for going out into the countryside, thanks to which the tick will not be able to move to the bare body (long trousers, high shoes, long-sleeved blouse, head and neck protection), (2) avoid the typical places of the tick occurrence (forest paths, animal tracks, forest edges, mid-forest clearings), places overgrown with tall grass, lush vegetation, (3) use special measures in the form of sprays, creams, which repel ticks, both in humans and animals (collars, drops), (4) check the body and clothing, during the walk/trip and a�er the treatm ent of attacks by the ticks ixodes ricinus in the selected areas in żyw iec and s ilesian b eskids s ilesian province – southern p oland) 64 a nn a k oc oń , m ag da le na n ow ak -c hm ur a returning home, whether the ticks are not accidentally on our body and clothing, we remove the planted parasite using tweezers or special pliers available in pet stores and pharmacies. remember also about looking at the body and fur of the pets. in recent years, the number of people su�ering from the tick-born diseases has signi�cantly increased. �e fauna scienti�c research dealing with broadening knowledge on the occurrence, 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(2014). rytm aktywności sezonowej kleszcza pospolitego ixodes ricinus (linnaeus 1758) na terenie miasta alwernia (województwo małopolskie). uniwersytet pedagogiczny im. komisji edukacji narodowej w krakowie. [in polish] the treatm ent of attacks by the ticks ixodes ricinus in the selected areas in żyw iec and s ilesian b eskids s ilesian province – southern p oland) 66 a nn a k oc oń , m ag da le na n ow ak -c hm ur a urbanowicz, a. (2000). rytm aktywności sezonowej kleszcza pospolitego ixodes ricinus (linnaeus, 1758) w uroczysku skała kmity koło krakowa. akademia pedagogiczna im. ken w krakowie. [in polish] wegner, z., racewicz, m., kubica-biernat, b., kruminis-łozowska, b., stańczak, j. (1997). występowanie kleszczy ixodes ricinus na zalesionych obszarach trójmiasta i ich zakażenie krętkami borrelia burgdorferi. przegląd epidemiologiczny, 51, 11–20. [in polish] wójcik-fatla, a., szymańska, j., buczek, a. (2009). choroby przenoszone przez kleszcze. część i. ixodes ricinus jako rezerwuar i wektor patogenów. zdrowie publiczne, 119(2), 213–216. [in polish] abstract ixodes ricinus (castor bean tick) is a ruthless parasite with the greatest medical and veterinary signi�cance out of all the ticks found in the polish fauna. �e research on the incidence and spread of this species was conducted in six selected research areas in the urban area of bielsko-biała and in the rural areas of ujsoły and złatna. another variable covered by the research was the seasonal activity of i. ricinus in three selected research areas in ujsoły. �e researched locations include recreational areas near rivers, forests, and hiking trails. �ey are frequented by tourists and cyclists. in studies faunal a  total of 60 tick specimens belonged to the i. ricinus and were collected with the �agging method. �e ticks were stored in test tubes �lled with 96% ethanol. �e analysis focused on the parasite’s taxonomic position and developmental stages. during the observation of the seasonal tick activity, the parasites were placed in dry tubes. once their taxonomic position and developmental stages were diagnosed, they were released into the wild (a total of 875 ticks collected belonged to the i. ricinus). �e observations and studies indicate the widespread and increasingly high incidence of this dangerous parasite, which means there is a high risk of tickborne diseases in both urban and mountain areas of the żywiec and silesian beskids. key words: ixodes ricinus, seasonal activity, silesian province, southern poland, żywiec and silesian beskids received: [2016.06.14] accepted: [2016.09.28] zagrożenie atakami kleszczy ixodes ricinus na wybranych obszarach w beskidzie żywieckim i śląskim (województwo śląskie – południowa polska) streszczenie ixodes ricinus (kleszcz pospolity) to bezwzględny pasożyt o największym znaczeniu medycznym i weterynaryjnym spośród wszystkich kleszczy występujących w polskiej faunie. badania nad występowaniem i rozprzestrzenianiem tego gatunku były przeprowadzane na sześciu wybranych obszarach badawczych na terenie miejskim w  bielsku-białej i  na terenie wiejskim w  miejscowościach ujsoły i  złatna w  województwie śląskim. badaniami objęto również aktywność sezonową i. ricinus na trzech wybranych obszarach badawczych w  ujsołach. wybrane do badań obszary reprezentują miejsca rekreacyjne, położone w pobliżu rzek, lasów, szlaków turystycznych, chętnie odwiedzane przez turystów i rowerzystów. w badaniach faunistycznych zastosowano metodę �agowania, dzięki której zebrano łącznie 60 okazów kleszczy należących do gatunku i. ricinus. kleszcze przechowywano w probówkach wypełnionych 96% alkoholem etylowym, a  następnie analizowano pozycję taksonomiczną i  stadia rozwojowe pasożyta. w trakcie obserwacji sezonowej aktywności kleszczy, pasożyty umieszczano w suchych probówkach i po rozpoznaniu pozycji taksonomicznej i  stadiów rozwojowych, wypuszczano je na wolność (łącznie zebrano 875 osobników kleszczy należących do gatunku i. ricinus). przeprowadzone badania i obserwacje wskazują na powszechne występowanie tego groźnego pasożyta, a zarazem wskazują na duże niebezpieczeństwo wystąpienia chorób odkleszczowych, zarówno na terenach miejskich, jak i  górskich beskidu żywieckiego i śląskiego. słowa kluczowe: ixodes ricinus, sezonowa aktywność, polska południowa, beskid śląski i żywiecki 67 information on the authors anna kocoń she is specializing in experimental and environmental biology at the pedagogical university of kraków. she is naturalist with passion, for a few years interested in study of ticks (acari: ixodida). magdalena nowak-chmura researcher and teacher at the pedagogical university of kraków, institute of biology, department of invertebrate zoology and parasitology. she is a parasitologist and acarologist. her scienti�c interests are: faunistics, taxonomy and biology of ixodida, migration of ticks (acari: ixodida) on the hosts, the medical and veterinary importance of ticks, epidemiology. the treatm ent of attacks by the ticks ixodes ricinus in the selected areas in żyw iec and s ilesian b eskids s ilesian province – southern p oland) 7 annales universitatis paedagogicae cracoviensis studia naturae, 3: 7–21, 2018, issn 2543-8832 doi: 10.24917/25438832.3.1 krystyna towpasz1, alina stachurska-swakoń2* 1institute of botany, jagiellonian university, kopernika 27 st., 31-501 kraków, poland 2institute of botany, jagiellonian university, gronostajowa 3 st., 30-387 kraków, poland, *alina.stachurska-swakon@uj.edu.pl the occurrence of alien species in the agriculture landscape: the case of the proszowice plateau (southern poland) introduction the phenomenon of the synanthropisation of flora and vegetation is an important issue according to the diversity of plants and animals. it can also influence a local economy (pimentel, 2011), especially when it refers to areas that neighbour large agglomerations and serve as an agricultural service for them. the appearance of alien species in local floras is a process dating back to the beginning of agriculture (e.g., kornaś, 1981). the european alien species database (daise) contains over 6.6 thousand plant species that have been recorded to date in europe. their origin is associated with intentional introductions connected with their use, but there is also a large group of plants unintentionally transferred. the proportion of non-native species in the floras of individual european countries is varied, and their number ranges from approx. 500 to 1.900 species (lambdon et al., 2008; tokarska-guzik et al., 2011). it is assumed that the number of alien species in poland is 939, which represents around 27% of the flora of poland (tokarska-guzik et al., 2012). in the studies on the causes and mechanism of the distribution of newcomers, the detailed cartographic studies of local floras cannot be overestimated (zając, zając, 2003). the proszowice plateau (342.23 by physico-geographical regionalisation of poland; małopolska upland, southern poland) represents a special area: neighbouring with kraków agglomeration, one of the biggest cities in poland and with an old tradition of agriculture dating back to the neolithic, it was one of the earliest, dated neolithic settlement centres in poland (kruk et al., 1996). nowadays, almost 80% of the area (of the 770 km2) is covered by arable fields, which is the consequence of the favourable natural conditions: rich soils, gentle landscape relief, and microclimatic conditions (towpasz, 2006). human settlements occupy a relatively small area, many of them are dispersed in the area, and a few are concentrated in several small towns. k ry st yn a to w pa sz , a lin a s ta ch ur sk as w ak oń 8 the botanical studies in this area were started in 1995, and they have revealed the occurrence of rare and endangered plant species and vegetation types (e.g., towpasz, 1994; 2004; towpasz, kotańska, 1999; 2001; trzcińska-tacik et al., 1998; kotańska et al., 2001; towpasz, cwener, 2002; towpasz, stachurska-swakoń, 2010; towpasz et al., 2001; 2011; 2017). the paper is devoted to the alien species occurring in the proszowice plateau. we concentrated on the kenophytes (species arrived in poland after 1500 a.d., kornaś, 1977), because the archaeophytes were analysed before (towpasz, kotańska, 2003). the aim of the analyses was the search for rules of the mechanism of the synanthropisation of local flora. methods the floristic analysis of the flora of the proszowice plateau was based on a detailed inventory with the use of the cartogram method of atpol (zając, 1978) with the base unit (square) of 2 km × 2 km. the results presented here are based on the published “flora of the proszowice plateau” (towpasz, 2006) with the addition of new findings from the last few years. the geographical-historical classification of the species was used (kornaś, 1977) and the terminology follows kornaś, medwecka-kornaś (2002). in this respect, the following terminology of anthropophytes (non-indigenous plant species) was used: archaeophytes – species alien to the natural indigenous flora which arrived and became permanently established before the end of the 15th century; kenophytes – species brought to the country intentionally or unintentionally from 16th century; epoecophytes – species occurred in fields or ruderal habitats; hemiagriophytes – species established in seminatural habitats like meadows; holoagriophytes – species established in naturally disturbed plant communities such as forests, marches, swamp, ponds; diaphytes – casual species; and, ergasiophytes – casual species from cultivations. the lists of kenophytes were established after zając et al. (1998) and mirek et al. (2002). the obtained group of kenophytes was analysed in terms of their distribution and frequency (number of squares in which the species was noted), origin (zając et al., 1998), life form (raunkiaer form – zarzycki et al., 2002), ellenberg indicator values (ellenberg et al., 1992), grime’s life strategies (klotz et al., 2002), and dispersion mode (klotz et al., 2002). the names of species used follow mirek et al. (2002). results the flora of proszowice plateau contained 1008 plant species, and 204 species of those were permanently established alien species, i.e. metaphytes and 27 temporally wild the occurrence of alien species in the agriculture landscape: the case of the p roszow ice p lateau (s outhern p oland) 9 diaphytes (ergasiophytes) (appendix 1 – table 1). among the established species of alien origin, archaeophytes (108 species) represent the largest group, and the list of kenophytes consists of 96 species. in the last group, 44 species were classified as epoecophytes, 46 as hemiagriophytes, and 6 as holoagriophytes (appendix 1 – table 2). almost half of the kenophytes are invasive species in poland (according to tokarska-guzik et al., 2012). the most frequent among kenophytes were species commonly known as weeds. they occurred in the arable fields or ruderal habitats in the investigated area. the following belong to the group of epoecophytes growing in the segetal habitats: galinsoga ciliata, g. parviflora, oxalis fontana, veronica persica, vicia dasycarpa. galinsoga parviflora was the species most frequently noted in the area, because it occurred in 85% of the atpol squares (in 253 of 296). the rarest weeds were chenopodium strictum and kochia scoparia, which had only one location. amaranthus retroflexus and chamomilla suaveolens were epoecophytes widespread both in segetal and ruderal habitats. the following species should also be mentioned, as they were not rare: amaranthus chlorostachys and sisymbrium loeselli. lycium barbarum, classified here as an epoecophyte, was noted rather frequently, i.e. in 47% of atpol squares. it created dense thickets on the steep field bounds. the group of rare epoecophytes included amaranthus lividus, salsola kali subsp. ruthenica, sisymbrium altissimum and sinapis alba. the following belong to the rare species growing in the ruderal habitats: cardaria draba, datura stramonium, diplotaxis muralis, geranium divaricatum, g. pyrenaicum, and lepidium densiflorum. in this group of epoecophytes, centaurea diffusa, silene dichotoma and tanacetum parthenium were found in few locations on roadsides. the group of species that were permanently established in natural and semi-natural habitats in poland was numerous in the proszowice plateau, together 52 species. the most frequent species, according to the number of atpol squares in the area, was conyza canadensis (71% of atpol squares) and solidago gigantea (50% of atpol squares). the last species was common in abandoned fields, unmanaged meadows, as well as in riparian communities along streams. solidago gigantea, like in other regions of our country, often creates large and single-species clusters. the following group of hemiagriophytes includes several species that have recently spread in poland, especially along river valleys: echinocystis lobata and species of the genus heracleum: h. mantegazzianum and h. sosnovskyi. acer negundo has to be also mentioned as the species with the expansive potential on the area. it was noted mainly in forests and thickets along streams; however, when planted along roads, it could easily overgrow larger areas. another threat to the local flora is parthenocissus inserta, which very easily expands due to vegetative dispersion. the following are species that are fairly often noted in the area: bidens frondosa, bromus carinatus, erigeron annuus, k ry st yn a to w pa sz , a lin a s ta ch ur sk as w ak oń 10 impatiens glandulifera, ligustrum vulgare, medicago sativa, onobrychis viciifolia, rudbeckia laciniata, and trifolium patens. in the group of holoagriophytes, six taxa were listed: acorus calamus, cerasus mahaleb, elodea canadensis, impatiens parviflora, quercus rubra, and robinia pseudoacacia. among alien tree species, robinia pseudoacacia were the most frequently noted in the area, and it was found in 81% of atpol squares. the species could be treated as a particularly expansive plant. it is already completely established in the forests, and it even creates single-species woodlands on steep slopes above the roads. the steep embankments of the vistula river are significantly overgrown by black locust. the undergrowth of these woodlands consisted of a mixture of forest and meadow plants. frequently, impatiens parviflora, also a holoagriophyte, covered the floors beneath the tree crowns. impatiens parviflora was also found in others forest and shrub communities. it was noted in 54% of atpol squares. other tree species, quercus rubra rarely occurred in the area, and it was found in eight atpol squares only. acorus calamus grew in the reeds and elodea canadensis in water reservoirs. these species were rarely found in the studied area, which is related to the local relief and the rarity of such habitats. the remaining four species were found in the forest communities. most alien species were found rarely in the studied area. 53 taxa (55% of all) were found very rarely, and they were noted in 1-10 atpol square (fig. 1). only six species occurred in more than 200 atpol squares: chamomila suaveolens (77% of atpol squares), conyza canadensis (71%), galinsoga ciliata (79%), g. parviflora (85%), robinia pseudoacacia (81%), and veronica persica (77%). fig. 1. the frequency of occurrence of kenophytes of the proszowice plateau (southern poland) in the atpol square (2 km × 2 km); a number of all squares: 296, a number of kenophytes: 96 0 10 20 30 40 50 60 1–10 11–30 31–60 61–100 101–150 151–200 > 200 % of species nu m be r of s qu ar es the occurrence of alien species in the agriculture landscape: the case of the p roszow ice p lateau (s outhern p oland) 11 concerning the origin of the kenophytes that occurred in the investigated area, species originating in northern america (33) and europe (22) make the highest contribution (appendix 1 – table 2). therophytes (39 species) predominate when considering the main plant life form among species. there is also a significant number of hemicryptophytes (36 species). even in the investigated areas that have an agriculture character, seventeen species had a form of phanerophyte. with the respect of the ecological indicators, most species preferred full light and a moisture indicator value between 4 and 6. concerning the nutrient soil condition, rich soil was favourable (n = 7–8 for 50% of species). the largest group of kenophytes produced anemochorous seeds (40% of species). 16% of species were autochorous. the vegetative mode of spreading predominates among perennials (16% of all species). the interesting issue is the allocation strategy of alien plants. 41% of species have been characterised as having the competitor strategy, while 27% had a cr strategy (fig. 2). however, when we take the species with an occurrence in more than 70% of atpol squares, most of them had the cr strategy. there were only two species with the sr strategy: impatiens parviflora, salsola kali subsp. ruthenicus. three species used the cs strategy: acorus calamus, hesperis matronalis subsp. matronalis, hyssopus officinalis. fig. 2. the percentage contribution of grime’s life strategy in the kenophytes of the proszowice plateau (southern poland); c – competitors, r – ruderals, s – stress tolerant; a number of kenophytes: 96 c cr r sr cs csr c cr r sr cs csr k ry st yn a to w pa sz , a lin a s ta ch ur sk as w ak oń 12 the analysis of the distribution of the kenophytes indicated the role of habitat differentiation and the land use method. the special role in the landscape diversity is played by river and stream valleys. the highest concentration of hemiagryophytes and holoagriophytes was found in the vistula river valley (fig. 3). fig. 3. the distribution of chosen kenophytes on the proszowice plateau (southern poland); atpol squares (2 km × 2 km) discussion detailed analyses of local floras provide a useful assessment of their synanthropisation (faliński, 1972; kornaś, 1977; kornaś, medwecka-kornaś, 2002) and provide a good base for further considerations, particularly connected with alien species that could significantly change the diversity (e.g., heger, trepl, 2003; bradley et al., 2010; tokarska-guzik et al., 2012; zając, zając, 2015). the proportion of anthropophytes in the local floras of poland is diversified, and it often depends on the accessibility and number of anthropogenic habitats. these kinds of studies are important for the monitoring of alien species (pyšek et al., 2008). in recent decades, different approaches have been discussed for analysing the causes and mechanisms of the encroachment the occurrence of alien species in the agriculture landscape: the case of the p roszow ice p lateau (s outhern p oland) 13 of alien species. one of them is to look for particular attributes that make if possible for species to invade (e.g., kornaś, 1990; faliński, 1998; fu dostatny, 1999; drescher, prots, 2003; tokarska-guzik, 2005; theoharides, dukes, 2007; towpasz, stachurska-swakoń, 2011; trzcińska-tacik, stachurska-swakoń, 2011; bartoszek, stachurska-swakoń, 2016). the other approach is to look for abiotic conditions that make sites resistant to invasion (e.g., pyšek et al., 2010; barabasz-krasny et al., 2018). the presented analysis of local flora gives a general view on the group of alien species occurring in the landscape that has been used intensively of a long time. the number of alien species is not very high, and they constitute about 20% of the flora of proszowice plateau. the main cause of this phenomenon is the lack of railroads and bigger cities or communication centres (towpasz, 2006). the long history of agriculture and the predomination of arable fields in the landscape are visible in the prevalence of archaeophytes among alien species (towpasz, kotańska, 2003). some of them are rare in poland. the rarity of archaeophytes is connected with global change known in farmlands, i.e. larger farms, mechanization, the use of herbicides, etc. (e.g., trzcińska-tacik et al., 1998; trzcińska-tacik, stachurska-swakoń, 2010; stachurska-swakoń, trzcińska-tacik, 2014). in general, some rules of occurrence of kenophytes and its features in the investigated area could be seen. similar to other regions in poland or adjacent countries, the alien species originated mainly from europe and north america (zając et al., 1998; tokarska-guzik et al., 2012). there are mainly therophytes and competitors (according to life strategy) predominates. this phenomenon was also noticed by tokarska-guzik (2003) when the alien flora of chosen polish cities was compared. the usefulness of life strategy to estimate anthropogenic areas was described by different authors (e.g., grime, 2002; lososová et al., 2006). vuković et al. (2014) pointed at the cr strategy that prevailed for invasive species in croatia. similarly, when we compare the list of species with a number of localities (the number of squares in which the species was recorded) and life strategy, it could be concluded that species with the cr strategy had a higher potential to invade. the distribution of kenophytes along rivers is perceived by many authors (e.g., dajdok, kącki, 2003; dajdok, tokarska-guzik, 2009). the river valleys serve as habitat diversification in the landscape and roads for dispersion of plant diaspores. the local character of the area is demonstrated by the spontaneous formation of woodlands with robinia pseudoacacia and the formation of dense thickets with lycium barbarum. on a local scale, the last species should be treated as a hemiagriophyte, if the tendency to occupy new patches will maintain. it can be also assumed that the long list of alien species with a low number of localities is the result of the intensive use of the landscape. both mentioned species are treated as invasive in poland. the number of invasive plant species in the investigated area is high; almost a half of the k ry st yn a to w pa sz , a lin a s ta ch ur sk as w ak oń 14 noticed kenophytes are found in the list of invasive plant species in poland. some of them are really aggressive in the area and a growing number of localities is observed. in this respect, the investigation of the local flora is still important. references barabasz-krasny, b., możdżeń, k., sołtys-lelek, a., stachurska-swakoń, a. 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(2002). ecological indicators of vascular plants in poland. kraków: w. szafer institute of botany, polish academy of science. k ry st yn a to w pa sz , a lin a s ta ch ur sk as w ak oń 18 appendix 1 tab. 1. the historical-geographical classification of the flora of proszowice plateau (southern poland) name of group number of species local flora [%] spontaneophytes 804 79.8 anthropophytes 231 22.9 metaphytes 204 20.2 archaeophytes 108 10.7 kenophytes 96 9.5 epoecophytes 44 4.4 hemiagriophytes 46 4.6 holoagriophytes 6 0.6 diaphytes (ergasiophytes) 27 2.7 tab. 2. kenophytes in the flora of the proszowice plateau; invasive species in poland (according to tokarska-guzik et al., 2012) are marked in bold species n um be r o f sq ua re s (m ax 2 96 ) 0r ig in lif e fo rm s lif e st ra te gy ep oe co ph yt es h em ia gr io ph yt es h ol oa gr io ph yt es acer negundo 72 am n m c + a. saccharinum 1 am n m + acorus calamus 7 asia c & s g, hy cs + aesculus hippocastanum 18 eur se m c + amaranthus chlorostachys 59 am s & c t cr + a. lividus 12 am s & c t cr + a. retroflexus 185 am n t cr + aster lanceolatus 1 am n h c + a. novae-angliae 1 am n h c + a. novi-belgii 20 am n h c + a. × salignus 1 am n h c + bidens frondosa 47 am n t cr + brassica nigra 3 eur sw t cr + b. rapa subsp. rapa 1 eur s t cr + bromus carinatus 45 am n t, h + b. japonicus 2 eur s, asia w h r + b. × pseudothominii 4 antrop h + bryonia alba 16 eur e, asia w h cr + bunias orientalis 3 eur se, asia w h c + cardaria draba 3 eur se, asia sw g, h csr + the occurrence of alien species in the agriculture landscape: the case of the p roszow ice p lateau (s outhern p oland) 19 centaurea diffusa 1 eur se, asia sw t, h csr + cerasus mahaleb 2 eur s, asia sw m c + chamomilla suaveolens 227 am n, asia e h r + chenopodium strictum 10 asia c t r + conyza canadensis 211 am n t, h cr + cuscuta campestris 1 am n [se] t, p + datura stramonium 14 am n [se] t cr + dianthus barbatus 1 eur c h + diplotaxis muralis 6 eur s & w t, h csr + echinocystis lobata 59 am n t, li cr + echinops sphaerocephalus 7 eur e, asia w h c + elodea canadensis 6 am n hy cr + elsholtzia ciliata 9 asia e t r + epilobium ciliatum 7 am n h + eragrostis minor 1 eur se t r + erigeron annuus 86 am n h c + galinsoga ciliata 233 am c t cr + g. parviflora 253 am s & c t cr + geranium divaricatum 5 eur s, asia c t r + g. pyrenaicum 1 eur s h csr + helianthus tuberosus 15 am n g c + heracleum mantegazzianum 3 asia c & e h c + h. sosnovskyi 1 asia sw h c + hesperis matronalis subsp. matronalis 13 eur s h cs + hyssopus officinalis 1 eur s & se ch cs + impatiens glandulifera 44 asia c t c + i. parviflora 161 asia c, asia e t sr + juglans regia 16 eur se m c + juncus tenuis 16 am n h csr + kochia scoparia 11 eur e, asia w t cr + lepidium densiflorum 2 am n t r + ligustrum vulgare 26 eur h cr + lolium multiflorum 127 eur sw, afr n, am sw h, t c + lonicera caprifolium 1 eur se n c + lupinus polyphyllos 4 am n h c + lycium barbarum 138 asia e n c + lycopersicon esculentum 3 am s t cr + lysimachia punctata 3 eur se h c + malus domestica 12 eur m c + medicago sativa 143 asia s h c + m. × varia 97 antrop h c + mentha spicata 2 antrop h + oenothera depressa 2 am n h cr + onobrychis viciifolia 50 eur s, se h cr + k ry st yn a to w pa sz , a lin a s ta ch ur sk as w ak oń 20 oxalis fontana 143 am n, asia e t r + padus serotina 4 am n[e] & ams[n] n, m c + parthenocissus inserta 38 am n[e] n, li c + physalis alkekengi 4 eur se, asia sw h c + prunus domestica subsp. domestica 10 asia m c + pyrus communis 65 eur m c + quercus rubra 10 am n m c + reynoutria japonica 30 asia e g c + robinia pseudoacacia 240 am n m c + rosa rugosa 3 asia e n c + rudbeckia laciniata 34 am n h c + rumex confertus 3 eur se, asia w h c + salsola kali subsp. ruthenica 1 eur se, asia c t sr + senecio vernalis 9 eur se, asia w t, h r + silene dichotoma 1 eur s, se h r + sinapis alba 6 eur s t cr + sisymbrium altissimum 4 eur se, asia c h, t cr + s. loeselii 30 eur se, asia c h, t cr + solidago canadensis 7 am n g, h c + s. gigantea 147 am n g, h c + symphoricarpos albus 16 am n n c + syringa vulgaris 19 eur se n c + tanacetum parthenium 5 eur se, asia sw h c + telekia speciosa 1 eur se, asia h c + trifolium patens 113 eur s t + typha laxmanii 1 eur, asia h, hy + veronica peregrina 1 am n t + v. persica 229 asia sw t cr + vicia dasycarpa 38 eur s t cr + v. grandiflora 3 eur s, asia sn t cr + xanthium albinum 3 am n [s] t cr + x. strumarium 1 am s t cr + life forms (raunkieur forms): ch – chamaephyte, g – geophyte, h – hemicryptophyte, hy – hydrophyte, li – liane, m – megaphanerophyte, n – nanophaneropyte, p – parasite, t – therophyte, life strategy (grime’s strategy): c – competitors, r – ruderal, s – stress tolerator abstract the synanthropisation of vegetation is presently a serious problem, both due to its impact on plant and animal diversity, as well as its significance through the impact on the global and local economy, both directly and indirectly. understanding the mechanisms of new arrivals is one of the important aspects of many types of research. the presented paper concerns the analysis of selected features of kenophytes occurring on the proszowice plateau (southern poland). on the basis of long-term floristic studies, a list of kenophytes was selected. the list was subject to a detailed analysis of the frequency, origin, ecological numbers, life forms, and life strategies. the results indicate that perennial species with a cr life strategy have the greatest chance of expansion in the agricultural area, although the annual species with the c strategy predominate. nowathe occurrence of alien species in the agriculture landscape: the case of the p roszow ice p lateau (s outhern p oland) 21 days, 42 plant species (from 96 kenophytes of the proszowice plateau flora) are listed as invasive species in poland. key words: flora, invasive species, kenophytes, life strategy, synanthropisation received: [2018.06.20] accepted: [2018.11.07] występowanie gatunków obcego pochodzenia w krajobrazie rolniczym: na przykładzie płaskowyżu proszowickiego (południowa polska) streszczenie synantropizacja szaty roślinnej jest współcześnie poważnym problemem, zarówno z powodu wpływu na różnorodność roślin i zwierząt, jak również ma znaczenie poprzez oddziaływanie ekonomiczne na gospodarkę w wymiarze bezpośrednim i pośrednim. poznanie mechanizmów wnikania nowych przybyszów jest jednym z ważnych aspektów badań nad inwazyjnością. prezentowana praca dotyczy analizy wybranych cech kenofitów występujących na płaskowyżu proszowickim (południowa polska). na podstawie długoletnich badań florystycznych wyłoniono listę kenofitów, którą poddano szczegółowej analizie względem częstości wystąpień, obszaru pochodzenia, liczb ekologicznych, form życiowych i strategii życiowych. wyniki wskazują, że największe szanse ekspansji w terenie rolniczym mają gatunki wieloletnie o strategii życiowej cr, chociaż liczebnie przeważają gatunki jednoroczne o strategii c. obecnie, 42 gatunki (spośród 96 kenofitów) występujące we florze płaskowyżu proszowickiego są wymienione jako inwazyjne we florze polski. słowa kluczowe: flora, gatunki inwazyjne, kenofity, strategie życiowe, synantropizacja information on the authors krystyna towpasz she is a retired professor at the institute of botany of the jagiellonian university in kraków. she is interested in geobotany, plant ecology, and plant protection. alina stachurska-swakoń https://orcid.org/0000-0003-0381-4520 she is interested in the vegetation ecology, particularly in the mechanisms of changes in plant communities and flora under natural and anthropogenic influence influence, plant ecology and plant protection. 186 annales universitatis paedagogicae cracoviensis studia naturae, 3: 186–190, 2018, issn 2543-8832 iv international xi interdisciplinary conference on “nature–human–culture” , kraków, june 14th–17th, 2018 iv międzynarodowa viii krajowa interdyscyplinarna konferencja „natura–człowiek–kultura”, kraków, 14–17 czerwca, 2018 iv international and xi interdisciplinary conference on “nature–human–culture”, organised by the self-government of phd students from pedagogical university of cracow, cracow association of knowledge and culture creators “episteme” and krakow.pl web portal, was held from the 14th to 17th of june under a patronage of hm rector prof. kazimierz karolczak and the minister of higher education and science phd jarosław gowin. the opening ceremony, held by deputy dean assoc. prof. katarzyna potyrała and assoc. prof. andrzej kornaś, prof. up from institute of biology took place before noon on the 15th of june 2018. after the ceremony and a short coffee break, organisers invited all attendees to the first scientific session entitled “art and architecture”. msc joanna galas from adam mickiewicz university of poznań (“within the city walls – geochemical description of sediments (poznan, the water gate area)”) and msc agata iżykowska from the university of wroclaw (“links between polish performative art of women artists and western art”) were among participants that presented their lectures. w  dniach 14–17 czerwca 2018 roku odbyła się iv międzynarodowa i  viii krajowa interdyscyplinarna konferencja „natura– człowiek–kultura”. głównymi organizatorami konferencji byli samorząd doktorantów uniwersytetu pedagogicznego w  krakowie, stowarzyszenie twórców nauki i  kultury „episteme” kraków, a także portal kraków.pl. patronat honorowy nad konferencją, w  tym roku, objęli jm rektor prof. dr hab. kazimierz karolczak oraz minister szkolnictwa wyższego i nauki dr jarosław gowin. w  godzinach przedpołudniow ych, 15 czerwca 2018 roku, uroczystego otwarcia konferencji dokonali prodziekan ds. studenckich dr hab. katarzyna potyrała, prof. up wraz z  dr hab. andrzejem kornasiem, prof. up z  instytutu biologii. po oficjalnym otwarciu i  krótkiej przerwie kawowej, organizatorzy zaprosili uczestników do udziału w pierwszej sesji naukowej zatytułowanej „sztuka i  architektura”. w  sesji tej wzięli udział, m.in. następujący prelegenci: mgr joanna galas uniwersytet adama mickiewicza w  poznaniu („wewnątrz kolegiaty – analiza litologicz187 r eportsparticipants of the second scientific session entitled “civilisation and society” included msc klaudia rosińska from cardinal stefan wyszyński university in warsaw (“fake news as a consequence of post-truth world”), msc izabella barbara janda from the pedagogical university of cracow (“is solitary life human life? jacques maritain’s philosophical view on solitude”), and phd krzysztof matuszek from the pedagogical university of cracow (“religiousness in societies of western civilisation – condition and perspectives”). the third and fourth scientific sessions entitled “cultural and historic legacy” and “law and political sciences” followed after a dinner break. the following lecturers, among others, appeared in the third session: msc natalia ryś from the pedagogical university of cracow (“historical development of hotels in cracow”), msc dawid gołąb from the pedagogical university of cracow (“digital databases as a tool for a modern historian: examples and potential”). the following lecturers were among attendees of the fourth scientific session: berenika sikora from the university of lodz (“taking into account social aspects in public procurement law”) and msc joanna picewicz from jesuit university ignatianum (“language from the point of view of the theory of sexual selection, or language as an element of courtship”) after a coffee break in the afternoon, a poster session was held. msc agnieszka czyżowska from the pedagogical university of cracow (“toxicity of silver nanoparticles in relation to plant and animal cells”), msc anna kocoń from the pedagogical university of cracow (“the ticks of cracow”), and msc arkadiusz na i  geochemiczna osadów zebranych we wschodniej części placu kolegiackiego”), mgr agata iżykowska uniwersytet wrocławski („powiązania między polską sztuką performaty wną kobiet-artystów a sztuką zachodnią”). w drugiej sesji „cywilizacja i społeczeństwo” referaty wygłosili: mgr klaudia rosińska uniwersytet kardynała stanisława wyszyńskiego w  warszawie („fałszywe informacje jako konsekwencja świata post-prawdy”), mgr izabella barbara janda uniwersytet pedagogiczny w krakowie („czy życie solaris jest życiem? – jacques maritains filozoficzne spojrzenie na samotność”), oraz dr krzysztof matuszek uniwersytet pedagogiczny w  krakowie („religijność w  społeczeństwach zachodniej cywilizacji – stan i perspektywy”). po przerwie obiadowej odbyły się wystąpienia w  dwóch panelach naukowych „dziedzictwo historyczne i  kulturowe” oraz „prawo i  nauki polityczne”. w  pierwszym panelu wystąpili, np.: mgr natalia ryś uniwersytet pedagogiczny w  krakowie („historyczny rozwój hotelu w  krakowie”, mgr dawid gołąb uniwersytet pedagogiczny w  krakowie („cyfrowa baza danych jako narzędzie dla współczesnego historyka – przykłady i  potencjał”). w  drugim panelu naukowym udział wzięli, m.in.: berenika sikora uniwersytet łódzki („uwzględnienie aspektów społecznych w  prawie zamówień publicznych”) oraz mgr joanna picewicz akademia ignatianum w  krakowie („język z  punktu widzenia teorii doboru płciowego, czyli język jako element zalotów”). w  godzinach popołudniowych, po przerwie kawowej, uczestnikami sesji posterowej byli, np.: mgr agnieszka czyżowska uniwersytet 188 r ep or ts gruca from the pedagogical university of cracow (“botanical-based microbial induced corrosion inhibitors”) were among the lecturers. in the forenoon of the next day, the 16th of june 2018, a session entitled “natural and medical sciences” took place and following participants were among lecturers: msc barbara dyba from the pedagogical university of cracow (“the langmuir technique: a useful tool for studying the properties of model and native membranes”), msc witold dzierzęga and msc arkadiusz gruca from the pedagogical university of cracow (“16s rrna sequencing method as a tool in determining the composition of microbial communities causing biocorrosion”). session entitles “physical chemistry” followed and was attended by, e.g., msc katarzyna wilkosz from agh university of science and technology (“the effect of the modification of zirconia-based nanopowders on benzo[a] pyrene sorption”) and michał figiel (“fractal analysis of pore space geometry”) from the same research unit. next, two scientific sessions entitled “science of life and environment” and “interdisciplinary social sciences” followed after a short coffee break. amongst lecturers that have attended the first session were the following: msc łukasz kołodziejczyk from the pedagogical university of cracow (“animal experimental models in the pathophysiology of reproduction: basic and applicative aspects”), iwona konieczna from the pedagogical university of cracow (“allelopathic effect of solidago canadensis l. on germination and early growth of trifolium pratense l.”), and msc rafał bielecki from the pedapedagogiczny w krakowie („toksyczność nanocząstek srebra w  odniesieniu do komórek roślinnych i  zwierzęcych”), mgr anna kocoń uniwersytet pedagogiczny w  krakowie („kleszcze krakowa”) oraz mgr arkadiusz gruca uniwersytet pedagogiczny w  krakowie („botaniczne, indukowane przez drobnoustroje, inhibitory korozji”). następnego dnia, tj. 16 czerwca 2018 roku, w godzinach przedpołudniowych w sesji „nauki przyrodnicze i  medyczne” referaty wygłosili, m.in.: mgr barbara dyba uniwersytet pedagogiczny w  krakowie („technika langmuira: przydatne narzędzie do badania właściwości modelu i  naturalnych membran”), mgr witold dzierzęga i  mgr arkadiusz gruca uniwersytet pedagogiczny w  krakowie („metoda sekwencjonowania rrna 16s jako narzędzie do określania składu środowisk drobnoustrojów powodujących biokorozję”). w  sesji „fizykochemia” zaprezentowali swoje wystąpienia, np.: mgr inż. katarzyna wilkosz akademia górniczo-hutnicza w  krakowie („wpływ modyfikacji nanoproszków na bazie tlenku cyrkonu na sorpcję benzo[a]pirenu”) oraz z tej samej jednostki michał figiel („analiza fraktalna geometrii przestrzeni porów”). po krótkiej przerwie na kawę odbyły się obrady w  dwóch kolejnych panelach naukowych: „nauki o  życiu i  środowisku” oraz „interdyscyplinarne nauki społeczne”. w  pierwszym panelu wystąpił, m.in.: mgr inż. łukasz kołodziejczyk uniwersytet pedagogiczny w  krakowie („modele eksperymentalne zwierząt w  patofizjologii rozrodu: aspekty podstawowe i  aplikacyjne”), iwona konieczna uniwersytet pedagogiczny w krakowie („allelopatyczne działanie solidago 189 r eportsgogical university of cracow (“sozological problems of the miechowski county”). in the second session, lectures were given by, e.g., phd maciej paszyn from the university of warsaw (“creating local paramilitary groups as a natural social need”) and msc sylwia sławińska from the pedagogical university of cracow (“police as the authority responsible for safety”). sessions named “languages and literature” and “economy and society” followed a dinner break. msc bogusław bogusz from jan dlugosz university in czestochowa (“figurative language and cognition”), msc weronika szota (“melic translation analysis”), msc karolina kucznik from nicolaus copernicus university in toruń (“the economic dimension of human rights to water”), and msc paulina mielnik from jan kochanowski university in kielce (“grand crown marshal attribute as a source of communication between authority and a society”). after a whole day of intense proceedings, organisers invited all attendees to participate in the integration meeting. on the last day of conference, the 17th of june 2018, participants were traditionally taken on a guided tour of cracow. the main aim of the “nature–human– culture” conference, since its beginning in 2007, is the integration of students and young scientists from a wide range of scientific fields. this year’s conference was honoured by almost 60 lecturers. there is a good chance that this number will be even higher next year. canadensis l. na kiełkowanie i  wczesny wzrost trifolium pratense l.”) oraz mgr rafał bielecki uniwersytet pedagogiczny w  krakowie („problemy sozologiczne powiatu miechowskiego”). w  drugiej sesji rezultaty swoich badań zaprezentowali następujący prelegenci: dr maciej paszyn uniwersytet warszawski („tworzenie lokalnych grup paramilitarnych, jako naturalna potrzeba społeczna”), mgr sylwia sławińska uniwersytet pedagogiczny w  krakowie („policja jako jeden z organów odpowiedzialnych za bezpieczeństwo”) i inni. po przerwie obiadowej w  sesjach o  tematyce „języki i  literatura” oraz „ekonomia i  społeczeństwo” wyniki badań naukowych przedstawili, m.in: mgr bogusław bogusz uniwersytet humanistyczno-przyrodniczy im. jana długosza w  częstochowie („język zfiguratywny a  poznanie”), mgr weronika szota („melic translation analysis”) oraz mgr karolina kucznik uniwersytet im. mikołaja kopernika w  toruniu („ekonomiczny wymiar prawa człowieka do wody”), a także mgr paulina mielnik uniwersytet jana kochanowskiego w  kielcach („atrybut marszałka wielkiego koronnego, jako źródło komunikacji między władzą a społeczeństwem”). wieczorem, tego samego dnia, po intensywnych obradach, organizatorzy zaprosili wszystkich uczestników do udziału w  spotkaniu integracyjnym, w jednym z krakowskich klubów młodzieżowych. natomiast ostatniego dnia konferencji, tj. 17 czerwca 2018, tradycyjnie atrakcją w  ramach tegorocznej konferencji było zwiedzanie z przewodnikiem miasta krakowa. od początku tych spotkań naukowych, tj. od 2007 roku, kiedy to po raz pierwszy od190 r ep or ts była się konferencja „natura–człowiek kultura”, ich celem jest integracja środowisk studentów, doktorantów i młodych naukowców, zajmujących się bardzo różnorodną tematyką badawczą. w tym roku w konferencji wzięło udział blisko 60 osób. można mieć nadzieję, że za rok spotkamy się w  jeszcze większym gronie. arkadiusz gruca institute of biology, pedagogical university of cracow, podchorążych 2 st., 30-084 kraków, arkadiusz.gruca@up.krakow.pl anatoliy a annales universitatis paedagogicae cracoviensis studia naturae, 8: xx–xx, 2023 issn 2543-8832 doi: 10.24917/25438832.8.x anna chrzan institute of biology and earth sciences, pedagogical university of krakow, podchorążych 2 st., 30–084 kraków, email anna.chrzan@up.krakow.pl biodiversity on one of the post-mining heaps in the silesian province (poland) introduction hazard to the environment is an inherent side effect of conducting the mining operations (sonter et al., 2018). hard coal mining and processing industries generate large quantities of mineral waste. despite substantial decline in hard coal production, the mining industry still remains one of the largest producers of industrial waste in poland (galos, szlugaj, 2014). in 2022, 115 million tons of industrial waste was generated (gus 2022). the main sources of waste were, as in previous years, mining and quarrying (61.3 million tons), industrial processing (21.3 million tons) and generation and supply of electricity, gas, steam, hot water 13.3 million tons (gus 2022). according to the central statistical office, the largest share of waste generated was waste generated from exploration, mining, physical and chemical processing of ores and other minerals (55%) and waste from thermal processes – 19% (gus 2022). the largest quantities of waste, i.e. a total of 58% of all generated waste, were generated in lower silesia and upper silesia provinces, which have significant concentrations of the mining industry (gus 2022). the dominant methods of handling waste generated in 2022 were recovery – 48.4% and landfilling 41.7% (gus 2022). the amount of waste disposed in the plants’ own facilities at the end of 2022 was 1,829 million tons. the uncultivated area of landfills (excluding municipal waste) was 8,000 hectares, of which landfills, mining waste facilities, including heaps accounted for 54.6%, and tailings ponds accounted for 45.4%. during the year, 57.2 hectares of waste disposal areas were rehabilitated (gus 2022). mining waste pose several environmental hazards, including contamination of soil, underground water, aquifers as well secondary contamination of the air (carlson, adriano, 1993; szczepańska, twardowska, 1999; tiwary, 2001; sułkowski et al., 2008; suponik, blanco, 2014; różański, 2019). emissions of aerosol contaminations to atmosphere as well as materials deposited in the waste heaps have all contributed to degradation of the natural environment and creation of new anthropogenic habitats (zając, zarzycki, 2013; sonter et al., 2018). soil organisms are an important factor in soil formation. they determine its functioning, productivity, its detoxification and the rate of remediation (richards, 1974; frouz et al., 2001, 2005, 2007, 2008, 2013; madej, 2002; siwek, 2008; manu et al., 2017; radosz et al., 2019; józefowska et al., 2020). the analyses conducted in post-mining areas included monitoring of contaminations, analyses of changes to and degradation of soils, observations of changes to plant compositions in the regions around the emissions sources as well as spontaneous natural succession on heaps and landfills (frouz et al., 2001, 2008; zając, zarzycki, 2013; pietrzykowski et al., 2014, 2015; józefowska et al., 2017; likuscieślik et al., 2023). among pollutants, heavy metals are particularly dangerous due to their toxicity, persistence in the environment and ability to bioaccumulate (kabata-pendias, pendias, 1999; krzaklewski et al., 2002; pietrzykowski et al., 2014). being persistent pollutants, heavy metals accumulate in the environment and consequently contaminate the food chains. accumulation of potentially toxic heavy metals in biota causes a potential health threat to their consumers including humans (kabata-pendias, pendias, 1999; ali et al, 2019). for the above reason, it is important to control metal concentrations in environments, also in post mining heaps. the aim of the studies was to determine the content of heavy metals, such as pb, cd, ni, zn and cu in the material collected at various distances from the peak of the mining waste heap in czerwionka-leszczyny and estimate their impact on quantities and diversity of soil organisms in those sites. material and methods the mining waste heap is located in the silesian province, on the territory of urban township of czerwionka-leszczyny (fig. 1; 50°08′56″n, 18°40′38″e). the waste heap with total area of 140 hectares was used from the start of the mining operations in the “dębieńsko” hard coal mine in 1898 until the mine’s shutdown in 2000. during that period, approximately 37 million tons of mining waste were deposited in that area (konior, 2006; stefaniak, twardowska, 2006). starting from 2003, mineral recovery works have been carried out on the two peaks of the heap. the purpose of these works is to recover minerals in an economically feasible way, which include mostly hard coal as well as materials to be used in road construction. a characteristic feature of waste minerals produced by the hard coal mining industry is that they are highly diverse from the mineral and petrographic standpoint. individual rocks have various physical and mechanical properties, which mostly determine how they could be utilized (góralczyk, baic, 2009). fig. 1. study area (source: https://docplayer.pl/109399706; changed). the remaining waste is once again stored on a flat waste heap. a biological phase of reclamation, which is used to initiate soil-forming processes, has been carried out on all the conical forms that make up the heap (krzaklewski, 2001; kasprzyk, 2009). the reclamation works, which are currently underway, aim at planting vegetation on the mining waste heap and preparing it for future development. in addition, the plants were observed to spontaneously grow on the heap. the mining waste heap is planned to serve as sports and recreation area, with construction of bicycle trails, cross country running trails as well as an observation deck (gawor et al., 2014). the forms created during the operation of the “dębieńsko” mine have the shape of cones with steep slopes (fig. 2). fig. 2. orthophotomap of a dumpin czerwionka-leszczyny (source: portal.gison.pl/czerwionka-leszczyny) due to specific shape of the slopes, the mining waste heap experiences significant erosion and denudation processes. the deposited mining waste mostly includes clayey shales, mainly comprised of kaolinite and illite, as well as carboniferous sandstone, claystones and hard coal. because the material has been laying on the heap for a long time, it is significantly eroded. its specific density is 2.24 g/cm 3 , and bulk density is 1.60–1.76 g/cm 3 . given the foregoing figures, the ground is highly water-permeable and susceptible to infiltration. moreover, the deposited mining waste is very susceptible to frost-induced erosion, which causes the material to split into fractions, i.e. sand and even dust, which is further susceptible to wind erosion. soil samples were collected in the township of czerwionka-leszczyny on five sites located at various distances from the peak of the highest mining waste heap: 1. approx. 11 meters from the peak, i.e. on the site where the grass was observed nearest the peak; 2. approx. 80 meters from the peak, on the site where the initial soil with loose structure begins to form; 3. at the bottom of the heap, approx. 230 meters from the peak, on the site where the mining waste material, which is deposited on the heap, is the oldest; 4. 300 meters from the peak, on the site located a short distance from the mining waste heap; 5. one kilometre from the peak, on the site where the waste heap’s impact on the soil is small. at that site, also grass was collected for the analysis of heavy metal content. the field research, encompassing soil temperature and ph measurements and collection of soil material, was conducted twice in 2015 (in the spring and in the autumn). soil samples were collected from the surveyed sites with soil frames, 25×25cm in size and 1m 2 in surface area (górny, grǖm, 1981). the frame was thrust into the soil on the depth of 10 cm. four soil samples were taken from each site on the same side of the heap. temperature and ph measurements of the soil were conducted with 330 wtw 330/set-1 ph-meter (wissenschaftlich – technische werkstätten 82362 weilheim) pocket ph meter equipped with an electrode for temperature measurements. in the laboratory, tullgren funnel (dynamic method) was used to extract living organisms from the soil samples (murphy, 1962; górny, grǖm, 1981). the preserved soil organisms were identified through keys (brauns, 1954, 1975; pławilszczikow, 1972) with use of binocular magnifiers. the identified organisms were classified to the level of orders or families. grass was also collected from a site located about 1km from the post-mining heap, in which the concentration of heavy metals was also determined. the aboveground part of the plants was cut from the plots from which soil was taken for testing. the grass cover of the meadow consisted mainly of meadow timothy phleum pratense l. with the addition of perennial ryegrass lolium perenne l. and smooth meadow-grass poa pratensis l. the analyses also involved measurements of heavy metal content in the soil and grass. heavy metal content in soils and grass was determined by faas after previous mineralization of the soil. for this purpose, soil samples were initially dried at 105°c, then the 2 g of dried soil was weighed from each location. mineralization of the soil was carried out with the velp scientifica dk-20 mineralizer, in concentrated nitric acid at 120°c. the resulting solutions were poured into measuring flasks and filled with distilled water to 10 cm 3 . in the samples thus prepared, concentration of heavy metals (cadmium, lead, nickel, copper and zinc) was determined by buck scientific 200a flame atomic absorption spectrophotometer faas. the aas limits of detection (lod) and quantification (loq) for pb were set at 0.027 ppm and 0.083 ppm (mg/cm 3 ), respectively, for cd 0.011 ppm (lod) and 0.033 ppm (loq), for ni 0.017 ppm (lod) and 0.05 ppm (loq), for cu 0.012 (lod) and loq=0.035 ppm and for zn: lod=0.023ppm and loq=0.069 ppm. statistica 13.3 software was used for statistical calculations. to determine the differences between the obtained experimental values, the standard deviation (±sd) was calculated for each parameter and duncan's test was used (n = 5, at p ≤0.05). pearson’s correlation coefficient was used to calculate the relationship between the variables. results and discussion results of soil temperature and ph measurements are presented in tab. 1. at the top of the heap, only soil temperature and ph were measured. the obtained results show that the temperature on the peak of the heap exceeded 50ºc, which makes it difficult for the plants to grow there (tab. 1). other authors also observed the surface temperature of the mining waste heaps to grow along with the increase in height (zając, zarzycki, 2013; surovka et al., 2017; radosz et al., 2019). due to high temperatures, no material was collected for testing. at the distance of approx. 11 meters from the peak, the soil temperature dropped to 30°c (tab. 1). tab. 1. characteristics the studied sites (temperature of soil, soil ph, density, diversity of pedofauna, percentage) parameters distance from the top of post-mining dumps pearson’s correlation coefficient the top of post-mining dumps 11m 80m 200m 300m 1km temperature of soil [ o c] >55 27.7–29.0 12.0–15.0 13–17.7 13.5–19.7 14–19 soil ph 5.38 6.51 6.69 6.55 6.79 7.06 0,619 diversity (number of soil fauna orders) no 8 9 9 11 12 0,883* density of pedofauna (n/m 2 ) no 7032 2056 3480 7152 17754 0,905* order of soil fauna (%): enchytraeidae 0 9.73 3.91 0.89 0.13 -0,412 collembola 53.01 13.23 25.29 7.16 15.73 -0,412 coleoptera l. 1.02 1.56 0.92 2.24 0.41 -0,367 formicidae 0.91 9.73 0.69 60.18 2.21 -0,066 homoptera 0.11 6.23 0.23 2.12 0.58 -0,304 acarina 43.46 49.80 63.45 24.38 76.92 0,623 *correlation is significant at the 0.05 level the thermophysical properties of the soil depend on several factors (pikoń, bugla, 2007; zając, zarzycki, 2013). the most important one of them is the solar radiation energy. the rate of heating up of the soil depends on the colour of the soil, its humidity and structure, as well as its granulometric composition, landscape, exposure of the area to elements and the plants that grow on it. in case of mining waste heaps, a significant role is also played by exothermic reactions which are continuously occurring in the deposited material (pikoń, bugla, 2007). the heating of coal and pyrite occurring in the waste material causes self-ignition and endogenous fires of the mining waste heaps. this process increases the temperature of the mining waste heap even further and generates smoke (zając, zarzycki, 2013; różański, 2019; sułkowski et al., 2008; łączny et al., 2012; surovka et al., 2017; fabiańska et al., 2018). therefore, to prevent fires, waste material from the mining waste heaps should be directly utilized through reclamation or other engineering projects (levelling the area, building embankments, etc.) (gawor et al., 2014). to avoid fire hazard and to improve the properties of the waste material as well as the options for its utilisation, it would be helpful to separate coal from the mining waste (różański, 2019). the ph of the analysed material ranged from slightly acidic to neutral (5.4–7.06) (tab. 1). the lowest average ph value was recorded at the top of the heap and the highest 1km from the top of the heap. development of soil on mining waste heaps is a long-term process with significant contribution of biotic elements such as plants, microorganisms as well as soil fauna (madej, 2002; manu et al., 2017; radosz et al., 2019). however, the analyses of processes taking place on mining waste heaps mostly involve plant formations, while not a lot of testing is done on the soil organisms which inhabit even the most degraded biotopes. radosz et al. 2019 points out that the participation of soil organisms in the soil-creation processes is just as important in the post-industrial areas as it is in natural and semi-natural ecosystems. by providing access to and ensuring circulation of elements, soil mesofauna and soil plants have a significant share in plant succession. research of quantitative and qualitative aspects of pedofauna occurring on areas affected by significant human footprint largely focuses on the groups of organisms that occur on such areas in greatest numbers, namely mites and ticks as well as springtails. the sites with the initial plant cover are characterised by low density and diversity of the species of mites of the oribatida order, however, after a few years, their numbers increase to several thousand per 1 square meter (madej, 2002). this is also confirmed by the analyses of the soil fauna in czerwionka-leszczyny (tab. 1). however, the correlation of the percentage of selected systematic groups with distance from the heap is not statistically significant. the number of organisms could be observed to steadily grow with the increase in the distance from the peak of the heap. the only exception was the site located 11 m from the peak, which may be attributable to a well-developed plant community in the higher part of the highest mining waste peak. a very high degree of correlation exists between distance from the heap and pedofauna density, while a high degree of correlation was noted between distance from the heap and pedofauna diversity (tab. 1). the highest abundance and diversity of soil invertebrates was observed at site 5 (1km from the post-mining heap). in addition to enchytraeidae, colembolla, coleoptera, formicidae, homoptera and acarina, nematoda, gastropoda, isopoda, myriapoda, diptera and aranea also occurred here. radosz et al. (2019) pointed out a positive impact of the plant cover on the number of nematodes and enchytraeidae worms in the soils in post-industrial areas. density and species richness in most of investigated groups of soil biota gradually increased with increasing succession age (frouz et al., 2001). also, frouz et al. (2008) emphasized that the greatest density of soil macrosaprophages, which are the most important organisms for decomposition of mulch and mixing of soil, was observed in the oldest mining waste heaps in and around the town of sokolov, czech republic (frouz et al., 2008). frouz et al. (2014) showed that the influence of tree species on soil development is substantially mediated by soil fauna activity. monitoring and analysis of heavy metal concentrations in the environment are necessary for pollution assessment and control (ali, khan, 2019). the analyses also involved measurements of heavy metal content in the soil. concentrations of heavy metals varied. the heavy metal contents ranged for cd from 1.18 to 1.54 mg/kg of dry mass (dm), for cu 20.82–66.20 mg/kg of dry mass (dm), for zn from 97.79 to 222 mg/kg dm, for pb 27.20– 50.18 mg/kg dm and for ni from 5.55 mg/kg dm to 56.23 mg/kg dm (tab. 2). according to the regulation (2016), the soils of the investigated dump are classified in group iv, which includes industrial areas, areas of active mining activities, communication areas, including roads, railroads, other communication areas, as well as areas intended for the construction of public roads or railroads. they were not found to be excessively contaminated with metals (regulation, 2016). also pietrzykowski et al. (2014) found that in forested areas affected by coal, sand, lignite and sulphur mining, there was no risk of trace element concentrations in mine soils. copper content ranged from 20 to 37 mg/kg, averaging at 28.32 mg (tab. 2). the content of copper in the soil was statistically insignificant. tab. 2. content of heavy metals in soil at various distances from the peak of the post distance from the top of post-mining dumps statistical parameters metal cu cd pb zn ni 11m average [mg kg -1 ] mediana [mg kg -1 ] sd 20.82 b 19.96 2.16 1.18 b 1.19 0.20 50.18 a 50.24 4.85 97.79 b 95.47 19.02 40.28 b 41.55 5.82 80m average [mg kg -1 ] mediana [[mg kg -1 ]] sd 34.29 ab 35.39 0.15 1.54 a 1.26 0.02 48.04 a 45.75 0.09 174.66 a 174.89 2.30 49.20 b 45.64 1.18 200m average [mg kg -1 ] mediana [[mg kg -1 ] sd 37.72 ab 37.60 2.11 1.54 a 1.52 0.10 38.01 b 35.86 5.37 177.05 a 173.90 5.75 56.23 b 54.57 9.42 300m average [mg kg -1 ] mediana [[mg kg -1 ]] sd 25.11 b 25.52 2.41 1.43 ab 1.39 0.11 27.20 c 27.08 2.39 183.53 a 184.00 18.98 41.77 b 41.09 5.11 1km soil average [mg kg -1 ] mediana [mg kg -1 ] sd 23.64 b 21.64 1.88 1.51 a 1.44 0.23 30.08 c 31.06 3.26 222.19 a 224.93 16.52 5.55 c 5.41 1.13 1 km grass average [mg kg -1 ] mediana [mg kg -1 ] sd 66.20 a 57.61 13.48 1.31 ab 1.37 0.49 27.49 c 28.81 5.73 203.89 a 185.80 27.27 150.06 a 146.83 18.46 bcf 2.80 0.87 0.91 0.92 27.04 sd – standard deviation, bcfbioconcentration factor mean values marked with different letters differ significantly according to duncan test p ≤ 0.05. elevated levels of copper were reported by bzowski (2013) in the mining waste heaps of the lubelskie coal basin as well as różański (2019) in the upper silesia coal basin (tab. 3). tab. 3. content of selected heavy metals in waste and permissible values metal bzowski (2013) lcb lublin coal basin różański gow (2019) upper silesian coal basin czerwionkaleszczyny (2016) the highest permissible value in grounds acc. ordinance me 2016, mg kg -1 (dm) group iv cd <2 no 1.2–1.5 15 cu 65 87–155 20.8–37.7 600 ni 52 58–111 5.55–56.2 500 pb 29 no 27.2–50.2 600 zn 137 110–201 97.8–183.5 1000 nickel content ranged from 5.5 mg/kg dm in the distance of 1km from the mining waste heap to 56.2 mg/kg dm in the distance of 200 m from the heap (tab. 2). higher contents of that metal were reported by różański (2019) on the przezchlebie mining waste heap in the upper silesia coal basin (tab. 3). bian et al. (2006) found the soil levels of cadmium, nickel, lead and zinc to be much lower at the distance of 150–180 m from the mining waste heap in the yanzhou coal basin in china. the content of ni in the soil was statistically lover at the distance of 1km from dump than at 11, 80, 200 and 300 m distances (tab. 2). the cadmium concentration ranged from approx. 1.2 to 1.5 mg dm (tab. 2). according to the findings, there was a significant decrease in the cd content at a distance of 11 meters compared to the other distances that were analysed. similar results were recorded in the lubelskie coal basin (tab. 3). https://dictionary.cambridge.org/pl/dictionary/english-polish/content https://dictionary.cambridge.org/pl/dictionary/english-polish/of https://dictionary.cambridge.org/pl/dictionary/english-polish/in https://dictionary.cambridge.org/pl/dictionary/english-polish/the https://dictionary.cambridge.org/pl/dictionary/english-polish/soil https://dictionary.cambridge.org/pl/dictionary/english-polish/was https://dictionary.cambridge.org/pl/dictionary/english-polish/at zinc concentration was increasing as the distance from the peak of the mining waste heap was growing, but the differences were not statistically significant (tab. 2). the highest concentration of 222 mg/kg of dry mass was recorded at the distance of 1km from the mining waste heap. the concentration of lead in the soil was statistically lower at the distance of 200m, 300m and 1000m from dump than at the 11m and 80m distances (tab. 2). at the distance of approx. 1km from the mining waste heap, it ranged from 25–32 mg/kg of dry mass. similar lead concentrations were recorded by bzowski (2013) in the mining waste heaps of the lubelskie coal basin (tab. 3). the studied soils were classified as pollution degree ii, i.e., moderately contaminated in respect of heavy metal content, according to iung guidelines (kabatapendias, 1995). in addition, the heavy metal content of plants collected at a distance of 1km from the post mining dump was determined. the results presented in table 2 indicate that the content of copper and nickel in the dried grass was significantly higher than in the soil. the content of copper in plants varies greatly depending on part of the plant and the plant’s development stage, variety and specie as well as its density in the habitat and climate conditions. plants growing in mining pits and mining waste heaps are exposed to excessive concentrations of that metal (kabata-pendias, pendias 1999). terelak et al. (2001) gives the average cu concentration in grass at 5.5 mg/kg. on the other hand, the paper shows much higher contents of copper, ranging from 36 mg/kg to 128 mg/kg of dry mass (average of 66.2 mg/kg of dry mass) (tab. 2). nickel is easily absorbed by plants. in most of the cases, this element is absorbed at the rate similar to its concentration in soil. in plants, nickel is very mobile and easily moves to above-ground parts of the plant, mostly to its seeds. plants growing in contaminated areas have much higher nickel contents (terelak et al., 2001; jasiewicz, antonkiewicz, 2004). according to research by pietrzykowski et al. (2014) in europe in afforested areas affected by hard coal, sand, lignite and sulphur mining, there is no risk of trace element concentrations in mine soils. the degree of accumulation of heavy metals in grasses is denoted by the bioaccumulation factor (bcf), which is defined as the ratio of metal concentration in plant biomass to that in the soil. in the analysed plant material, the bioaccumulation factor (bcf) was the highest for nickel, followed by copper. other metals had lower accumulation rates of bcf<1 (tab. 2). conclusions industrial and post-industrial areas are characterised by significant changes to the environment and local spatial planning for the given territorial unit. the mining waste heap located in czerwionka-leszczyny significantly affected not only that city’s landscape but it also impacted the quality of certain elements of nature. several land reclamation projects are carried out in order to mitigate the waste heap’s adverse impact on the environment. reclamation procedures were carried out on all conical forms comprising the mining waste heap. such procedures involved planting trees and allowing the plants to spontaneously grow on the heap. such procedures involved planting trees and allowing the plants to spontaneously grow on the heap. in this study, correlations were found between the distance from the heap and the density and diversity of pedofauna. a very high degree of correlation was noted between density and a high degree of correlation between distance from the heap and pedofauna diversity. the contents of the analysed metals did not exceed their permitted levels in soil and earth as defined by the regulation of the minister of environment of 2016 in the matter of the procedure for conducting the assessment of soil surface contamination. no clear regularities were observed in the content of heavy metals in the soil at various distances from the top of the heap. the obtained results indicate that the content of copper and nickel in the dried grass was significantly higher than in the soil. conflict of interest the author declares no conflict of interest related to this article. references ali, h., khan, e., ilahi, i. 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(2001). trace element content (cd, cu, ni, pb, zn) in farm-land soils in poland. archiwes of environmental protection, 27(4), 159–174. tiwary, r.k. (2001). environmental impact of coal mining on water regime and its management. water, air and soil pollution, 132(1–2), 185–199. https://doi.org/10.1023/a:1012083519667 zając, e., zarzycki, j. (2013). the effect of thermal activity of colliery waste heap on vegetation development. annual set the environment protection, 15, 1862–1880. abstract waste generated by the hard coal mining and processing industries pose significant environmental hazard through, among other things, impacting water and soil. the process that is particularly dangerous is trace element accumulation. excessive quantities of heavy metals pose grave threat to plants, humans and soil organisms. the purpose of the studies was to determine the content of heavy metals, such as pb, cd, ni, zn and cu in the material collected at various distances from the peak of the mining waste heap in czerwionkaleszczyny and estimate their impact on quantities and diversity of soil organisms in those sites. studies have shown a high degree of correlation between the distance from the top of the heap and the density and diversity of pedofauna. the highest abundance and diversity of soil invertebrates was observed at site 5 (1km from the postmining heap). the content of heavy metals in the tested soils ranged for cd from 1.18 to 1.54 mg/kg of dry mass (dm), for cu 20.82–66.20 mg/kg dm, for zn from 97.79 to 222 mg/kg dm, for pb 27.20–50.18 mg/kg dm and for ni from 5.55 mg/kg dm to 56.23 mg/kg dm. the contents of the analysed metals did not exceed their permitted levels in soil and earth as defined by the regulation of the minister of environment of 2016 in the matter of the procedure for conducting the assessment of soil surface contamination. the obtained results indicate that the content of copper and nickel in the dried grass was significantly higher than in the soil. key words: heavy metals, soil fauna, waste heaps received: [2023.06.29] accepted: [2023.08.09] bioróżnorodność na jednej z hałd pokopalnianych w województwie śląskim (polska) streszczenie odpady powstające w przemyśle wydobywczym i przetwórczym węgla kamiennego stanowią poważne zagrożenie dla środowiska, między innymi poprzez oddziaływanie na wody i gleby. szczególnie niebezpiecznym procesem jest akumulacja pierwiastków śladowych. nadmierne ilości metali ciężkich stanowią poważne zagrożenie dla roślin, ludzi i organizmów glebowych. celem badań było określenie zawartości metali ciężkich, takich jak pb, cd, ni, zn i cu, w materiale pobranym w różnych odległościach od szczytu hałdy w czerwionce-leszczynach oraz oszacowanie ich wpływu na liczebność i różnorodność organizmów glebowych na tych stanowiskach. badania wykazały wysoki stopień współzależności między odległością od szczytu hałdy a zagęszczeniem i różnorodnością pedofauny. największą liczebność i różnorodność bezkręgowców glebowych stwierdzono na stanowisku 5 (1km od hałdy poeksploatacyjnej). zawartości metali ciężkich na badanych stanowiskach wahały się dla cd w przedziale od 1.18 do 1.54 mg/kg s.m., dla cu 20.82–66.20 mg/kg s.m., dla zn od 97.79 do 222 mg/kg s.m., dla pb 27.20–50.18 mg/kg s.m. i dla ni od 5.55 mg/kg s.m. do 56.23 mg/kg s.m. zawartości analizowanych metali nie przekroczyły dopuszczalnych poziomów w glebie i ziemi, określonych w rozporządzeniu ministra środowiska z 2016 r. w sprawie sposobu prowadzenia oceny zanieczyszczenia powierzchni ziemi. uzyskane wyniki wskazują, że zawartość miedzi i niklu w wysuszonej trawie była istotnie wyższa niż w glebie. słowa kluczowe: fauna glebowa, metale ciężkie, odpady pogórnicze information on the author anna chrzan; https://orcid.org/ 0000-0002-2186-3126 assistant professor at the department of ecology and geoinformation of the institute of biology and earth science of the pedagogical university of krakow. she deals with environmental monitoring and soil ecology. he also conducts soil biomonitoring research in environments under various anthropopressure. 135 annales universitatis paedagogicae cracoviensis studia naturae, 3: 135–141, 2018, issn 2543-8832 doi: 10.24917/25438832.3.10 saikat kumar basu research division, ps, lethbridge ab canada t1j 4b3; skt.basu@gmail; https://orcid.org/0000-0001-7305-4817 canada goose branta canadensis (linnaeus, 1758): a majestic water bird from the continent of north america canada goose branta canadensis (linnaeus 1758) (synonym: anas canadensis linnaeus 1758) is a major water bird, native to the continent of north america (basu, 2015a). the species is sexually non-dimorphic, and males are slightly larger in size than the females (appendix 1 – fig. 1a). the avian member represents an iconic symbol of the majestic beauty of wild north america. the species is distributed across the continent from the pacific coast in the west to the atlantic in east; making the distribution of the species as pan american. the species is distributed across the length and breadth of the continent, stretching from northern canada, across the united states into northern mexico (basu, 2015b). it has been reported from eastern siberia and eastern china into japan as the eastern most global range of the species. outside the continental range of north america and eurasia, canada geese has been introduced in uk, some parts of eu; and also in new zealand and argentina. canada geese, being a migratory species (appendix 1 – fig. 1b), travels to the warmer part of the north american continent to southern united states, puerto rico as well as northern mexico to avoid harsh north american winter (collias, jahn, 1959; madge, burn, 1988). they have also been reported to travel to parts of caribbean latin america, such as cuba, belize, bermuda, jamaica, trinidad and tobago, dominican republic, grenada, saint pierre, and miquelon (basu, 2015c). the birds are reported to travel at a speed of 60–80 km per hour and are known to cover as much as 800-900 km per day during their classic migration flight (madge, burn, 1988). the sight of v-shaped migration pattern of canada geese flocks with their characteristic honks during autumn in search of their winter homes down south is an outstanding moment of natural beauty. each such flock is led by an experienced leader bird guiding the entire flocks to their preferred destination like a highly coordinated squadron (collias, jahn, 1959; mowbray et al., 2002). however, recently, a number of resident flocks are being reported from the us and canada (maccarone, cope, 2004). s ai ka t k um ar b as u 136 the species is associated with the folklores, poems, songs, art work, and socio-cultural traditions of several north american and eastern siberian aboriginal communities (appendix 1 – fig. 1c) (basu, 2015a,b,c). according to an online survey by the canadian geographic magazine in 2015, the canada goose was voted among the top five preferred avian species to be considered as the national bird of canada (basu et al., 2016). life cycle and other characteristics canada geese inhabit a wide range of habitats from mountainous lakes and rivers to the canadian prairies (appendix 1 – fig. 1d) and the great northern plains of the us, boreal forests, woodlands, parks and meadow grassland, flat river basins as well as coastal areas and dry southern plains of southern us and northern mexican plains to preferred caribbean island habitats (basu, 2017a; mowbray et al., 2002). the specie varies in weight between 4–10 kg with length varying between 0.76–1.1 m and a wingspan in the range of 1.4–1.8 m. the majestic species is characterised by the presence of a black head and a long extended neck with a distinct white face and body plumage varying between brown, black, white, and grey and webbed feet (appendix 1 – fig. 1e). around seven to ten different subspecies of canada goose is reported from north america (tab. 1). but it is important to mention that there are differences of opinions among ornithologists regarding the exact number of subspecies of canadian goose across north america. some of the smaller canada geese (b. canadensis ssp. minima ridgway 1885) subspecies are often confused with some related species, like cackling goose b. hutchinsii richardson 1832. some subspecies like the lesser canada goose is believed to be of a hybrid nature and not a true subspecies (hanson, 1997; mowbray et al., 2002). however, more taxonomical, molecular and biochemical studies needed to be conducted before all the current canada goose subspecies can be clearly distinguished and is subjected to change in future (madge, burn, 1988; mowbray et al., 2002; basu et al., 2016; basu, 2017). the canada goose is strictly monogamous and highly gregarious in nature. they assemble in large numbers in water bodies and agricultural fields during their annual migration (collias, jahn, 1959). the species is known to be extremely territorial, and male birds are aggressive in defending their territories and mate. both parents are extremely dedicated towards their parental duties and aggressively protect their foraging space, nesting ground, nests, eggs, and goslings from other competing geese as well as their natural predators. the species is predominantly vegetarian in nature and depend on aquatic vegetation in water bodies like ponds, pools, lakes, swamps, bogs, as well as fresh germinating sprouts or young seedlings in a farmer fields or fresh grass tips in yards, lawns, city gardens and parks, golf courses, and roadside ditches. 137 c anada goose branta canadensis (linnaeus, 1758): a m ajestic w ater bird from the continent of n orth a m erica tab. 1. the most frequent subspecies of canada goose noticed from north america no. name of subspecies range of occurrence 1. atlantic canada goose (branta canadensis ssp. canadensis linnaeus 1758) nesting and breeding in newfoundland and labrador (eastern canada) and wintering in southern us 2. dusky canada goose (b. canadensis ssp. occidentalis. sf. baird 1858) breeding around south western alaska and wintering in the us 3. giant canada goose (b. canadensis ssp. maxima delacour 1951) restricted to southern canada and northern us 4. interior canada goose (b. canadensis ssp. interior todd 1938) breeding in canada and wintering in southern us and northern mexico 5. lesser canada goose (b. canadensis ssp. parvipes cassin 1852) breeding from central alaska (us) to the canadian prairie and wintering in southern us 6. moffitt’s canada goose (b. canadensis ssp. moffitti aldrich 1946) breeding around southern canada, great northern plains and the great basin of us and wintering in southern us and northern mexico 7. vancouver canada goose (b. canadensis ssp. fulva delacour 1951) is a predominantly resident species distributed from southern alaska (us) to the province of british columbia (canada) occasionally they also forage on aquatic insects and crustaceans. goose droppings on fields and turf as well as in parks and gardens could turn into a challenging concern if huge flocks stop for temporary or permanent shelter in these places, and this serves as a potential source of coliform bacteria. canada goose is also known to be notoriously dangerous in colliding with aircrafts and are a serious threat to safe aviation at some places where their populations explode due to abundant food supply and the lack of natural predators checking their numbers (mowbray et al., 2002). the number of eggs produced per nest can vary between 2-18 depending upon on safety of the nesting site and the local predation pressure (appendix 1 – fig. 1f, 2a). in a good year, a pair can raise up to 8–16 chicks (goslings) depending upon availability of food, good weather conditions and low predation pressures (collias, jahn, 1959; mowbray et al., 2002). the size of the canada goose egg is almost 2–2.5 x times that of an average poultry chicken egg (appendix 1 – fig. 2b, c), and the yolk colour of canada goose egg is pale orange compared to the bright yellow coloured yolk of poultry chicken (appendix 1 – fig. 2d). the volume of yolk is almost 3x in the case of canada goose compared to that of a poultry chicken egg. among the most known natural predators of canada goose are mammals like coyotes, raccoons, foxes, badgers, and bears, plus birds like the american crow, carrion crow, common ravens, gulls, and raptors (mowbray et al., 2002). they mostly target eggs, goslings as well as injured, sick and aged birds struggling to defend themselves (appendix 1 – fig. 2e). in addition, the canada goose is treated as a traditional game bird and extensively hunted by both professional and amateur hunters as well as members of aboriginal communities across north america. young chicks (goslings), jus ai ka t k um ar b as u 138 veniles, sub adults, and even adults also die due to accidental road kills across north america (appendix 1 – fig. 2f ). due to population explosion and the lack of natural predation pressure, the bird has been transforming into a local pest in some parts of north america. canada goose is an iconic member and symbol of the north american wildlife. it currently has a least concern iucn conservation status with an estimated 4–5 million strong population across different north america habitats. although, due to population explosion and low predation pressure, the species has turned into a nuisance in some parts of the continent reducing the values of properties and serving as a potential source of coliform bacteria and a threat to safe aviation. however, it still serves as an important constituent of the natural ecosystems. the migratory species with a number of sub species distributed across the continent serves as a socio-cultural heritage of several aboriginal communities and wildlife enthusiasts of north american continent. references basu, s.k. (2015a). canada goose (branta canadensis l.): an iconic symbol of wilderness in the continent of north american. scientific india, 3(6), 45. basu, s.k. (2015b). feature: canada goose. the global new light of myanmar, 2(181), 8. basu, s.k. (2015c). feature: nature and wildlife: north american canada goose. sikkim express, gangtok, sikkim, india, 39(332), 4. basu, s.k (2017a). canada geese. nesa newsletter, 20(3), 5. basu, s.k. (2017b). call of the wild (opinion/column): canada goose: the most majestic waterfowl species in the world. manoram, kottayam, kerala, india. august 26, 2017. http://english.manoramaonline.com/news/columns/call-of-the-wild/2017/08/26/canada-goose-majestic-waterfowl-species-saikat-kumar-basu.html basu, s.k., zandi, p., chalaras, s.k. (2016). canada goose (branta canadensis l.): an iconic symbol of wild continental north america. book of proceedings of the seventh international scientific agriculture symposium. agrosym 2016, jahorina, bosnia and herzegovina, october 6–9, 2016. section 1. plant production. pp. 1233–1236 (p–178). collias, n.e., jahn, l.r. (1959). social behaviour and breeding success in canada geese (branta canadensis) confirmed under semi-natural conditions. the auk: ornithological advance, 76, 478–509. hanson, h.c. (1997). the giant canada goose (2nd ed.). southern illinois university press. maccarone, a.d., cope, c. (2004). recent trends in the winter population of canada geese (branta canadensis) in wichita, kansas: 1998–2003. transact kansas academy of science, 107(1, 2), 77–82. madge, s., burn, h (1988). waterfowl: an identification guide to the ducks, geese, and swans of the world. boston: houghton mifflin. mowbray, t.b., ely, c.r., sedinger, j.s., trost, r.e. (2002). canada goose (branta canadensis). in: a. poole (ed.), the birds of north america. ithaca: cornell lab of ornithology. 139 appendix 1 fig. 1. canada goose male (a); a migrating flock of canada geese across the canadian prairie at the end of fall (b); a flock taking their flight beside an irrigation canal (c); a canada geese family thriving in a local water body in the canadian prairies (d); footprints of canada geese captured on clayey soil (e); a typical canada geese nest made up of grass, dried leaves, stem and other easily available plant materials, soil clods and feathers for maintaining internal nest temperature (f ) (photo. s.k. basu) c anada goose branta canadensis (linnaeus, 1758): a m ajestic w ater bird from the continent of n orth a m erica s ai ka t k um ar b as u 140 fig. 2. canada geese nest with eggs (a); size comparison between eggs of canada geese (left) and poultry chicken (right) (b); a canada goose egg shell found outside a nest (left) and an unhatched egg found in an abandoned nest (right) (c); comparison between egg yolks: chicken (left) and canada geese (right) (d); predation by local mammals (like badgers, coyote and foxes) or birds (like raptors) act as a natural balance for regulating wild canada geese populations (e); accidental road kills are a common cause of death of young, juvenile and adult canada geese (f ) (photo. s.k. basu) 141 bernikla kanadyjska branta canadensis (linnaeus, 1758): majestatyczny ptak wodny z kontynentu ameryki północnej streszczenie bernikla kanadyjska branta canadensis (linnaeus, 1758) jest ikoną dzikiej ameryki i należy, zarówno do naturalnego, jak i społeczno-kulturowego, dziedzictwa ameryki północnej. ten monogamiczny, gromadny i migrujący gatunek majestatycznego ptaka wodnego jest szeroko rozpowszechniony w ameryce północnej. występuje w 8-10 różnych podgatunkach w szerokim spektrum naturalnych siedlisk i ekosystemów. gatunek jest rozpowszechniony od alaski przez kanadę, usa aż do meksyku. aby uniknąć trudnych warunków zimowych, migruje z północy do południowych stanów usa i północnego meksyku. z powodu eksplozji wielkości jego populacji, jak również braku presji drapieżników, ptak ten w niektórych częściach kontynentu, zmienił się w lokalnego szkodnika. to krótkie opracowanie opisuje jego cykl życiowy rozmieszczenie, reprodukcję i zachowanie w środowisku. key words: canada goose, branta canadensis, north america, bird, sub species, canada, the us, mexico c anada goose branta canadensis (linnaeus, 1758): a m ajestic w ater bird from the continent of n orth a m erica 80 annales universitatis paedagogicae cracoviensis studia naturae, 3: 80–89 2018, issn 2543-8832 doi: 10.24917/25438832.3.6 gracjana budzałek, sylwia śliwińska-wilczewska*, adam latała institute of oceanography, university of gdansk, gdynia, poland, *s.sliwinska@ug.edu.pl allelopathic effect of ulva intestinalis l. on the baltic filamentous cyanobacterium nostoc sp. introduction field evidence and laboratory studies indicate that allelopathy occurs in all aquatic habitats (marine, brackish, and freshwater) and that all primary producing organisms (cyanobacteria, microand macroalgae as well as angiosperms) are capable of producing and releasing allelopathically active compounds (tang, 2011). molisch (1937) was the first who defined the term allelopathy in a broad sense to describe either positive or negative biochemical interactions among plants. after a few decades, inderjit and dakshini (1995) gave an overview of allelopathic activities in aquatic habitats with particular emphasis on algae. currently, it is believed that allelopathy is a prevalent natural phenomenon in aquatic ecosystem (omezzine et al., 2009). macroalgae from the genus ulva are cosmopolitan organisms, and in nutrient-rich coastal waters, they are often dominant and even bloom-forming species (rybak, 2018a,b; rybak, gąbka, 2018). several species of ulva have distributions that extend into the baltic sea (leskinen, 2004). ulva intestinalis l. (synonym: enteromorpha intestinalis (l.) nees) is the principal marine and benthic macroalga growing in isolated rockpools, on rocks, and even on artificial substrates (breakwaters, jettys, etc.) (björk, 2004). some researchers have demonstrated that ulva can reduce eutrophication and promote the productivity, survival rate, and feeding coefficient of the culture species, e.g., prawn and shrimp, by means of polyculture (wang et al., 2001). many papers have reported that ulva can take up nutrients from mariculture waters and improve water quality (jin, dong, 2003). harmful cyanobacterial blooms (cyanohabs) are a significant threat to fisheries, economies around the world, and public health (paerl, 2018). on the other hand, allelopathy in aquatic environments may provide a competitive advantage of selected macroalgae relative to other primary producers, including cyanobacteria (gross, 2010). 81 a llelopathic effect of u lva intestinalis l. on the b altic filam entous cyanobacterium n ostoc sp. thus, allelopathic macroalgae may be a promising mitigation strategy for cyanohabs (tang, 2011). however, we have still little knowledge about the allelopathic interactions between the macroalgae and cyanobacteria. in the present study, we performed a series of laboratory experiments under controlled conditions to examine the allelopathic interactions between the ulva intestinalis and nostoc sp. in order to verify the hypothesis of allelopathy between macroalgae and cyanobacteria. material and methods the experiments were conducted with the strain ba-81 of cyanobacterium nostoc sp. (fig. 1). this strain was isolated from the coastal zone of the gulf of gdańsk (southern baltic sea) and was maintained as monocultures in the culture collection of baltic algae (ccba) at the institute of oceanography, the university of gdańsk, poland (latała et al., 2006). brackish water adapted macroalgae u. intestinalis was selected, based on allelopathic potential of species from the order ulvales (e.g., nan et al., 2004). analysed green alga was collected manually from the coastal zone of the gulf of gdańsk (54°30ʹ16ʹʹn 18°33ʹ26ʹe) and immediately and carefully washed with distilled water to remove attached organisms. the cyanobacteria culture used in the experiments was maintained in 25-ml glass erlenmeyer flasks at 17°c and a 16:8 h of light : dark cycle at a irradiance of 20 μmol photons (par) m–2s–1. the culture was acclimated to these conditions for 7 days, and these growth conditions were used for the experiments. fluorescent lamps (cool white 40w, sylvania, usa) were used as a source of irradiance. the intensity of photosynthetically active radiation (par) was measured using a quantum-meter (licor, nebraska, usa) with a cosine collector. the culture medium employed was f/2 (guillard, 1975). culture media were prepared with baltic sea water filtered through fig. 1. nostoc sp. strain ba-81 used in this study: left panel depicts cyanobacterial filaments from the light microscope (a), whilst right panel illustrates target species under an epifluorescence microscope (b). scale bars = 10 µm (photo. s. śliwińska-wilczewska) g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a, a da m l at ał a 82 glass fibre filters (whatman gf/c) and autoclaved. the salinity was 8 psu as measured with a salinometer (inolab cond level 1, weilheim in oberbayern, germany). allelopathic effects were tested according to a method proposed by ghobrial et al. (2015) with modifications. the cyanobacteria cultures were exposed to the macrophyte extracts obtained from u. intestinalis. dried plant materials were homogenised in a mortar grinding machine. for the bioassay experiment, 2 g dry weight of dried plants was extracted with 40 ml of f/2 medium for 10 minutes. extracts were filtered through glass fibre filters (whatman gf/c) using a vacuum pump (400 mbar) to remove plant particles for bioassay experiments. the concentrations of major nutrients in the controls and all treatments were adjusted to the same level as in the f/2 growth medium. therefore, the effects of major nutrients, microelements, and vitamin limitations in the control and allelochemical treatments can be excluded. sterilised erlenmeyer flasks 25-ml contained 10 ml f/2 medium with a cyanobacterial initial inocula (the final chlorophyll a (chl a) concentration in the experimental cultures was 0.4 µg chl a ml–1) and different volumes of green alga extract treatments. experimental treatments were prepared by adding 100, 500, and 1000 µl of these extracts to 25-ml erlenmeyer flasks containing 10 ml of cell suspensions of the targeted cyanobacteria. the final concentrations of extract were 10, 50, and 100 µl ml–1. the selection of these concentrations was based on previous introductory experiments to determine the effective-range broadly. controls consisted in the addition of 100, 500, and 1000 µl of filtrated f/2 medium to 25-ml erlenmeyer flasks containing 10 ml of cell suspensions of the same cyanobacteria species. the aliquots of the target species inoculated in the experimental flasks came from exponentially growing cultures. the flasks with cyanobacteria were swirled daily. tests were conducted in triplicate. the experiments lasted 7 days. the number of cells (n) in nostoc sp. cultures was estimated with previously determined linear correlations between cell abundance (n ml–1) and optical density (od). n was counted using a bürker chamber (48 squares per count) and a light microscope (lm) following a procedure according to guillard and sieracki (2005), and the od was measured spectrophotometrically at 750 nm with a multiskan go uv-vis spectrophotometer (thermo scientific, massachusetts, usa). the linear correlation between n and od for nostoc sp. was y = 39.8·106x 1.1·104; (r2 = 0.95), where y = n (ml–1) and x = od. od measurements were performed on the 0th, 1st, 3rd, and 7th days of experiment and control. chlorophyll a fluorescence was measured with a pulse amplitude modulation (pam) fluorometer (fms1, hansatech), using a 594 nm amber modulating beam with a 4-step frequency control as a measuring light. samples were taken for chlorophyll fluorescence analysis after the 1st, 3rd, and 7th days of experiment. samples were filtered through 13-mm glass fibre filters (whatman gf/c). before measurement, the filtered 83 sample was kept in the dark for approximately 5 min. the maximum psii quantum efficiency (fv/fm) was calculated (campbell et al., 1998). repeated measures anova was used to test the effect of macroalgae extract on the growth and fluorescence of the targeted cyanobacteria during the following days of experiment. a post-hoc tukey’s test was used to determine significant differences between the control and the other treatment levels. data are reported as the means ± standard deviations (sd). the statistical analyses were performed using statistica® 13.1 software. results our experiment demonstrated that the dry powder of u. intestinalis contains water-soluble allelochemical(s) and is capable of restricting the growth and fluorescence of filamentous cyanobacterium nostoc sp. the addition of 10, 50, and 100 µl ml–1 of extracts obtained from ulva intestinalis significantly decreased the number of cells of nostoc sp. (anova, f3,16 = 11.2, p < 0.001, anova, f3,16 = 25.4, p < 0.001 and anova, f3,16 = 26.3, p < 0.001, respectively), whereas the control sample showed acfig. 2. the number of nostoc sp. cells (106 cell ml–1) for controls (c) and experiments (ex) with addition of extracts: 10, 50 and 100 (µl ml–1) obtained from ulva intestinalis l. after 0, 1, 3, and 7 days of exposure; the values refer to means (n = 3, mean ± sd); asterisks indicates statistically significant difference compared with control obtained with the anova and tukey’s post hoc test; levels of significance were: * p < 0.05; ** p < 0.01; *** p < 0.001 a llelopathic effect of u lva intestinalis l. on the b altic filam entous cyanobacterium n ostoc sp. g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a, a da m l at ał a 84 tive growth (fig. 2). after the addition of extracts obtained from u. intestinalis, the highest decline in growth for nostoc sp. was observed on the third and seventh day of the experiment. the growth of target cyanobacterium after the third day and the addition of 10 and 50 µl ml–1 of extracts obtained from u. intestinalis was reduced to 59% (tukey, p < 0.001) and 71% (tukey, p < 0.01), respectively, compared to the control. moreover, the addition of 50 and 100 µl ml–1 extracts inhibited the growth of cyanobacterium; and, after seven days of exposition, the reduction was 35% (tukey, p < 0.001) and 81% (tukey, p < 0.01), respectively, of the initial amount of nostoc sp. the effect of macrophytes extracts on chlorophyll fluorescence parameter fv/fm after 1, 3, and 7 days of incubation is shown in figure 3. the cyanobacterium showed statistically significant different responses to 10, 50, and 100 µl ml–1 of extract additions obtained from u. intestinalis (anova, f2,12 = 6.9, p < 0.001, anova, f2,12 = 0.9, p > 0.05 and anova, f2,12 = 44.5, p < 0.0001, respectively). it was found that, on the third day of the experiment, the values of fv/fm of target cyanobacterium after the addition of 10 µl ml–1 extracts was reduced to 75% (tukey, p < 0.05), compared to control treatment. the highest decrease in fv/fm for nostoc sp. was observed after the first, third, and seventh day of experiment, after the addition of 100 µl ml–1 extracts obtained from u. inestinalis with a magnitude of 69% (tukey, p < 0.01), 59% (tukey, p < 0.001) and 49% (tukey, p < 0.001), respectively, compared to the control. discussion aquatic macroalgae and macrophytes have long been suspected of suppressing phytoplankton growth through the excretion of chemical substances that inhibit phytoplankton growth (hutchinson, 1975). in addition, the production and excretion of allelochemicals by aquatic macroalgae and macrophytes could be an effective defence strategy against other photosynthetic organisms competing for nutrients and light (wium-andersen et al., 1982; gopal, goel, 1993; elakovich, wooten, 1995). some of the results confirmed that ulva sp. is able to suppress the growth of different species by allelopathy (friedlander et al., 1996). nan et al. (2008) demonstrated that green macroalga ulva sp. has allelopathic effects on the three species of red tide microalgae. although toxic properties are rarely associated with the bloom-forming green macroalgae, there is evidence that ulva sp. produce chemical defences against herbivores (van alstyne et al., 2001), and their extracts have allelopathic properties on other organisms, such as larval oyster, fucus gardneri p.c. silva zygote and epiphytic diatom (nelson et al., 2002). on the other hand, nan et al. (2004) demonstrated that in the nutrient replete semicontinuous co-cultures, ulva sp. may have continuously released growth-inhibiting allelochemicals throughout the cultivation period; whereas, in the initial addition culture solution, the allelochemicals may inhibit growth only 85 at the start of cultivation, because they are quickly degradable. furthermore, nakai et al. (1999) showed that initial addition culture solution might underestimate the allelopathic inhibitory effects as compared with coexistence assays. moreover, allelochemicals could exhibit positive effects on the target organism at lower concentrations and exhibit negative effects at higher concentrations (van aller, 1985). other studies have shown that macrophytes may also exhibit allelopathic effects on different phytoplankton species, including cyanobacteria (van donk, gulati, 1995; mjelde, faafeng, 1997). mjelde and faafeng (1997) found that ceratophyllum demersum l. hampers phytoplankton development in some small norwegian lakes over a wide range of phosphorus concentrations and geographical latitude. moreover, van donk and gulati (1995) suggested that c. demersum can inhibit the growth of both epiphytes and phytoplankton species. additionally, the impact of different submerged macrophytes or their extracts on natural phytoplankton assemblages was studied under experimental conditions by jasser (1995). this author also concluded that the release of organic compounds by c. demersum apparently contributed to a decline of cyanobacteria oscillatoria limnetica lemm. by changing the phytoplankton dominance fig. 3. the fluorescence parameter fv/fm of nostoc sp. cells for controls (c) and experiments (ex) with addition of extracts: 10, 50 and 100 (µl ml–1) obtained from ulva intestinalis l. after 1, 3, and 7 days of exposure; the values refer to means (n = 3, mean ± sd); asterisks indicates statistically significant difference compared with control obtained with the anova and tukey’s post hoc test; levels of significance were: * p < 0.05; ** p < 0.01; *** p < 0.001 a llelopathic effect of u lva intestinalis l. on the b altic filam entous cyanobacterium n ostoc sp. g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a, a da m l at ał a 86 structure. natural phytoplankton assemblages, in which cyanobacteria were abundant, were incubated in the field in bags together with intact plants or plant extracts. live c. demersum, myriophyllum spicatum l., potamogeton lucens l., statiotes aloides l., and chara fragilis desv. and its extract, had quite similar effects on the phytoplankton; whereas, the biomass and percentage contribution of cyanobacteria to total algal biomass declined, and those of green algae increased. wium-andersen et al. (1983) isolated a sulphur compound with allelopathic properties from c. demersum. moreover, gross and sütfeld (1994) found that m. spicatum l. was able to release allelopathic polyphenols into the surrounding water and thereby strongly suppressed the growth of cyanobacterium anacystis sp. some authors noted that the identified allelochemicals belong to different chemical classes, such as sulphur compounds, polyacetylenes, polyphenols, and oxygenated fatty acids; however, most of the allelochemicals are still not identified (gross, 1999). in this study, it has been demonstrated that u. intestinalis can release some kind of allelopathic substances and effectively inhibit the growth and fluorescence parameter of cyanobacterium nostoc sp. our results not only provide insight into the interactions between macroalgae and cyanobacteria but also lead us to isolate and characterize these allelopathic substances in the future. moreover, our results may provide new insight into the ecological role of macroalgae, such as u. intestinalis, in the occurrence of cyanohabs. acknowledgements the authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. this study was supported by bmn grants, poland, no. 538-g245-b116-18. references björk, m., axelsson, l., beer, s. 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(1995). transition of a lake to turbid state six years after biomanipulation: mechanisms and pathways. water science and technology, 32, 197–206. doi: 10.1016/02731223(95)00699-0 wang, j.q., jin, c.l., zhang, x., liu, g. (2001). polyculture of experiment penaeus chinensis with various biomass of ulva pertusa. journal of fisheries of china, 25, 32–38. wium-andersen, s., anthoni, u., houen, g. (1983). elemental sulphur, a possible allelopathic compound from ceratophyllum demersum. phytochemistry, 22, 2613. doi: 10.1016/0031-9422(83)80178-2 wium-andersen, s., christophersen, c., houen, g. (1982). allelopathic effects on phytoplankton by substances isolated from aquatic macrophytes (charales). oikos, 39, 187–190. doi: 10.2307/3544484 abstract allelopathy is a prevalent natural phenomenon in aquatic ecosystems. we reported the effects of the green macroalga ulva intestinalis l. collected from estuaries of the baltic sea (poland) on the growth and chlorophyll fluorescence of common filamentous cyanobacterium nostoc sp. it was found that the addition of 50 µl ml–1 extracts obtained from u. intestinalis inhibited the growth of cyanobacterium, and, after one week of exposition, the reduction was 35% of initial amount of nostoc sp. in addition, we demonstrated that, on the seventh day of the exposition, the values of fv/fm of target cyanobacterium after the addition of 100 µl ml–1 extracts obtained from u. intestinalis was reduced to 49%, compared to control treatment. these results showed for the first time the allelopathic activity of u. intestinalis on baltic filamentous cyanobacteria nostoc sp. key words: allelopathy, chlorophyta, cyanobacteria, extract, fluorescence, green algae, growth, macroalgae, ulvophyceae received: [2018.03.04] accepted: [2018.10.30] zjawisko oddziaływania allelopatycznego ulva intestinalis l. na bałtycką sinicę nitkowatą nostoc sp. streszczenie allelopatia jest powszechnym zjawiskiem występującym w  ekosystemach wodnych. w  artykule opisano wpływ zielenicy bentosowej ulva intestinalis l. (ulwa kiszkowata), zebranej z  morza bałtyckiego (zatoka gdańska); na wzrost i fluorescencję chlorofilu u nitkowatej sinicy nostoc sp. na podstawie uzyskanych 89 wyników badań stwierdzono, że po dodaniu ekstraktów uzyskanych z  u. intestinalis, najmniejszy wzrost nostoc sp. obserwowano w  siódmym dniu eksperymentu. dodanie 50 µl ml–1 ekstraktu z  u. intestinalis zahamowało rozwój sinicy nostoc sp. i po 7 dniu ekspozycji jej wzrost osiągnął poziom 35% w stosunku do warunków kontrolnych. ponadto największy spadek wartości parametru fluorescencji fv/fm dla nostoc sp. obserwowano po tygodniu trwania eksperymentu. stwierdzono, że po 7 dniach ekspozycji wartość fv/fm dla badanej sinicy po dodaniu 100 µl ml–1 ekstraktu z u. intestinalis spadła o 49%. przeprowadzone badania po raz pierwszy wykazały, że u. intestinalis wpływa allelopatycznie na bałtyckie sinice nitkowate z rodzaju nostoc. słowa kluczowe: allelopatia, chlorophyta, ekstrakt, fluorescencja, makroglony, sinice, ulvophyceae, wzrost, zielenice information on the authors gracjana budzałek the field of her interest is allelopathic interactions between macroalgae and cyanobacteria. in her research, she is focusing mostly on baltic species. she is investigating the influence of allelopathic compounds produced by macroalgae on bloom-forming cyanobacteria. sylwia śliwińska-wilczewska https://orcid.org/0000-0002-3147-6605 she is interested in the allelopathy of cyanobacteria and microalgae, in particular, of picocyanobacteria synechococcus sp. allelopathy plays an important role in interspecific competition and contributes to cyanobacterial bloom maintenance. in her study, the influence of allelochemicals on the growth, chlorophyll fluorescence, and photosynthesis irradiance curves of different phytoplankton species was investigated. she is also investigating the influences of environmental factors on produced allelopathic compounds on algae and cyanobacteria. adam latała https://orcid.org/0000-0001-6092-887x he has wide experience in ecophysiology and ecotoxicology of marine benthic and planktonic algae, and the influence of the main environmental factors, such as salinity, temperature, and light on the photosynthesis, photoacclimation, fluorescence, respiration, and the growth of algae from natural communities and cultured under laboratory conditions. he studies the use of fluorescence techniques to determine algal and cyanobacterial ecophysiology and ecotoxicology. he is the curator of culture collection of baltic algae (ccba) at the institute of oceanography, university of gdańsk. actually, ccba maintains more than 100 baltic strains from three taxonomic groups: blue-green algae, green algae, and diatoms. a llelopathic effect of u lva intestinalis l. on the b altic filam entous cyanobacterium n ostoc sp. 47 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 47–53, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.6 renata muchacka1*, ewa sosnówka-czajka2, iwona skomorucha2, edyta kapusta1, agnieszka greń1, zofia goc1 1department of animal physiology and toxicology, institute of biology, pedagogical university of cracow, podbrzezie 3, 30-054 kraków, poland, *renata.muchacka@up.krakow.pl 2department of poultry breeding, national research institute of animal production, krakowska 1, 32-083 balice n. kraków, poland antioxidant enzymes activity, gsh and mda level in eggs from heritage breeds introduction in eggs, as in any raw food material, there are a number of di�erent quality changes. �e moment of laying the egg is a  speci�c beginning of these changes. from that moment, the eggs begin to undergo metabolic processes that contribute to the gradual deterioration of the quality of the raw material until the loss of consumer and technological usefulness (jankowska, 2010). on the farm, there is a  group of factors that have an impact a�er the egg is laid and modi�es their quality. �ese include hygienic conditions, and the manner and conditions for the collection and distribution of raw egg product. �e temperature and distribution time between the producer and the consumer as well as the storage conditions are decisive here (calik, 2011). another group are factors that a�ect the quality of eggs even before they are laid depend on the laying hens and, to a large extent, on the farmer. �e quality of eggs is in�uenced by the origin of laying hens (i.e. breed, line). it a�ects, among other things, the size of the egg, the colour of the eggshell, and the nutritional value (singh et al., 2009; calik, 2013). moreover, the following factors are also important: the age of the laying hens (older hens laid larger eggs, but of inferior quality shells), the rearing system, their feeding (batkowska, brodacki, 2017), and veterinary prevention (calik, 2013). �e aim of the study was to determine the di�erences in antioxidant enzyme activities and di�erences in the level of reduced glutathione and malondialdehyde in eggs from three heritage breeds. r en at a m uc ha ck a, e w a s os nó w ka -c za jk a, iw on a s ko m or uc ha , ed yt a k ap us ta , a gn ie sz ka g re ń, z of ia g oc 48 material and methods hens and experimental design �e experiment was carried out with 99 hens, approximately 48 weeks of age, of three heritage breeds, ‘greenleg partridge’ (z-11), ‘sussex’ (s-66), and ‘rhode island red’ (r-11), which were assigned to groups i to iii (33 hens per group), respectively. each group was subdivided into 3 subgroups, each having 11 birds. hens were reared in the litter system at a stocking density of 9 birds · m-2 and had outdoor access with green paddocks (2.5 m2/hen). �e birds were fed ad libitum with standard diets based on concentrates for laying hens, which contained 17.5% protein and 11.6 mj me/ kg feed (fig. 1). �e diet consisted of wheat, maize, soybean expeller, dried alfalfa, rapeseed oil, limestone, di-calcium phosphate, common salt, dl-methionine, and dj pre�x. �e birds had free access to water throughout the experiment. all the groups were managed under uniform environmental (air humidity and temperature, lighting programme) and feeding conditions. a  lighting schedule of 16 h of light and 8 h of darkness (16l : 8d) was used. sample collection and laboratory analyses at 48 weeks of age, 6 eggs from each group were collected for the next three days. eggs were evaluated a�er 24 h of refrigerated storage at 4°c and 55% humidity. in the samples of egg yolks and albumens, the concentration of glutathione (gsh) and malondialdehyde (mda), and the activity of superoxide dismutase (sod), glutathione peroxidase (gpx), and catalase (cat) were estimated. �e collected yolks and albumens were homogenised in 50 mm phosphate bu�er cooled to 4°c and containing k2hpo4 (dipotassium hydrogen phosphate) + kh2po4 (potassium dihydrogen phosphate) + fig. 1. nutritive value of the diet 17.5 0.41 0.86 3.7 0.4 0 5 10 15 20 crude protein methionine lysine calcium phosphorus [%] components a ntioxidant enzym es activity, g s h and m d a level in eggs from heritage breeds 49 0.1 mm edta (ethylenediaminetetraacetic acid disodium salt) (ph = 7.0) + 0.1% bsa (bovine serum albumin) and then centrifuged. �e obtained supernatants of the yolks and albumens were used for further investigations. �e activity of sod in the supernatants of homogenates of yolks and albumens was determined according to spectrophotometric cytochrome method by diplock et al. (1991). �e activity of cat was determined according to the spectrophotometric method by aebi (1983), and the activity of gpx was estimated by the modi�ed method of lück (1963). �e activity of the studied antioxidative enzymes in yolk and albumen extracts was expressed in u · mg-1 of protein. mda reacts with thiobarbituric acid (tba) in an acid medium and the produced coloured tba-complex that could be measured colorimetrically according to the method of ohkawa et al. (1979). mda concentration was expressed in nm · mg-1 protein. total protein concentration in the supernatants of homogenates of the yolk and albumen was determined according to the bradford method (1976) with bovine serum albumin as a standard. in the supernatants of yolks and albumens, obtained a�er deproteinisation and centrifugation, gsh was determined according to the method of ellman (1959). �e concentration of gsh was estimated in the yolk and albumen in μm · g-1 of protein. statistical analysis all data was analysed by the analysis of variance procedures (anova) and duncan’s multiple-range test using the computer program statistica version 10.0 pl. �e data was expressed as means ± sd and was considered as signi�cant when p values were less than 0.05. tab. 1. antioxidant enzymes activity and the level of mda and gsh in yolk; values shown as di�erent letters within the line di�er signi�cantly according to the duncan test at p ≤ 0.01 item group sd i ii iii sod [u/mg] 3.0240a 7.5730b 3.0130a 1.0810 gpx [u/mg] 0.0044 0.0065 0.0048 0.0034 cat [u/mg] 0.0470 0.0650 0.0890 0.0620 gsh [µm/g] 0.3410b 0.3850b 0.1250a 0.0170 mda [µg/g] 2.4490ab 2.2500a 2.4680b 0.0460 results �e highest activity of sod, both in yolks (tab. 1) and albumens (tab. 2), was found in eggs of sussex hens (p ≤ 0.01). gpx and cat activity was in all eggs at a similar level. �e highest level of gsh was in egg yolks of ‘greenleg partridge’ and ‘sussex’ hens (p ≤ 0.01) r en at a m uc ha ck a, e w a s os nó w ka -c za jk a, iw on a s ko m or uc ha , ed yt a k ap us ta , a gn ie sz ka g re ń, z of ia g oc 50 (tab. 1) and in egg albumens of ‘greenleg partridge’ hens (p ≤ 0.01) (tab. 2). �e lowest level of mda was found in egg yolk and egg albumen of ‘sussex’ hens (p ≤ 0.05) (tab. 1–2). tab. 2. antioxidant enzymes activity and the level of mda and gsh in albumen; values shown as di�erent letters within the line di�er signi�cantly according to the duncan test at p ≤ 0.01 item group sd i ii iii sod [u/mg] 1.1320a 3.1870b 1.1290a 0.4580 gpx [u/mg] 0.0049 0.0062 0.0079 0.0032 cat [u/mg] 0.0640 0.0650 0.0940 0.0660 gsh [µm/g] 0.3250b 0.1500a 0.1460a 0.0160 mda [µg/g] 1.8790ab 1.6080a 1.9130b 0.0220 discussion in optimal conditions, there is a  balance between oxidants and antioxidants in the body. �e increase in free radical production or decrease of antioxidant activity causes a  disturbance of balance in the oxidation reaction, which is called oxidative stress. during oxidative stress, stationary concentrations of reactive oxygen species (ros) increase signi�cantly. free radicals oxidize row fatty acids (lipids), structural proteins, and enzymatic proteins (surai, 2015). �erefore, it is important that the body’s antioxidant system functions properly. �e quality characteristics of eggs and also the antioxidant enzymes and gsh found in the eggs are synthesised and formed in the hen’s body during eggs formation. according to our research, the activity of antioxidant enzymes (mainly sod) and the levels of gsh and mda found in eggs di�er depending on the breed of hens. �e highest activity of sod, the highest level of gsh, and the lowest level of mda were characterised by eggs from sussex hens, which may indicate they have the best quality. similar results were obtained in the previous studies by muchacka et al. (2016; 2018). it is known that the origin of the laying hens (i.e. breed, line) a�ects the quality of eggs. it a�ects egg size, shell colour, and the nutritional value (singh et al., 2009; calik, 2013) and probably the activity of antioxidant enzymes and levels of gsh (muchacka et al., 2016; 2018). di�erences in the activity of antioxidant enzymes and gsh levels may be caused by di�erences in the metabolism of the tested hens. it may be important because these substances can a�ect the quality of the poultry products, e.g., eggs and protect the germ cell and growing embryo against ros in the �rst days a�er the egg is laid. based on the obtained results, it can be concluded that the activity of antioxidant enzymes (mainly sod) and the levels of gsh and mda which occur in eggs may depend on the breed of hens and may indicate their quality; however, further studies on this topic are necessary. a ntioxidant enzym es activity, g s h and m d a level in eggs from heritage breeds 51 references aebi, h.e. (1983). catalase. in: h.u. bergmeyer (ed.), methods of enzymatic analysis. new york: academic press, pp. 273–286. batkowska, j., brodacki, a. (2017). selected quality traits of eggs and the productivity of newly-created laying hens dedicated to extensive system of rearing. archives of animal breeding, 60, 87–93. bradford, m.m. (1976). a  rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle for protein-dye binding. analytical biochemistry, 72, 248–254. calik, j. (2011). ocena jakości jaj sześciu rodów kur nieśnych w zależności od ich wieku. żywność nauka technologia jakość, 5(78), 85–93. [in polish] calik, j. (2013). zmiany cech jakościowych jaj pochodzących od kur nieśnych żółtonóżka kuropatwiana (ż-33), w zależności od warunków ich przechowywania. żywność nauka technologia jakość, 2(87), 73–79. [in polish] diplock, a.t., symons, m.c.r., rice-evans, c.a. (1991). techniques in free radical research. london: elsevier. ellman, g.l. (1959). tissue sul�ydryl groups. archives of biochemistry and biophysics, 82, 70–77. doi: 10.1016/0003-9861(59)90090-6 jankowska, k. (2010). od zniesienia do konsumpcji – jaja kurze (spożywcze). zachowanie higieny w produkcji jaj. polskie drobiarstwo, 2, 52–54. [in polish] lück, h. (1963). peroxidases. in: h.u. bergmeyer (ed.), methods of enzymatic analysis. new york: academic press, pp. 895–897. muchacka, r., sosnówka-czajka, e., skomorucha, i. (2018). antioxidant enzymes activity, gsh and mda level in eggs from heritage breeds reared under organic conditions. 8th international scienti�c symposium „poultry days 2018“, 21–22. june 2018, brno, czech republic, 30. muchacka, r., sosnówka-czajka, e., skomorucha, i., kapusta, e., greń, a., goc, z. (2016). lipid peroxidation, antioxidant enzymes activity and glutathione level in eggs from heritage breed and commercial crosses hens. proceedings of 12th international scienti�c conference “animal physiology 2016”, may 13th–15th 2016, bořetice, czech republic, 191–196. ohkawa, h., ohishi, n., yagi, k. (1979). assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. analytical biochemistry, 95, 351–358. singh, r., cheng, k.m., silversides, f.g. (2009). production performance and egg quality of four strains of laying hens kept in conventional cages and �oor pens. poultry science, 88(2), 256–264. doi: 10.3382/ps.2008-00237 surai, p. (2015). antioxidant systems in poultry biology: superoxide dismutase. journal of animal research and nutrition, 1(1), 1–8. doi: 10.21767/2572-5459.100008 abstract �e objective of the study was to determine the activity of antioxidant enzymes and the level of gsh and mda in the yolk and albumen of hen’s eggs. �e experiment was carried out with 99 hens of three heritage breeds (‘greenleg partridge’ – group i, ‘sussex’ – group ii, ‘rhode island red’ – group iii). layers were reared in the litter system and had outdoor access. birds were fed ad libitum standard diets based on concentrates for laying hens and had free access to water throughout the experiment. all the groups were managed under uniform environmental (air humidity and temperature, lighting programme) and feeding conditions. at 48 weeks of age, 6 eggs from each group were collected. in the samples of egg yolks and albumens, the concentration of gsh and mda, and activity of sod, gpx, and cat were estimated. �e results were statistically analysed by one-way analysis of variance, and signi�cant di�erences were estimated with duncan’s multiple range tests. �e highest activity of sod, the highest level of gsh, and the lowest level of mda were r en at a m uc ha ck a, e w a s os nó w ka -c za jk a, iw on a s ko m or uc ha , ed yt a k ap us ta , a gn ie sz ka g re ń, z of ia g oc 52 characterised by eggs from ‘sussex’ hens, which may indicate that they have the best quality. based on the obtained results, it can be concluded that the activity of antioxidant enzymes (mainly sod) and the levels of gsh and mda that occur in eggs may depend on the breed of hens. key words: antioxidant enzymes, eggs, gsh, hens, mda received: [2018.05.15] accepted: [2018.07.30] aktywność enzymów antyoksydacyjnych, poziom gsh i mda w jajach pochodzących od kur ras zachowawczych streszczenie celem doświadczenia było zbadanie aktywności enzymów antyoksydacyjnych oraz poziomu gsh i mda w żółtkach i białkach jaj kurzych. materiał doświadczalny stanowiło 99 kur trzech ras zachowawczych (‘zielononóżka kuropatwiana’ – i grupa, ‘sussex’ – ii grupa i ‘rhode island red’ – iii grupa). nioski utrzymywano w systemie ściółkowym z możliwością korzystania z trawiastych wybiegów. ptaki karmiono ad libitum standardową paszą przygotowaną na bazie koncentratów dla kur niosek. w trakcie doświadczenia nioski miały swobodny dostęp do wody. wszystkie grupy były utrzymywane w jednakowych warunkach środowiskowych (wilgotność powietrza i temperatura, program świetlny) i żywieniowych. w 48. tygodniu życia, z każdej grupy zebrano po 6 jaj. w próbkach żółtek i białek oznaczono poziom gsh i mda oraz zbadano aktywność sod, gpx i cat. wyniki zostały opracowane statystycznie za pomocą jednoczynnikowej analizy wariancji, szacując istotności różnic testem duncana. najwyższą aktywnością sod, najwyższym poziomem gsh i najniższym poziomem mda charakteryzowały się jaja od kur rasy ‘sussex’, co może wskazywać na ich najlepszą jakość. na podstawie uzyskanych wyników można stwierdzić, iż aktywność enzymów antyoksydacyjnych (głównie sod) oraz poziom gsh i mda występujących w jajach mogą zależeć od rasy kur. słowa kluczowe: enzymy antyoksydacyjne, jaja, gsh, kury, mda information on the authors renata muchacka https://orcid.org/0000-0002-8255-006x she is an assistant professor at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. she is interested mainly in animal physiology, oxidative stress in animals, and the quality of food products of animal origin. ewa sosnówka-czajka https://orcid.org/0000-0003-3720-1685 she is an assistant professor at the department of poultry breeding in the national research institute of animal production in cracow. she is interested mainly in poultry welfare and the technology of poultry production. iwona skomorucha https://orcid.org/0000-0003-1241-7703 she is an assistant professor at the department of poultry breeding in the national research institute of animal production in cracow. she also focuses on poultry welfare and the technology of poultry production. edyta kapusta https://orcid.org/0000-0002-4350-5514 she is a laboratory specialist at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. she is interested in animal physiology, mainly in oxidative stress in animals. agnieszka greń https://orcid.org/0000-0003-2383-1096 she is a  professor at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. she is interested mainly in animal physiology, diabetes, and natural products, predominantly in diabetes prevention and therapy. a ntioxidant enzym es activity, g s h and m d a level in eggs from heritage breeds 53 zo�a goc https://orcid.org/0000-0001-9626-5895 she is an assistant professor at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. she is interested in research on animal cells in vitro and animal physiology. 142 annales universitatis paedagogicae cracoviensis studia naturae, 3: 142–151, 2018, issn 2543-8832 doi: 10.24917/25438832.3.11 zuzana frajtová, marcela kocianová-adamcová* department of biology and ecology, faculty of natural sciences, matej bel university, tajovského 40, 974 01 banská bystrica, slovakia, *marcela.adamcova@umb.sk canistherapy and its application to the teaching process at a grammar school the positive effect of animal on humans has been known since the beginning of the domestication of animals. with the help of animals, it is possible to influence a wide range of human physical and mental problems, which are caused especially by the modern way of life and by the disconnexion from nature. this paper presents an overview of one of the forms of zootherapy – canistherapy, which exploits the positive influence of dogs on both physical and mental health of humans. since the dog is the oldest domesticated animal, during its cohabitation with man, it has learned to read the body language of humans and is also able to feel men´s conditions. canistherapy is currently slowly getting into the public awareness; however, in slovakia, this process is hampered mainly due to the lack of information and scientific research but also because of the lack of willingness or distrust of its application in practice. this work compares the changes in the behaviour and level of knowledge of 12 to 13 year-old students at a grammar school in slovakia after the implementation of the canistherapy into the teaching process. canistherapy sessions were focused on the development and changes in social behaviour, emotional expressions, and the knowledge of the students. these areas were continuously monitored and finally evaluated by questionnaires and interviews. we analysed the effect of animal assisted education (aae) form of canistherapy, which helped us to establish the dog’s positive impact on the mental conditions of 12 to 13 year-old students. the experimental group (class a) where canistherapy was applied consisted of 28 students (fig. 1a), while the control group (class b), where the training was conducted without the presence of dogs, consisted of 27 students. canistherapy in the selected class was realised by canistherapist mgr. martina michalková with her four therapy dogs, eila, a 15-year-old west white highland terrier (wwht), kloe, a 3-year-old wwht breed, keri, an 11-year-old wwht breed, and elfin, a 10-year-old samoyed female. all dogs are certified and have previously completed regular canistherapeutic trials (fig. 1b). c anistherapy and its application to the teaching process at a gram m ar school 143 fig. 1. the students of the experimental class a with the therapy dogs (a); martina michalková and her therapy dogs from a civic association “the meaning of life” (b) (photo. t. čontofalský) zu za na f ra jto vá , m ar ce la k oc ia no vá -a da m co vá 144 tab. 1. overview of the topics that have been taught during the research in class a month topics september basic information about civic association "the meaning of life" establishment of the rules during a work with animlas information about choosing and traing a dog october vital expressions and animal behaviour november animal reproduction january canistherapy february welfare of the dogs march protection of the wildlife animals april revision of the knowledge and checking the feedback tab. 2. the comparison of the acquired knowledge from biology of class a (with canistherapy) and class b (control group) during the analysed research period no. question/order answer class an = 25 class b n = 24 1. write examples of inappropriate human behavior towards the dogs 3 examples and more 3 examples and more 2 examples and less 25 25 0 5 5 19 2. which protected animal is directly threatened by thermal insulation of panel houses? correct incorrect 25 0 7 17 3. how can dogs communicate with each other? 2 examples and more 1 example 25 0 10 14 4. have you heard about the concept of canistherapy? if yes, briefly characterise it. yes, i have no, i have not 25 0 14 10 5. which senses are well developed in carnivores? correct incorrect correct incorrect 24 1 24 0 20 4 20 46. who has more bones in the body – a dog or a man? 7. write at least 3 examples of protect-ed animals in slovakia 3 examples and more 2 examples and less 22 3 16 8 8. what types of behavior can we ob-serve in mammals? 3 examples and more 2 examples and less 20 5 16 8 9. after how many weeks can be a puppy taken from his mother? correct incorrect 18 7 10 14 10. what is the difference in a set of teeth between the herbivores and carnivores? correct incorrect 18 7 10 14 the research was conducted twice a month from september 2015 to april 2016. after consulting with the biology teacher, the topics that were taught in class a were covered within the curriculum for the 7th year of a grammar school (tab. 1). the initial lessons were focused on basic information about the civic association “the meaning of life”, the canistherapeutical team and its work. to compare the acquired knowledge from the biology of class a and class b during the research period, a short test consistc anistherapy and its application to the teaching process at a gram m ar school 145 ing of 10 questions was used (tab. 2). a total of 25 students participated in class a and 24 students in class b. the content of the lesson was the same in both classes, but the method of presentation was different. while in class b there was a classic explanation, in class a there was an experience learning in which the students were actively involved. to determine the impact of canistherapy on class a students, a questionnaire method was used which consisted of five open-ended questions and 23 students attended it (tab. 3). the aim was to find out the subjective attitudes of students and their perception of canistherapy. to demonstrate the impact of canistherapy on inadaptable students, we interviewed the biology teacher and the canistherapist. the questionnaire showed that the experimental group (class a) had different results than the control group (class b). the exact numbers of the answers can be found in table 2. however, the positive influence of dogs was not only didactic but also psychological. evidence is provided by questionnaires filled in by students from class a and by the teacher, which clearly showed the positive effect of canistherapy on the psychic aspect of the students. a summary of the students’ answers can be found in table 3. the students stated that, thanks to the lessons with the dogs, they were able to better remember the new information (44%). learning with dogs helped the students to realise how to behave correctly to animals and nature (35%), and it also brought new experiences and knowledge (48%). they also understood and memorised the curriculum (17%). all the students said that they would very much like to continue with this form of teaching. they were most interested in the following topics: animal care (especially information about the bears and bats – 31%), canistherapy (demonstration of positioning – 25%) (fig. 2a, b), animal reproduction (21%) and others. in the questionnaire, the teacher stated that her opinion on the usage of the dogs in the educational process is definitely positive. at the beginning of the meetings, the teacher was afraid that students would be disrespectful and undisciplined when the dogs would suddenly appear in the classroom. however, the result was opposite. during the meetings, she watched her students closely, and she was very surprised that they were much quieter, more impressed with the curriculum, and engaged in the discussions and activities, even though they had to be forced into such matters before. according to the teacher, a great contribution of these lessons was the arousal of empathy, of interest in nature, and the possibility of physical contact with the animal (tab. 3). in her opinion, the physical contact is absent among today’s youth. since the parents are often busy, their absence is compensated by gifts, and not by real contact and time spent with their children. the meetings with animals at least partially replaced the absence of such contact. the dog‘s presence had both direct and indirect effects, which means that children were directly involved when dogs were present at the lesson but also when dogs were not there, because the students were looking forward to the next meeting and were discussing their experiences with them. zu za na f ra jto vá , m ar ce la k oc ia no vá -a da m co vá 146 tab. 3. the impact of canistherapy on students of class a no. question/order answer type [%] n = 23 1. how do you perceive the fact that the dogs were making you a company during the lesson? it helped me overcome my own fear of dogs 4 i liked it, it was a nice dulcification of the day 17 the meetings were very reassuring 35 using the dogs during the lesson helped me to better remember the new information 44 2. what has the learning with the dogs given to you? new experiences and knowledge 35 finding out how to behave right to animals and nature 48 better understanding and memorizing of the curriculum 17 4. would you like the dogs to continue to be part of the class? the answers of the students showed that all of them would very much like to continue with this form of teaching 100 5. write in brief what you learned during these lessons animal welfare (especially information about the bears and bats) 31 canistherapy (and demonstration of positioning) 25 animal reproduction 21 how do animals behave and how we should treat them 16 the students learned that the foreknowledge of nature is very important 7 during the meetings, the teacher has informed us about an inappropriate student in the experimental class (class a). this student has no proper diagnosis from an expert because of lack of willingness from his parents to pursue the matter, and that is why our results are based only on observations. we decided to find out whether the sessions will be beneficial for him and will improve his performance during classes. his behaviour was carefully monitored during the sessions and at the end evaluated. before the meetings, the student was very sensitive and moody. he was usually absent-minded and unable to concentrate for a longer time. it was necessary to lecture and warn him and, as a consequence, the teaching process was slowed down for the whole classroom. the results of both teachers and canistherapists evaluations showed that there was a marked improvement in his behaviour before and during the canistherapy. in spite of the initial nervousness, the student quickly established a very strong relationship with the dogs. he was in contact with them constantly. from the time they came until they left the classroom, he was holding, hugging, and caressing them. he engaged in the activities and, after the end of the lesson, he continued asking the canistherapist for her personal experiences with dogs. suddenly, he was able to focus more on the curriculum as well as on the overall happenings in the classroom. c anistherapy and its application to the teaching process at a gram m ar school 147 his improvement has been beneficial for the whole classroom, so the achieved results could be evaluated as positive. one of the intentions of the research was to verify how the presence of dogs influences the ability of students to better memorise the curriculum. it was about targeted contacts of students with the animals, particularly focusing on the modification of education and upbringing. the animal itself played the role of some kind of supportive medium which motivates to engage in various educational activities (velemínsky, 2007). de anda (2001) points out the fact that special education in the form of a visitor program offers students, among other things, the possibility of a regular contact with another model of an adult, which can lead to many educational, social, and emotional benefits for students. levinson (1997) claims that the dog does not act as a co-therapist during canistherapeutic activities but as a therapist itself that is in good agreement with our observations. this creates a strong bond between the client and the canistherapeutical dog, which is based on emotions, understanding, and help, whether emotional or physical. according to tichá (2002), dogs in the canistherapy process can be directly involved in the activities, being a motivational factor. for some students, attending a school represents a significant emotional stress, which can be demonstrated not only in their behaviour, but also in their ability to receive and process the curriculum. for this reason, a dog can serve as a suitable medium for releasing and eliminating stress. we are inclines to the opinion of tichá (2002), because, during the canistherapeutic sessions, we observed that the dog was a very strong motivating element for fulfilling the activities and a great motivation for learning. it was proved by the results of the comparative sciental test between classes a and b, where class a students overtopped their class b classmates with higher number of the correct answers (tab. 2). students declared themselves that the lessons with dogs were much more interesting for them than typical lessons. that is why we assumed better results in the test were caused by students´ higher motivation in the teaching process. as piper (2014) claims, students are often stressed by teachers, which may be reflected in their learning achievements. however, in the presence of a dog, this stress disappears, and it is shown that dogs can help the students to improve their knowledge by up to two degrees. in the second part of the research, we dealt with the influence of canistherapy on the mental state of the students. it is possible to assess that the results from the research during canistherapeutic meetings and from the questionnaires confirmed the results of other experts dealing with canistherapy. wilson (1993) discusses the importance of a bond between man and animal, which is natural from historical and biological points of view, but it is often forgotten these days. the author emphasizes that contact with animals enriches us and helps us to develop, and as children are naturally fascinated by nature, and contact with the animal is a good way to raise their interest. sitková zu za na f ra jto vá , m ar ce la k oc ia no vá -a da m co vá 148 fig. 2. demonstration of the positioning (a); demonstration of a work and communication with the canistherapeutical dog (b) (photo. z. frajtova) c anistherapy and its application to the teaching process at a gram m ar school 149 (2010) documented the progress made by zootherapists during their practice. as one of the greatest advances, she clearly marked the overcoming of fear of the animals, the improved communication skills, and the total relaxation of children. in general, all the components of children‘s personality improved, and they were able to keep attention longer. our results were in good agreement with these findings, as the canistherapy helped several students to overcome anxiety and fear of dogs. according to the biology teacher’s assessment, who is also the class teacher of class a, there was an obvious development of the social behaviour, emotions, communication skills, and knowledge of all the students. students whom she usually sees as withdrawn and quiet were suddenly communicative and livelier than ever during the lessons with the dogs assistance. the presence of a dog in the class automatically decreases the shyness that could be felt by the students against the canistherapist in the absence of the canistherapeutical dogs (ellis et al., 2001). it is natural that children are reserved in a presence of an unknown person. thus, it was a great surprise to see how friendly and immediate the students behave to the canistherapist. we attribute that to the presence of the dogs in the classroom. the third part of our survey provides information about the impact of animal assisted education on an inadaptable student. there were visible changes in the behaviour of the inadaptable student, manifested in improved communication skills, longer concentration, calmer behaviour, and interest in the classroom happening. a dog can help to the inadaptable students to improve their speaking skills, memory or vocabulary (kollus, 1999). according to hilbertová (2009), the presence of an animal helps to link up the immediate contact between the teacher and the child. moreover, it can create the atmosphere of confidence and helps to reduce the student’s inappropriate behaviour. students who are often negativist and passive are influenced by canistherapy by improving interhuman relationships, reducing aggression, and developing communication skills (odendaal, 2007). frank (2001) discusses the fact that teachers have strict rules on physical contact with students. however, this contact may be needed from time to time. for this reason, the presence of a dog during the teaching process is beneficial, because the child can caress and hug the dog. our research confirmed that canistherapy is an effective way to strengthen the boy’s mental state in the conditions of the school environment, to dismantle sadness in the short term, to help improving concentration, to damp problematic behaviour, to raise interest in the lesson, and to improve interhuman relationships. conclusion the work was focused on canistherapy and its usage at school facilities. the main aim was to find out and describe the influence of education with assistance dogs on students of a selected class of a grammar school through direct observation, questionzu za na f ra jto vá , m ar ce la k oc ia no vá -a da m co vá 150 naires, and test. a sciental test that compared the knowledge of the experimental class a (with canistherapy) and the control class b (without canistherapy) students at the end of the research period confirmed the presumption that canistherapeutical sessions positively influenced the memorisation of curriculum and also acted as a motivation for students to learn. questionnaires filled by class a students and their biology teacher clearly demonstrated the positive effect of canistherapy on the mental aspect of students. after the end of the canistherapeutical sessions, a positive change of behaviour occurred in the students. they were quieter, more active, and more focused on work tasks, and some individuals were able to overcome their fear of dogs. observing class a students revealed that the canistherapeutical sessions were important, not only for the group, but also for individuals. in the behaviour of the inadaptable student, there were visible changes, such as better communication, longer concentration, calmer behaviour, and interest in classroom happening. subsequent analysis of the results confirmed that canistherapy has a positive impact on individuals and is a valuable supportive method for improving social behaviour, mental health, and communication abilities in children. however, canistherapy is inhibited from increased development and wider use because of a non-existent uniform methodology for canistherapy performance, the lack of professional literature, outdated legislation, and mistrust in application in practice. we believe that the obtained results and findings could act as a tool for other school institutions which are considering canistherapeutical sessions. references de anda, d. (2001). a qualitative evaluation of a mentor program for at-risk youth: the participants perspective. child and adolescent social work journal, 18, 97–117. ellis, j., small-mcginley, j., de fabrizio, l. (2001). caring for kids in communities: using mentorship, peer support, & student leadership programs in schools. new york: counterpoints. hilbertová, l. (2009). felinoterapie a canisterapie v zařizení pro výkon ústavní výchovy. in: pravda o zooterapiích: sborník příspěvků z odborné konference s mezinárodní účastí. české budějovice. [in czech] kollus, b. (1999). the feline gift. in cats magazine, roč. 55: 10. doi: 0008-8544/ 36. levinson, b. (1997). pet-oriented psychotherapy. springfield, illinois: charles c. thomas, publisher. odendaal, j. (2007). zvířata a naše mentální zdraví. praha: nakladatelství brázda, s.r.o., p. 173. [in czech] piper, l. (2014). the practice of animal-assisted psychotherapy: an innovative modality for facilitating mental wellness. usa: e street lane publications llc, p. 31. sitková, t. (2010). využití canisterapie u dětí. olomouc, univerzita palackého v olomouci, 2010. [in czech] tichá, v. (2002). otazníky kolem canisterapie. pes přítel člověka. 3: 32. [in czech] velemínský, m. (2007). zooterapie ve světle objektivních poznatků. české budějovice: nakladatelství dona, pp. 32–33. [in czech] wilson, e.o. (1984). biophilia: the human bond with other species. england: harvard university press, cambridge. c anistherapy and its application to the teaching process at a gram m ar school 151 canisterapia i jej zastosowanie w procesie nauczania w szkole średniej streszczenie analizy koncentrują się na zastosowaniu i  wdrożeniu canisterapii (kynoterapii) w  procesie nauczania w  szkole średniej. głównym celem badań było porównanie wpływu edukacji uczniów z  udziałem i  bez udziału psów. narzędziami badawczymi były ankiety i test oraz bezpośrednia obserwacja. populacja badawcza obejmowała uczniów w wieku 12–13 lat. porównywano efekt terapii między dwiema grupami uczniów: eksperymentalną objętą canisterapią (klasa a, 28 uczniów), kontrolną (klasa b, 27 uczniów), bez canisterapii. badania przeprowadzono dwa razy w okresie od września 2015 roku do kwietnia 2016 roku. ankiety wykazały, że sesje canisterapii pozytywnie wpłynęły na zapamiętywanie programu nauczania, a także były motywacją do nauki. ponadto zauważalny był pozytywny wpływ canisterapii na mentalną stronę uczniów. uczniowie byli bardziej aktywni i skoncentrowani na zadaniach dydaktycznych. niektórzy uczniowie przezwyciężyli również wrodzony strach przed psami. bezpośrednie obserwacje uczniów na zajęciach wykazały, że sesje z użyciem psów były ważne nie tylko dla całej grupy, ale także dla poszczególnych osób. key words: canistherapy, animaltherapy, animal assisted education, teaching process, grammer school received: [2018.02.02] accepted: [2018.09.24] 33 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 33–38, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.4 łukasz m. kołodziejczyk*, magdalena puzik, agnieszka greń, marta batoryna, grzegorz formicki, edyta kapusta department of animal physiology and toxicology, institute of biology, pedagogical university of cracow, podbrzezie 3, 31-054 kraków, poland; *lukasz.kolodziejczyk@up.krakow.pl does benzo[a]pyrene affect the embryonic development of the heart? introduction �e chicken embryo and the avian in ovo model are one of the prominent experimental procedures in several tests for toxins and the preclinical testing of drugs. it is also the oldest known embryological protocol of basic developmental research (davey, tickle, 2007). it is well documented that many milestone discoveries on the embryology of vertebrates and on the organogenesis of brains or hearts were made using this model organism (le douarin, 1998; pardanaud et al., 2001). benzo[a]pyrene is a polycyclic aromatic hydrocarbon that is a well-known carcinogen, teratogen, and neurotoxin widely present in urban air pollution, cigarette smoke, and certain kinds of foods, i.e. smoked �sh, smoked meat, etc. �e problem of intoxication with this substance is actually one of the most urgent in big developing cities, such as cracow (european environment agency…, 2016). �ere are many reports con�rming its toxicity for mice and other mammals; however, knowledge on its impact on birds is still limited. whereas, birds are an important element of the typical urbicenosis, and basic knowledge on the biology of this group of vertebrates suggests that they may be a useful biotest for this stress factor (lee, shim, 2007). �e e�ect of benzopyrenes on the developing heart of vertebrates is unknown and requires e�ective update. �e embryos of birds exhibit several opportunities to perform physiological experiments on the heart (tazawa et al., 1994). �e aim of presented paper is to determine the main action of benzo[a]pyrene on selected parameters of the heart muscle of chicken embryos in the in ovo developmental model, with special attention to the antioxidative defence mechanisms and the bioelectric properties of heart rhythm. łu ka sz m . k oł od zi ej cz yk , m ag da le na p uz ik , a gn ie sz ka g re ń, m ar ta b at or yn a, g rz eg or z fo rm ic ki , e dy ta k ap us ta 34 material and methods we used chicken embryos of the race ‘ross 308’ to verify the in�uence of benzo[a] pyrene on physiological and biochemical parameters of the heart muscle. fertilised chicken eggs were obtained from a certi�ed farm (łężkowice, poland) and incubated in an automated incubator (heka, germany) at 37.5oc. �e benzo[a]pyrene in an organic oil solution (sigma aldrich, usa) was injected in ovo on the 6th day of the incubation into the yolk at the following doses of 1 mg/kg weight of eggs; 0.5 mg/kg w. e. and 0.1 mg/kg w. e. �e intact eggs and eggs injected with the organic oil were used as control groups. on the 14th day of the incubation, eggs were opened in order to examine embryos and achieve tissues for further analyses. we performed the electrocardiography of embryos using ascard amber equipment (aspel, poland) and 4 extremital copper electrodes. we also estimated the weight of hearts post mortem using a laboratory balance (rad wag, poland). finally, we determined the concentration of reduced glutathione (gsh) according to ellman’s method and malonyldialdehyde (mda) using tbars mda assay in the heart tissue. to perform all spectrophotometric measurements, we used a sunrise absorbance reader (tecan, austria). �e quantitative data was analysed statistically using shapiro-wilk tests and student tests with the signi�cance level at p < 0.05. results and discussion �e electrocardiography performed on the 14th day of incubation does not show any important changes in the heart rate and rhythm in relation to controls (fig. 1). in the electrocardiogram basal sinus rhythm without evident qrs intervals was visualised, which is typical for immature bird hearts (yoshiyama, kanke, 2005). �e increased weight of the heart muscle was observed in the embryos treated with a dose of 1 mg/kg w. e. in this group, the average heart weight accounted 211 mg. in contrast, the average heart weight in control groups was approximately equal to 120 mg. (fig. 2a). �is result may suggest that the higher doses of benzo[a]pyrene increase blood retention in the systemic circulation, which results in the heart hypertrophy. a  similar e�ect was described in several pathologies connected with increased vascular resistance (pardanaud et al., 2001). in groups contaminated with lower doses of benzo[a]pyrene, heart weight did not di�er signi�cantly from the control and intact eggs. we determined a statistically signi�cant increase of the gsh concentration in the heart tissue from the group injected with 1 mg/kg w. e. in the case of the lower benzo[a]pyrene doses, the level of gsh was similar to the controls (fig. 2b). 35 d oes benzo[a]pyrene affect the em bryonic developm ent of the heart? fig. 1. examples of electrocardiograms of chicken embryos on the 14th day of incubation. c – control; bap – an individual contaminated with benzo[a]pyrene di�erences in the mda concentrations in all experimental groups were not statistically signi�cant in relation to the controls (fig. 2c). łu ka sz m . k oł od zi ej cz yk , m ag da le na p uz ik , a gn ie sz ka g re ń, m ar ta b at or yn a, g rz eg or z fo rm ic ki , e dy ta k ap us ta 36 fig. 2. a – the e�ects of benzo[a]pyrene on the heart weight post mortem of chicken embryos; b – the e�ects of benzo[a]pyrene on the concentration of reduced glutathione in the heart tissue; c – the e�ects of benzo[a]pyrene on the concentration of malonyldialdehyde in the heart tissue; int – intacts; c – control; b1 – 1 mg/kg w. e. of benzo[a]pyrene; b0.5 – 0.5 mg/kg w. e. of benzo[a]pyrene; b0.1 – 0.1 mg/kg w. e. of benzo[a]pyrene; signi�cant di�erences between the experimental and control groups are indicated with asterisks (*p < 0.05, **p < 0.01); the error bars denote the standard deviation of the mean value; n = 6 37 �ese results may suggest that benzo[a]pyrene is a toxicological stress factor, which activates the glutathione synthesis in the organism in response to the acute poisoning and may activate other mechanisms of the glutathione-dependent antioxidative defence. it has been proven that some neurotoxins (i.e. acrylamide) involve varied disturbances and defence responses in the antioxidative system of the chicken embryo’s brain (batoryna et al., 2017; 2018). conclusion we conclude that the subacute dose of benzo[a]pyrene is a stress factor, which strongly activates the glutathione-dependent antioxidative defence and probably do not affect the heart conducting system of the chicken embryo; however, the in�uence of this substance on the morphology and biochemistry of the developing heart requires further examination. references batoryna, m., lis, m.w., formicki, g. (2017). acrylamide-induced disturbance of the redox balance in the chick embryonic brain. journal of environmental science and health, part b, 52(8), 600–606. doi: 10.1080/03601234.2017.1316158. batoryna, m., lis, m.w., formicki, g. (2018). antioxidant defense in the brain of 1-d-old chickens exposed in ovo to acrylamide. british poultry science, 59(2), 198–204. doi: 10.1080/00071668.2017.1415427. davey, m.g., tickle, c. (2007). �e chicken as a model for embryonic development. cytogenetic and genome research, 117, 231–239. european environment agency (2016). air quality in europe – 2016 report, eea report no 28/2016. publications o�ce of the european union, luxembourg. le douarin, n.m. (1998). les chimères de caille et de poulet pour étudier l’embryogenèse. pour la science, 252, 46–54. [in french]. lee, b.m., shim, g.a. (2007). dietary exposure estimation of benzo[a]pyrene and cancer risk assessment. journal of toxicology and environmental health, part a, 70(15–16), 1391–1394. pardanaud, l., moyon, d., eichmann, a. (2001). l’embryologie des vaisseaux. médecine sciences, 5(17), 543–551. [in french] tazawa, h., watanabe, w., burggren, w.w. (1994). embryonic heart rate in altricial birds, the pigeon (columba domestica) and the bank swallow (riparia riparia). physiological zoology, 67(6), 1448– 1460. yoshiyama, y., kanke, m. (2005). toxic interactions between �uconazole and disopyramide in chick embryos. biological and pharmaceutical bulletin, 28(1), 151–153. abstract benzo[a]pyrene is a polycyclic aromatic hydrocarbon with a well-proven toxic e�ect on animal cells and tissues. we used a chicken in ovo developmental model to verify its in�uence on selected parameters of the heart rhythm and on the antioxidative defence in the heart tissue. we determined that the dose of 1 mg/kg weight of eggs of benzo[a]pyrene strongly activates the glutathione-dependent antioxidative system, but it did not signi�cantly a�ect the heart conducting system of the chicken embryo. we postulate that further d oes benzo[a]pyrene affect the em bryonic developm ent of the heart? łu ka sz m . k oł od zi ej cz yk , m ag da le na p uz ik , a gn ie sz ka g re ń, m ar ta b at or yn a, g rz eg or z fo rm ic ki , e dy ta k ap us ta 38 study on the benzo[a]pyrene action during embryonic development of birds is recommended. key words: benzo[a]pyrene, ecg, embryo, gsh, heart, mda received: [2018.07.11] accepted: [2018.12.10] czy benzo[a]piren wpływa na rozwój zarodkowy serca? streszczenie benzo[a]piren jest wielopierścieniowym węglowodorem aromatycznym o  dobrze znanym toksycznym działaniu na komórki i tkanki zwierząt. wykorzystaliśmy model rozwoju zarodków kury in ovo do wery�kacji wpływu tej substancji na rytm serca oraz wybrane parametry układu antyoksydacyjnego w mięśniu sercowym. wykazaliśmy, że dawka 1 mg/kg masy jaj silnie aktywuje zależny od glutationu mechanizm antyoksydacyjny, ale jak się wydaje nie wpływa znacząco na układ przewodzący serca zarodków kury. konieczne są dalsze badania wpływu benzo[a]pirenu na rozwój zarodkowy ptaków. słowa kluczowe: benzo[a]piren, ekg, zarodek, gsh, serce, mda information on the authors łukasz m. kołodziejczyk he is a phd student at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. his scienti�c interests are developmental biology and comparative anatomy. magdalena puzik she is a graduate of bioinformatics in the institute of biology, pedagogical university of cracow. agnieszka greń https://orcid.org/0000-0003-2383-1096 she is a professor at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. marta batoryna she is a phd student at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. grzegorz formicki https://orcid.org/0000-0001-9964-6132 he is a professor at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. edyta kapusta https://orcid.org/0000-0002-4350-5514 she is a laboratory specialist at the department of animal physiology and toxicology in the institute of biology, pedagogical university of cracow. 7 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 7–16, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.1 mateusz ciepliński1*, mariusz kasprzak1, monika grandtke1, aleksandra steliga2, piotr kamiński1,3, leszek jerzak1 1faculty of biological sciences, university of zielona góra, prof. z. szafrana str., 65-516 zielona góra, poland, *m.cieplinski@wnb.uz.zgora.pl 2department of health sciences, pomeranian university of słupsk, 64 bohaterów westerplatte str., 76-200 słupsk, poland 3department of medical biology and biochemistry, department of ecology and environmental protection, faculty of medicine, collegium medicum in bydgoszcz, nicolaus copernicus university in toruń, 9m. skłodowskiej-curie, 85-094 bydgoszcz, poland the impact of udn on selectwed blood parameters of female sea trout salmo trutta m. trutta l. spawners introduction sea trout – salmo trutta morpha trutta l. (fig. 1) is an anadromous form of brown trout – salmo trutta (bouza et al., 1999). it belongs to salmonidae family which includes atlantic salmon (salmo salar l.), arctic charr (salvelinus alpinus l.) and rainbow trout (oncorhynchus mykiss walbaum) (frank-gopolos et al., 2015). anadromous �sh a�ect many other species in their environment and have a profound e�ect on the stability of aquatic and terrestrial ecosystems in which they occur; therefore, they are considered as a ‘keystone’ species (willson, halupka, 1995). main threats to anadromous �sh populations in poland are migration barriers, over�shing, poaching, and environmental changes linked with water pollution (helcom, 2015; kazuń et al., 2011; radtke et al., 2012; “�e valley of słupia”…, 2017). another important factor limiting sea trout populations in poland is ulcerative dermal necrosis (udn), which is like the disease observed in the słupia river from 2007 (grudniewska et al., 2011; grudniewska et al., 2012; kazuń et al., 2011). udn is an infectious skin disease of unknown etiology a�ecting adult, wild, anadromous salmonids migrating from open seas to fresh water during the spawning season (bruno et al., 2013; harris et al., 2011). first documented mentions of the disease originate from late 19th century great britain (roberts, 1993). in the second half of the xx century, outbreaks were reported in many european countries: austria, belgium, france, luxembourg, germany, great britain, switzerland, sweden, and canada (grudniewska et al., 2012; johansson et al., 1982). �e highest mortalities are observed in november and december. udn m at eu sz c ie pl iń sk i, m ar iu sz k as pr za k, m on ik a g ra nd tk e, a le ks an dr a s te lig a, p io tr k am iń sk i, le sz ek j er za k 8 fig. 1. sea trout with author for scale (photo. m. ciepliński) occurrence is linked to low water temperature during these months. large �sh concentrations during spawning season create perfect conditions for contagious diseases to spread (munro, 1970). �e �rst visible signs of udn are small, grey lesions on the operculum, �ns, and head (bruno et al., 2013). �ese rapidly ulcerate and frequently become infected with opportunistic pathogens, mainly the oomycete saprolegnia diclina humphrey, which has expansive growth that makes skin damage even greater (roberts, 1993). such extensive epidermal damage induces osmotic haemodilution leading to circulatory failure and the death of infected �sh (bruno et al., 2013). �e słupia river is one of the �rst rivers where occurrence of sea trout spawners with udn-like symptoms was reported (bartel et al., 2009). since then, the disease occurrence is observed on a yearly basis (“�e valley of słupia”, management personal communication). tab. 1. number of healthy, sick and agonal (n) female sea trout salmo trutta m. trutta l. blood samples used in the present study in each and all years of research health status year 2014 2015 2016 2017 2014–2017 healthy (1) 15 8 1 32 56 sick (2) 8 31 19 5 63 agonal (3) 8 6 17 0 31 total 31 45 37 37 150 9 the im pact of u d n on selectw ed blood param eters of fem ale sea trout salm o trutta m . trutta l. spaw ners blood tests are a very valuable tool in human and veterinary medicine. due to its properties, blood is virtually the easiest tissue to obtain and to test. blood tests are also one of the most comprehensive ways to assess the status of animal health. despite rapid development of laboratory techniques, the haematology of �sh is still underestimated in the assessment of �sh welfare. many factors in�uence �sh blood results, inter alia, age, sex, reproductive and nutritional status, water temperature, and oxygen concentration (řehulka, adamec, 2004; witeska, 2013). blood tests reference values found in literature are scarce and o�en vary signi�cantly between reports. in the present study, the authors decided to investigate the impact of udn on selected haematological (red blood cell count – rbc, haemoglobin concentration – hgb, haematocrit hct) and biochemical (total protein, albumin and bun concentration) parameters of female sea trout spawners from the słupia river during four (2014–2017) consecutive spawning seasons. we assume that, along with health deterioration, selected blood parameters, i.e. rbc, hgb, hct, and concentrations of total protein, albumin, and blood urea nitrogen (bun) will change signi�cantly. material and methods fish mature, spawning sea trout (salmo trutta m. trutta) females (n = 150) (mean mass 2.02 ± 0.90 kg, mean length 60.01 ± 7.64 cm) were caught during four consecutive spawning seasons in november 2014–2017 (tab. 1) on a polish angling association trapping point (54°27ʹ37.4ʹʹn; 17°02ʹ21.1ʹʹe) on the słupia river, słupsk, northern poland. water temperature during catches ranged between 4.6–8.6°c (data provided by “�e valley of słupia” landscape park). all animals with evident signs of a disease were a bycatch of a targeted trapping of healthy spawners necessary for arti�cial spawning. all specimens were euthanized with blunt force trauma to the cranium region which was followed by pithing by paa workers. healthy specimens were euthanized a�er arti�cial spawning. all �sh used in the present research were donated by the paa. all applicable international and national guidelines for the care and use of animals were followed by the authors. blood and other tissues for further examination, not presented in this article, were collected. photographic documentation of �sh skin infection patterns covered both sides of the body. photographs were made with a nikon d80 camera. health status assessment on the basis of photographic documentation, infection schemes were prepared for each udn a�ected specimen in the gimp 2.8.18 so�ware (�e gimp team, www.gimp.org, 1997–2016). all visible areas of damaged skin were mapped on an outline depicting trout contour (fig. 2a). salmo trutta m. fario l. illustration (böhmig, brauer, 1909) was m at eu sz c ie pl iń sk i, m ar iu sz k as pr za k, m on ik a g ra nd tk e, a le ks an dr a s te lig a, p io tr k am iń sk i, le sz ek j er za k 10 fig. 2. salmo trutta m. trutta l.: a – sample (le� body side) infection scheme made on the basis of photographic documentation; areas of damaged skin were mapped on an outline depicting trout contour, b – healthy salmo trutta m. trutta female specimen,; c – sick female specimen, d – agonal female specimen (photo. m. ciepliński) 11 chosen due to its simplicity. for each specimen, two schemes were prepared (for both sides of the body). obtained images were then analysed with imagej 1.50i (rasband, 1997) so�ware to determine the percentage of skin damage. obtained results for both sides of the body were then averaged, due to high bilateral symmetry of lesions. skin damage as little as 10% is known to result in nearly 50% mortality (noga, 2000). �is fact being known, the authors decided to group �sh into three categories (fig. 2): (2b) healthy specimens with no visible signs of udn; (2c) sick, where from 0.01% up to 10% skin was damaged (mean = 2.77%); and, (2d) agonal, where lesions covered more than 10% of the body surface (mean = 23.94%). laboratory analysis blood for haematological analysis was drawn from caudal vein by means of a 1.2 × 44 mm needle and 5 ml syringe. about 8–10 ml of blood in total was obtained from each specimen. we used a minimum two 5 ml syringes, and not one 10 ml, because the high vacuum created by such syringe could damage erythrocytes. immediately a�er collection, 2 ml of blood was then transferred to standard test tubes containing k2edta (1.8 mg/ml k2edta for 2ml of blood) (medlab-products) for basic haematological analysis. �e remaining amount of blood was le� to clot in 7 ml conical centrifuge tubes. clot was then centrifuged to obtain serum for biochemical analysis (total protein, albumin, and bun concentrations). in 2014, serum samples were not collected. basic haematological analysis (rbc, hgb, hct) was performed immediately a�er collection with use of manual methods. red blood cell count (rbc) was determined with use of bürker hemocytometer and natt and herrick (1952) stain. drabkin’s (drabkin, 1945) cyanmethemoglobin method was used to assess haemoglobin concentration (hgb). hematocrit (hct) was determined according to turgeon (2012). biochemical analysis (total protein, albumin, and blood urea nitrogen concentration (bun)) was performed with use of an architect c 4000 clinical chemistry analyser (abbott diagnostics). unfortunately, on a few samples, concentrations of selected biochemical parameters were beyond the measuring range of the analyser. statistics analysis statistical analysis was performed in search for di�erences in blood parameters between di�erent health status groups. due to the lack of normal distribution in all presented parameters, kruskal-wallis tests for independent groups and were used. analysis was made with statistica 12.5 so�ware (statso�, 2006). the im pact of u d n on selectw ed blood param eters of fem ale sea trout salm o trutta m . trutta l. spaw ners m at eu sz c ie pl iń sk i, m ar iu sz k as pr za k, m on ik a g ra nd tk e, a le ks an dr a s te lig a, p io tr k am iń sk i, le sz ek j er za k 12 results a comparison of selected sea trout (salmo trutta m. trutta, female spawners) hematological parameters in relation to health status is presented in table 2. signi�cantly lower rbc counts (p < 0.05), hgb (p < 0.01) and hct (p < 0.01) were observed in agonal �sh than in the remaining two health status groups, between which no statistically signi�cant di�erences were observed. in total protein (p < 0.0001) and albumin (p < 0.0001) concentrations, all groups di�ered signi�cantly. in these parameters, a clear value drop is visible along with health status deterioration. agonal specimens had signi�cantly higher urea concentrations (p < 0.0001) than the other two groups. tab. 2. selected haematological parameters of healthy, sick and agonal sea trout salmo trutta m. trutta l.; values are mean ± sd; means in parameters marked with di�erent superscript are signi�cantly di�erent (p < 0.05) health status parameter rbc [t/l] hgb [g/dl] hct [%] total protein [g/dl] albumin [g/dl] bun [mg/dl] healthy (1) 1.16a ± 0.23 n = 56 9.79a ± 1.86 n = 56 46.93a ± 7.52 n = 56 5.34a ± 1.87 n = 40 1.81a ± 0.41 n = 41 4.63a ± 1.48 n = 40 sick (2) 1.18a ± 0.26 n = 63 10.20a ± 1.64 n = 63 46.28a ± 6.98 n = 63 3.35b ± 1.13 n = 52 1.52b ± 0.45 n = 52 4.74a ± 1.70 n = 63 agonal (3) 1.02b ± 0.27 n = 31 8.95b ± 3.87 n = 31 38.61b ± 1.87 n = 31 1.31c ± 0.46 n = 17 0.64c ± 0.25 n = 16 6.74b ± 1.65 n = 31 note: rbc – red blood cell count, hgb – haemoglobin concentration, hct – haematocrit, bun – blood urea nitrogen discussion skin integrity is vital for the maintenance of �sh homeostasis. in a fresh water environment, it prevents water intake, which is consistent with concentration gradient. �e damage caused by infection development disturbs this property (noga, 2000). uncontrolled water uptake leads to the decrease in hematologic parameters. a similar e�ect is observed in total protein and albumin concentrations, both of which help to maintain proper colloid osmotic pressure (harr, 2006). although this parameter was not determined in the current study, globulin concentration, which is a di�erence between total protein and albumin concentrations, would also decrease indicating immunode�ciency (�rall et al., 2012), and elevated blood urea nitrogen in �sh is a sign of gill dysfunction (nelson et al., 1999). in the present study, results show that skin damage of more than 10% leads to severe hemodilution and a signi�cant drop in investigated haematological parameters 13 (rbc, hgb, hct). a slight increase in hgb observed in sick �sh, as compared to healthy �sh, may suggest that, during the �rst stages of udn-like infection, the �sh organism is trying to defend itself from hypoxia caused by developing respiratory failure (a�onso et al., 2002). �e decrease of htc, in relation to disease development, is the consequence of excessive water uptake via damaged skin and drop in red blood cell count (weiss, wardrop, 2010). even slight, super�cial skin damage (less than 10% of body surface) leads to a signi�cant decrease in total serum protein and albumin concentrations directly caused by water in�ux (harr, 2006). �is e�ect is even greater in severely diseased �sh. �e a�ermath of gill damage is an increase in bun concentration (nelson et al., 1999). due to many factors in�uencing �sh blood results (i.e. age, sex, reproductive and nutritional status, water temperature, oxygen concentration) (řehulka, adamec, 2004; witeska, 2013), normal values that have been presented could serve as reference only for healthy female sea trout during the spawning season. acknowledgements �e research was entirely funded by faculty of biological sciences, university of zielona góra. we would like to thank the management and sta� of “�e valley of słupia” landscape park for help and support during �eldwork, as well as the polish angling association of the district of słupsk for providing essential research material. references a�onso, e.g., polez, v.l.p., corrêa, c.f., mazon, a.f., araújo, m.r.r., moraes, g., rantin, f.t. (2002). blood parameters and metabolites in the teleost �sh colossoma macropomum exposed to sul�de or hypoxia. comparative biochemistry and physiology part c: toxicology & pharmacology, 133(3), 375–382. doi: 10.1016/s1532-0456(02)00127-8 bartel, r., bernaś, r., grudniewska, j., jesiołowski, m. (2009). furunculosis in salmon (salmo salar) and sea trout (salmo trutta trutta) in poland in 2007 and 2008. komunikaty rybackie, 110(3), 7–13. böhmig, l., brauer, a. (1909). die süsswasserfauna deutschlands: eine exkursionsfauna he� 1: mammalia, aves, reptilia, amphibia, pisces. germany, jena: verlag von gustav fischer. https://archive.org/ details/diessswasserfa01brau [in german] bouza, c., arias, j., castro, j., sanchez, l., martinez, p. (1999). genetic structure of brown trout, salmo trutta l., at the southern limit of the distribution range of the anadromous form. molecular ecology, 8(12), 1991–2001. doi: 10.1046/j.1365-294x.1999.00794.x bruno, d., noguera, p.a., poppe, t.t. (2013). a colour atlas of salmonid diseases (2nd ed.). netherlands: springer science & business media. drabkin, d.l. (1945). crystallographic and optical properties of human hemoglobin – a proposal for the standardization of hemoglobin. american journal of the medical sciences, 209, 268–270. frank-gopolos, t., bekkevold, d., guyomard, r., de innocentiis, s., martínez portela, p., fernández, c., vera, m. (2015). aquatrace species lea�et – brown trout (salmo trutta). https://aquatrace.eu/ lea�ets/trout grudniewska, j., bartel, r., bernaś, r., ciżmowski, l., jesiołowski, m., kacperska, b., siwicki, a.k. 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(1997). image j, u. s. national institutes of health (version 1.50i). maryland, usa: bethesda. https://imagej.nih.gov/ij/ řehulka, j., adamec, v. (2004). red blood cell indices for rainbow trout (oncorhynchus mykiss) reared in cage and raceway culture. acta veterinaria brno, 73(1), 105–114. doi: 10.2754/avb200473010105 roberts, r.j. (1993). ulcerative dermal necrosis (udn) in wild salmonids. fisheries research, 17(1–2), 3–14. doi: 10.1016/0165-7836(93)90003-p statso� (2006). elektroniczny podręcznik statystyki pl. kraków, poland. http://www.statso�.pl/textbook/stathome.html [in polish] “�e valley of słupia” landscape park (2017). protection of migratory fish. http://dolinaslupi.pl/ czynna-ochrona/ochrona-ryb-wedrownych/ �rall, m.a., weiser, g., allison, r., campbell, t. (2012). veterinary hematology and clinical chemistry (2nd ed.). ames, usa: wiley-blackwell. turgeon, m.l. (2012). clinical hematology: �eory and procedures (5th ed.). baltimore, usa: lippincott williams & wilkins. weiss, d.j., wardrop, k.j. (2010). schalm’s veterinary hematology (6 edition). ames, usa: wiley-blackwell. willson, m.f., halupka, k.c. (1995). anadromous fish as keystone species in vertebrate communities. conservation biology, 9(3), 489–497. doi: 10.1046/j.1523-1739.1995.09030489.x witeska, m. (2013). erythrocytes in teleost �shes: a review. zoology and ecology, 23(4), 275–281. doi: 10.1080/21658005.2013.846963 15 abstract blood tests were performed on 150 female sea trouts salmo trutta m. trutta l. during four spawning seasons (2014–2017). fish were caught on a polish angling association trapping point, the słupia river, słupsk (northern poland). �e blood for analysis was drawn from caudal vein of 56 healthy and 94 udn (ulcerative dermal necrosis) infected females. fish were divided into three groups: (1) healthy, with no visible signs of udn; (2) sick, with up to 10% skin damage; and, (3) agonal, where more than 10% of body surface was infected. a statistically important decrease in red blood cell count (rbc), haemoglobin concentration (hgb) and haematocrit (hct) were found between sick and agonal �sh groups. �e concentration of total plasma protein and albumin decreased in relation to �sh health deterioration. blood urea nitrogen (bun) had an inverse proportionality to total plasma protein and albumin concentration. based on the decrease of rbc, hgb, and hct due to the development of udn symptoms, a decrease in �sh condition was observed. signi�cantly higher urea concentrations observed in agonal �sh may indicate respiratory and excretory systems failure. key words: blood, haematology, sea trout, ulcerative dermal necrosis received: [2018.06.29] accepted: [2018.12.10] wpływ udn na wybrane parametry krwi samic troci wędrownej salmo trutta m. trutta l. w czasie tarła streszczenie w latach 2014 do 2017 w czterech sezonach rozrodczych przeprowadzono badania krwi na 150 samicach troci wędrownej salmo trutta m. trutta l. ryby odłowiono na punkcie odłowu troci na rzece słupi w słupsku (północna polska). z żyły ogonowej pobrano krew od 56 zdrowych oraz 94 chorych na udn (ulcerative dermal necrosis – wrzodziejąca martwica skóry) samic. badane osobniki podzielono na 3 grupy: (1) zdrowe bez widocznych objawów udn, (2) chore ze zmianami do 10% powierzchni ciała oraz (3) agonalne, gdzie więcej niż 10% powierzchni ciała zostało uszkodzone. istotne statystycznie zmniejszenie wartości liczby krwinek czerwonych (rbc), stężenia hemoglobiny (hgb) oraz hematokrytu (hct) stwierdzono pomiędzy osobnikami chorymi i  agonalnymi. stężenie białka całkowitego oraz albumin zmniejszało się istotnie statystycznie wraz z pogorszeniem stanu zdrowia ryb. stężenie mocznika zmieniało się odwrotnie proporcjonalnie do stężenia białka i albumin. wraz ze wzrostem intensywności rozwoju udn stwierdzono osłabienie kondycji wyrażające się zmniejszeniem rbc, hgb jak i  hct. znacznie wyższe stężenie bun u ryb agonalnych może wskazywać na niewydolność układu oddechowego i wydalniczego. słowa kluczowe: krew, hematologia, troć wędrowna, wrzodziejąca martwica skóry udn information on the authors mateusz ciepliński http://orcid.org/0000-0002-3386-9744 he is a phd student and a scienti�c-technical worker on the faculty of biological sciences, university of zielona góra. his research interests focuses on vertebrate haematology and its practical application in veterinary medicine. mariusz kasprzak http://orcid.org/0000-0001-9088-8098 he is focused on the relationship between the quality of the environment and the condition of the animals, and is an assistant professor in the department of zoology at the faculty of biological sciences, university of zielona góra. monika grandtke http://orcid.org/0000-0001-9472-252x she is a phd student at the faculty of biological sciences at the university of zielona góra. she works at the university of zielona góra as a scienti�c and technical assistant. she is interested in the in�uence of the environment with varying degrees of pollution on the white and red blood cell parameters of storks. the im pact of u d n on selectw ed blood param eters of fem ale sea trout salm o trutta m . trutta l. spaw ners m at eu sz c ie pl iń sk i, m ar iu sz k as pr za k, m on ik a g ra nd tk e, a le ks an dr a s te lig a, p io tr k am iń sk i, le sz ek j er za k 16 aleksandra steliga http://orcid.org/0000-0002-2153-9414 she is a phd student at gdański uniwersytet medyczny in gdańsk. she works at the department of health sciences on pomeranian university in słupsk and is interested in the haematology of vertebrates and human glial cells. piotr kamiński http://orcid.org/0000-0003-1978-6018 he is an assistant professor in the department of biotechnology at the faculty of biological sciences, university of zielona góra and nicolaus copernicus university in toruń, collegium medicum in bydgoszcz. he is fascinated by animal ecophysiology. leszek jerzak http://orcid.org/0000-0001-5332-279x he is full professor in nature protection at the university of zielona góra, poland (faculty of biological sciences). he studies the biology and ecology of animals (especially the white stork and the magpie) and co-operates with research centres in germany, usa, ireland, and russia. 54 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 54–59, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.7 terézia pošiváková1*, rudolf hromada1, jozef švajlenka2, ján pošivák3 1department of the environment, veterinary legislation and economy, university of veterinary medicine and pharmacy of kosice, komenského 73, 041 81 košice, slovak republic, *terezia.posivakova@uvlf.sk 2department of technology and management, technical universities of kosice, vysokoškolská 4, 042 00 košice, slovak republic 3clinic for ruminants, university of veterinary medicine and pharmacy of kosice, komenského 73, 041 81 košice, slovak republic assessing mouflon biochemical parameters depending on genders introduction �e mou�on (ovis aries musimon pallas) is the only species of the genus ovis l. living in the wild in europe. �ey come from the islands of sardinia and corsica, following their likely migration from asia minor thousands of years ago (bazer et al., 2012). �ey are not native to slovakia. instead, they were introduced to slovakia from the island of corsica (ciuti et al., 2009). mou�on’s habitat in our geographical conditions is predominantly temperate forests (ciberej et al., 2010). czech and slovak mou�on are world leaders for value, trophies, and stature amongst hunters (hell et al., 2008; frojtek, 2012). mou�on numbers are ever-growing, so it is important to extend our knowledge regarding their physiology. biochemical markers are important factors in the body re�ecting the animal’s response to various stressors or negative e�ects of the environment (sherwood et al., 2013; kimáková et al., 2018). negative biochemical balance may disrupt the body, resulting in a variety of pathological changes, physical strain, or infectious disease (lehocký, kuric, 2007). biochemical markers may be a sign of disease or pathological conditions within the body noted in research in di�erent �elds (ciberej, 2013). our study provides the assessing of mou�on biochemical parameters depending on genders, which can be used to manage the health status of these animals. material and methods �e mou�on used in this research came from a game reserve in the eastern part of the slovak republic. water intake from natural water sources was unrestricted and the a ssessing m ouflon biochem ical param eters depending on genders 55 animals were fed hay during winter. �e animals showed no signs of disease. between 2015 and 2016, a total of 60 mou�ons (ovis aries musimon pallas), n = 30 males and n = 30 females, were analysed. blood samples were taken by a veterinarian from the jugular vein from ��een males and ��een females. �e experimental group consisted of 4 to 5 year-old mou�ons. samples were collected in winter using blood collection tubes and heparin tubes. �e following biochemical variables were analysed: albumin – alb, alkaline phosphatase – alp, alanine aminotransferase – alt, aspartate aminotransferase – ast and lactate dehydrogenase – ldh. biochemical indicators were measured using an automatic analyser with a �exible system for consolidating routine examinations (lehocký, kuric, 2007). �e biochemical variables were evaluated in relation to the sex of the animals in the experimental groups. �e obtained results were analysed via the statistical method of mann-whitney’s u test using statistica 12 so�ware for comparing biochemical variables of males and females, depending on year and biochemical variables. results and discussion �e results of the biochemical analysis are shown in tables 1 to 4. �e analysed biochemical parameters are grouped into tables according to statistical signi�cance in the context of the years being compared and depending on sex. for the sake of comparing various biochemical variables of males and females in 2015 and 2016, we used the mann-whitney’s u test, which is utilised in comparing two or more groups with a low number of measurements. by comparing 2015 and 2016, we recorded similar statistically signi�cant �ndings in the case of both sexes, i.e. in the case of alb and ast parameters. �e aforementioned parameters were signi�cantly higher in 2015 compared to 2016. �e average values of these parameters in the monitored years were approximately the same in the case of both sexes except ast, where these values were markedly higher in males than in females. by comparing the individual sexes regardless of monitored years, we recorded statistically signi�cant di�erences in the case of alb, alt, ast, and ldh parameters. all of these parameters were higher in males than in females. lower values of alb, alt, and ast in females may have been caused by the fact that they were in their rutting period, during which there is a change in the values of intermediary metabolism in terms of enzyme reduction (ciberej, 2014). higher values of ldh in males may have been associated with stress during handling and subsequent increased muscle activity (krauss, nies, 2014). when handling the animals, the males resisted more than the females, which ultimately shows in the higher values of the aforementioned parameters. 56 te ré zi a p oš iv ák ov á , r ud ol f h ro m ad a, j oz ef š va jle nk a, j án p oš iv ák ta b. 1 . s ig ni �c an t d i� er en ce s f or e ac h se le ct ed se ru m b io ch em ic al v ar ia bl e of m al es in th e co m pa re d ye ar s o f 2 01 5 an d 20 16 ; n = 1 5 m al e ye ar s s 20 15 20 16 va ri ab le m ea n m ed ia n ± sd m in m ax c on f. in t. m ea n m ed ia n ± sd m in m ax c on f. in t. a lt [i u /l ] 31 .9 8 32 .6 3 10 .8 4 10 .1 2 48 .7 4 25 .9 9– 37 .9 6 21 .4 4 19 .4 0 11 .6 8 9. 70 48 .1 4 14 .9 7– 27 .9 0 ** a st [i u /l ] 10 2. 75 79 .5 8 45 .9 9 59 .6 4 21 6. 00 77 .3 1– 12 8. 20 66 .1 1 54 .1 3 32 .7 5 32 .1 6 16 3. 41 47 .9 0– 84 .1 9 ** a lb [g /l ] 11 .7 2 11 .3 2 2. 02 9. 14 14 .9 9 10 .6 0– 12 .8 4 9. 66 9. 13 2. 04 6. 84 14 .2 2 8. 52 –1 0. 79 ** n ot e: ± s d – st an da rd d ev ia tio n, m in – m in im um , m ax – m ax im um , c on f. in t. – 95 % c on �d en ce in te rv al , * *p < 0 .0 1, a lt – a la ni ne a m in ot ra ns fe ra se , a st – a sp ar ta te a m in ot ra ns fe ra se , a lb – a lb um in , s – si gn i� ca nc e ta b. 2 . n on -s ig ni �c an t d i� er en ce s o f e ac h se le ct ed se ru m b io ch em ic al v ar ia bl e of m al es in c om pa re d ye ar s 2 01 5 an d 20 16 ; n = 1 5 m al e ye ar s s 20 15 20 16 va ri ab le m ea n m ed ia n ± sd m in m ax c on f. in t. m ea n m ed ia n ± sd m in m ax c on f. in t. a lp [i u /l ] 37 .9 0 35 .3 3 13 .6 5 16 .7 60 .4 0 30 .3 6– 45 .4 5 26 .6 5 20 .3 6 16 .1 1 4. 79 49 .1 0 17 .6 6– 35 .5 1 n s ld h [i u /l ] 49 6. 80 45 6. 00 13 8. 00 32 5 72 0 42 0. 4– 57 3. 2 49 4. 7 41 9 18 8. 6 28 7 81 6 39 0. 3– 59 9. 2 n s n ot e: ± s d – st an da rd d ev ia tio n, m in – m in im um , m ax – m ax im um , c on f. in t. – 95 % c on �d en ce in te rv al , n s – no t s ig ni �c an t, a lp – a lk al in e ph os ph at as e, l d h – la ct at e de hy dr og en as e, s – si gn i� ca nc e a ssessing m ouflon biochem ical param eters depending on genders 57 ta b. 3 . s ig ni �c an t d i� er en ce s f or e ac h se le ct ed se ru m b io ch em ic al v ar ia bl e of fe m al es in th e co m pa re d ye ar s o f 2 01 5 an d 20 16 ; n = 1 5 fe m al e ye ar s s 20 15 20 16 va ri ab le m ea n m ed ia n ± sd m in m ax c on f. in t. m ea n m ed ia n ± sd m in m ax c on f. in t. a st [i u /l ] 64 .3 7 59 .6 4 16 .0 5 41 .2 0 94 .6 0 55 .4 5– 73 .2 3 48 .2 0 43 .3 5 15 .8 1 26 .2 0 75 .8 1 39 .4 0– 56 .8 9 ** a lb [g /l ] 10 .8 7 9. 42 3. 09 7. 94 17 .0 0 9. 16 – 12 .5 7 8. 99 7. 77 3. 93 4. 73 17 .1 3 6. 81 – 11 .1 7 * n ot e: ± s d – st an da rd d ev ia tio n, m in – m in im um , m ax – m ax im um , c on f. in t. – 95 % c on �d en ce in te rv al , * p < 0. 05 , * *p < 0 .0 1, a st – a sp ar ta te a m i no tr an sf er as e, a lb – a lb um in , s – si gn i� ca nc e ta b. 4 . n on -s ig ni �c an t d i� er en ce s f or e ac h se le ct ed se ru m b io ch em ic al v ar ia bl e of fe m al es in th e co m pa re d ye ar s o f 2 01 5 an d 20 16 ; n = 1 5 fe m al e ye ar s s 20 15 20 16 va ri ab le m ea n m ed ia n ± sd m in m ax c on f. in t. m ea n m ed ia n ± sd m in m ax c on f. in t. a lp [i u /l ] 27 .3 7 25 .1 5 7. 31 16 .7 43 .1 23 .2 9– 31 .3 8 25 .6 9 23 .3 5 13 .7 7 5. 99 43 .7 1 18 .0 2– 33 .2 3 n s a lt [i u /l ] 17 .9 0 18 .2 0 7. 49 7. 01 27 .3 13 .7 7– 22 .0 4 16 .1 7 15 .9 3 9. 16 5. 87 39 .3 4 11 .0 8– 21 .2 0 n s ld h [i u /l ] 38 0. 47 36 5 12 9. 80 22 1 61 0 30 8. 6– 45 2. 4 37 4. 8 30 3 17 0. 0 16 3 71 4 28 0. 7– 46 9. 0 n s n ot e: ± s d – st an da rd d ev ia tio n, m in – m in im um , m ax – m ax im um , c on f. in t. – 95 % c on �d en ce in te rv al , n s – no t s ig ni �c an t, a lp – a lk al in e ph os ph at as e, a lt – a la ni ne a m in ot ra ns fe ra se , l d h – la ct at e de hy dr og en as e, s – si gn i� ca nc e te ré zi a p oš iv ák ov á , r ud ol f h ro m ad a, j oz ef š va jle nk a, j án p oš iv ák 58 conclusion �e dynamics of the observed selected biochemical variables indicates their importance when maintaining homeostasis in animals. in our research, we investigated the biochemical status of mou�ons of both sexes as the current physiological status for this type of wild game. based on our monitoring of the biochemical status of mou�on, we recorded signi�cant di�erences between sexes in the case of alb, alt, ast, and ldh parameters. all of these parameters in our study were higher in males than in females. acknowledgement �is work was supported from grant: kega 003 uvlf-4/2016. references bazer, f.w., spencer, t.e., �atcher, w.w. (2012). growth and development of the ovine conceptus. journal of animal science, 90(1), 159–170. doi: 10.2527/jas.2011-4180 ciuti, s., pipia, a., grignolio, s., ghiandai, f., apollonio, m. (2009). space use, habitat selection and activity patterns of female sardinian mou�on (ovis orientalis musimon) during the lambing season. european journal of wildlife research, 55(6), 589–595. doi: 10.1007/s10344-009-0279-y ciberej, j., lazar, p., halász, j. (2010). game caring and diseases. zvolen: technical university, p. 108–222. hell, p., slamečka, j., gašparík, j. (2008). fallow deer and mou�on in slovakia. bratislava: papress. frojtek, p. (2012). mou�on from south moravia, history, management and trophy. modrý kameň, p. 18–26. sherwood, l., klandorf, h., yancey, p. (2013). animal physiology. from genes to organisms. belmont, usa: ca brooks/cole cengage learning. kimáková, t., kuzmová, l., nevolná, z., bencko, v. (2018) fish and �sh products as risk factors of mercury exposure. annals of agricultural and environmental medicine, 25(3), 488–493. doi: 0.26444/ aaem/84934 lehocký, m., kuric, p. (2007). management in huntings and breeding of wild game. zvolen: national forest centre. ciberej, j. (2013). hunters zoology and biology. zvolen: technical university. ciberej, j. (2014). indents and teeth. hunting and �shing, 66, 8–11. krauss, j.g., nies, d.h. (2014). ecological biochemistry, environmental and interspecies interactions. usa: wiley-blackwell. abstract �e aim of our study was to evaluate the selected biochemical parameters of mou�on depending on gender. �irty mou�ons of both sexes with the same approximate age in the winter season were used for research. blood samples for biochemical analysis were taken from vena jugularis for determination of selected biochemical parameter. biochemical indicators were measured using the standard automatic analyser. �e results of statistical testing of selected biochemical parameters in the experimental group of animals con�rmed di�erences between the genders and at the selected biochemical parameters. key words: biochemical status, female, male, laboratory analyses, mou�on received: [2018.04.08] accepted: [2018.12.10] a ssessing m ouflon biochem ical param eters depending on genders 59 ocena parametrów biochemicznych muflona w zależności od płci streszczenie celem badań była ocena wybranych parametrów biochemicznych u mu�ona, zależnych od płci. w sezonie zimowym, do badań wykorzystano trzydzieści mu�onów obu płci, o przybliżonym wieku. próbki krwi do analizy biochemicznej pobrano z vena jugularis, w celu określenia wybranego parametru biochemicznego. wskaźniki biochemiczne mierzono za pomocą standardowego automatycznego analizatora. wyniki badań statystycznych wybranych parametrów biochemicznych w  grupie zwierząt doświadczalnych potwierdziły różnice między płciami i wybranymi parametrami biochemicznymi. słowa kluczowe: status biochemiczny, samica, samiec, analizy laboratoryjne, mu�on information on the authors terézia pošiváková she currently works as an educator in the �elds of ecology and environment. in her �eld of scienti�c work, she is focusing on monitoring the biochemical, haematological, and immunochemical conditions of animals, including interspecies breeding and the sex di�erences of animals. her work focuses on problem-solving animal hygiene due to the strategically necessary improvement of food security and production. rudolf hromada he is a specialist and expert in veterinary medicine. he has professional experience in the �elds of hygiene, sanitation, and welfare in animal management and public health, ecology, issues of water and wastewater contamination, and the composting of organic wastes. he currently works as an educator in the �elds of ecology and environment and animal hygiene. he is the author of several university textbooks. jozef švajlenka he is currently working as an educator in the �elds of technology and building management, ecology, and the environment. in his �eld of scienti�c work, he is focusing on the indoor and outdoor environmental quality of buildings, farm building quality, and the relation of functional and spatial processes with agricultural production systems, which refers to the quality of the health and wellbeing of those who occupy space within it. ján pošivák he is a specialist and expert in veterinary medicine. he is currently working as an educator in the �eld of veterinary medicine and animal reproduction. he applies his teaching and research knowledge in education and scienti�c practice. he has extensive experience in the �eld of animal physiology, animal reproduction, and assisted reproductive farming, and in wild animals. he deals with the study of biochemical parameters in relation to the reproduction of animals. 39 annales universitatis paedagogicae cracoviensis studia naturae, 2: 39–56, 2017, issn 2543-8832 doi: 10.24917/25438832.2.3 valerián franc*, michal fašanga department of biology and ecology, faculty of natural sciences, matej bel university, tajovského 40, 97401 banská bystrica, *valerian.franc@umb.sk spiders (araneae) of the abandoned pasture near the village of malé kršteňany (western slovakia) introduction our research site is located on the se slope of the hill of drieňový vrch (cadaster of the village of malé kršteňany). it is the southernmost edge of the strážovské vrchy mountains (mts) (48°55ʹ46ʹʹn; 18°26ʹ05ʹʹe), separated from the central massif by the river �ow nitrica. �is area is considerably in�uenced by human activity: in the past, it had massive deforestation and agricultural use (mainly as pasture), recently, it is dominated by mining activities (several quarries). �e whole area is out of the territorial protection, with the exception of the little nature reserve veľký vrch, surrounded by two quarries, and the le� one is more or less abandoned. in the past, this area was used mainly for grazing, but this is currently very limited. our research site is an abandoned pasture; therefore, ecological succession is carried out intensively here. forgotten a�er-utility areas (abandoned quarries, pastures, industrial sites) are usually considered to be ‘sterile’ and unattractive for zoological research, but this may not always correspond to reality. even in our research site, we have carried out several rare and surprising �ndings. we would like to present the results of our research in this paper. it is sad, but a large amount of abandoned pastures is scattered throughout slovakia. �is contradicts the fact that traditional methods of farming (especially grazing) are most suitable for the sustainable development of suburban and rural partially agricultural land. grazing helps to preserve steppe character of the habitats in central europe. grazing restriction, evident almost in the whole of europe, is unnecessary and counterproductive also in protected areas. on the other hand, it should be supported. it does not concern intensive and concentrated grazing in a small plot of course – it causes destruction of the soil, including fauna. scattered grazing in the meadows, in sparse forests and xerothermic slopes, is the most desirable from the gene-pool point of view; and it does not only concern spiders and insects. v al er iá n fr an c, m ic ha l f aš an ga 40 material and methods �e research of spiders of the nature reserve veľký vrch had been carried out already in 1994, and the results were published later (gajdoš, 1996). �e author used di�erent sampling methods (mainly pitfall trapping in the mentioned protected territory) while we dealt especially with individual collecting on the secondary habitat outside the nature reserve, and then the faunal comparison between these two sites may be methodologically inadequate. our study area is west of the reserve veľký vrch, above the more-or-less abandoned quarry (fig. 1–2). coordinates of the centre of the research site: 48°38ʹ49.41ʹʹn; 18°26ʹ44.37ʹʹe, the altitude 270–385 m. �e research of spiders was carried out during the vegetation season of 2015 and in 1 sampling excursion in march 2017. we applied current methods of sampling, especially si�ing of leaf litter and detritus, sweeping the vegetation, and shaking down the spiders from tree branches. we also dealt with individual exploration under the bark and in rotten wood of old trunks and stumps. we took merely a small number of individuals, and easily identi�able species were mostly noted down only. �e spiders were identi�ed according the keys by miller (1971) and heimer, nentwig (1991) and the specialised web-site: https://araneae.unibe.ch/. several documented species are o�en cited in various red lists of european countries. later will be evaluated and compared their ecosozological status according to the following red lists: slovakia (gajdoš, svatoň, 2001), czech republic (kůrka et al., fig. 1. research site above the quarry (red line), green line indicates nr veľký vrch (source: https://www. google.sk/maps/@48.6486548,18.4538526,1464m/data=!3m1!1e3) s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 41 2015), austria – the carinthia county (komposch, steinberger, 1999), poland (staręga et al., 2002), and germany (blick et al., 2016). �e further old and undated records will be not mentioned. coordinates of the sites are added only in the case of thoroughly localised records. �e code dfs (databank of fauna of slovakia) is cited only in several inaccurately mentioned records, where the coordinates are not exactly detectable. additional data, if possible, are listed in chronological order. results and discussion a total of 146 spider species belonging to 25 families were collected in the studied territory (appendix 1 – tab. 1). some records deserve special note (marked by ”◄“ in table 1): 1. acartauchenius scurrilis – a scarce species of xerothermic meadows and pastures. notable due to its ecology: it occurs sporadically in the colonies of ants tetramorium caespitum, especially under stones. a larger number of recent records is available. 2. centromerus albidus – a rare species of well-preserved forest habitats, little known due to its hidden way of life. formerly, it had been considered to be utmost rare (miller, 1971). usually found under larger stones and in rock �ssures. several recent records are available: tríbeč mts – nature reserve (later only ‘nr’) zoborská lesotep (dfs 7674), april 13, 1978, 1 ♀ (gajdoš, 1985); krupinská planina plateau – plášťovce (48°10ʹ15.63ʹʹn; 19°0ʹ24.78ʹʹe, 279 m a. s. l. – later only ‘m’), may 23, 1995, 1 ♀ (franc, 1999); cerová vrchovina mts – nr steblová skala (48°4ʹ44.61ʹʹn; 19°58ʹ46.96ʹʹe, 442 m), may 6, 1995, 1 ♀; and nr pohanský hrad – the nyári cave fig. 2. panoramic view on the veľký vrch hill; study site is indicated by red arrow. it is nearly unbelievable that this nature reserve (green arrow) is in the immediate vicinity of the active quarry (photo. m. fašanga) v al er iá n fr an c, m ic ha l f aš an ga 42 (48°11ʹ54.71ʹʹn; 19°55ʹ22.77ʹʹe, 570 m), may 29, 1995, 10 ♀ (franc, hanzelová, 1995); muránska planina plateau – nr šiance (48°46ʹ24.60ʹʹn; 20°05ʹ22.66ʹʹe, 898 m), june 22, 2001; and nr šarkanica (48°42ʹ34.55ʹʹn; 19°58ʹ35.37ʹʹe, 710 m), june 25, 2001 (franc, 2014); bystrická vrchovina mts – stará kopa mt (48°43ʹ38.05ʹʹn; 19°10ʹ53.19ʹʹe; 486 m), may 11, 2002, 1 ♀ (franc, 2005); krupinská planina plateau – príbelce (48°12ʹ14.70ʹʹn; 19°14ʹ49.88ʹʹe, 362 m), between rocks in shady oak forest, june 21, 2008 (franc, 2010a); cerová vrchovina mts – nature monument belinské skaly (48°13ʹ45.83ʹʹn; 19°51ʹ39.74ʹʹe, 471 m), june 6, 2006, p. gajdoš lgt. (svatoň et al., 2009). note: its conspicuously pale colouration re�ects that it tends to underground way of life. despite not documented in neighbouring countries, its discovery is expectable, especially in the czech republic and austria. 3. frontinellina frutetorum – a scarce species of xerothermic habitats and edges of open deciduous forests. a larger number of recent records is available. 4. mermessus trilobatus – an allochthonous species, originally known from north america, where it is quite frequent. he was brought into europe by tra�c, probably with us army soldiers, and it now occurs in several western and central european countries (dolanský et al., 2009). it prefers semi-natural and disturbed habitats. 5. �eonina cornix – occurs locally and rarely in rocky and sandy steppes and open dry deciduous forests. it is also known from the site located 8.5 km on the north: strážovské vrchy mts – horné vestenice (48°43ʹ19.96ʹʹn; 18°25ʹ37.71ʹʹe, 436 m), april 28, 2002, 2 ♂ (franc, 2004). several further recent records are accessible; hitherto known from 13 grid mapping squares of the dfs (gajdoš, in verb.). it may be locally more abundant in the southwestern part of slovakia: malé karpaty mts – svätý jur (dfs 7769), old vineyards, undated, 16 specimens! (dankaninová, gajdoš, 2012). 6. trichoncus a nis – a scarce species of rocky steppes. more than 50 recent records from slovakia are accessible. it always indicates well-preserved warm habitats. 7. phycosoma inornatum – a scattered and very rare species of xerothermic (often karst) grasslands and open forests. known only from three further recent records: strážovské vrchy mts – dolné vestenice: záviničie (48°42ʹ01ʹʹn; 18°23ʹ17ʹʹe, 230 m), pitfall trap, 2001, 4 specimens (gajdoš et al., 2009); starohorské vrchy mts – jakub protected site (48°46ʹ0.16ʹʹn; 19°08ʹ34.32ʹʹe, 434 m), april 9, 2004, 1 ♂ p. gajdoš rev. (franc et al., 2009); krupinská planina plateau – príbelce (48°12ʹ16.35ʹʹn; 19°15ʹ0.89ʹʹe, 328 m), forest steppe above the shooting range, june 21, 2008, 2 ♂ (franc, 2010a). very rarely found also in the neighbouring czech republic (buchar et al., 2002), and apparently elsewhere. note: it ranks among little known species, because the species of the former genus dipoena are di�cult to identify. 8. altella biuncata – a very rare species of forest steppes. known only from a few isolated records: krupinská planina plateau – plášťovce (dfs 6879), december 15, s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 43 year is not mentioned, probably the 50s, f. miller lgt. – it is apparently the �rst record from the territory of slovakia! (kůrka, 1994); malá fatra mts – the starhradská valley, surely it is contemporary nr krivé (dfs 6879), open deciduous forest, pitfall trap, august 23, 1974, 1 ♀ (svatoň, 1981); nr hradová (48°40ʹ47.58ʹʹn; 19°55ʹ21.40ʹʹe, 795 m), forest steppe with the south exposition, 1979–1980, date unavailable (svatoň, 1985) – the last two records from mountain altitudes are highly notable; malé karpaty mts – nr devínska kobyla (dfs 7868), undated (gajdoš, 2005a); burda mts – nr burdov (dfs 8178), undated, 1 ♀ j. svatoň lgt. (gajdoš, 2016). 9. brommella falcigera – another very rare species of xerothermic habitats. only a small amount of records is available: malé karpaty mts – nr devínska kobyla (dfs 7868), undated (gajdoš, 2005a); nr ostrovné lúčky (48°02ʹ25.40ʹʹn; 17°10ʹ31.23ʹʹe, 129 m), september 19, 1993, 1 ♂ and 4 ♀ lgt. p. gajdoš (unpublished); national park poloniny – nr hrúnok (49°0ʹ50.79ʹʹn; 22°14ʹ9.70ʹʹe, 350 m), september 21, 1998, 1 ♀ (svatoň et al., 2003); cerová vrchovina mts – protected site vinohrady (48°16ʹ33.92ʹʹn; 20°10ʹ8.51ʹʹe, 238 m), june 8, 2006, 1 ♀ lgt. s. korenko (svatoň et al., 2009); strážovské vrchy mts – dolné vestenice (48°42ʹ44.69ʹʹn; 18°23ʹ56.88ʹʹ e, 348 m), xerothermic karst slope, march 24, 2011, 2 ♂ v. franc lgt. (previously unpublished). 10. mastigusa arietina – found under the stone in the colony of camponotus ligniperdus (latreille, 1802). a rare species of warmer grasslands and xerothermic habitats, living in or near ant colonies; nevertheless, its relationship with ants is not clear (the same concerns all myrmecophilous spiders). referred �nding is remarkable because the occurrence in association of this ant species has not hitherto been published. �e following records from slovakia are available: nr rohy near the town of detva (48°32ʹ53.27ʹʹn; 19°21ʹ11.58ʹʹe, 577 m), in an under-stone colony of lasius niger, march 3, 1991, 1 ♂; krupinská planina plateau – plášťovce (48°10ʹ15.63ʹʹn; 19°0ʹ24.78ʹʹe, 279 m), may 23, 1995, 1 ♀ in the colony of messor structor on a xerothermic slope, april 1, 1994, 2 ♀ – together with the very rare leiodid-beetle attaephilus arenarius (hampe, 1852)! (franc, 1999); krupinská planina plateau – rykynčice (48°12ʹ21.70ʹʹn; 18°57ʹ40.40ʹʹe, 267 m, the same circumstances, april 9, 1994, 2 ♂, 1 ♀, all records v. franc and a. hanzelová lgt.; ostrôžky mts – nedelište (48°23ʹ30.62ʹʹn; 19°25ʹ8.63ʹʹe, 457 m), in the colony of lasius alienus in a xerothermic oak forest, april 22, 2000, 1 ♂ v. franc lgt. (all records franc, 2007); nr zoborská lesostep near the town of nitra (dfs 7674), april 29, 1978 (gajdoš, krumpál, 1987); national park poloniny – ruské (49°07ʹ7.37ʹʹn; 22°19ʹ51.91ʹʹe, 568 m), semi-xerothermic pasture, pitfall trap, 1 specimen undated (žila, gajdoš, 2014); ostrôžky mts – praha (48°22ʹ07.69ʹʹn; 19°30ʹ26.27ʹʹe, 504 m), xerothermic shrubby pasture, in the colony of messor structor, 3 ♀ (franc, 2010b). its abundance is recently probably increasing due to the global warming. 11. diaea livens – a rare species of warmer deciduous forests and groves, formerly had been ranked among utmost rare species. �e �rst record from slovakia was pubv al er iá n fr an c, m ic ha l f aš an ga 44 lished from protected site gavurky (48°27ʹ51.53ʹʹn; 19°07ʹ53.90ʹʹe, 466 m), may 22, 1992, 1 ♂ (franc, hanzelová, 1996). �e second �nding has been carried out in the site jasenov – hôrka (dfs 7097), june 15, 1994, 1 ♂, and published as a new species for the slovakian fauna (�omka, 1996), because the author did not have information on the �rst record from gavurky. �e further records: cerová vrchovina mts – obručná (dfs 7885), 20. 5. 1999, 1 ♂, lgt. eva svatoňová, det. j. svatoň (svatoň et al., 2009); poľana mts – forest steppe above the bátovský boulder (48°39ʹ39.88ʹʹn; 19°22ʹ47.46ʹʹe, 736 m), may 13, 2005, 1 ♂, v. franc lgt., j. svatoň rev. (franc, 2013); ostrôžky mts – lysec (48°20ʹ52.41ʹʹn; 19°27ʹ40.54ʹʹe, 633 m), june 25, 2007, 2 ♂ (franc, 2010b); krupinská planina plateau – príbelce (48°12ʹ16.35ʹʹn; 19°15ʹ0.89ʹʹe, 328 m), forest steppe above the shooting range, june 21, 2008, 1 ♂ (franc, 2010b); nr šúr (dfs 7769), salty grassland, may 28, 2009, 1 ♂, lgt. o. majzlan (gajdoš, 2010). recent records are also accessible from the slovenský kras mts – surroundings of the domica cave (dfs 7588), shrubby and tree margin of the dry calcareous grassland, but undated (gajdoš, 2005b). maybe, its abundance is recently increasing due to the global warming as well. 12. tmarus stellio – a rare thermophilous species of forest steppes and edges of open deciduous forests. several records from slovakia are accessible: krupinská planina plateau – medovarce (48°14ʹ13.54ʹʹn; 18°59ʹ18.12ʹʹe, 242 m), swept from the vegetation of forest steppe, may 16, 1992, 1 ♀ (franc, 1999); poľana mts – forest steppe above the bátovský boulder (48°39ʹ39.88ʹʹn; 19°22ʹ47.46ʹʹe, 736 m), june 1, 2004, 1 ♂ (franc, 2013); poľana mts – hrochoť, forest steppe above the beňova valley (48°39ʹ12.94ʹʹn; 19°19ʹ44.08ʹʹe, 660 m), june 19, 2005, 1 ♂ and 1 ♀ (franc, 2013); ostrôžky mts – lysec (48°20ʹ51.41ʹʹn; 19°27ʹ41.74ʹʹe, 622 m), june 14, 2007, 2 ♂ (franc, 2010b); malé karpaty mts – svätý jur (dfs 7769), old vineyards, undated, 1 specimen (dankaninová, gajdoš, 2012); burda mts – burdov (dfs 8178), june 19, 2005 (gajdoš, 2016). 13. chalcoscirtus brevicymbialis – a local and rare species, known from a few records in the warmest sites. only a few recent records from slovakia are accessible: nr turecký vrch (dfs 7273), july 12, 1978, 1 ♀ p. gajdoš lgt. – the �rst record for the territory of slovakia (gajdoš et al., 1984)*; strážovské vrchy mts – nr kňaží stôl (48°48ʹ15.70ʹʹ; 18°17ʹ30.35ʹʹe, 542 m), june 3, 1984, 1 ♀ (gajdoš, 1986a)*; pohronský inovec mts – nr včelár (48°27ʹ01.48ʹʹn; 18°28ʹ41.17ʹʹe, 383 m), june 28, 1984, 1 ♀ (gajdoš, 1987)*; strážovské vrchy mts – nr veľký vrch (48°8ʹ54.07ʹʹn; 18°27ʹ13.82ʹʹe, 385 m), summer 1985, several specimens (gajdoš, 1986b)*. krupinská planina plateau – medovarce (48°14ʹ13.54ʹʹn; 18°59ʹ18.12ʹʹe, 242 m), between rocks of a forest steppe, may 27, 1995, 1 ♂ (franc, 1999). strážovské vrchy mts – dolné vestenice (48°42ʹ44.69ʹʹn; 18°23ʹ56.88ʹʹe, 348 m), xerothermic karst slope, july 2, 2002, 1 ♂ (franc, 2004); burda mts – burdov (dfs 8178), 1 specimen j. buchar lgt. (gajdoš, 2016). *note: cited as chalcoscirtus in�mus (simon, 1868), despite it has not been hitherto documented from slovakia. �ese species may be confused; nevertheless, the male s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 45 palpal organ of ch. brevicymbialis is apparently shorter than in ch. in� mus (it re� ects in its name). faunistic records on these tiny jumping spiders may be sometimes debatable, because the taxonomic situation in this genus may be not clear, especially in older identi� cation keys. 14. pellenes nigrociliatus – a rare species of rocky steppes. a larger number of recent records is available. it always indicates well-preserved warm habitats. in our introductory research, we found out 146 spider species. � is number is not de� nitive, of course. some of documented species rank among infrequent or up to rare ones, including � eonina cornix, phycosoma inornatum, altella biuncata, brommella falcigera, mastigusa arietina, diaea livens, tmarus stellio, chalcoscirtus brevicymbialis, pellenes nigrociliatus and haplodrassus kulczynskii. several � ndings in this anthropogenous habitat are surprising. later, we will deal with instructive analysis of the thermopreference (fig. 3a) and the originality of habitats (fig. 3b) from the perspective of individual species. we assumed that the share of thermophilous species will be prevailing – it is more than half: 52.83%. � e species of warm and moderate habitats together represent more than 97% of the spider fauna. � e analysis of the spider faunal composition according to originality of habitat is even more interesting. � e share of climax species is surprisingly high – 54.83%, despite this habitat is not at all ‘pristine’. experts opinions on the climax, especially those of a conservative nature, require revision and comprehensive view of modern ecology. we suppose that some particular stages of the on-going succession process can be interpreted as ‘partial climax stages’ tending towards the ‘� nal’ arrangement of the biota. it is necessary to emphasize that the climax is not at a ‘de� nitive’ and ‘constant’ stage, on the contrary, it is a highly dynamic and variable process. fig. 3. spider faunal composition according to thermopreference – a): 1 – thermophilous, 2 – mesophilous, 3 – oreophilous species; spider faunal composition according to originality of habitat – b): 1 – climax, 2 – semi-natural, 3 – disturbed, 4 – arti� cial habitats 1 2 3 64.50 44.18% 77.15 52.83% 4.35 2.99% 1 2 3 451.80 35.48% 80.05 54.83% 4.17 2.85% 9.98 6.84% a) b) v al er iá n fr an c, m ic ha l f aš an ga 46 references blick, t., finch o.-d., harms, k.-h., kiechle, j., kielhorn, k.-h., kreuels, m., malten, a., martin, d., muster, c., nährig, d., platen, r., rödel, i., scheidler, m., staudt, a., stumpf, h., tolke, d. 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[in slovak] s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 49 appendix 1 tab. 1. spiders (araneae) of the abandoned pasture near the village of malé kršteňany family / species codes of records �ermopreference originality of habitat ecosozological status sk cz ak g pl pholcidae pholcus opilionoides (schrank, 1871) c-/1 + t (m) c sn d dysderidae harpactea hombergi (scopoli, 1763) c-/1 + t m c sn h. rubicunda (c.l. koch, 1839) a1/-+ t m c sn a uloboridae hyptiotes paradoxus (l. koch, 1834) h1/+ m sn araneidae araneus diadematus (clerck, 1757) e1/+ t m o c sn d a.(= atea) triguttatus (fabricius, 1793) b1/1 (t) m c sn nt a. sturmi (hahn, 1831) c-/1 t m c sn nt araniella cucurbitina (clerck, 1757) e3/1 + t m c sn d a. opisthographa (kulczyński, 1905) d1/t m c sn cyclosa conica (pallas, 1772) b-/1 d-/1 (t) m c sn gibbaranea bituberculata (walckenaer, 1802) c-/1 d-/2 t (m) c (sn) nt g. gibbosa (walckenaer, 1802) d3/3+ t m c sn vu mangora acalypha (walckenaer, 1802) b1/+ t m c sn d nuctenea umbratica (clerck, 1757) b-/1 e-/2 + (t) m c sn a zilla diodia (walckenaer, 1802) b-/1 c-/1 d1/1+ m c sn nt mimetidae ero aphana (walckenaer, 1802) j-/1s t c (sn) lc nt tetragnathidae metellina (= meta) merianae (scopoli, 1763) f-/1 t m o c sn a pachygnatha degeeri (sundevall, 1830) b1/d1/t m (o) c sn d tetragnatha montana (simon, 1874) e1s/(t) m c sn t. pinicola (l. koch, 1870) e1/t m c sn linyphiidae abacoproeces saltuum (l. koch, 1872) e-/1 (t) m c sn v al er iá n fr an c, m ic ha l f aš an ga 50 acartauchenius scurrilis (o.p.cambridge, 1872)◄1 c-/1 t m c sn lc vu nt vu agyneta (= meioneta) rurestris (c.l. koch, 1836) j2/1 gd t m o c sn d centromerus albidus (simon, 1929)◄2 c-/1 m c (sn) nt ∅ ∅ ∅ ∅ diplostyla concolor (wider, 1834) b-/1 t m o c sn d frontinellina frutetorum (c.l. koch, 1834)◄3 e2/t c en nt vu vu linyphia triangularis (clerck, 1757) h-/3 t m c sn d maso sundevalli (westring, 1851) e-/1 t m (o) c sn mermessus trilobatus (emerton, 1882)◄4 j2/2 gd alien sp. sn d minicia marginella (wider, 1834) e-/1 t m c sn vu nt nt neriene clathrata (sundevall, 1830) b2/t m c sn neriene peltata (wider, 1834) b-/1 m c sn porrhomma microphthalmum (o.p.-cambridge, 1871) j1/1 gd t m c sn d vu tapinocyba insecta (l. koch, 1869) b-/1 t m c sn tenuiphantes tenuis (blackwall, 1852) j-/1 gd t m c sn d �eonina cornix (simon, 1881)◄5 j-/1gr t c nt vu nt vu vu �yreosthenius parasiticus (westring, 1851) f-/1 m o c sn d trematocephalus cristatus (wider, 1834) e-/1 (t) m c sn trichoncus a nis (kulczyński, 1894)◄6 b-/1 e-/1 t m c sn vu en en t. auritus (l. koch, 1869) a1/t c vu er walckenaeria corniculans (o.p.cambridge, 1875) f-/1 m c sn walckenaeria cucullata (c.l. koch, 1836) b-/1 m (o) c sn w. dysderoides (wider, 1834) b1/(t) m c sn �eridiidae asagena (= steatoda) phalerata (panzer, 1801) e1/+ t m c sn vu cryptachaea (= achaearanea) riparia (blackwall, 1834) e2/(t) m c sn dipoena melanogaster (c.l. koch, 1837) d1/1 e1/2 + t (m) c sn vu enoplognatha ovata (clerck, 1757) e1/t m c sn d e. thoracica (hahn, 1833) c-/1 d-/2e1/2+ t m c sn d s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 51 episinus truncatus (latreille, 1809) e1/f1/t c sn nt euryopis �avomaculata (c.l. koch, 1836) e2/t m c sn nt heterotheridion nigrovariegatum (simon, 1873) e2/+ t c sn vu nt vu lasaeola (= dipoena) tristis (hahn, 1833) e1/2 f-/2 (t) m c sn nt nt neottiura bimaculata (linnaeus, 1767) e2/t m c sn d parasteatoda (= achaearanea) lunata (clerk, 1757) f1/(t) m c sn phycosoma inornatum (o.p.cambridge, 1861)◄7 (= dipoena inornata) b1/t m c sn en cr ? vu vu phylloneta (= �eridion) impressa (l.koch, 1881) e3/+ t m (o) c sn d platnickina (= keijia) tincta (walckenaer, 1802) d1/e2/2 + t m c sn robertus arundineti (o.p.cambridge, 1871) b-/1 d1/1 (t) m c sn d r. lividus (blackwall, 1836) b2/1 t m o c sn sardinidion (= �eridion) blackwalli (o.p.-cambridge, 1871) e1/t m c sn a vu steatoda albomaculata (de geer, 1778) j-/1s t m c sn nt nt s. triangulosa (walckenaer, 1802) d1/e1/t d a �eridion mystaceum (l. koch, 1870) b-/1s m c sn t. pinastri (c.l. koch, 1872) e2/t m c sn dictynidae altella biuncata (miller, 1949)◄8 -/1gr t c vu en er en dd dictyna (=brigittea) latens (fabricius, 1775) e1/t c vu nt vu dictyna uncinata (�orell, 1856) j-/1 (t) m c sn d brommella falcigera (balogh, 1935)◄9 d-/1 gd t c en en en en lathys humilis (blackwall, 1855) b1/c1/t c vu mastigusa arietina (�orell, 1871)◄10 b-/1 t m (o) c vu vu dd vu titanoecidae titanoeca quadriguttata (hahn, 1833) h-/1 + t m c sn nt vu amaurobiidae amaurobius fenestralis (ström, 1768) b-/1 + m o c sn v al er iá n fr an c, m ic ha l f aš an ga 52 agelenidae eratigena (= tegenaria) agrestis (walckenaer, 1802) h-/2 t m c sn d textrix denticulata (olivier, 1789) h-/1+ t c nt vu urocoras (= coelotes) longispinus (kulczyński, 1897) d-/1 + t c (sn) ∅ ∅ zodariidae zodarion germanicum (c.l. koch, 1837) b-/1s c1/+ t m c sn vu vu vu zodarion rubidum (simon, 1914) e-/2 t c sn lycosidae alopecosa cuneata (clerck, 1757) e-/1 t m (o) c sn d a. fabrilis (clerck, 1757) d-/1 m c dd cr vu alopecosa farinosa (herman, 1879) (= accentuata (latreille, 1817)) b1/1 t m sn nt a. sulzeri (pavesi, 1873) c1/t c vu en en pardosa alacris (c.l. koch, 1833) b1/t (m) sn d dd trochosa terricola (�orell, 1856) b-/1 e-/2+ t m (o) c sn d pisauridae pisaura mirabilis (clerck, 1757) b-/1 e-/1+ t m c sn d �omisidae coriarachne depressa (c.l. koch, 1837) h-/1s e-/1s t m c sn nt cozyptila blackwalli (simon, 1875) b2/+ t c vu vu en diaea livens (= pictilis) (simon, 1876)◄11 d1/t m c cr en dd ebrechtella (= misumenops) tricuspidata (fabricius, 1775) d2/t (m) c sn misumena vatia (clerck, 1757) d-/1+ t m c sn d ozyptila claveata (walckenaer, 1837) b2/t m c nt pistius truncatus (pallas, 1772) c1s/e1/t m c sn nt synema globosum (fabricius, 1775) b1/+ t m c sn nt nt vu tmarus piger (walckenaer, 1802) d2/1 h-/3s+ t (m) c sn vu vu vu t. stellio (simon, 1875)◄12 e-/1 t c lc en ∅ xysticus audax (schrank, 1803) d1/(t) m (o) c sn x. cristatus (clerck, 1757) d2/-+ t m (o) c sn d x. erraticus (blackwall, 1834) e2/(t) m c sn nt x. kochi (�orell, 1872) f-/1 t m c sn (d) x. lanio (c.l. koch, 1835) e1/t m c sn nt anyphaenidae anyphena accentuata (walckenaer, 1802) d-/1 e1/-+ t m c sn s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 53 clubionidae clubiona comta (c.l. koch, 1839) c-/1 e-/3 + t m c sn c. brevipes (blackwall, 1841) j-/1 t (m) c sn nt philodromidae philodromus aureolus (clerck, 1757) e-/1 t m c sn d p. buchari (kubcová 2004) e2/t m c sn nt p.margaritatus (clerck, 1757) c-/1s t m c sn vu �anatus formicinus (clerck, 1757) b2/h1s/t m c sn nt nt nt tibellus oblongus (walckenaer, 1802) b1/t m c sn salticidae ballus chalybeius (walckenaer, 1802) e1/+ t m c sn carrhotus xanthogramma (latreille, 1819) c1/t c vu vu dendryphantes rudis (sundevall, 1833) h-/1 (t) m c sn euophrys frontalis (walckenaer, 1802) d1/e1/t m c sn evarcha arcuata (clerck, 1757) b1/-+ t m c sn e. falcata (clerck, 1757) f-/1 (t) m c sn evarcha laetabunda (c.l. koch, 1846) e2/t (m) c vu nt heliophanus cupreus (walckenaer, 1802) d3/e3/+ t m c sn chalcoscirtus brevicymbialis (wunderlich, 1980)◄13 b2/+ t c vu en leptorchestes berolinensis (c.l. koch, 1846) c1/d1/+ t c (sn) vu en vu macaroeris (= eris) nidicolens (walckenaer, 1802) d1/t c (sn) vu marpissa muscosa (clerck, 1757) b1/d-/1+ t m c sn nt nt pellenes nigrociliatus (simon, 1875)◄14 c1/1 t c vu en vu pseudeuophrys lanigera (simon, 1871) b1/t m sn a p. obsoleta (simon, 1868) c2/2 d1/1 t c vu nt en vu pseudicius encarpatus (walckenaer, 1802) e1/t m c sn lc nt en salticus scenicus (clerck, 1757) e1/t m c sn a s. zebraneus (c.l. koch, 1837) d2/t m c sn sitticus pubescens (fabricius, 1775) e3/1 + m c sn a nt miturgidae zora nemoralis (blackwall, 1861) e-/1+ (t) m c sn nt v al er iá n fr an c, m ic ha l f aš an ga 54 z. pardalis (simon, 1878) c1/1 t c sn cr z. spinimana (sundevall, 1833) a1/b1/t m o c sn d eutichuridae cheiracanthium elegans (�orell, 1875) e-/1 t c en en ch. mildei (l. koch, 1864) d-/1 t (m) sn a nt gnaphosidae callilepis schuszteri (herman, 1879) (appendix 2 – fig. 4) e-/2 t c vu nt vu drassodes lapidosus (walckenaer, 1802) d1/e1/t m c sn drassyllus (= zelotes) prae�cus (l. koch, 1866) f-/2 t m c sn vu d. pusillus (c.l. koch, 1833) c1/t m c sn (d) d. villicus (�orell, 1875) e-/2 t c vu nt nt gnaphosa lucifuga (walckenaer, 1802) c1/h1/1 t c nt vu nt g. montana (l. koch, 1866) (appendix 2 – fig. 5) e-/2 m o c sn nt vu vu en g. opaca (herman, 1879) f-/1 t c vu en en haplodrassus kulczynskii (lohmander, 1942)◄14 c-/1 t c lc vu vu nt dd micaria fulgens (walckenaer, 1802) b1/t m c sn nt nt scotophaeus quadripunctatus (linnaeus, 1758) f-/2 t m c sn a s. scutulatus (l. koch, 1866) a-/1 h-/1 t m c sn a zelotes hermani (chyzer, 1897) d-/1 t c ∅ nt ∅ z. petrensis (c.l. koch, 1839) b1/2 c-/1+ t m c sn phrurolithidae phrurolithus festivus (c.l. koch, 1835) e 1/1 t m c sn codes of records: a – march 28, 2015, b – april 17, 2015, c – april 24, 2015, d – may 12, 2015, e – june 6, 2015, f – june 18, 2015, g – june 26, 2015, h – october 2, 2015, i – october 30, 2015, j – march 10, 2017; 1/one male, 1/2 – one male and two females, 1/-+ – one male, but more specimens observed, s – subadult specimen; gd – p. gajdoš det., gr – p. gajdoš rev. �ermopreference: t – thermophilous, m – mesophilous, o – oreophilous. originality of habitat: c – climax, sn – semi-natural, d – disturbed, a – arti�cial habitat. ecosozological status (ess): sk – slovakia, cz – czech republic, ak – austria (the carinthia county), pl – poland, g – germany. categories of ess: cr – critically endangered, en – endangered, vu – vulnerable, nt – near threatened, lc – least concern, dd – data de�cient, er – extremely rare, ∅ – not documented in this country till now, ? – the species is surprisingly missing in referred red list. s piders (a raneae) of the abandoned pasture near the village of m alé k ršteňany (w estern s lovakia) 55 appendix 2 fig. 5. gnaphosa montana (l. koch, 1866) – scarce species on edges of open deciduous forests (photo. ľ. černecká) fig. 4. callilepis schuszteri (herman, 1879) – quite abundant species on xerothermic habitats (photo. ľ. černecká) v al er iá n fr an c, m ic ha l f aš an ga 56 abstract abandoned pastures are frequent phenomenon throughout central europe due to the reduction of grazing. �is also concerns the abandoned pasture near the village of malé kršteňany (western slovakia). we dealt with the research of spiders in this site in 2015 and spring of 2017. despite it is a secondary anthropogenous habitat, the fauna of spiders is relatively rich, including infrequent or up to rare species, including �eonina cornix, phycosoma inornatum, altella biuncata, brommella falcigera, mastigusa arietina, diaea livens, tmarus stellio, chalcoscirtus brevicymbialis, pellenes nigrociliatus and haplodrassus kulczynskii. �e species of warm and moderate habitats are highly prevailing here – together it is 97%. �e analysis of the spider faunal composition according to originality of habitat is even more interesting. �e share of climax species is surprisingly high – 54.83%, despite this habitat is not at all ‘pristine’. experts opinions on the climax, especially those of a conservative nature, require revision and a comprehensive view of modern ecology. we suppose that some particular stages of the on-going succession process can be interpreted as “partial climax stages” tending towards the ‘�nal’ arrangement of the biota. it is necessary to emphasize that the climax is not at the ‘de�nitive’ and ‘constant’ stage, on the contrary, it is a highly dynamic and variable process. key words: abandoned pasture, araneae, malé kršteňany, slovakia, spiders, succession received: [2017.05.18] accepted: [2017.10.30] pająki (araneae) odłogowanych pastwisk w pobliżu miejscowości malé kršteňany (zachodnia słowacja) streszczenie ze względu na zmniejszenie wypasu, odłogowanie pastwisk jest zjawiskiem częstym w europie środkowej. dotyczy to również zaniedbanych gospodarczo pastwisk w pobliżu miejscowości malé kršteňany (zachodnia słowacja). w 2015 roku oraz wiosną 2017, podjęto badania pająków w okolicach wyżej wymienionej miejscowości. mimo, że jest to wtórne siedlisko antropogeniczne, fauna pająków jest tu stosunkowo bogata. obejmuje ona, zarówno nieczęste, jak i  rzadkie gatunki, w  tym: �eonina cornix, phycosoma inornatum, altella biuncata, brommella falcigera, mastigusa arietina, diaea livens, tmarus stellio, chalcoscirtus brevicymbialis, pellenes nigrociliatus i haplodrassus kulczynskii. dominują tutaj gatunki siedlisk ciepłych i umiarkowanych (łącznie 97%). ciekawostką jest analiza składu pająków według oryginalności siedliska. udział gatunków klimaksowych jest tu zaskakująco wysoki – 54.83%, mimo, że siedlisko to nie jest „pierwotne”. w  nowoczesnej ekologii, opinie specjalistów na temat klimaksu, zwłaszcza te konserwatywne, wymagają rewizji i kompleksowego spojrzenia. przypuszczamy, że pewne konkretne etapy postępującego procesu sukcesji można interpretować jako „częściowe etapy klimaksu”, zmierzające w kierunku „ostatecznego” rozmieszczenia bioty. należy podkreślić, że klimaks nie jest etapem „ostatecznym” i „stałym”, a wręcz przeciwnie, jest to bardzo dynamiczny i zmienny proces. słowa kluczowe: odłogowane pastwisko, araneae, malé kršteňany, słowacja, pająki, sukcesja information on the authors valerián franc lecturer of general zoology and systematic invertebrate zoology. he has deal with the research of beetles and spiders for more than 30 years, with special regard to the nature conservation and the problems of biological indication and the factors of endangerment of separate animal taxa. michal fašanga he is a student of biology and geography at the faculty of natural sciences (matej bel university in banska bystrica, slovakia) with a special interest in entomology and arachnology. 7 annales universitatis paedagogicae cracoviensis studia naturae, 1: 7–23, 2016, issn 2543-8832 raúl alvarez-mora1, william cetzal-ix2*, saikat kumar basu3, eliana noguera-savelli4, noel a. gonzález-valdivia2, jesús f. martínez-puc2, peiman zandi5, katarzyna możdżeń6 1 jardín botánico xoxoctic, cuetzalan del progreso, puebla 2 instituto tecnológico de chiná, calle 11 entre 22 y 28, colonia centro chiná 24050, campeche, méxico, *rolito22@hotmail.com 3 department of biological sciences, university of lethbridge, lethbridge, ab canada t1k 3m4 4 catedrática conacyt, colegio de postgraduados campus campeche. carretera haltunchén-edzná km. 17.5, sichochac, champotón, campeche. mexico. c.p. 24450 5 institute of crop science, chinese academy of agricultural sciences, beijing 100081, people’s republic of china 6 department of plant physiology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland orchid diversity at cuetzalan del progreso, puebla, mexico – anthropogenic threats and potential for organic production for conservation purposes introduction �e puebla state, mexico has not performed a comprehensive survey of the local orchid �ora for a considerably long period. �ere are still fragments of tropical montane cloud forest and tropical rain forest in puebla between the biogeographical provinces of the sierra madre oriental and the gulf of mexico (veracruz state border) which have not been totally explored. �e current orchid species reported for the state can be attributed to random studies conducted in the past two decades. soto-arenas (1988) recorded 127 species of orchids for puebla and a decade later espejo-serna and lópez-ferrari (1998a, 1998b) recorded 146 species. recently, pérez-bravo et al. (2010) conducted collection of orchids in the pine-oak forest in the north-east part of puebla in the xochiapulco and zacapoaxtla municipalities, recording 12 species that increased the total number of orchid members for the state to 158. additionally, various taxonomic (e.g. solano-gómez, 1993; soto-arenas et al., 2007; hágsater, soto-arenas, 2003, 2008) and �oristic studies (e.g. romero-giordano, 2000) recorded through photographs and herbarium material another 14 species for the state, increasing the current number of species to 172. �e largest number of the species reported for puebla is distributed mostly in the north-east and south-east parts of the state (conabio, 2008), where the diversity of r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 8 the vegetation types of the gulf of mexico lowlands and the mountainous parts of the sierra madre oriental makes these municipalities ecologically rich with high orchid diversity. �is area is also considered as priority in conservation for its tropical montane cloud forest and tropical rain forest by the comisión nacional para el conocimiento y el uso de la biodiversidad of mexico (conabio, 2008). however, no comprehensive study of the orchid �ora of the region has been previously conducted to assess the number of orchid species and their conservation status in cuetzalan del progreso municipality. our objective has been to explore this habitat, collect and record with photographs di�erent species of orchidaceae and also to identify any anthropogenic factors impacting the survival and existence of the local orchids in this fragile and highly fragmented ecosystem. furthermore, we were to establish a model for the sustainable management and to promote conservation action of species at some level of threat and in the restoration of areas with fragmented forests. a viable approach will be to allow the introduction of in vitro orchid germination program and organic production of orchids by local residents and forest fringe communities; that will improve the local economy, reduce their dependence on the scarce forest resources and prevent illegal harvesting and tra�cking of wild orchids, and at the same time provide economic development for the local communities. materials and methods study area cuetzalan del progreso is one of the municipalities of the state of puebla in mexico, that still has conserved fragments of tropical montane cloud forest (fig. 1b) and tropical rain forest (fig. 1c). �is municipality is located in the north-eastern puebla between the limits of the eastern sierra madre and the gulf of mexico provinces (veracruz border) (morrone, 2001, fig. 2). �is ecosystem is located at an altitude ranging between 180–1600 m, with semi-warm to warm humid climate and rainfall throughout the year (instituto nacional de estadística…, 1987, 2009). �e ecosystem also constitutes the pine-oak and tropical deciduous forests. however, these forests have been severely fragmented, unfortunately reducing them to minor fragments of vegetation due to drastic changes in land use pattern for cash rich co�ee production and for establishing grazing areas for the local livestock and dairy herds (instituto nacional de estadística..., 1985). botanical collections as part of our study of the orchid �ora of mexico and as an e�ort to contribute to the knowledge of the �ora of the puebla and cuetzalan del progreso, photographic o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 9 documentations were made between december 2010 and november 2013. �e specimens were photographed in their natural habitats. we have included the photos of the new records only (appendix 1, fig. 3–4). some species were collected vegetatively and cultured in the xoxoctic botanical garden in cuetzalan del progreso (20°2’22.63” n, 97°30’32.16” w) and when the plants later �owered, they were photographed and identi�ed. �e identi�cation of specimens was made based on the digital catalog of the orchids of mexico (soto-arenas et al., 2007). �e map of the study area was downloaded from the free relief layers for google maps (2013) and edited with the adobe photoshop 6.0.1. (adobe systems inc, san jose, california). �e line that divides the municipality into two biogeographic provinces is based on morrone (2001) (fig. 2). data collection of species of plants illegally traded in tianguis of cuetzalan del progreso �e “tianguis” in cuetzalan del progreso are informal markets that are established on the major streets of the town on saturdays and sundays between 7 am to 5 pm. �roughout 2013 we conducted weekly visits to record and identify species of orchids that are being sold there. results and discussion floristic inventory, new records of orchids for puebla, conservation status and illegally trade a total of 93 species were recorded in cuetzalan del progreso and it is quite exciting to report that 25 of these species are new reports for the puebla �ora (appendix 1, fig. 3–4). with our latest �ndings the number of orchid species in this speci�c habitat has now increased to 197. it is noteworthy that about half (47%) of the species present in puebla (3 430 600 ha) are in the cuetzalan del progreso (13 522 ha), which demonstrates the wide diversity of orchidaceae of this municipality. nine species are listed in the nom-059-semarnat-2010 (semarnat, 2010) and one in documents of international union for the conservation of nature (iucn red list, 2016) (appendix 1). of the new records, two species are listed as subject to special protection and one as threatened. �e habitats where those species were photographed are quite close (ca. 500–1000 m of distance) to the surrounding human settlements and have been found to be severely deforested for indiscriminate agricultural activities and/or illegal timber collection of �rewood by the local settlers. while the survey identi�ed 25 new records from the region, it also identi�ed potential pitfalls in the e�orts for the conservation of the rich biodiversity of the region, including the spectacular orchid species biodiversity for the �rst time. �e species are extremely vulnerable to exploitation by local and indigenous populations. irregular and r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 10 fig. 1. cuetzalan del progreso, puebla, mexico. a – mountains of the eastern sierra madre, b – tropical montane cloud forest, c – tropical rain forest (photo. r. alvarez-mora) repeated harvesting of the species by local populations for illegal nursery trade could result in serious depletion of several species in the long term if careful attention is not paid immediately (cetzal-ix et al., 2014). �e other threats for the orchid species are: lack of monitoring and increased encroachments in the forested areas over the decades o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 11 fig. 2. study area and distribution of new records in cuetzalan del progreso, puebla, mexico. 1 – acianthera angustifolia, 2 – anathallis abbreviata, 3 – aspydogyne querceticola, 4 – campylocentrum micranthum, 5 – chysis bractescens, 6 – corymborchis forcipigera, 7 – dichaea trichocarpa, 8 – encyclia gravida, 9 – epidendrum di�usum, 10 – erycina pusilla, 11 – gongora truncata, 12 – habenaria �oribunda, 13 – heterotaxis maleolens, 14 – jacquiniella equitantifolia, 15 – leochilus labiatus, 16 – lophiaris lurida, 17 – ornithocephalus in�exus, 18 – polystachya lineata, 19 – prescottia stachyodes, 20 – sacoila lanceolata, 21 – scaphyglottis fasciculata, 22 – scaphyglottis lindeniana, 23 – specklinia alata, 24 – stelis rubens, 25 – vanilla planifolia r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 12 fig. 3. study area and locations of new records of orchidaceae species in cuetzalan del progreso, puebla, mexico; a  – acianthera angustifolia, b – anathallis abbreviata, c – aspydogyne querceticola, d – campylocentrum micranthum, e – chysis bractescens, f – corymborchis forcipigera, g – dichaea trichocarpa, h – encyclia gravida, i – epidendrum di�usum, j – erycina pusilla, k – gongora truncata, l – habenaria �oribunda, m – heterotaxis maleolens, n – jacquiniella equitantifolia, o – leochilus labiatus, p – lophiaris lurida (photo. r. alvarez-mora) o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 13 fig. 4. study area and locations of new records of orchidaceae species in cuetzalan del progreso, puebla, mexico; a – ornithocephalus in�exus, b – polystachya lineata, c – prescottia stachyodes, d – sacoila lanceolata, e – scaphyglottis fasciculata, f – scaphyglottis lindeniana, g – specklinia alata, h – stelis rubens, i – vanilla planifolia (photo. r. alvarez-mora) by local populations; cattle and livestock grazing in sensitive habitats; collection of food, fuel, fodder and fertiliser resources from the forests are adding serious anthropogenic interferences within the sensitive and extremely fragile ecosystem due to absence of any legal restrictions and comprehensive forest management policy. many species of orchids present in puebla state are con�scated from tianguis and private collections by the federal attorney for environmental protection (profepa – procuraduría federal de protección al ambiente) (un1ón, 2013). in 2013, species rescued by profepa from illegal trade estimated to over 1000 specimens r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 14 of orchids in puebla alone (islas, 2014). in this sense, 37 out of 93 species present in cuetzalan del progreso were recorded from tianguis of this municipality, involved in illegal trade of wild orchids (appendix 1). surprisingly, four new records presented here were observed on sale in di�erent tianguis of cuetzalan del progreso (chysis bractescens, dichaea trichocarpa, encyclia gravida and epidendrum di�usum). also, �ve taxa under the category of “threatened” (camaridium densum, chysis bractescens, mormodes maculata var. unicolor, oncidium incurvum, stanhopea oculata and stanhopea tigrina) and two species subject to “special protection” (prosthechea vitellina and vanilla planifolia) were also recorded from there (appendix 1). �ose species were sold for between 250–280 pesos (approximately us$ 18.85–21.11) (barragán, 2013). organic production of local orchids based on our study, the risks of commercial exploitation of the local orchid species, as well as illegal harvesting and trade on them by the local human populations have been substantial. �e average annual income of the majority of the family groups living around those biodiversity hotspots has been quite minimal in economic terms, barely su�cient to support them and to respond to their daily needs. as a consequence such remote rural residents and fringe forest dwellers have become increasingly dependent on whatever local forest resources are available for their sustenance. slowly and steadily they have been encroaching illegally into the protected forested zones out of poverty and due to absence of any viable alternative sources of income. �ey are also getting involved in illegal trade and the� of forest resources (with local orchids becoming a primary target for ruthless exploitation) and slowly turning into an impending danger towards the destruction of the orchid biodiversity of this region. we sincerely believe that the situation will not improve unless a  sustainable approach is adapted to cater for the economic needs of the local resident populations in the region from a humanitarian perspective. if alternative sources of income, education, and health care are introduced in a timely fashion for those helpless communities, the bene�ts would come directly in the form of e�ective conservation of the local orchids and other �oral and faunal biodiversity of the puebla ecoregion. we humbly suggest the introduction of in vitro orchid germination program and organic production of local orchids in the small community run cooperative nurseries and development of small in vitro and plant tissue culture laboratories in the region. �e local administration, the forest, and social services departments must cooperate to train enthusiastic residents into in vitro germination programme and organic production of orchids in those proposed cooperatives. some interested and enthusiastic local and international non-government organizations (ngos) dedicated to the cause of biodiversity conservation and helping remote rural communities in becoming self-sufo rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 15 �cient and economically sustainable could be included as important stakeholders for further facilitating of the process. �e plants, cut �owers and seeds generated in those government and/or ngo monitored cooperatives could be bought back by the forest department and they could be easily sold to bigger commercial nurseries in the nearby cities that have high demand for orchids. �e plants and the seeds could be used in replenishing the reduced stocks of several populations of threatened orchid species in the local forests, thereby supporting the e�ort towards conservation of the local orchid biodiversity. because there is a high demand for the local orchids, the factor can be rather explored to provide alternative employment for the needy local communities, as well as useful for conservation and biodiversity management too. if successful, the project could be extended to other local �oral groups available in the local forests, for which there is high demand. furthermore, as local communities are traditionally and predominantly agrarian societies, it would be easier to teach them the practices of organic production of orchids rather than trades of modern agriculture heavily tilted towards over application of agro-chemicals. commercial organic productions are suggested to avoid using excessive agro-chemicals in the production system that may in turn cause contamination and pollution of the locality, and hence indirectly impact the local forests, as well as to keep the production costs under control. �e bene�ts of sustainable organic production will be higher economic returns for the community cooperatives, as well as reduced chances of undesirable pollution of the local environment. another important concern and factor favoring the use of organic production of orchids by local cooperatives in this proposed model is that the adult plants and seedlings used as stocking and replenishment populations for conservation purposes in the remote forests are completely free of any external chemicals. �e precaution is essential to avoid contaminating the local forest ecosystem as less as possible. all cares must be taken so that it does not transmit to other �oral and faunal communities in the forest via food chains and food webs thorough the process of biomagni�cation, particularly by means of any biological pollinators. although the initial cost of establishing training centers, nurseries, greenhouses, and o�ces will be covered, in the end, if the project runs successfully, it could well turn out into a  successful model that can be extended to other parts of mexico and latin america with similar and related anthropogenic threats for both economic bene�ts of the local communities, as well as conservation of local biodiversity. it will be extremely important to note that in the initial or founding years complete monitoring and surveillance of the production system will be absolutely essential. strong government support, initiatives, training, funding, as well as political willingness and determination at the beginning to carry the project forward would be important in making the organic production of local orchids sustainable, viable and successful. r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 16 according to our humble evaluation, the �rst step in establishing this proposed model and platform will be to reach the local residents with empathy and patience, communicate with them through friendly dialogues, explain them the issues in simple terms and in the next stage slowly make them participants and stakeholders in the entire process. as they will realize and understand the bene�ts of the project, they will also reciprocally become respectful towards their local environment and they are expected to become partners in the conservation process in the long term. based on this proposed model we sincerely believe that the promotion of commercial production of local orchids in the cooperative nurseries developed for the local rural communities and fringe dwellers could make signi�cant sustainable changes both in the life of the resident communities and in the conservation of local orchid biodiversity. conclusions we anticipate that if proper measures are not adopted by the local administration and by the forest department, several of the majestic species of orchids existing in the region will soon be threatened with extinction. since we consider orchids to be �agship plant species highlighting the rich biodiversity of this unique ecosystem, the loss of the orchids from the region will also indicate indirect loss of several other key species from this sensitive and fragile ecosystem. climate change and global warming are causing widespread impact on the survival of several plant species including orchids and their primary and secondary pollinators (hegland et al., 2009). similar impacts are also possibly a�ecting the orchid species at puebla. in addition, the excessive anthropogenic stress in the region could have severe long-term impact on the survival and natural propagation of the orchid species of this region. �e majority of the new records reported here are also found to be distributed in veracruz and in some cases in other states of south-eastern mexico, such as oaxaca and chiapas. however, they represent an important contribution to the knowledge of the biodiversity of puebla and even more when you consider the degree of fragmentation the forests of the state are now exposed too. it also suggests the importance of botanical explorations in this municipality and in other boundaries of the sierra madre oriental and the gulf of mexico to the north-western portion of puebla. �is will possibly help to increase the existing number of taxa for orchids, as well as other plant groups present within this area and should be considered a conservation priority for its tropical montane cloud forest and tropical rain forest by the comisión nacional para el conocimiento y el uso de la biodiversidad (arriaga et al., 2009). o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 17 acknowledgements �e senior author would like to thank justo alvarez and flora mora galicia for funding the �eld work. special thanks to cristina garcía juárez, owner of the jardín botánico xoxoctic for the facilities provided for growing the orchids and miguel alvarez soto for providing excellent assistance during the �eld work. references arriaga, l., espinoza, j.m., aguilar, c., martínez, e., gómez, l., loa, e. (2000). regiones terrestres prioritarias de méxico. comisión nacional para el conocimiento y uso de la biodiversidad. conabio, méxico. http://www.conabio.gob.mx/conocimiento/regionalizacion/doctos/tmapa.html. [in spanish] barragán, s. (2013). mercado negro hace negocio con las orquídeas. http://www.unionpuebla.mx/articulo/2013/10/04/empresas/puebla/mercado-negro-hace-negocio-con-las-orquideas. [in spanish] conabio. (2008). red mundial de información sobre biodiversidad (remib). http://www.conabio.gob. mx/remib/doctos/remibnodosdb.html?. [in spanish] cetzal-ix, w., alvarez-mora, r., basu, s.k., cosme-pérez, j., noguera-savelli, e. 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(1998b). las monocotiledóneas mexicanas, una sinopsis �orística. 1. lista de referencia. parte viii. orchidaceae ii. méxico, d.f., méxico: consejo nacional de la flora de méxico, universidad autónoma metropolitana y comisión nacional para el conocimiento y uso de la biodiversidad. [in spanish] free relief layers for google maps. (2013). shaded relief mashup of the world for google maps. http:// www.maps-for-free.com hágsater, e., soto-arenas, m.a. (2003). icones orchidacearum fascicle 5 & 6, orchids of méxico part 2 & 3. méxico, d.f., méxico: herbario amo. http://www.herbarioamo.org/index_archivos/fascicles5&6.pdf hágsater, e., soto-arenas, m.a. (2008). icones orchidacearum fascicle 10, orchids of méxico part 4. méxico, d.f., méxico: herbario amo. http://www.herbarioamo.org/index_archivos/fascicle10.pdf hegland, s.j., nielsen, a., lazaro, a., bjerknes, a.l., totland, ø. (2009). how does climate warming a�ect plant-pollinator interactions? ecology letters, 12, 184–195. doi: 10.1111/j.1461-0248.2008.01269.x instituto nacional de estadística, geografía e informática. (1985). cuetzalan del progreso, estado de puebla: cuaderno estadístico municipal. aguascalientes, méxico: instituto nacional de estadística y geografía. [in spanish] instituto nacional de estadística, geografía e informática. (1987). síntesis geográ�ca, nomenclátor y anexo cartográ�co. aguascalientes, méxico: instituto nacional de estadística y geografía. [in spanish] instituto nacional de estadística, geografía e informática. (2009). prontuario de información geográ�ca municipal de los estados unidos mexicanos. cuetzalan del progreso, puebla. clave geoestadística 21043. http://www.inegi.org.mx/default.aspx. [in spanish] islas, l. (2014). ma�a de las orquídeas “azota” puebla. http://www.unionpuebla.mx/articulo/2014/03/24/ medio-ambiente/ma�a-de-las-orquideas-azota-puebla. [in spanish] r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 18 iucn red list. (2016). guidelines for using the iucn red list categories and criteria: version 10. prepared by the standards and petitions subcommittee in february 2016. http://intranet. iucn.org/web�les/doc/ssc/redlist/redlistguidelines.pdf morrone, j.j. (2001). biogeografía de américa latina y el caribe. m&t-manuales & tesis sea, 3. zaragoza, spain: gor�sa. http://www.bio-nica.info/biblioteca/morrone2001caribe.pdf. [in spanish] pérez-bravo, r., salazar, g.a., mora-guzmán, e. (2010). orquídeas de las lomas-la manzanilla, sierra madre oriental, puebla, méxico. boletín de la sociedad botánica de méxico, 87, 125–129. [in spanish] romero-giordano, c. (2000). orquídeas de la sierra norte de puebla. in: m. sarmiento-fradera, c. romero-giordano (eds.), orquídeas mexicanas. méxico, d.f.: editorial porrúa, 129–144. [in spanish] semarnat. (2010). norma o�cial mexicana nom-059-semarnat-2010, protección ambiental – especies nativas de méxico de �ora y fauna silvestres – categorías de riesgo y especi�caciones para su inclusión, exclusión o cambio. – lista de especies en riesgo. diario o�cial de la federación, 2ª sección, 30 de diciembre de 2010. secretaría de medio ambiente y recursos naturales, méxico. http://www.profepa.gob.mx/innovaportal/�le/435/1/nom_059_semarnat_2010.pdf. [in spanish] solano-gómez, r. (1993). el género stelis sw. (orchidaceae: pleurothallidinae) en méxico. orquídea (méxico city), 13, 1–112. [in spanish] soto-arenas, m.a. (1988). listado actualizado de las orquídeas de méxico. orquídea (méxico city), 11, 233–277. 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[in spanish] o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 19 appendix 1 list of orchidaceae species of cuetzalan del progreso, puebla state, mexico. growth form: e – epiphytic, t – terrestrial, l – litophytic. vegetation type: tmcf – tropical montane cloud forests, trf – tropical rain forest, pof – pine–oak forest. photographer: ram: raúl alvarez-mora; conservation status: according to nom-059-semarnat-2010: a – threatened, pr – subject to special protection; iucn red list (2016): lc – least concern. new records for the state (*). species plants illegally traded in “tiangui” of cuetzalan del progreso, puebla (†) no. species characteristics 1. acianthera angustifolia (lindl.) luer (e; trf; 272 m; ram 2 photo) (*) 2. anathallis abbreviata (schltr.) pridgeon & m.w. chase (e; tmcf; 886 m; ram 3 photo) pr (*) 3. a. sertularioides (sw.) pridgeon & m.w. chase (e; trf; 272 m; ram 93 photo) 4. aspydogyne querceticola (lindl.) meneguzzo (t; trf; 1600 m; j. santana et al. 10 uamiz)(*) 5. bletia purpurea (lam.) dc. (t; tmcf; 1381 m; ram 26 photo) (†) 6. brassia verrucosa bateman ex lindl. (e; trf; 690 m; ram 27 photo) lc 7. camaridium cucullatum (lindl.) m.a. blanco (e; trf; 557 m; ram 28 photo) (†) 8. c. densum (lindl.) m.a. blanco (e; trf; 713 m; ram 29 photo) a (†) 9. c. meleagris (lindl.) m.a. blanco (e; tmcf; 1187 m; ram 30 photo) 10. campylocentrum micranthum (lindl.) rolfe (e; trf; 389 m; ram 4 photo) (*) 11. catasetum integerrimum hook. (e; trf; 466 m; ram 31 photo) (†) 12. chysis bractescens lindl. (e; trf; 256 m; ram 1 photo) a (*) (†) 13. c. laevis lindl. (e; tmcf; 1377 m; ram 32 photo) (†) 14. coelia macrosthachya lindl. (l; pof; 1429 m; ram 33 photo) (†) 15. c. triptera (sm.) g. don ex steud. (l; tmcf; 901 m; ram 34 photo) (†) 16. corymborchis forcipigera (rchb.f. & warsz.) l.o. williams (e; trf; 379 m; ram 5 photo) (*) 17. cyclopogon comosus (rchb.f.) burns-bal. & e.w. greenw. (t; pof; 1533 m; ram 35 photo) 18. c. luteo-albus (a. rich. & galeotti) schltr. (t; pof; 1552 m; ram 36 photo) 19. dichaea glauca (sw.) lindl. (e; trf; 594 m; ram 37 photo) (†) 20. d. intermedia ames & corell (e; tmcf; 1029 m; ram 38 photo) 21. d. muricatoides hamer & garay (e; trf; 688 m; ram 39 photo) (†) 22. d. trichocarpa (sw.) lindl. (e; tmcf; 1068 m; ram 6 photo) (*) (†) 23. dinema polybulbon (sw.) lindl. (e; trf; 929 m; ram 40 photo) 24. elleanthus cynarocephalus (rchb.f.) rchb.f. (e, l; tmcf; 1079 m; ram 41 photo) 25. encyclia candollei (lindl.) schltr. (e; tmcf; 1385 m; ram 42 photo) (†) r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 20 26. e. gravida (lindl.) schltr. (e; tmcf; 948 m; ram 7 photo) (*) 27. epidendrum atroscriptum hágsater (l; tmcf; 1028 m; ram 44 photo) (†) 28. e. cardiophorum schltr. (e; trf; 549 m; ram 43 photo) 29. e. di�usum sw. (e; trf; 663 m; ram 8 photo) (*) (†) 30. e. eustirum ames, f.t. hubb. & c. schweinf. (e; trf; 1034 m; ram 46 photo) 31. e. longipetalum a. rich. & galeotti (e; tmcf; 1390 m; ram 47 photo) (†) 32. e. melistagum hágster (e; trf; 732 m; ram 48 photo) 33. e. polyanthum lindl. (e; tmcf; 1073 m; ram 49 photo) (†) 34. e. ramosum jacq. (e; trf; 735 m; ram 50 photo) 35. e. tuxtlense hágsater, garcía-cruz & l. sánchez (l; tmcf; 616 m; ram 51 photo) (†) 36. e. veroscriptum hágsater (e; trf; 1092 m; ram 52 photo) (†) 37. erycina pusilla (l.) n.h. williams & m.w. chase (e; trf; 377 m; ram 9 photo) (*) 38. eulophia alta (l.) fawc. & rendl (t; trf; 330 m; ram 53 photo) 39. gongora galeata (lindl.) rchb. f. (e; trf; 738 m; ram 54 photo) 40. g. truncata lindl. (e; tmcf; 1065 m; ram 10 photo) (*) 41. habenaria �oribunda lindl. (t; tmcf; 1429 m; ram 11 photo) (*) 42. heterotaxis maleolens (schltr.) ojeda & carnevali (e; tmcf; 1199 m; ram 12 photo) (*) 43. h. sessilis (sw.) f. barros (e; tmcf; 1199 m; ram 56 photo) 44. isochilus latibracteatus a. rich. & galeotti (e; trf; 726 m; ram 57 photo) (†) 45. i. major schltdl. & cham. (e; tmcf; 991 m; ram 58 photo) (†) 46. jacquiniella equitantifolia (ames) dressler (e; trf; 814 m; ram 13 photo) (*) 47. j. teretifolia (sw.) britton & p. wilson (e; tmcf; 949 m; ram 59 photo) 48. leochilus labiatus (sw.) kuntze (e; trf; 417 m; ram 14 photo) (*) 49. lophiaris lurida (lindl.) braem (e; trf; 211 m; ram 15 photo) (*) 50. lycaste aromatica (graham) lindl. (e; trf; 846 m; ram 60 photo) (†) 51. l. deppei (lodd.) lindl. (e; tmcf; 1435 m; ram 61 photo) (†) 52. malaxis excavata (lindl.) kuntze (t; pof; 1517 m; ram 62 photo) 53. masdevallia �oribunda lindl. (e; trf; 592 m; ram 63 photo) 54. maxillariella tenuifolia (lindl.) m.a.blanco & carnevali (e; trf; 731 m; ram 64 photo) 55. m. variabilis (bateman ex lindl.) m.a. blanco & carnevali (e; tmcf; 1080 m; ram 65 photo) (†) 56. mormodes maculata var. unicolor (hook.) l.o. williams (e; tmcf; 1205 m; ram 66 photo) a (†) 57. nidema boothii (lindl.) schltr. (e; trf; 864 m; ram 67 photo) (†) 58. notylia barkeri lindl. (e; trf; 903 m; ram 68, 16 photo) 59. oncidium incurvum barker ex lindl. (e; tmcf; 1222 m; ram 69 photo) a (†) 60. o. sphacelatum lindl. (e; trf; 489 m; ram 70 photo) (†) 61. ornithocephalus in�exus lindl. (e; trf; 545 m; ram 17 photo) (*) 62. o. iridifolius rchb.f. (e; trf; 386 m; ram 94 photo) 63. platystele stenostachya (rchb.f.) garay (e; trf; 735 m; ram 71 photo) 64. pleurothallis cardiothallis rchb.f. (e; trf; 763 m; ram 72 photo) o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 21 65. polystachya lineata rchb.f. (e; trf; 365 m; ram 18 photo) (*) 66. ponera juncifolia lindl. (e; pof; 1434 m; ram 73 photo) 67. prescottia stachyodes (sw.) lindl. (t; tmcf; 1069 m; ram 19 photo) (*) 68. prosthechea cochleata (l.) w.e. higgins (e, l; trf; 963 m; ram 74 photo) (†) 69. p. ochracea (lindl.) w.e. higgins (e; trf; 706 m; ram 75 photo) 70. p. pseudopygmaea (finet) w.e. higgins (e; trf; 1050 m; ram 76 photo) 71. p. pygmaea (hook.) w.e. higgins (e; tmcf; 816 m; ram 77 photo) 72. p. radiata (lindl.) w.e. higgins (e; trf; 777 m; ram 78 photo) (†) 73. p. rhynchophora (a. rich. & galeotti) w.e. higgins (e; trf; 611 m; ram 79 photo) 74. p. vitellina (lindl.) w.e. higgins (e; tmcf; 1340 m; ram 80 photo) pr (†) 75. sacoila lanceolata (aubl.) garay (t; trf; 385 m; ram 20 photo) (*) 76. scaphyglottis fasciculata hook. (e; trf; 886 m; ram 21 photo) (*) 77. s. lindeniana (a. rich. & galeotti) l.o. williams (e; trf; 517 m; ram 22 photo) (*) 78. sobralia macrantha lindl. (e, l; tmcf; 1077 m; ram 81 photo) (†) 79. specklinia alata (a. rich. & galeotti) soto arenas (e; trf; 753 m; ram 23 photo) (*) 80. stanhopea oculata (g. lodd.) lindl. (e; tmcf; 1419 m; ram 82 photo) a (†) 81. s. ruckeri lindl. (e; tmcf; 997 m; ram 83 photo) (†) 82. s. tigrina bateman ex lindl. (e; tmcf; 909 m; ram 84 photo) a (†) 83. stelis emarginata (lindl.) soto arenas & r. solano (e; trf; 736 m; ram 85 photo) (†) 84. s. nagelii solano (e; trf; 882 m; a. espejo 7173 uamiz) 85. s. ornata (rchb.f.) pridgeon & m.w. chase (e; tmcf; 1476 m; ram 86 photo) 86. s. pachyglossa (lindl.) pridgeon & m.w. chase (e; tmcf; 961 m; ram 87 photo) 87. s. platystylis (schltr.) solano & soto arenas (e; tmcf; 961 m; ram 88 photo) 88. s. rubens schltr. (e; tmcf; 1002 m; ram 24 photo) (*) 89. s. veracrucensis solano (e; trf; 667 m; ram 89 photo) (†) 90. stenorrhynchos speciosum (jacq.) rich. ex spreng. (e; pof; 1431 m; ram 90 photo) 91. trichocentrum candidum lindl. (e; trf; 582 m; ram 91 photo) 92. trichosalpinx ciliaris (lindl.) luer (e; trf; 882 m; ram 92 photo) 93. vanilla planifolia andrews (h; trf; 277 m; ram 25 photo) pr (*) (†) r aú l a lv ar ez -m or a, w ill ia m c et za l-i x, s ai ka t k um ar b as u, e lia na n og ue ra -s av el li n oe l a . g on zá le zv al di vi a, j es ús f . m ar tín ez -p uc , p ei m an z an di , k at ar zy na m oż dż eń 22 abstract �e orchid �ora of puebla state represents 16% of the total species present in mexico demonstrating rich biodiversity. however, several municipalities in the north-east of puebla area are located within a conservation priority area for its tropical montane cloud forest and tropical rain forest due to decision of the comisión nacional para el conocimiento y el uso de la biodiversidad of mexico. we have recorded 93 species from cuetzalan del progreso with 25 new records of the �ora of the state. �ese new additions thereby currently increase the orchid �ora of puebla to 197 species. of these, six species are classi�ed as threatened, three as subject to special protection by the nom-059-semarnat-2010 and one classi�ed as least concern by the international union for the conservation of nature. moreover, we recorded 37 species illegally traded in “tianguis” (informal markets) of cuetzalan del progreso. �e orchid diversity in the fragmented forests can be harnessed in establishing a model for the sustainable management and to promote conservation action of species at some level of threat and in the restoration of areas with fragmented forests. unfortunately, the orchid diversity has been seriously endangered by several anthropogenic factors. we have also recorded signi�cant anthropogenic threats in this municipality for the long-term existence of the local orchid members and species with potential for commercial production. a viable approach will be to allow the introduction of in vitro germination program and organic production of orchids by local and fringe communities; this will reduce inhabitants’ dependence on the scarce forest resources, lessen illegal harvesting and tra�cking of wild orchids, and at the same time provide economic development for the local communities. key words: anthropogenic factors, biodiversity, conservation, cuetzalan del progreso, mexico, orchids received: [2016.07.18] accepted: [2016.10.26] różnorodność storczyków w cuetzalan del progreso, puebla, meksyk – antropogeniczne zagrożenia i potencjał do produkcji ekologicznej w celach ochronnych streszczenie flora storczyków stanu puebla charakteryzuje się dużą różnorodnością i stanowi 16% ogólnej liczby gatunków występujących w  meksyku. kilka gmin w  północno-wschodniej części puebla, położonych w  obrębie górskiego tropikalnego lasu mglistego i lasów tropikalnych deszczowych, jest szczególnie chronionych przez krajową komisję ds. wiedzy i użytkowania różnorodności biologicznej meksyku. w cuetzalan del progreso odnotowano 93 gatunki storczyków, wśród których 25 było nowych dla �ory tego stanu. powiększyło to liczebność �ory storczykowej puebla do 197 gatunków. spośród nich, 6 gatunków zaliczono do zagrożonych, 3 do podlegających szczególnej ochronie przez nom-059-semarnat-2010 oraz 1 sklasy�kowano jako gatunek najmniejszej troski wg międzynarodowej unii ochrony przyrody. ponadto odnotowano 37 gatunków w nielegalnym handlu „tianguis” (na nieformalnym rynku) w cuetzalan del progreso. różnorodność storczyków we fragmentarycznych lasach można wykorzystać w tworzeniu modelu zrównoważonego zarządzania i  promocji działań ochronnych gatunków na tym samym poziomie zagrożenia oraz w odtwarzaniu tych obszarów. niestety, różnorodność storczyków jest poważnie zagrożona przez kilka czynników antropogenicznych. w badanych gminach stwierdzono istotne zagrożenia antropogeniczne dla długoterminowej lokalnej egzystencji przedstawicieli storczykowatych oraz gatunków z  potencjałem dla produkcji komercyjnej. stosownym rozwiązaniem będzie możliwość wprowadzenia programu in vitro do kiełkowania i produkcji ekologicznej storczyków przez lokalne społeczności, co przyczyni się do poprawy miejscowej gospodarki, zmniejszenia jej zależności od ograniczonych zasobów leśnych oraz nielegalnego pozyskiwania i handlu dzikimi storczykami. słowa kluczowe: czynniki antropogeniczne, bioróżnorodność, ochrona, cuetzalan del progreso, meksyk, storczyki o rchid diversity at c uetzalan del p rogreso, p uebla, m exico – anthropogenic threats and potential for organic production for conservation purposes 23 information on the authors raúl alvarez-mora interested in taxonomy and conservation of orchids of puebla, méxico. currently involved in designing botanical gardens with a focus on growing di�erent orchid species. william cetzal-ix interested in systematics, taxonomy and conservation of neotropical orchids and ferns. also, in �oristic studies of indicator species (epiphytes) conservation of forests of south-eastern mexico, as well as knowledge and conservation of plants with potential uses (melliferous, medicinal and ornamental). saikat kumar basu traditionally trained in botany (plant sciences) and specializing in microbiology, works actively in various areas of plant sciences and environmental conservation. �e author works extensively on forage crops with particular reference to annual forage legume and medicinal herb and spice, fenugreek. currently he is working in biomolecular sciences dealing with plant biotechnology and genetic engineering application in small grain cereals like wheat. eliana noguera-savelli interested mainly in systematics, taxonomy, �oristic and anatomy of neotropical vascular plants, developing research to generate knowledge on biodiversity, exploration of timber and non-timber forest resources and training of human resources to support knowledge, the conservation and sustainable use of natural resources. noel a. gonzález-valdivia professor and researcher at the instituto tecnológico nacional, sni level 1; an agronomist specializing in the rescue, assessment and management of native germplasms adapted to the tropics. also interested in ethnobiology studies, ecology and sustainable development, which have recently resulted in innovative applications of traditional knowledge in obtaining human products such as biopesticides, medicinal and food products based on �torrecursos utility. participated in the training of human resources; and in the development of initiatives and projects with positive impacts on the rural environment. jesús f. martínez-puc extensionist, researcher, and professor interested in the tropical apiculture. currently developing projects on pest control bees, nutritional quality of nectar and mellifera �ora. peiman zandi deeply trained in agronomy (crop science) and specializing in stress physiology, biotic/abiotic stresses and agroecology. he is also interested in working in di�erent areas of plant developmental biology, agroecology, plant nutrition, botany, plant breeding and genetics. katarzyna możdżeń her scienti�c interests concentrate on the e�ects of di�erent environmental factors (light, ozone, heavy metals, allelopathic extracts) on the morphology and physiology of plants cultivated, protected and invasive species. 81 annales universitatis paedagogicae cracoviensis studia naturae, 2: 81–96, 2017, issn 2543-8832 doi: 10.24917/25438832.2.6 hamid dorosti,1 katarzyna możdżeń,2 peiman zandi3*, morteza siavoshi4 1rice research institute of iran (rrii), p.o. box 1658, rasht, iran 2department of plant physiology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland 3institute of environment and sustainable development in agriculture, chinese academy of agricultural sciences, p.o. box 10081 beijing, p. r. china; *z_rice_b@yahoo.com 4assistant professor, department of agricultural sciences, payame noor university, i.r. of iran the influence of foliar “feeding” of urea on yield and its components of iranian hybrid rice oryza sativa l. ‘bahar 1’ introduction rice (oryza sativa l.) has become a highly strategic and priority crop in approximately half of the global population with regard to procuring energy, proteins, and vitamins (irri, 2008). it is mainly cultivated in asian countries under a hot humid climate in both tropical and temperate regions. rice is regarded as the major source of food in iran (population 77 million; 46 kg per capita/year), with an average annual domestic production of 2.35 million tons of paddy rice grown on 539.000 hectares and an annual consumption of 3.5 million tons (ahmadi et al., 2014). it is projected that iran needs to produce 5 million tons of rice by 2035 to achieve self-su�ciency (akhgari, 2010). �e recent rice crisis proves again that a sustainable increase in rice production is crucial for food security in the rice-growing countries of asia (irri, 2008). hybrid rice technology is a key strategy for increasing rice production and maintaining self-su�ciency and food security (dorosti, 2009). �ese rice varieties give about 15–25% more yield than improved inbred rice in farmers’ �elds (xie, hardy, 2009). its potential for enhancing rice production and productivity has motivated many countries, like iran, to exploit this technology. �e hybrid rice program in iran was launched at the rice research institute of rasht at the beginning of 1987 through establishing a collaborative project together with the international rice research institute (irri, 2008), philippines, as the major factor contributing to the remarkable success of hybrid rice technology in iran (dorosti, 2009). providing e�cient quantities of nutrients needed by plants, in particular the essential elements, is one of the most important factors of crop management (youseh am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 82 �, zandi, 2012). crop production can be increased through application of fertilisers containing these elements (shen et al., 2013). increasing the concentration of essential elements in plants assigns an important role in improving the quality of food products and improves the health of society (tavakoli et al., 2014). dwindling water resources, intermittent droughts, low fertility of agricultural lands, the lack of available nutrients, and their low accessibility to the organic resources are among barriers to accessing proper operation in the �eld of agriculture (shirani rad, zandi, 2014). fageria (2013) reported that nitrogen is one of the essential elements required for plant growth. crop plants assimilate nutrients not only through their leaves but as well as the other shooting parts, like young panicles, stems, and �owers. fresh and swi�ly expanding leaves are more streamlined in absorption of foliar-sprayed nutrients from their upper/ lower surfaces than those of fully matured ones (mondal, al-mamun, 2011). nitrogen contributes to grain �lling through elevating this element content in leaves and to sink size through increasing the hull size and mitigating the number of degenerated spikelets (singh et al., 2014). liu et al. (2014) declared that consuming nitrogen fertiliser in a proper time sequence helps to eliminate excessive use of it. due to the high solubility of nitrogen fertilisers, proper timing of nitrogen application is assumed to be very pivotal in better e�ciency of nitrogen use and less loss of this element to the environment (fageria, 2013). in their study on aman rice, bhuyan et al. (2012) demonstrated that foliar application of n fertiliser in a bed planting method increased yield attributes, such as grain yield, the number of panicles per square meter, the number of grains per panicle, 1000-grain weight, and water use e�ciency for biomass and grain production much more than the conventional methods. singh et al. (2014) suggested that foliar feeding of rice by inputting higher doses of nitrogen fertiliser enhanced the growth dynamic, biomass partitioning, and cha�y grain. however, in their study, nitrogen use e�ciency remained una�ected. furthermore, the yield of rice and components were found to have less attribution to foliar application of urea fertilisation (hasanuzzaman et al., 2009). �e highest crop in canola was recorded by foliar spraying of nitrogen at the stem elongation or right before the �owering stage (tousikehal et al., 2011). azarpour et al. (2011), in their study on rice crops, reported a consecutive enhancement in yield as a result of adding nitrogen to the soil. although foliar spraying during mead-season had a positive e�ect on the grain yield of rice crop (brown, petrie, 2006; asadi et al., 2014), the same result in grain protein was noticed exclusively when spraying was done at active tillering and booting stages (asadi et al., 2015). in recent decades, agricultural products have mostly relied on the utilisation of chemical inputs, which in itself has detrimental e�ects on the environment (zandi, basu, 2016). comparatively, the soil application of nitrogenous fertiliser does not appear to be a better method, because it causes the plants to absorb the nutrients 83 much slower, and large quantities will be required for normal growth in contrast to the smaller ones of the same generally required for foliar application (mondal, al-mamun, 2011). in particular, the e�ectiveness of foliar feeding is augmented when the canopy microclimate is occupied by high levels of leaf sources (akhgari, 2010). due to the limited or inconclusive information on the e�ciency of the foliar application of urea fertiliser on yield attributes and the yield of hybrid rice grown under moderate conditions, it was decided to conduct this research. �e aim of the study was to examine the in�uence of foliar application of urea fertiliser at active tillering and booting stages on the yield of hybrid rice o. sativa ‘bahar 1’ and �nding out the optimum concentration of urea fertilisation for the maximum yield of rice under moderate climatic conditions. it was hypothesised that di�erent concentrations of foliar application of urea fertiliser would receive more palatable nitrogen fertiliser use e�ciency than conventional fertilising method. materials and methods �e �eld experiment was conducted in talesh county, guilan-iran in 2014, at latitude 48°37ʹn and longitude 54°48ʹe, with 45 m of altitude. �e climate, according to köeppen classi�cation, is mediterranean with mild winters and warm-humid summers. most precipitation occurs in late summer and early spring. soil ph was determined using glass electrode in a 1: 2.5 soil/water suspension. available phosphorus (p) in the soil samples was computed by leaching the soil with 0.002 n sulfuric acid (1 soil: 200 h2so4 suspension, w/v) and agitating it for at least half an hour and �ltering it through whatman �lter paper no. 42. available p in the extract was determined spectrophotometrically at the wavelength 690 nm (ravikumar, somashekar, 2013). leaching the soil sample with 1n ammonium acetate at ph 7.0 (w/v), keeping it for overnight, �ltering it through whatman �lter paper no. 42, and brining the volume up to 100 ml with distilled water, the exchangeable k+ in soil could be extracted a�er the method followed by britzke et al. (2012). �e �ltrate (sample) containing k was then used for the �ame photometric determination of potassium. �e soil analysis showed that the texture of the experimental site (0–25 cm) was silty clay with 0.71 ds m-1 ec; 6.6 ph (h2o); 2.21% organic matter; 20.1 mg kg -1 available phosphorus; 127 mg kg-1 exchangeable potassium, and 0.238% total nitrogen. �e experiment was laid out in a two-factor randomised complete block design (rcbd) with three nitrogen treatments (n0, n10, n20 kg n ha -1) and two application times (at tillering and booting stages), and it was replicated three times. each plot was 3×4 m in size. urea fertiliser (co(nh2)2), as the best source of actual nitrogen (46% n) for aerial application (norton, 2011), was considered for foliar spraying (control or no n fertilisation, 10 kg n ha-1 ≈ 22 kg urea ha-1, 20 kg n ha-1 ≈ 44 kg urea ha-1) the influence of foliar “feeding” of urea on yield and its com ponents of iranian hybrid rice o ryza sativa l. ‘b ahar 1’ h am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 84 using a single-nozzle hand sprayer. �e urea spray volumes were prepared by mixing 5 kg of urea in 100 l of water (i.e. 5% urea solution) as per treatment (asadi et al., 2014). �e plots were sprayed during late a�ernoon hours when the wind speed was less than 12 km hr−1. �e hybrid rice variety of ‘bahar 1’ was used that was bred through combining a cytoplasmic male sterile line and a restorer line (ir58025a / ir42686r) (dorosti, 2009). fertiliser recommendations for experimental plots were based on soil surveys. basal fertilisers of nitrogen (50 kg n ha-1), triple superphosphate (160 kg tsp ha-1), and potassium chloride (50 kg kcl ha-1) were applied right before transplanting. an additional amount of potassium chloride (50 kg kcl ha-1) was applied at the maximum tillering stage. crop cultivation was carried out on april 14th, and when the seedlings height was about 25 cm in early may (on may 5th), they were transferred (three seedlings per hill and 25×25 cm spacing) to the main �eld of transplantation. weeds (butachlor 4 l ha-1) and pests (diazinon 5% granule) were controlled by adopting conventional methods of the region. all records were taken during the plant growth. at maturity, a�er eliminating the marginal lines on both sides of each plot, all plants in the harvest area (6 m2) were selected and cut at above ground level. �e crop was harvested at a grain moisture content of about 20–25% wet basis. grain yield was calculated as kg ha-1 at 14% moisture content. �e rice panicles in the �nal-harvesting area were counted, and their average was reported as the number of panicles per square meter. �e number of grains per panicle was computed from the randomly selected plants in each plot. for the same samples, twenty panicles from each plot were randomly chosen. �e main panicle fertility percentage was measured based on the grain number in the main panicle and the number of sterile (non-fertile) �owers. �e �lled/fertile grains were then randomly selected from the grain samples corresponding to each plot and the mean weight of 10 replications of 100 grains multiplied by 10 reported as 1000-grain weight (g). �e experimental data were subjected to statistical veri�cation with fishers analysis of variance (anova) using the sas 9.1 statistical so�ware package. means were separated based on multiple range test of duncan (msrt) at a 0.05 probability level. bar graphs were depicted using excel (microso�, redmond, wa, united states). correlation coe�cients of pearson were calculated on the traits studied for the correlation analysis. �e r package ‘corrgram’ was employed to display the correlations between the selected traits by using a ‘correlogram’ (asters et al., 2014). a correlogram was a direct visual display of the matrix of pearson coe�cients that were estimated from the experimental data. by this method, correlations between traits were displayed by grouping traits that have similar characteristics, and the values and signs of the correlations were visualised schematically in numbers and 85 tab. 1. plant parameters of iranian hybrid rice oryza sativa l. ‘bahar 1’ as a�ected by foliar spraying of nitrogen fertiliser and timing of application mean square df source of variation grain yield (kg ha-1) panicle number per unit area grain number per panicle panicle fertility rate [%] 1000 grain weight [g] 202632.62 ns31.26 ns222.94 ns7.03 ns0.03 ns2r 5126514.14**9151.98**1999.83**364.38**0.26 ns2a 10076519.95**16642.17**3957.53**671.36**0.15 ns1linear a 176562.32 ns1661.78**42.13 ns57.39*0.36 ns1quadratic a 1256327.58*421.16 ns23.36 ns45.6*0.04 ns2b 2266943.83*650.4 ns11.20 ns47.46*0.07 ns1linear b 245711.34 ns191.91 ns35.52 ns11.74 ns0.001 ns1quadratic b 84789.78 ns24.80 ns24.77 ns7.32 ns0.16 ns4a×b 320634.79128.83136.2111.930.1716error 9.594.596.885.191.94-cv [%] experimental meanstreatments c 5101.10c 211.30c 154.01a 73.70a 21.26a1foliar application at tillering (a) kg ha-1 b 6020.90b 258.35b 171.49b 64.50a 21.10a2 a 6597.60a 272.11a 183.67b 41.67a 21.44a3 b 5484.20b 293.36a 169.74b 63.99a 21.34b1foliar application at booting (b) kg ha-1 ab 6041.00a 251.02a 168.01a 67.47a 21.26b2 a 6194.00a 251.38a 171.32a 68.19a 21.21b3 notes: * – p < 0.05, ** – p < 0.01, ns – p > 0.05; df – degrees of freedom, r – replication, a – tillering e�ect, b – booting e�ect; a×b – represent interaction terms between the treatment factors; cv [%] – coe�cient of variation, means in each column, down-parts of table 1, followed by the di�erent letters are signi�cantly di�erent (p < 0.05) according to duncan test; a1, b1: control or no n fertilisation, a2, b2: 10 kg n ha-1 ≈ 22 kg urea ha-1, a3, b3: 20 kg n ha-1 ≈ 44 kg urea ha-1 the influence of foliar “feeding” of urea on yield and its com ponents of iranian hybrid rice o ryza sativa l. ‘b ahar 1’ colour-coded pie graphs. �e pie graphs were �lled in proportion to the pearson coe�cient values, counter clockwise for negative correlations (in red) and clockwise or positive correlations (in blue). results and discussion panicle number per unit area in this study, the number of panicles per unit area, being the most important components of grain yield, was strongly in�uenced by the time of fertilising, in a way that the foliar spraying of urea fertiliser at the tillering stage showed a highly significant e�ect (p < 0.01), while it le� no e�ect at the booting stage (tab. 1). �e result would likely re�ect the superior impact of urea fertiliser at the time of tillering in improving the number of reproductive units per unit area. generally, the number of h am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 86 productive panicles per plant is determined during the reproductive phase (singh et al., 2014). �is indicates the hypothesis that most of the nitrogen foliar-sprayed at booting was utilised for improving the fertility rate of panicles rather than their quantity (dorosti, 2009). �e interaction e�ect of foliar feeding of urea at di�erent stages of tillering and booting was found to be non-signi�cant on the number of panicles. a comparison of means associated with the tillering stage showed that, by gradual increasing of urea fertiliser application, the number of panicles per unit area increased as well. �e highest (272.1) and the lowest (211.3) number of panicles belonged to those of plots enriched with 20 and 0 kg of n per hectare (fig. 1, tab. 1). it is suggesting that the plant behaviour is completely proportional to the amount of urea being applied at tillering, and that the relationship between the number of panicles per unit area and the rate of urea fertiliser is slightly of a quadratic equation. in other words, nitrogen application up to level of 10 kg ha-1 drastically enhanced the number of panicles by 22%, and any further application contributed to only a 5% increase in panicle number, indicating that an extra n input was utilised for other processes (liu et al., 2016). in support to our �ndings, kazemeini and ghadiri (2005), peng et al. (2010) and more recently cao et al., (2013) documented a noticeable decrease in photosynthetic n use e�ciency under an elevated use of n fertiliser. fig. 1. in�uence of foliar applied n fertiliser (0 kg n ha-1, 10 kg n ha-1 ≈ 22 kg urea ha -1, 20 kg n ha-1 ≈ 44 kg urea ha-1) on panicle number per square meter; urea was applied in solution with 5% dry matter basis; signi�cant di�erences between the treatments in tillering and booting stages are indicated with asterisks (* – p < 0.05, ** – p < 0.01, ns – p > 0.05, duncan multiple range test); �e error bars denote the standard deviation (± sd) of the mean value; n = 6 87 grain number per panicle foliar spraying of urea in the tillering stage signi�cantly (p < 0.01) and linearly a�ected the number of grains generated in each panicle, while it had no e�ect on this trait during the booting stage. in other words, the number of grains in panicles of those plants receiving foliar spray of n, by the booting stage, did not di�er signi�cantly from that of untreated check plants, suggesting that the n fertiliser was exploited for other processes, such as improving panicles quality. �e number of grains per panicle is mainly associated with the factors in�uencing the growth parameters right before the initiation of pollination process. accordingly, any defect in these requisite factors could likely reduce the grain number per panicle (dorosti, 2009). �us, the availability of nitrogen through foliar spraying of urea prior to the formation of panicle could be the main reason for its e�ectiveness in increasing the grain number per panicle. a comparison of mean values shows that the highest number of grains per panicle (183.6) was achieved by the application of 20 kg n ha-1 at the tillering stage, followed by 10 kg n ha-1, and the lowest value (154) was recorded in the untreated (non-sprayed) check plants (fig. 2, tab. 1). liu et al. (2016), in their study on yield of lowland rice receiving n fertilisation treatment, concluded that the increase in the number of grains per panicle was in extremely close association with nitrogen application rate under constant submerged condition. the influence of foliar “feeding” of urea on yield and its com ponents of iranian hybrid rice o ryza sativa l. ‘b ahar 1’ fig. 2. in�uence of foliar applied n fertiliser (0 kg n ha-1, 10 kg n ha-1 ≈ 22 kg urea ha-1, 20 kg n ha-1 ≈ 44 kg urea ha-1) on grain number per panicle; urea was applied in solution with 5% dry matter basis; signi�cant di�erences between the treatments in tillering and booting stages are indicated with asterisks (* – p < 0.05, ** – p < 0.01, ns – p > 0.05, duncan multiple range test); �e error bars denote the standard deviation (± sd) of the mean value ; n = 6 h am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 88 panicle fertility rate �e data shows that the panicle fertility percentage (seed setting) was signi�cantly di�erent according to the timing of urea spraying (fig. 3). a comparison of means regarding the levels of nitrogen fertiliser at the active tillering stage shows that the number of �lled grains in the panicles decreased by increasing the amount of urea fertiliser. �e maximum value of fertile panicles were recorded in untreated check plants (73.7), followed by 10 kg n ha-1 (64.5), which was on par with 20 kg n ha-1 (61.47) (tab. 1). a notable reduction in the fertility percent of panicles was observed in return for enhanced n application in the tillering stage, which might be due to an increased number of infertile (or non-productive) tillers and panicles per unit area (akhgari, 2010). srividya et al. (2010) pointed out that an excess application of n input could increase the number of cha�y (un�lled) grains or sterile spikelets in a relatively high responsive manner to n fertilisation in comparison with optimum n dose. urea spraying at the booting stage also had a signi�cant e�ect on the percentage of fertile panicles (p < 0.05). �e number of �lled grains in the panicles was increased in line with the increased level of urea fertiliser. urea application that augment plant nitrogen before heading (at booting stage) can be highly e�ective in giving rise to the number of �lled grains and/ or spikelets (kamiji et al., 2011). fertiliser levels of 10 and 20 kg n ha-1, being on par with each other, was better than no spraying and revealed the appropriate timing for n application at this stage. interaction between foliar feeding of n and developmental stages of tillering and booting was found to be non-signi�cant. mingotte et al. (2013) believed that, if the n fertiliser supplementation is applied in the period that the spikelets initiated di�erentiation in the panicle, the plant itself does not encounter any problem regarding n de�ciency, especially when initiating �ower primordia, and hence it will produce more �lled grains. singh et al. (2014) demonstrated that the number of un�lled (sterile spikelets) rice panicles could be multiplied with the excess application of nitrogen as compared with the optimum n level. indeed, a further increase in n levels decreases the physiological nitrogen-use e�ciency by reducing the nitrogen content in leaves to be remobilized during grain feeling (cao et al., 2013; kant et al., 2011). �ousand grain weight �ousand grain weight (tgw) is a complex quantitative genetic trait for rice and is one of the three key factors that in�uence the grain yield (wei et al., 2014). �e grain weight is a highly stable varietal character (fageria et al., 2011) that is determined during the ripening phase. �ere was no in�uence of urea fertilisation on tgw (p > 0.05) at both the growth stages (tillering and booting). under urea fertiliser treatment, tgw varied from 21.1 to 21.44 g (tillering stage; average value of 21.3 g) and from 20.26 to 21.34 g (booting stage; average value of 21 g) (tab. 1). fageria et al. (2011) re89 ported tgw of lowland rice genotype ‘brsgo guar’ varied from 21.1 to 24.6 g, with an average value of 23.0 g. similar results were also documented in those of previous studies that reported the lack of in�uence of soil (azarpour et al., 2011) and foliar (asadi et al., 2011, 2014) application of nitrogen on tgw between the genotypes tested. sarwa et al. (2011) also stated that thousand-grain weight depends on the genetic constitution and is less a�ected by growing conditions. �e natural conditions and environment are believed to have more interference in determining grain quantity (seed size) rather than inherent factors (wei et al., 2014). grain yield in the present study, the grain yield was signi�cantly and linearly a�ected by various foliar n fertiliser levels at both the tillering (p < 0.01) and booting (p < 0.05) stages, while no interactive e�ect was noticed among the di�erent combination treatments (fig. 4, tab. 1). �e di�erences in the grain yield were largely because of variations in the yield components, including the number of grains per panicle, the number of panicles per unit area, panicle fertility percent, and thousand-grain weight (hasanuzzaman et al., 2009). �e comparison of means at the tillering stage revealed that the grain yield was increased by applying the amount of n fertiliser from 0 to 20 kg ha-1. �e treatment 20 kg n ha-1 resulted in the highest rice grain yield (6597 kg ha-1), which was followed by 10 kg n ha-1 (6020 kg ha-1). �is dose was found to be superior to 10 kg n ha-1 treatment and control. �e lowest grain yield was observed in untreatthe influence of foliar “feeding” of urea on yield and its com ponents of iranian hybrid rice o ryza sativa l. ‘b ahar 1’ fig. 3. in�uence of foliar applied n fertiliser (0 kg n ha-1, 10 kg n ha-1 ≈ 22 kg urea ha-1, 20 kg n ha-1 ≈ 44 kg urea ha-1) on panicle fertility percentage; urea was applied in solution with 5% dry matter basis; significant di�erences between the treatments in tillering and booting stages are indicated with asterisks (* – p < 0.05, ** – p < 0.01, ns – p > 0.05, duncan multiple range test); �e error bars denote the standard deviation (± sd) of the mean value ; n = 6 h am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 90 ed/ unsprayed check plants. �e linear increase in grain yield with increasing foliar n fertiliser rates may be associated with more grain, panicle, and/or tiller number produced per plant (tab. 1). furthermore, the positive and signi�cant relationship between the grain yield and the number of panicles per unit land area at harvest time (xu et al., 2015) might be an indicative of this result. �ere was a signi�cant and linear increase in grain yield with increasing n rates with urea in the booting stage. in other words, grain yield was signi�cantly improved in a linear fashion when n rates increased from 0 to 20 kg ha-1 by urea fertilisation. maximum grain yield was achieved at 20 kg n ha-1 (6194 kg ha-1), which was on par with 10 kg n ha-1 treatment (6041 kg ha-1). �e increase in grain yield may be attributed to the high fertility percentage of panicles in return for increasing n rates during the booting stage. mingotte et al. (2013), working with nitrogen topdressing, obtained the highest rice yield when foliar n application was made before the crop was headed (panicle di�erentiation). our results are consistent with recently released investigations in terms of the existence of a noticeable increase in grain yield a�er foliar spraying of n fertiliser in maximum tillering (asadi et al., 2011, 2014) and booting stages (asadi et al., 2014, 2015). correlations between the examined traits to determine if there were associations between all the tested traits, correlation analysis was performed on mean values derived out of anova analysis, and the fig. 4. influence of foliar applied n fertiliser (0 kg n ha-1, 10 kg n ha-1 ≈ 22 kg urea ha-1, 20 kg n ha-1 ≈ 44 kg urea ha-1) on grain yield as kg ha-1; urea was applied in solution with 5% dry matter basis; significant differences between the treatments in tillering and booting stages are indicated with asterisks (* – p < 0.05, ** – p < 0.01, ns – p > 0.05, duncan multiple range test); the error bars denote (± sd); n = 6 91 correlation matrices were visualised schematically by using r package ‘corrgram’ (fig. 5). correlations displayed in a correlogram were organised in the order that traits having similar characteristics were grouped together. �e colour-coded pie graphs were re�ected in the upper triangle, and the values and signs of the pearson coe�cients were represented schematically with the correlation coe�cients and the 95% con�dence intervals displayed in the lower triangle. yield related traits, i.e. grain yield (gy), and grain number per panicle (gn.p) were found positively correlated to each other with a coe�cient being at least 0.9 (p < 0.05). gn.p were also noticed to be correlated to panicle number per square meter (pn.sm; r: 0.72; p > 0.05) and thousand grain weight (tgw; r: 0.39; p > 0.05), and negatively correlated with panicle fertility rate (pfr; r: -0.88; p < 0.05). no signi�cant association was recorded for tgw. as per correlation coe�cients results associated with pfr, our study showed a close/ direct association between the number of cha�y grains and gn.p, which suggested that the mitigation of grain number per panicle is likely involved in enhancement of fertility rate. conclusions nitrogen is believed to be the most constraining factor on crop production in many of the world’s agricultural regions, and its e�cacious adoption is indispensable for the frugal sustainability of cropping systems. moreover, the dynamic nature of this element and its inherent tendency for loss from soil-plant systems creates a challenging and peerless space for its optimal management. foliar fertilisation technique may also be a good alternative to the convectional soil application to avoid the risk of �xation or leaching of nutrients. �ese results clearly showed that foliar-applied urea at active tillering could have a better performance once applied at higher concentrations (≈ 44 kg urea ha-1). as an instance, the grain number per panicle and panicle number per unit area increased to about 19.26 and 28.78%, respectively, compared with those of untreated check plots. plants fertilised with a lower dose of urea (10 kg n ha-1≈ 22 kg urea ha-1) in the booting stage gave the more appropriate performance for both the panicle fertility rate and grain yield. in other words, high n fertiliser application exceeding 10 kg ha-1 in the boot stage has likely resulted in a considerable decline in n use e�ciency and could lead to an increased n loss risk. �ousand-grain weight was not appreciably a�ected by urea fertilisation timing, since it is mostly governed by genetic constitution. although, the di�erent concentrations of urea fertiliser at two stages of growth were used, further investigations relying on timing and other concentrations are needed to be undertaken in di�erent agro-ecological regions. the influence of foliar “feeding” of urea on yield and its com ponents of iranian hybrid rice o ryza sativa l. ‘b ahar 1’ h am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 92 fig. 5. correlogram display of correlation matrices for experimental data (grain yield: gy, �ousand grain weight: tgw, panicle fertility rate: pfr, panicle number per square meter: pn.sm; gran number per panicle: gn.p); �e pie graphs are �lled in proportion to the pearson’s coe�cient values, clockwise for positive correlations (in blue) and anti-clockwise for negative correlations (in red); �e numbers are pearson coe�cients with 95% con�dence intervals references ahmadi, k., gholizadeh, h., ebadzadeh, h.r., hosseinpour, r., hatami, f., fazli, b., kazemian, a. ra�ei. m. 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(2016). role of plant growth-promoting rhizobacteria (pgpr) as biofertilizers in stabilizing agricultural ecosystems, chapter 3. in: d. nandwani (ed.), organic farming for sustainable agriculture. germany: springer international publishing, 71–87. doi: 10.1007/978-3-319-26803-3_3 abstract because of waterlogged conditions in rice cultivation, much of the surface-broadcasted urea dissolves in paddy water and is unreachable to the rice crop for this reason. a �eld experiment was conducted to estimate the in�uence of urea fertilisation on yield and yield components of hybrid rice ‘bahar 1’. �ree doses (n0, n10, n20 kg n ha -1) of nitrogen from urea sources were foliar-sprayed once at active tillering and booting stages. grain yield, the number of panicles per unit area, and the number of grains per panicle in the tillering 95 the influence of foliar “feeding” of urea on yield and its com ponents of iranian hybrid rice o ryza sativa l. ‘b ahar 1’ stage of rice were signi�cantly (p < 0.01) increased in a linear fashion when n rates increased from 0 to 20 kg ha-1. �e rate of panicle fertility was negatively/positively in�uenced with increasing n rates in the tillering and booting stages, respectively, indicating the creation of more number of non/partial productive tillers per hill in the vegetative stage than in the reproductive stage. in line with panicle fertility (%), the grain yield was also signi�cantly a�ected by n treatments with urea fertilisation in the booting stage. maximum grain yield was obtained with the application of 20 kg n ha-1 at both the tillering and booting stages. foliar spraying of urea at tillering (20 kg n ha-1) and booting (10 kg n ha-1) stages had a pronounced e�ect on achieving higher yields as compared to other combinations. �e study suggests that foliar application of urea for hybrid rice cultivation might have a potential role in improving nitrogen use e�ciency. key words: oryza sativa l. ‘bahar 1’, foliar spraying, urea fertilisation, grain yield received: [2017.07.11] accepted: [2017.10.31] wpływ „dożywiania” dolistnego mocznikiem na plon i jego składniki irańskiego mieszańca ryżu oryza sativa l. ‘bahar 1’ streszczenie ze względu na podmokłe warunki upraw ryżu, większość powierzchniowej transmisji mocznika rozpuszcza się w wodzie gruntowej i z tego powodu nie jest osiągalna dla uprawy ryżu. przeprowadzono doświadczenie polowe w  celu oszacowania wpływu nawożenia mocznikiem na wydajność i  składniki plonu mieszańca ryżu oryza sativa l. ‘bahar 1’. trzy dawki azotu (n0, n10, n20 kg n ha -1), pochodzące z zasobów mocznika, były rozpryskiwane na liście ryżu jednorazowo, przed stadiami krzewienia i tworzenia liścia �agowego. plon ziarna, liczba wiech na jednostkę powierzchni i liczba ziaren przypadających na wiechę w fazie krzewienia ryżu wzrastały istotnie (p < 0,01) w sposób liniowy, gdy dawki azotu (n) wzrastały od 0 do 20 kg ha-1. poziom płodności wiechy był odpowiednio uzależniony od wzrostu dawek azotu (n) w stadiach krzewienia i tworzenia liścia �agowego; wskazując na utworzenie większej liczby częściowo nieproduktywnych na wierzchołku źdźbeł w stadium wegetatywnym niż w stadium generatywnym. z kolei, z płodnością wiech (%), plon ziarna był także istotnie uzależniony od azotu (n) poprzez traktowanie mocznikiem w stadium tworzenia liścia �agowego. maksymalna wydajność plonu ziarna uzyskano przy aplikacji 20 kg n ha-1 w obydwu stadiach: krzewienia i tworzenia liścia �agowego. opryskiwanie liści mocznikiem w fazach krzewienia (20 kg n ha-1) i tworzenia liścia �agowego (10 kg n ha-1) miało wyraźny wpływ na uzyskiwanie wyższych plonów, w porównaniu z innymi kombinacjami. badania wskazują, że dolistne zastosowanie mocznika w uprawie mieszańców ryżu może odgrywać potencjalną rolę w  poprawie efektywności wykorzystania azotu. słowa kluczowe: oryza sativa l. ‘bahar 1’, spryskiwanie dolistne, nawożenie mocznikiem, plon ziarna information on the authors hamid dorosti he obtained his phd from gorgan university of agricultural sciences and natural resources in 2015. before then, having master degree in plant breeding, he joined rrri (rasht rice research institute, guilan prov., iran). he had been involved in several projects performed at rrri, rasht, iran as scientist fellow for over 30 years. katarzyna możdżeń her scienti�c interests concentrate on the e�ects of di�erent environmental factors (light, ozone, heavy metals, allelopathic extracts) on the morphology and physiology of cultivated, protected, and invasive species of plants. peiman zandi he was deeply trained in agronomy (crop science) and specialising in stress physiology, biotic/abiotic stresses, and agroecology. he is also interested in working in di�erent areas of plant developmental bioloh am id d or os ti, k at ar zy na m oż dż eń , p ei m an z an di , m or te za s ia vo sh i 96 gy, agroecology, plant nutrition, botany, plant breeding, and genetics. his previous research project, which was funded by payame noor university (ganaveh, iran), was ‘foliar application of ascorbate on the physiological and biochemical attributes of iranian fenugreek (trigonella foenum-graecum l.) landraces under drought stress’. currently, he joined a research group at chinese academy of agricultural sciences (beijing, china) attempting to discern the possible function of iron plaque on chromium acquisition, accumulation and translocation in selected rice (oryza sativa l.) cultivars grown in di�erent planting media. �eir main aim is to verify ip role in wetland plants subjected to si fertilisation as an e�ective strategy to decrease cr accumulation in o. sativa grown in cr-contaminated environments. morteza siavoshi he is an assistant professor and faculty member of agricultural science department in payame noor university, iran. he was awarded his phd degree in agronomy in pune university, pune, india. now he is a college principal in ganaveh payame noor college, iran. 60 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 60–66, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.8 iwona skomorucha*, ewa sosnówka-czajka department of poultry breeding, national research institute of animal production, 32-083 balice n. kraków, poland, *iwona.skomorucha@izoo.krakow.pl the effect of mixed herb extracts on selected stress parameters in broiler chickens of three genetic lines introduction stress is present in every living animal organism as a result of stimulation from environmental factors. it is an integral part of life, vital to life, and the normal adaptation of the body (selye, 1936). stress most o�en occurs due to unfavourable living conditions or a persisting situation that causes it (stressor), where the problem cannot be solved. although stress is a response assisting the body in adapting to ambient conditions, it reduces welfare and thus productivity of animals, and o�en has a negative e�ect on the quality of animal products (puvadolpirod, �axton, 2000b). in states of stress, the sympathetic-adrenal-medullary, hypothalamo-pituitary-adrenal, and hypothalamo-pituitary-thyroid systems are stimulated. �e levels of catecholamines (post et al., 2003), corticosterone (puvadolpirod, �axton, 2000a,b) and the heterophil to lymphocyte ratio increase in blood (scanes, 2016). intensive livestock production contributes to stress, behavioural and physiological abnormalities, as well as many injuries and diseases of animals. �at is why, in many countries, notably the european union, e�orts are made to make extreme systems of livestock production more welfare friendly. currently, particular interest is given to medicinal plants and herbal preparations, which show broad and diverse biological and health properties. scienti�c literature reports many studies concerning the bene�cial e�ect of dried herbs as a biocomponent of concentrate feeds or as extracts and infusions on the gastrointestinal, endocrine, and immune systems (nasir, grashorn, 2010; wallace et al., 2010). furthermore, they also have bacteriostatic, antiviral, sedative, and oxidation-reduction action (wang et al., 2008; wallace et al., 2010; voljč et al., 2013). �e increased interest in medicinal plants and their potential use in animal husbandry is also due to the fact that they are relatively safe and require no withdrawal period. skomorucha and sosnówka-czajka (2013) and zhang et al. (2013) reported that some the effect of m ixed herb extracts on selected stress param eters in broiler chickens of three genetic lines 61 herbs and herbal extracts may reduce the body’s physiological response to stress associated with adverse housing conditions. �erefore, the aim of the study was to determine the e�ect of adding a mixture of anti-stress herbs to drinking water on the level of stress hormones (noradrenaline, adrenaline, corticosterone) and the heterophil to lymphocyte ratio in the blood of intensively reared broiler chickens of three genetic lines. material and methods �e experiments were performed at a poultry farm of the national research institute of animal production in aleksandrowice in poland with 360 broiler chickens of three genetic groups. on the �rst day of life, broiler chicks were weighed, tagged, and divided into group i – ‘ross 308’, group ii – ‘cobb 500’, and group iii – ‘hubbard flex’ broilers. each group had three subgroups. birds were kept for 42 days on litter at a stocking density less than 33 kg/m2. all groups had uniform environmental (temperature, air humidity, light regime) and feeding conditions. birds were fed ad libitum starter (me 3000 kcal, cp 21.0%), grower (me 3100 kcal, cp 19.8%), and �nisher diets (me 3100 kcal, cp 18.5%) based on concentrates at 1–3, 4–5 and 6 weeks of age, respectively. birds had free access to water drinkers throughout the study. in each group from 21 to 35 days of rearing, water drinkers were supplemented for 5 h/day (08:00–13:00 hours) with a tincture from mixed herbs at 2 ml/l water. �e herb mixture contained 30% chamomile (matricaria chamomilla l.), 10% oregano (origanum vulgare l.), 10% yarrow (achillea millefolium l.), 10% knotgrass (poligonum aviculare l.), 10% valerian (valeriana o�cinalis l.), and 20% in�orescence of large-leaved lime (tilia platyphyllos l.). �is mixture was selected based on the present authors’ earlier research. �e tincture from mixed herbs was made by a professional herbal company and has been certi�ed to conform to the company’s quality standards. at 21, 28, 35, and 42 days of age, blood was collected from 10 birds per group to determine the levels of corticosterone (by enzyme immunoassay), noradrenaline, and adrenaline (by radioimmunoassay). blood smears were made from the collected blood to count lymphocytes and heterophils and to calculate the heterophil to lymphocyte ratio (h:l). lymphocytes and heterophils were counted under a nikon ys 100 microscope a�er may-grünwald and giemsa staining. �e results were statistically analysed by an analysis of variance, and signi�cant di�erences were estimated with duncan’s test. �e statistical calculations were performed with statgraphics plus 6.0. e�ects were considered signi�cant at a probability of p ≤ 0.05 and p ≤ 0.01. iw on a s ko m or uc ha , e w a s os nó w ka -c za jk a 62 tab. 1. �e level of stress hormones in the blood of broiler chickens; values shown as di�erent letters within the line di�er signi�cantly according to the duncan test at p ≤ 0.05; n = 10 hormones [ng/ml] days of rearing excrement group ross 308 cobb 500 hubbard flex sem noradrenaline 21 121.56 132.16 146.68 27.94 28 64.54 70.58 118.34 18.83 35 87.94 101.49 122.68 16.12 42 99.93a 50.31b 98.78a 16.69 adrenaline 21 53.18 146.44 133.34 39.66 28 78.96 60.67 127.39 25.93 35 78.38 154.85 164.72 30.50 42 76.19 34.03 64.17 17.90 corticosterone 21 11.85 14.03 12.14 2.56 28 17.10 17.33 15.58 4.33 35 14.93 18.74 15.98 2.15 42 19.65 19.41 20.09 2.39 note: group i – ‘ross 308’, group ii – ‘cobb 500’, and group iii – ‘hubbard flex’ broilers; sem – standard error of the mean results on day 42 of the experiment, the noradrenaline concentration was lowest in the blood of ‘cobb 500’ compared to ‘ross 308’ and ‘hubbard flex’ broilers (p ≤ 0.05) (tab. 1). no statistically signi�cant di�erences in the adrenaline and corticosterone levels were found between the experimental groups. on day 21 of age, ‘cobb 500’ broiler chickens were characterised by the highest heterophil percentage and lowest lymphocyte percentage compared to the other genetic groups (p ≤ 0.01), which translated into a di�erence in the h:l ratio between these groups (p ≤ 0.01) (tab. 2). on day 42 of age, the di�erence in the percentage of heterophils and lymphocytes, and also in the h:l ratio became apparent between ‘ross 308’ and ‘cobb 500’ chickens (p ≤ 0.01). discussion a decisive role in the stress mechanism, or the body’s response to environmental stimuli, is played, among others, by the stress hormones corticosterone, adrenaline, and noradrenaline (puvadolpirod, �axton, 2000a; pohle, cheng, 2009). catecholamines (adrenaline and noradrenaline) are involved in numerous physiological and pathological processes, and their release into the bloodstream may increase blood pressure, heart beat, and muscle tension, in addition to increasing blood glucose levels by stimulating the conversion of hepatic glycogen to glucose (virden et al., 2007; kober et al., 2010). the effect of m ixed herb extracts on selected stress param eters in broiler chickens of three genetic lines 63 tab. 2. heterophil and lymphocyte levels (%) in the blood of broiler chickens; values shown as di�erent letters within the line di�er signi�cantly according to the duncan test at p ≤ 0.01; n = 10 item days of rearing group ross 308 cobb 500 hubbard flex sem heterophils 21 33.90a 47.30b 34.70a 2.23 lymphocytes 61.00a 46.30b 60.80a 1.85 h:l 0.57a 1.04b 0.58a 0.06 heterophils 28 29.20 31.80 34.80 2.61 lymphocytes 64.90 60.50 58.70 3.18 h:l 0.47 0.56 0.64 0.07 heterophils 35 41.90 37.70 35.30 3.99 lymphocytes 52.80 55.00 58.50 4.11 h:l 1.35 0.71 0.65 0.37 heterophils 42 37.00a 21.10b 27.90 3.17 lymphocytes 56.10a 71.10b 62.60 3.47 h:l 0.74a 0.31b 0.49 0.09 note: group i – ‘ross 308’, group ii – ‘cobb 500’, and group iii – ‘hubbard flex’ broilers; sem – standard error of the mean cheng et al. (2001) and pohle and cheng (2009) report that changes in adrenaline and noradrenaline blood levels may serve as an indicator of welfare and coping with stress. in our study, ‘cobb 500’ broiler chickens responded most favourably to drinking water with a mixed herb extract, which was re�ected in the lowest level of noradrenaline and a tendency for lower blood adrenaline compared to ‘ross 308’ and ‘hubbard flex’ chickens. �e bene�cial e�ect of adding mixed herbs to water on ‘cobb 500’ chickens was also re�ected in the lower h:l ratio, especially when compared with ‘ross 308’ chickens. likewise, naja� and torki (2010) found a signi�cant decrease in the amount of heterophils and an increase in the amount of lymphocytes in the blood of broiler chickens in response to a diet with herbs. in turn, yesilbag et al. (2012) showed increased heterophil percentage as well as increased h:l percentage ratio in the blood of birds fed a diet supplemented with an extract from oregano and rosemary essential oils compared to the control group. long-term stress results in the corticosterone stress hormone being released into the blood of birds (virden et al., 2007). a  high level of this hormone adversely a�ects the body and may induce cardiovascular diseases, hypercholesterolemia, damage to the digestive tract, changes in immune function, as well as metabolic changes (puvadolpirod, �axton, 2000c; virden et al., 2007). kannan et al. (1997) report that a lower blood corticosterone concentration is generally indicative of the body’s lower susceptibility to stressors. however, in our study, we did not observe any e�ect of supplementing the herb mixture on a lower level of this hormone in the blood in any of the chickens from the experimental groups. in summary, the herb extract had the most favourable e�ect on relieving the body’s physiological response to stress, and thus on improving the welfare of ‘cobb 500’ comiw on a s ko m or uc ha , e w a s os nó w ka -c za jk a 64 pared to ‘ross 308’ and ‘hubbard flex’ broilers. it can therefore be considered that active substances in herbs have varying e�ects on the body of broiler chickens of different origin. references cheng, h.w., dillworth, g., singleton, p., chen, y., muir, w.m. (2001). e�ects of group selection for productivity and longevity on blood concentrations of serotonin, catecholamines, and corticosterone of laying hens. poultry science, 80, 1278–1285. doi: 10.1093/ps/80.9.1278 kannan, g., heath, j.l., wabeck, c.j., souza, m.c.p., howe, j.c., mench, j.a. (1997). e�ects of crating and transport on stress and meat quality characteristics in broilers. poultry science, 76, 523–529. doi: 10.1093/ps/76.3.523 kober, a.k.m.h., aoyama, m., sugita, s. (2010). immunohistochemical localization of catecholamine biosynthetic enzymes in the adrenal gland of the domestic fowl (gallus domesticus). poultry science, 89, 1709–1715 . doi: 10.3382/ps.2009-00588 naja�, p., torki, m. (2010). performance, blood metabolites and immunocompetence of broiler chicks fed diets included essential oils of medicinal herbs. journal of animal and veterinary advances, 9(7), 1164–1168. doi: 10.3923/javaa.2010.1164.1168 nasir, z., grashorn, m.a. (2010). e�ects of intermittent application of di�erent echinacea purpurea juices on broiler performance and some blood parameters. archiv fur ge�ügelkunde, 74(1), 36–42. pohle, k., cheng, h.w. (2009). comparative e�ects of furnished and battery cages on egg production and physiological parameters in white leghorn hens. poultry science, 88, 2042–2051. doi: 10.3382/ ps.2009-00171 post, j., rebel, m.j., huurne, a.a.h.m. (2003). physiological e�ect of elevated plasma corticosterone concentrations in broiler chickens. an alternative means by which to assess the physiological e�ects of stress. poultry science, 82, 1313–1318. doi: 10.1093/ps/82.8.1313 puvadolpirod, s., �axton, j.p. (2000a). model of physiological stress in chickens. 1. response parameters. poultry science, 79, 363–369. doi: 10.1093/ps/79.3.363 puvadolpirod, s., �axton, j.p. (2000b). model of physiological stress in chickens.2. dosimetry of adrenocorticotropin. poultry science, 79, 370–376. doi: 10.1093/ps/79.3.370 puvadolpirod, s., �axton, j.p. (2000c). model of physiological stress in chickens. 4. digestion and metabolism. poultry science, 79, 383–390. doi: 10.1093/ps/79.3.383 scanes, c.g (2016). biology of stress in poultry with emphasis on glucocorticoids and the heterophil to lymphocyte ratio. poultry science, 95, 2208–2215. doi: 10.3382/ps/pew137 selye, h. (1936). a syndrome produced by diverse nocuous agents. nature, 138, 32. doi:10.1038/138032a0 skomorucha, i., sosnówka-czajka, e. (2013). e�ect of water supplementation with herbal extracts on broiler chicken welfare. annals of animal science, 13(4), 849–857, doi: 10.2478/aoas-2013-0057 virden, w.s., �axton, j.p., corzo, a., dozier, iii w.a., kidd, m. (2007). evaluation of models using corticosterone and adrenocoticotropin to induce conditions mimicking physiological stress in commercial broilers. poultry science, 86, 2485–2491. doi: 10.3382/ps.2006-00215 voljč, m., levart, a., žgur, s., salobir, j. (2013). �e e�ect of α-tocopherol, sweet chestnut wood extract and their combination on oxidative stress in vivo and the oxidative stability of meat in broiler. british of poultry science, 54(1), 144–156. doi: lo.l080/00071668.2012.760l90 wallace, r.j., oleszek, w., franz, c., hahn, i., baser, k.h.c., mathe, a., teichmann, k. (2010). dietary plant bioactives for poultry health and productivity. british of poultry science, 51(4), 461–187. doi: 10.1080/00071668.2010.506908 the effect of m ixed herb extracts on selected stress param eters in broiler chickens of three genetic lines 65 wang, l., piao, x.l., kim, s.w., piao, x.s., shen, y.b., lee, h.s. (2008). e�ects of forsythia suspensa extract on growth performance, nutrient digestibility, and antioxidant activities in broiler chickens under high ambient temperature. poultry science, 87, 1287–1294. doi: 10.3382/ps.2008-00023 yesilbag, d., gezen, s.s., biricik, h., bulbul, t. (2012). e�ect of rosemary and oregano volatile oil mixture on performance, lipid oxidation of meat and hematological parameters in pharaoh quails. british of poultry science, 53(1), 89–97. doi: 10.1080/00071668.2012.654763 zhang, h.y., piao, x.s., zhang, q., li, p., yi, q., liu, j.d., li, q.y., wang, g.q. (2013). �e e�ect of forsythia suspensa extract and berberine on growth performance, immunity, antioxidant activities, and intestinal microbiota in broilers under high stocking density. poultry science, 92, 1981–1988. doi: 10.3382/ps.2013-03081 abstract �e aim of the study was to determine the e�ect of adding a mixture of anti-stress herbs to drinking water on stress hormone levels and the heterophil to lymphocyte ratio in the blood of intensively reared broiler chickens of three genetic lines. �e experiment was performed with ‘ross 308’, ‘cobb 500’, and ‘hubbard flex’ broiler chickens, which were divided into three experimental groups. birds were maintained on litter for 42 days at a stocking density less than 33 kg/m2. all groups were provided with the same environmental and feeding conditions. in each group from 21 to 35 days of rearing, water drinkers were supplemented for 5 h/day (08:00–13:00 hours) with an alcoholic extract from mixed herbs (30% chamomile, 10% oregano, 10% yarrow, 10% knotgrass, 10% valerian, 20% in�orescence of large-leaved lime) at 2 ml/l water. at 21, 28, 35, and 42 days of rearing, blood was collected from 10 birds per group to determine corticosterone, noradrenaline, and adrenaline levels. �e heterophil to lymphocyte ratio (h:l) was also calculated. on day 42 of the study, 500 broiler chickens were characterised by the lowest noradrenaline concentration and a lower h:l ratio compared to ‘ross 308’ and ‘hubbard flex’ (p ≤ 0.05) and ‘ross 308’ chickens (p ≤ 0.01), respectively. it was concluded from the study that the herb extract had the most favourable e�ect on relieving the body’s physiological response to stress, and thus on improving welfare in ‘cobb 500’ compared to ‘ross 308’ and ‘hubbard flex’ broilers. it can therefore be considered that active substances in herbs have varying e�ects on the body of broiler chickens of di�erent origin. key words: adrenaline, broiler chickens, corticosterone, h:l, mixed herb extract, noradrenaline received: [2018.06.26] accepted: [2018.12.12] wpływ ekstraktu z mieszanki ziół na wybrane parametry stresu kurcząt brojlerów trzech linii genetycznych streszczenie celem badań było określenie wpływu dodatku do wody pitnej ekstraktu z mieszanki ziół wykazujących właściwości antystresowe na poziom hormonów stresu oraz stosunek hetero�lii do limfocytów we krwi kurcząt brojlerów trzech linii genetycznych utrzymywanych w systemie intensywnym. doświadczenie przeprowadzono na kurczętach brojlerach: ‘ross 308’, ‘cobb 500’ oraz ‘hubbard flex’, które przydzielono do trzech grup doświadczalnych. ptaki utrzymywano przez okres 42 dni, na ściółce o obsadzie nie przekraczającej 33 kg/m2. wszystkie grupy miały ujednolicone warunki środowiskowe oraz żywieniowe. w każdej grupie od 21 do 35 dnia odchowu przez 5 h/dobę (od 8.00–13.00) dodawano do poideł z wodą ekstrakt spirytusowy z mieszanki ziół (30% rumianku pospolitego, 10% ziela lebiodki pospolitej, 10% ziela krwawnika pospolitego, 10% ziela rdestu ptasiego, 10 % kozłka lekarskiego, 20% kwiatostanu lipy szerokolistnej) w ilości 2 ml/l wody. w 21, 28, 35 oraz 42 dniu odchowu pobrano krew od 10 ptaków z grupy i oznaczono poziom: kortykosteronu, noradrenaliny i adrenaliny a także obliczono stosunek hetero�lii do limfocytów (h:l). kurczęta brojlery ‘cobb 500’ charakteryzowały się w 42 dniu doświadczenia najniższym poziomem noradrenaliny oraz węższym stosunkiem h:l w porównaniu odpowiednio z kurczętami ‘ross 308’ i ‘hubbard flex’ przy iw on a s ko m or uc ha , e w a s os nó w ka -c za jk a 66 p ≤ 0,05 i kurczętami ‘ross 308’ przy p ≤ 0,01. na podstawie badań stwierdzono, że najkorzystniejszy wpływ podawanego ekstraktu z mieszanki ziół na łagodzenie �zjologicznej reakcji organizmu na stres, a tym samym na poprawę dobrostanu stwierdzono w przypadku kurcząt brojlerów ‘cobb 500’ w porównaniu z kurczętami ‘ross 308’ i ‘hubbard flex’. można uznać, że substancje czynne zawarte w ziołach mają zróżnicowany wpływ na organizm kurcząt brojlerów różnego pochodzenia. słowa kluczowe: adrenalina, ekstrakt z mieszanki ziół, h:l, kortykosteron, kurczęta brojlery, noradrenalina information on the authors iwona skomorucha https://orcid.org/0000-0003-1241-7703 she is an assistant professor at the department of poultry breeding in the national research institute of animal production in cracow. she focuses also on poultry welfare and the technology of poultry production. ewa sosnówka-czajka https://orcid.org/0000-0003-3720-1685 she is an assistant professor at the department of poultry breeding in the national research institute of animal production in cracow. she is interested mainly in poultry welfare and the technology of poultry production. 90 annales universitatis paedagogicae cracoviensis studia naturae, 3: 90–99, 2018, issn 2543-8832 doi: 10.24917/25438832.3.7 peiman zandi1, beata barabasz-krasny2, alina stachurska-swakoń3, joanna puła4, katarzyna możdżeń2* 1 institute of environment and sustainable development in agriculture, chinese academy of agricultural science, beijing 100081, p. r. china 2 institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland, *katarzyna.mozdzen@up.krakow.pl 3 institute of botany, jagiellonian university, gronostajowa 3, 30-387 kraków, poland 4 faculty of agriculture and economics, university of agriculture, mickiewicza 21, 31-120 kraków, poland allelopathic effects of stellaria media (l.) vill. on germination and early stages of growth of raphanus sativus var. radicula introduction chickweed (stellaria media (l.) vill., caryophyllaceae) is a cosmopolitan plant species known as a particularly troublesome weed in field crops and gardens. the plant may produce several generations of seeds throughout the year. the seeds could germinate at low temperatures and they are long-lived. the greatest damage by the species to crops is noticed at seedling stage of the crop plants due to the competition for soil nutrients and light. chickweed germinates and grows fast and causes the shading of crop seedlings (turkington et al., 1980). the high specialisation of chickweed as a crop weed results from the necessity to develop mechanisms that allow it to survive in unfavourable conditions. s. media produces a lot of chemical compounds, including the following: apigenin and its c-glycosides (bouillant et al., 1984; dong et al., 2007), vitexin (ma et al., 2012), isovitexin (budzianowski et al., 1991), luteolin glycosides, orientin, isoorientin (hu et al., 2009), isoscutellarein c-glycoside (yasukawa et al., 1982) and chrysoeriol c-glycoside (sharma, arora, 2012). there has been only one reference about the presence of isoflavonoids in the stellaria genus (kitanov, 1992) and a second concerning an isoflavonoids in another representative of the caryophyllaceae family (liu et al., 2007). the presence of o-glycosides of apigenin, luteolin, and other aglycones, and glycosides was also documented (mikšátková et al., 2014). radish (raphanus sativus l. var. radicula pers., brassicaceae) is an edible root vegetable well-known and popular throughout the world. probably, it originates from southeast asia, and it was known for greek and roman agriculturalists of the first 91 century ad. it has numerous cultivars, varying in size, flavour, colour, and length of time they take to mature. radishes owe their sharp flavour to the various chemical compounds produced by the plants, including glucosinolate, myrosinase, and isothiocyanate (kunachowicz et al., 2017). the aim of the experiment was to investigate the allelopathic effect of aqueous extracts from aboveground organs of stellaria media on germination, the early stages of growth and electrolyte leakage through cell membranes of radish (raphanus sativus var. radicula) that are commonly cultivated in poland three cultivars: ‘rowa’, ‘krakowianka’, and ‘półdługa’. material and methods fresh aboveground parts of chickweed (stellaria media (l.) vill.) were collected in nature from the the southern part of poland (suchoraba 49°58ʹ37ʹʹn  20°11ʹ49ʹʹe). the seeds of radish were purchased in the ordinary market from polan co. three common cultivars of radish were used in the experiment: ‘rowa’, ‘krakowianka’, and ‘półdługa’. the aqueous extracts from fresh aboveground organs of s. media in the form of decoction, infusion, and macerate were prepared according to czerwińska et al. (2015). the seeds of radish cultivars were rinsed under running and distilled water and placed on sterile 9 cm petri dishes (100 seeds per dish, in 5 repetitions with 2 series). they were placed in a growth chamber at constant temperature conditions of 20°c, a relative humidity of about 90%, with 12-hour periods of light intensity for 8 days. during the experiment, seeds were watered with adequate aqueous extract from s. media every day and the control group was watered with distilled water. the same content of extract or water was always used. the experiment was repeated five times with two series. the germination rate of radish seeds, seedling lengths, the fresh and dry mass of seedlings, and the degree of destabilisation of cell membranes were determined. the number of germinated seeds was counted every 24 h. based on the results, the germination rate was calculated: the number of germinated seeds/total seeds number for the germination experiment × 100%. the average speed of germination was measured according to chapisuio et al. (1997). the seedling length was measured at the end of the experiment as mean of lengths of whole seedlings (cm) and as the length of underground and aboveground parts. after eight days, the fresh and dry mass of seedlings was determined. the degree of the destabilisation of cell membranes was determined using the method described as in skrzypek et al. (2015), barabasz-krasny et al. (2017; 2018). a llelopathic effects of stellaria m edia (l.) v ill. on germ ination and early stages of grow th of r aphanus sativus var. radicula p ei m an z an di , b ea ta b ar ab as zk ra sn y, a lin a s ta ch ur sk as w ak oń , j oa nn a p uł a, k at ar zy na m oż dż eń * 92 the significance of differences were examined with inter-facility parametric statistical test – anova (test one, simple), using duncan’s test (hsd) for homogeneous groups, at the level of p < 0.05. the calculations were performed using statistica 13.0 from statsoft, inc. (2018) for windows. results and discussion the common opinion that plants considered to be weeds are harmful, unwanted, and should be removed from the crops as soon as possible is not entirely true. wild plants create perfect conditions for the life of many insects and other organisms. biodiversity is conducive to maintaining the proper balance in the environment and prevents pest gradation. some weed species are indicators of the soil type and soil conditions and can serve as soil cover against erosion during winter. weeds could produce different chemical compounds that could stimulate or inhibit the development or growth of various organisms (skrzypek et al., 2015) and could be also used in organic farming. in the light of the mentioned significance of the weeds, studies on the biology of crop weeds and their interaction with crop species are desirable and important. tab. 1. cumulative percentage of seed germination of raphanus sativus l. var. radicula pers. cultivars: ‘rowa’, ‘krakowianka’, and ‘półdługa’ under influence of stellaria media (l.) vill. aqueous extracts treatment time of germination [day] 1 2 3 4 5 6 7 8 ‘rowa’ control 15.80 26.00 74.40 95.60 99.60 99.60 99.60 99.60 decoction 7.40* 15.00 30.60* 48.40* 55.00* 57.80* 59.60* 62.60 infusion 27.80 54.00* 70.20 81.80 86.40 90.40 90.80 91.40 macerate 8.00* 30.20 33.00* 37.60* 41.00* 42.40* 45.00* 45.40* ‘krakowianka’ control 79.80 90.00 97.00 98.00 98.40 98.60 99.80 100.00 decoction 86.60 95.00 98.20 99.00 99.80 99.80 99.80 99.80 infusion 72.20 94.00 95.00 95.40 95.40 95.40 95.40 95.40 macerate 69.80 71.60 73.80 76.40 78.80 79.60 79.60 79.60 ‘półdługa’ control 24.80 40.40 45.60 66.80 95.80 99.60 99.80 100.00 decoction 10.80* 35.20 44.60 55.40 58.60* 66.40 70.00 75.25 infusion 12.40* 37.40 44.00 45.80* 54.00* 60.60 63.60* 66.20* macerate 23.00 52.20 59.20* 63.80 65.80 68.80 71.20 71.20 values * differ significantly according to duncan’s test p < 0.05 in the presented experiment, the germination rate of radish seeds watered with extracts from s. media was lower than the control group, independently of the type 93 of used extracts. the highest germination rate and capacity for ‘krakowianka’ cultivar on decoction, and the lowest for ‘rowa’ on macerate were observed (tab. 1–2). the inhibition of germination could be attributed to the biochemical effects caused by the extracts. for example, jezierska-domaradzka and kuźniewski (2007), in biotests, showed that 2.5% of s. media extracts negatively affected origanum majorana l. and ocimum basilicum l. the growth of ‘rowa’ and ‘krakowianka’ seedlings (measured as length) was stimulated by infusion and slightly inhibited by decoction and macerate. the growth of ‘półdługa’ seedlings was inhibited by each of the extracts, relative to the control group (fig. 1). the lack of negative effects of the extracts on seeds germination was most likely caused by the low concentration of allelopathic substances (duer, 1996). fig. 1. the length of seedlings (whole, underground, aboveground parts) raphanus sativus l. var. radicula pers. cultivars: ‘rowa’, ‘krakowianka’, and ‘półdługa’ under influence of stellaria media (l.) vill. aqueous extracts. values a, b, c differ significantly according to duncan’s test p < 0.05 a llelopathic effects of stellaria m edia (l.) v ill. on germ ination and early stages of grow th of r aphanus sativus var. radicula p ei m an z an di , b ea ta b ar ab as zk ra sn y, a lin a s ta ch ur sk as w ak oń , j oa nn a p uł a, k at ar zy na m oż dż eń * 94 tab. 2. average speed of germination raphanus sativus l. var. radicula pers. cultivars: ‘rowa’, ‘krakowianka’, and ‘półdługa’ under influence of stellaria media (l.) vill. aqueous extracts treatment time of germination [day] 1 2 3 4 5 6 7 8 ‘rowa’ control 37.40 35.98 68.24 80.33 79.68 74.70 74.70 66.40 decoction 20.70 19.76* 30.80* 41.92* 44.61* 43.50* 44.95* 41.73* infusion 45.20 52.98 63.34 69.32 69.70 67.83 68.15 60.93 macerate 18.58* 28.23 29.83* 32.25* 33.14* 32.02* 33.78* 30.27* ‘krakowianka’ control 85.51 80.45 83.47 81.86 78.82 74.05 74.87 66.67 decoction 90.56 83.99 84.41 82.63 79.84 74.85 74.85 66.53 infusion 79.82 82.51 81.50 79.50 76.32 71.55 71.55 63.60 macerate 72.21 63.79 64.10 64.13 63.15 59.70 59.70 53.07 ‘półdługa’ control 43.09 43.01 46.65 60.84 77.17 74.72 74.87 66.67 decoction 27.09* 35.34* 41.80 47.83 48.36* 50.10 52.94 50.17 infusion 26.33* 36.02 39.88* 40.42* 44.36* 45.70* 47.92* 44.13* macerate 36.88 48.28 52.29 53.95 53.16 51.80 53.40 47.47* values * differ significantly according to duncan’s test p < 0.05 the fresh mass of radish seedlings was lower in all used type of extract, compared to the control group. the exception was the ‘rowa’ cultivar, for which the fresh mass increased on macerate. there were no statistical differences between the dry mass of ‘krakowianka’ cultivar watered with extracts and distilled water. however, the dry mass of ‘rowa’ and ‘półdługa’ was lower than in the control group (tab. 3). the products of the breakdown of glucosinolates can inhibit and delay seed germination by inhibiting seeds and protein synthesis in the seedlings (leblová-svobodová, kostír, 1962). the lower effects on the germination rate and dry mass may be attributed to the low persistence of glucosinolate hydrolysis products in the soil (haramoto, gallandt, 2004). compared to the control, the s. media extracts decreased the electrolyte leakage through the cell membranes of all radish seedlings. the exception was the macerate, which, in the ‘rowa’ cultivar, caused the increase of the destabilisation of cell membranes (tab. 3). lower membrane damage can be correlated with an increased capacity to accumulate sugars at the leaf level during stress (bajji et al., 2000). in fact, it was hypothesized that sugars, particularly non-reducing disaccharides, interact with cellular membranes to increase the stability of the lipid layers (nilsen, orcutt, 1996). this may prevent lateral phase transition and the formation of lipid domains which have the potential of forming inverted micelles and thus increasing membrane leakage (bajji et al., 2000). 95 the different effect of s. media extracts on radish seeds may result from the various concentrations of chemical compounds contained in them (mikšátková et al., 2014; skrzypek et al., 2015; barabasz-krasny et al., 2017). according to inderjit and dakshini (1998), both young and adult chickweed species release soil-soluble phenols that inhibit plant growth. in addition, the size of tested seeds is important, because small seeds, such as radish, are more sensitive to the effects of allelopathic substances than larger seeds (riece, 1984). tab. 3. fresh and dry mass and electrolyte leakage of raphanus sativus l. var. radicula pers. cultivars: ‘rowa’, ‘krakowianka’, and ‘półdługa’ under influence of stellaria media (l.) vill. aqueous extracts treatment name of cultivars ‘rowa’ ‘krakowianka’ ‘półdługa’ fresh mass [g] control 0.0832 0.1244 0.0872 decoction 0.0600 0.1104 0.0702 infusion 0.0766 0.1036 0.0690* macerate 0.1134* 0.1138 0.0866 dry mass [g] control 0.0068 0.0154 0.0090 decoction 0.0048 0.0114 0.0062 infusion 0.0066 0.0114 0.0054* macerate 0.0116* 0.0114 0.0068 electrolyte leakage [%] control 40.94 49.33 37.89 decoction 36.13 43.86 31.67 infusion 38.79 33.89* 31.31* macerate 44.46 34.05* 29.46* values * differ significantly according to duncan’s test p < 0.05 the use of an alternate crop is a basic, very effective, and well-known method that limits the occurrence of weeds in crop fields. a few-year-old cultivation of the same species or plants from the same group causes that the weeds accompanying these crops intensively increase their number and cover, increase the soil seed bank, and thus increase the cover in the field next year. on the other hand, cultivation of other species the following year could change the soil conditions; and as a consequence, the weeds from previous year could not have favourable conditions for germination and further development, or they die soon after germination (sturm et al., 2016). it is also important to introduce new cultivars (or varieties) and check their resistance to allelopathic weeds. a llelopathic effects of stellaria m edia (l.) v ill. on germ ination and early stages of grow th of r aphanus sativus var. radicula p ei m an z an di , b ea ta b ar ab as zk ra sn y, a lin a s ta ch ur sk as w ak oń , j oa nn a p uł a, k at ar zy na m oż dż eń * 96 conclusion the aqueous extracts from fresh aboveground organs of stellaria media (l.) vill. have allelopathic potential. they can stimulate or inhibit germination and early stages of radish growth, depending on the form of extract and, as we suppose, the type and concentration of chemical compounds in extracts. the obtained results provide the basis for further studies on the allelopathic influence of chickweed on radish cultivars or other vegetables in further stages of their development. references 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(1982). studies on the constituents in the water extracts of crude drugs. 4: on the roots of stellaria dichotoma l. var. lanceolata bge. yakugaku zasshi, 102, 292–294. abstract the aim of the study was to investigate the allelopathic influence of stellaria media (l.) vill. on the germination and early stages of growth of commonly grown radish (raphanus sativus l. var. radicula pers.) in three cultivars: ‘rowa’, ‘krakowianka’, and ‘półdługa’. the aqueous extracts from fresh shoots of s. media in the form of: decoction, infusion, and macerate were prepared. the control group was radish seeds watered with distilled water. the germination capacity of radish seeds, seedlings growth, the fresh and dry mass of seedlings, and the degree of destabilisation of cell membranes were determined. this experiment showed that the germination of radish seeds was lower, independently of the type of s. media extracts used, compared to the control group. the lowest germination capacity was observed for the ‘rowa’ cultivar on macerate, and the highest germination capacity was for ‘krakowianka’ on decoction. on each of the extracts, the growth of ‘półdługa’ seedlings was inhibited in relation to the control. in the other two cultivars, the growth of radish seedlings was stimulated by the infusion, and it was slightly inhibited by the decoction and macerate. the fresh mass of radish seedlings was lower on each extracts, compared to the control group. the exception a llelopathic effects of stellaria m edia (l.) v ill. on germ ination and early stages of grow th of r aphanus sativus var. radicula p ei m an z an di , b ea ta b ar ab as zk ra sn y, a lin a s ta ch ur sk as w ak oń , j oa nn a p uł a, k at ar zy na m oż dż eń * 98 was the ‘rowa’ cultivar, for which the fresh mass of seedlings increased on macerate. the dry mass values for ‘rowa’ and ‘półdługa’ cultivars were lower than in the control, and, no significant differences were found for ‘krakowianka’. compared to the control group, the applied extracts reduced the outflow of electrolytes through the cell membranes of radish seedlings of all the analysed cultivars. the exception was macerate, which caused an increase in the electrolyte leakage in the ‘rowa’ cultivar. key words: chickweed, electrolyte leakage, germination rate, radish, seedling growth, weeds received: [2018.07.02] accepted: [2018.11.15] allelopatyczny wpływ stellaria media (l.) vill. na kiełkowanie i wczesne etapy wzrostu raphanus sativus var. radicula streszczenie w  pracy podjęto próbę zbadania allelopatycznego oddziaływania stellaria media (l.) vill. na kiełkowanie i wczesne etapy wzrostu powszechnie uprawianej rzodkiewki (raphanus sativus l. var. radicula pers.) w trzech odmianach: ‘rowa’, ‘krakowianka’ i ‘półdługa’. sporządzono wodne ekstrakty ze świeżych pędów s. media w postaci: wywaru, naparu i maceratu. grupę kontrolną stanowiły nasiona rzodkiewki podlewane wodą destylowaną. w doświadczeniu określono zdolność i szybkość kiełkowania nasion rzodkiewki, wzrost siewek, przyrost ich mas, stopień destabilizacji błon komórkowych. eksperyment ten pokazał, że w porównaniu z kontrolą nasiona rzodkiewki kiełkują słabiej niezależnie od rodzaju zastosowanego wyciągu ze s. media. najmniejszą szybkość kiełkowania zaobserwowano dla odmiany ‘rowa’ na szalkach z maceratem, a największą dla odmiany ‘krakowianka’ na wywarze. hamowanie wzrostu siewek stwierdzono u odmiany ‘półdługa’ dla każdego z ekstraktów, w stosunku do próby kontrolnej. u pozostałych dwóch odmian napar stymulował wzrost siewek rzodkiewki, a  wywar i  macerat nieznacznie ograniczały przyrost na długość. świeża masa siewek badanych odmian rzodkiewki była mniejsza dla każdego rodzaju wyciągu, w odniesieniu do kontroli. odstępstwo od tego stanowi odmiana ‘rowa’, dla której odnotowano przyrost świeżej masy siewek podlewanych maceratem. wartości suchej masy dla odmian ‘rowa’ i ‘półdługa’ były mniejsze niż w kontroli, a dla ‘krakowianka’ nie wykazano istotnych różnic. w porównaniu z kontrolą, zastosowane wyciągi zmniejszały wypływ elektrolitów przez błony komórkowe siewek rzodkiewki wszystkich analizowanych odmian. wyjątkiem okazał się macerat, który u odmiany ‘rowa’ powodował wzrost destabilizacji błon komórkowych. słowa kluczowe: gwiazdnica pospolita, wypływ elektrolitów, kiełkowanie, rzodkiewka, wzrost nasion, chwasty information about authors peiman zandi https://orcid.org/0000-0003-3520-3994 he was intensely trained in agronomy (crop science) and specialising in stress physiology, biotic/abiotic stresses, and agroecology. he is also interested in working in different areas of plant developmental biology, agroecology, plant nutrition, botany, plant breeding, and genetics. his previous research project which was funded by payame noor university (ganaveh, iran) entitled ‘foliar application of ascorbate on the physiological and biochemical attributes of iranian fenugreek (trigonella foenum-graecum l.) landraces under drought stress’. currently, he joined a research group at the chinese academy of agricultural sciences (beijing, china) attempting to discern the possible function of iron plaque on chromium acquisition, accumulation, and translocation in selected rice (oryza sativa l.) cultivars grown in different planting media. their main aim is to verify the ip role in wetland plants subjected to si fertilisation as an effective strategy to decrease cr accumulation in o. sativa grown in cr-contaminated environments. beata barabasz-krasny https://orcid.org/0000-0002-5800-6953 her main scientific interests include floristics and phytosociology of non-forest plant communities with particular emphasis on the course of succession processes in the areas where agricultural activities were 99 discontinued, the transformation of the plant cover of thermophilic swards, and the active protection of non-forest plant communities. alina stachurska-swakoń https://orcid.org/0000-0003-0381-4520 she is interested in the vegetation ecology, particularly in the mechanisms of changes in plant communities and flora under natural and anthropogenic influences. joanna puła her research is connected with agrotechnology in plant cultivation and plant ecology. presently, she is interest to use of biomass of plants and other organic fertilisers, like biochar, in agriculture. katarzyna możdżeń https://orcid.org/0000-0002-5695-4474 her scientific interests concentrate on the effects of different environmental factors (light, ozone, heavy metals, and allelopathic extracts) on the morphology and physiology of cultivated, protected, and invasive species of plants. a llelopathic effects of stellaria m edia (l.) v ill. on germ ination and early stages of grow th of r aphanus sativus var. radicula 24 annales universitatis paedagogicae cracoviensis studia naturae, 1: 24–41, 2016, issn 2543-8832 pavel širka1*, ingrid turisová1, anna petrášová2 1 department of biology and ecology, faculty of natural sciences, matej bel university, tajovského 40, banská bystrica 974 01, slovakia, *pavel.sirka@umb.sk 2 clementisa 214, 981 01 hnúšťa, slovakia bryophytes of cu-mine heaps in the vicinity of banská bystrica (central slovakia) introduction mine heaps and wastes created by mining industry are one of the extreme habitats made by human activity. for plants mine heaps are distinctive habitats with speci�c ecological conditions: the lack of soil as well as nutrients and moisture. mine heaps are characterized by a skeletal substrate from 52.90%–87.38% and a lack or complete absence of humus layer (banásová, 1976). in addition, compared to natural soils, mine heaps contain a higher content of heavy metals. all these conditions signi�cantly limit the list of plant species and vegetation types that can grow on such habitats (baker et al., 1988; banásová, hajdúk, 2006). in slovakia, a land with rich mining history, there are numerous old mine heaps – remainders of exploitation of di�erent ores. on mine heaps, we encounter plant succession which takes place in strange and complex conditions consequently forming new, unknown or very little known plant communities (banásová, 1976). because mine heaps originate from di�erent space of time, the vegetation formed on them is present in di�erent successional stages (banásová, 1983). banásová (1976) stated that plants on mine heaps mostly colonize depressions or parts with more disintegrated material that are more humid and where dead plant material accumulates, forming raw humus. �us a mosaic vegetation cover is formed. on mine heaps perennial plants predominate, while annual and biannual plants occur sparsely. on the rock surface and among grass and herbs grow tolerant species of bryophytes and lichens. mine heaps with anomalous metal content are like ecological islands because in comparison to their surroundings they have a very speci�c vegetation. �roughout many decades some plants (immigrants) that had no ability to adapt were excluded by natural evolution. a result of this evolution is b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 25 fig. 2. location of ľubietová, staré hory and špania dolina within slovakia (source: by courtesy of turisová et al., 2014) fig. 1. initial succesional stage with bryophytes (photo. i. turisová) p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 26 a small group of plants with strange species combination capable of existing in these phytotoxical conditions. bryophytes represent a very signi�cant group of organisms that sensibly react and indicate changes in the natural environment, especially changes of anthropogenic character (kubinská et al., 2001). in recent decades they have been successfully used as biomonitors of heavy metal accumulation throughout europe (harmens et al., 2013). bryophytes play an important role in primary successional stages when colonizing anthropogenic substrate (fig. 1), where they signi�cantly participate in soil cover formation. some bryophyte communities may be particularly important as a successive stage to some other vegetation types. bryophytes are o�en omitted in botanical studies, especially those of mine heaps. �e aim of this article is to present basic information on bryophyte species composition in the chosen metal-contaminated areas in central slovakia. materials and methods study area �ree mine heaps near the city of banská bystrica in central slovakia were studied: podlipa (cadaster ľubietová), richtárová (cadaster staré hory) and maximilián in the village of špania dolina (fig. 2). mine heap ľubietová – podlipa (fig. 3) is situated in the north-eastern part of slovenské stredohorie mts which include the northern part of neovolcanic massif poľana and the northern part of veporské vrchy (mazúr, lukniš, 2002). �e mine heap lies approximately 1 km to the east of the center of the village on the southern slope of vysoká (995 m a.s.l.). �e podlipa deposit was dug by 18 tunnels ranging from 570–700 m a.s.l. (bergfest, 1951). in the dump material the content of copper was stated to be from 0.9–2.4 mass percent. �e mine heap represents an area visibly changed by the historic exploitation of copper ore especially from 15th and 16th century until the end of the 19th century (koděra, 1990). small mining activities persisted until the beginning of the 20th century when the last mining works were shut down in april 1915 during the first world war (andráš, 2009). �e whole body of the mine heap is surrounded by forest and is drained in a thalweg of zelená valley by a brooklet which draws on the water from the hillsides and also water percolating in the mine heap sediment into a  shallow depression. from here the water is led to a  �ood pool on the base of the mine heap through a system of wooden and tinny channels (andráš et al., 2007). on this mine heap the substrate mostly has a high proportion of rock and vegetation cover is poorly developed (only on about 10% of the dump-�eld area). in a mosaic of plant communities, especially on less steep slopes with shallow soil, dominate species-poor grassland islets with agrostis capillaris and acetosella vulgaris or only islets b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 27 of bryophytes with lichens in steep areas and places with a thin soil layer. flat areas are colonized by groups of pioneer tree species dominated by pinus sylvestris, sporadically by quercus petraea and picea abies. černý (2015) recorded here a total of 74 taxa of vascular plants. mining region špania dolina – staré hory is geographically divided into two parts: northern (staré hory) and southern (špania dolina). �ey are named a�er two most distinguished sites where copper and silver ore mining was carried out throughout several centuries. �e border between the northern and southern part of the mining area is formed by a  narrow ridge dividing the valley of richtársky potok from veľká zelená valley, with zelený potok crossing through it (mazúrek, 1989). area of špania dolina ore �eld represents one of the historically most distinguished copper mining deposits in europe. �e mineralization forms a  4 km long and 1.5 km wide vein stretching between panský diel (1100 m a.s.l.) on the south and staré hory on the north to the east of starohorský potok (michňová, ozdín, 2010). �e �rst written reports of ore mining in the area of staré hory and špania dolina are from the 11th century (from the year 1006), although on the deposit piesky copper is proven to be mined already in the late stone age (točík, bublová, 1985). at �rst, copper and silver ore was mined in staré hory (historical deposit haliar). later, the mining expanded further south in richtárová and špania dolina (koděra et al., 1990). a�er depleting the surface portions the mining was gradually transferred to underground mining from the mid-14th century. maximum development of copper ore mining with an extremely valuable silver content was in the years 1496–1546. since the 17th century, mining gradually declined until it completely disappeared in the early 20th century (jeleň et al., 2009). during the years 1963–1964, old mine heaps (originating from the 16th – 19th century) in the area of village richtárová were re-mined by surface mining, which signi�cantly changed the original con�guration (mazúrek, 1989). mine heaps in staré hory – richtárová part (fig. 4) �ll the richtársky potok valley lenghtwise and they are mine heaps in a natural valley, unlike the mine heap in špania dolina which represents a sloping mine heap (dobríková, 2011). turisová et al. (2014) recorded a total of 147 taxa of vascular plants and 13 taxa of lichens from the mine heap richtárová and 83 taxa from maximilián. �e representation of higher plants in waste dump-�eld richtárová is relatively low and they cover about 30% of the area. vegetation is concentrated mainly in �eld depressions, on even surfaces or in the areas with �ne substrate. vegetation is incoherent and forms a mosaic pattern. on considerably exposed parts of the heap grow only lichens and mosses. dominant plant species on this mine heap are agrostis capillaris (sometimes forming a relatively continuous cover), silene dioica, acetosella vulgaris and arabidopsis arenosa. dominant trees are picea abies and pinus sylvestris (štrba et al., 2014). similarly, on the mine heap maximilián (fig. 5) vegetation is present individually and in islets and a total vegetation p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 28 fig. 3. mine heap podlipa (photo. p. širka) fig. 4. mine heap richtárová (photo. i. turisová) cover reaches 30%–40%. on a relatively �at top part of the mine heap a forest habitat is established in which the highest plant diversity was recorded. on surfaces without humus and �ne earth grow only lichens – the most dominant group of organisms on the mine heap, represented mostly by genera rhizocarpon, cladonia, cetraria, lecanora and peltigera. b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 29 �e most wide-spread grass is also agrostis capillaris which o�en forms monocultural overgrowth among lichens. among herbs the most frequent species are arabidopsis arenosa, acetosella vulgaris and silene dioica. pioneer tree species such as betula pendula, salix caprea, picea abies, pinus sylvestris and abies alba are present sporadically. �ey o�en have a  dwar�sh appearance, deformed shape or weakened vitality (aschenbrenner et al., 2011). methods �e �eld survey of chosen mine heaps was conducted in november and december of 2013, using the method of the zürich-montpellier school (braun-blanquet, 1964) adjusted to bryophytes. a total of 54 relevés were made. a standard surface size of 1 m2 was chosen for all relevés. �e sites were chosen randomly. bryophytes were determined according to pilous, duda (1960) and smith (2004). nomenclature follows hill et al. (2006) and ros et al. (2007). species cover-abundance was recorded using the extended ninegrade braun-blanquet scale (westho�, van der maarel, 1973): r – 1–2 individuals with insigni�cant cover-abundance; + – cover-abundance not higher than 1%; 1 – 1%–5% cover-abundance; 2 m – cover-abundance about 5%; 2a – 5%–15% cover-abundance; 2b – 15%–25% cover-abundance; 3 – 25%–50% cover-abundance; 4   – 50%–75% cover-abundance; 5 – 75%–100% cover-abundance. �e list of all relevés is presented in appendix 1, the information contains respectively: relevé number, short habitat description, geographical coordinates, altitude, slope, terrain exposure (using a gps device garmin – oregon 600), tree layer cover (e3), shrub layer cover (e2), herb layer fig. 5. mine heap maximilián (photo. i. turisová) p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 30 cover (e1), moss layer cover (e0), total number of recorded bryophyte species and date of collection. results and discussion a total of 45 bryophytes species were identi�ed in 54 relevés in the invesigeted areas (tab. 1). tab. 1. �e list of recorded bryophyte species; p – mine heap podlipa, r – mine heap richtárová, m – mine heap maximilián; number of relevés and cover (shown in brackets) name of species localities abietinella abietina (hedw.) m. fleisch. p: 3(1), 4(2a), 5(1), 8(1); r: 28(2a) amblystegium serpens (hedw.) schimp. p: 4(1); r: 24(1), 25(1), 3w1(1) aulacomnium palustre (hedw.) schwägr. p: 20(1) barbilophozia barbata (schmidel ex schreb.) loeske p: 5(1) barbula unguiculata hedw. r: 22(2m), 26(2a), 30(1) brachytheciastrum velutinum (hedw.) ignatov & huttunen r: 30(2a); m: 39(1), 47(1) brachythecium albicans (hedw.) schimp. p: 2(2a), 4(1), 8(1), 18(2m); r: 34(r), 36(1); m: 53(+) b. rivulare schimp. r: 23(2a) b. rutabulum (hedw.) schimp. p: 11(1), 14(2a), 15(2b) b. salebrosum (ho�m. ex f. weber & d. mohr) schimp., nom. cons. p: 2(1), 11(2a); r: 24(2a), 25(2m), 26(1), 29(1), 30(1); m: 43(2a), 51(1) bryum caespiticium hedw. p: 3(2m), 6(1); r: 25(1), 26(1); m: 49(1) b. capillare hedw. p: 20(2a); r: 21(2b), 23(2b), 30(2a) b. moravicum podp. p: 4(+), 18(1); r: 32(1); m: 39(1), 47(1) calliergonella cuspidata (hedw.) loeske p: 20(+) campylopus intro�exus (hedw.) brid. m: 48(3), 49(1), 50(2a) ceratodon purpureus (hedw.) brid. p: 4(r), 6(1), 12(2a); r: 24(1), 26(2a), 27(2a), 28(2a), 29(2a), 31(2a), 33(2a), 34(2m), 36(1); m: 38(2a), 39(2a), 40(+), 41(1), 43(3), 45(1), 47(2a), 48(1), 49(1), 51(2m), 52(2a), 54(2a) cirriphyllum piliferum (hedw.) grout r: 35(1) climacium dendroides (hedw.) f. weber & d. mohr p: 2(1), 5(2m), 7(1), 8(2a); r: 28(1) dicranella heteromalla (hedw.) schimp. p: 20(1) dicranum montanum hedw. p: 13(1); m: 38(+) d. scoparium hedw. p: 6(2m), 7(1), 9(1); r: 30(3), 31(2m); m: 40(2a), 50(2a), 51(1), 52(2a), 53(2a), 54(2a) didymodon fallax (hedw.) r.h. zander r: 32(1) drepanocladus aduncus (hedw.) warnst. r: 24(2a) grimmia pulvinata (hedw.) sm. m: 37(+) hylocomium splendens (hedw.) schimp. p: 6(1), 7(2m), 17(1); r: 30(1), 31(1), 34(2m), 36(1); m: 53(2m) b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 31 hypnum cupressiforme hedw. p: 4(2m), 5(1), 6(2a), 9(1); m: 39(2a), 40(2a), 44(3), 47(+), 51(2m), 53(3), 54(2a) lophocolea bidentata (l.) dumort. p: 11(1) oxyrrhynchium hians (hedw.) loeske r: 29(1) plagiomnium a�ne (blandow ex funck) t.j. kop. p: 7(2a), 11(1), 19(r); r: 30(1), 33(1), 35(2a); m: 53(2a) p. cuspidatum (hedw.) t.j. kop. p: 4(1), 5(1), 6(r), 8(2b), 14(r) pleurozium schreberi (willd. ex brid.) mitt. p: 2(2a), 4(2a), 6(2a), 7(1), 9(1), 17(2a), 18(2m); r: 30(2m), 31(1), 34(2a), 36(2m) pohlia cruda (hedw.) lindb. p: 9(1), 10(2b), 13(1), 19(2a); r: 21(1), 23(1), 24(1), 25(1), 28(2m), 30(2a), 31(2m), 36(2a); m: 39(1), 41(2a), 42(1), 46(2b), 49(1) p. drummondii (müll. hal.) a.l. andrews p: 1(3), 2(1), 16(2a), 20(1) polytrichastrum formosum (hedw.) g.l. sm. p: 2(2m), 9(1), 17(1); r: 28(2a), 31(1), 36(2m); m: 51(1), 53(2m), 54(2a) pterigynandrum �liforme hedw. m: 40(1) racomitrium canescens (hedw.) brid. p: 8(2m), 19(1); r: 24(1), 25(1), 26(1), 27(1); m: 38(2a), 41(1), 43(1) r. lanuginosum (hedw.) brid. p: 5(1); m: 39(1) rhytidiadelphus squarrosus (hedw.) warnst. p: 5(2a), 17(2a), 20(1); r: 30(1), 34(2a), 35(r), 36(2m) r. triquetrus (hedw.) warnst. r: 34(2a) sciuro-hypnum populeum (hedw.) ignatov & huttunen r: 33(1) s. re�exum (starke) ignatov & huttunen p: 5(2a) schistidium apocarpum (hedw.) bruch & schimp. r: 26(1); m: 51(1) syntrichia ruralis (hedw.) f. weber & d. mohr p: 5(2a), 8(1); r: 27(1), 28(+) �uidium delicatulum (hedw.) schimp. p: 6(1), 8(+); r: 26(1) tortella tortuosa (hedw.) limpr. m: 38(1), 39(1), 45(3), 47(1) on the mine heap ľubietová – podlipa a total of 31 bryophyte species were recorded (29 mosses and 2 liverworts). �is area is the species-richest among studied. �e highest number of species (10) was noticed in one relevé (no. 5), while only one species was recorded in 5 relevés. �e highest frequency on this mine heap had species pleurozium schreberi (present in 7 relevés) and plagiomnium cuspidatum (5). ceratodon purpureus as a characteristic species for this type of habitat was present in 3 relevés. species with the highest cover (25%) was pohlia drummondii in relevé no. 1, followed by pohlia cruda (20% – relevé no. 10) and brachythecium rutabulum (20% – relevé no. 15). on the mine heap staré hory – richtárová a  total of 29 species of mosses were recorded. �e highest number of species (10) was present in one relevé (no. 30), one species was recorded only in one relevé. to the most frequently occured species belong ceratodon purpureus (9) and pohlia cruda (8). �e species with the highest cover are: dicranum scoparium (30% – relevé no. 30) and bryum capillare (25% – relevé no. 21 and 20% – relevé no. 23). p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 32 species-poorest mine heap was maximilián in špania dolina where we found only 20 species of mosses. �e highest number of species present in a relevé was 7 (relevé no. 39) and one species was recorded in 4 relevés. few species had the highest cover compared to other studied areas: hypnum cupressiforme (40% – relevé no. 53 and 30% – relevé no. 44), ceratodon purpureus (35% – relevé no. 43), tortella tortuosa (30% – relevé no. 45) and campylopus intro�exus (30% – relevé no. 48). �e most frequent species was ceratodon purpureus (present in 12 relevés) and hypnum cupressiforme (7). a total of 11 moss species were present on all three mine heaps: brachythecium albicans, b. salebrosum, bryum caespiticium, b. moravicum, ceratodon purpureus, dicranum scoparium, hylocomium splendens, plagiomnium a�ne, pohlia cruda, polytrichastrum formosum and racomitrium canescens, while 9 bryophyte species were found only on podlipa (aulacomnium palustre, barbilophozia barbata, brachythecium rutabulum, calliergonella cuspidata, dicranella heteromalla, lophocolea bidentata, plagiomnium cuspidatum, pohlia drummondii and sciuro-hypnum re�exum), 8 on richtárová (barbula unguiculata, brachythecium rivulare, cirriphyllum piliferum, didymodon fallax, drepanocladus aduncus, oxyrrhynchium hians, rhytidiadelphus triquetrus and sciuro-hypnum populeum) and 4 on maximilián (campylopus intro�exus, grimmia pulvinata, pterigynandrum �liforme and tortella tortuosa). �e most frequent species on all studied sites was ceratodon purpureus, recorded in 24 relevés altogether, followed by pohlia cruda (17) and dicranum scoparium, hypnnum cupressiforme and pleurozium schreberi (all present in 11 relevés). �e genus with the highest number of species was brachythecium (b. albicans, b. rivulare, b. rutabulum and b. salebrosum). hypnum cupressiforme was the species with the highest overall cover (40% – relevé no. 53), but was not recorded on mine heap richtárová. species with the highest cover that were present on all three studied sites (at least 2a – 5%–15% cover-abundance) were brachythecium salebrosum, ceratodon purpureus, plagiomnium a�ne and pohlia cruda. according to kubinská et al. (2001) none of the species recorded in our survey belongs to the “red list of bryophytes of slovakia”. according to kubinská, janovicová (2001) species campylopus intro�exus (found in our �eld survey only on maximilián) belongs to invasive species in slovakia. as a general rule in our studied areas, relevés with the highest number of species were situated in sites with deeper soil layer such as the ecotone between the mine heap and forest habitat or on sites with lower inclination. �is is evident by the presence of species typical for humid and shady habitats such as climacium dendroides, dicranum scoparium, hylocomium splendens, rhytidiadelphus squarrosus etc. (pilous, duda, 1960). on the contrary, relevés with o�en only one species present lie more or less on bare rocky sites with species such as ceratodon purpureus, pohlia cruda, p. drummondii, racomitrium canescens, tortella tortuosa, or species from genus bryum. b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 33 štrba et al. (2014) but also dobríková (2011) reported several bryophyte species from mine heap richtárová in staré hory: abietinella abietina, ceratodon purpureus, dicranum scoparium, hylocomium splendens, plagiomnium a�ne, p. undulatum, pleurozium schreberi, pohlia cruda, polytrichastrum commune, p. formosum, racomitrium canescens, r. lanuginosum, rhytidiadelphus squarrosus, r. triquetrus and �uidium tamariscinum. on the mine heap maximilián in špania dolina genera such as dicranum, hylocomium, plagiomnium, pleurozium, polytrichastrum, rhytidiadelphus and �uidium had the highest abundance (aschenbrenner et al., 2011). data on bryophytes from the mine heap podlipa in ľubietová have not been published yet. a high heavy metal content in soil has a strong selection pressure on vegetation. most plant species are not capable of adapting. �ere is, however, a  small group of plants – specialists, that could tolerate these soils (ernst, 1974; ernst et al., 1992). studies have shown that tolerant ecotypes are formed within a certain species that are adapted to these habitats. in recents years it has been discovered that some species of grasses, herbs and also lichens form tolerant ecotypes (bačkor et al., 1998; bačkor, váczi, 2002). among bryophytes such tolerant ecotypes were reported in ceratodon purpureus, pohlia drummondii and pleurozium schreberi from mine heaps in staré hory, gelnica and smolník (banásová, 2006; banásová et al., 2007) or brachythecium albicans in banská štiavnica (banásová et al., 2012). all these species were noted in the investigated areas. a tolerant ecotype was also reported in ceratodon purpureus (jules, shaw, 1994), funaria hygrometrica hedw. (shaw, 1988) and in two liverworts, marchantia polymorpha l. (briggs, 1972) and solenostoma crenulata (sm.) mitt. (brown, house, 1978). shaw (1990) stated that funaria hygrometrica forms copper-tolerant ecotypes, although tolerance to other metals (e.g. zinc and cadmium) appears to be due mostly to cross-tolerance and generally vigorous growth. shacklette (1967) stated that some bryophyte species have been known to occupy substrates with greater than the normal content of copper or other metals. �ese species are commonly known in literature as copper mosses (limpricht, 1895; morton, gams, 1925; persson, 1956; schatz, 1955; brooks et al., 1985), although some are associated with metals other than copper and some are liverworts, not mosses. schatz (1955) considered copper mosses to be more properly termed “sulfur mosses” because of their frequent linkage with sulfur compounds of copper, lead, zinc, and iron, as well as with sulfur deposits at mineral springs. persson (1956) proposed that the controlling factor for the distribution of the copper mosses was the low ph (3.4–4.3) and observed that copper mosses were never found over ultrama�c substrates (high ph) despite the high concentrations of heavy metals in these rocks and soils. johnson-groh (1987) also pointed out the importance of microclimate. širka (2014) reported a signi�cant amount of cu accumulated in the species pohlia drummondii from the mine heap podlipa in ľubietová (4010.3 mg/kg) compared to p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 34 �uidium delicatulum (8.5 mg/kg), and the mean value of cu concentration in the substrate (3253 mg/kg). since bryophytes do not have a  well developed cuticle and root system like vascular plants and rely mostly on atmospheric deposition for nourishment, there is much controversy as to whether copper itself (or other heavy metal for that matter) is indeed the controlling factor for the distribution of bryophytes on metal-contaminated sites. fernández (2013) and brown (1995) concluded that mosses remain in a state of unstable equilibrium between inputs and outputs of contaminants and do not integrate metals from deposition and that this equilibrium of the concentrations of elements in moss cannot be studied by only considering the total concentrations of contaminants in the tissues as it is known that contaminants may be located in di�erent cell compartments. according to brooks et al. (1985) it is possible that the “preference” shown by some mosses for sites contaminated by heavy metals is a result of two factors: an acquired resistance to toxic ions, and a low tolerance to competition from other species outside contaminated sites. results by jules, shaw (1994) give some indication that there might be a biological cost associated with metal-tolerance in ceratodon purpureus. in their study smelter plants produced fewer stems and fewer gametangia on the control treatment than on the contaminated treatment and although this result may be, in fact, due to other di�erences between the studied sites (e.g. nutrient content), their evidence is suggestive of a  cost. similarly, shaw (1990) demonstrated that metal-tolerant funaria hygrometrica individuals form stems more slowly on normal (uncontaminated) treatments than non-tolerant individuals. �e tendency of these mosses to grow on mineralized substrates might be a species characteristic, not a generic one (shacklette, 1967). according to jules, shaw (1994) bryophytes can better adapt to heterogeneous environments than angiosperms as they have higher levels of phenotypic plasticity, rather than small-scale genetic responses. strong conclusions cannot be made as to lack of evidence at present. acknowledgements �e article was �nancially supported by grant scheme vega 1/0538/15 conclusions a total of 45 species of bryophytes (43 mosses and 2 liverworts) in 54 releves were found in three mine heaps with copper-rich substrate in central slovakia that had a similar bryophyte species composition. species-richest mine heap was podlipa (31 recorded species), while species-poorest mine heap was maximilián (20 recorded species). most frequent species were ceratodon purpureus, pohlia cruda, dicranum scoparium, hypnum cupressiforme and pleurozium schreberi. hypnum cupressiforme was the species with the highest overall cover. we con�rmed that bryophytes play an b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 35 important role in the initial stages of the succession process on anthropogenically created habitat types, as with lichens they formed a major component of the vegetation cover on mine heaps where any kind of mining activity was ended centuries ago. �eir detailed study in relation to phytoextraction or phytostabilization of heavy metals may be used in succession management process aimed at restoring the environment, particularly the soil and air. references andráš, p., jeleň, s., križáni, i. 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(1973). �e braun-blanquet approach. in: r.h. whittaker (ed.), ordination and classi�cation of communities, handbook of vegetation science, 5th ed. �e hague: dr. w. junk b. v. publishers, 617–726. p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 38 appendix 1 �e list of all relevés (explanation in the “methods”). ľubietová – podlipa (p) 1. bare coarse scree; 48°44’45.63” n, 19°23’4.04” e; 611 m; 0°; -; e0 25%; 1; 8.11.2013 2. shrubland with pioneer tree species and moderately developed soil; 48°44’45.84” n, 19°23’2.94” e; 617 m; 5°; ne; e2 35%; e1 15%; e0 25%; 6; 8.11.2013 3. dry �ne scree with slightly developed soil; 48°44’45.54” n, 19°23’3.48” e; 611 m; 0°; -; e1 5%; e0 7%; 2; 8.11.2013 4. sparse vegetation on medium-sized scree with moderately developed soil; 48°44’47.90” n, 19°23’5.69” e; 628 m; 15°; ne; e3 25%; e1 15%; e0 35%; 8; 8.11.2013 5. shrubland vegetation with developed soil; 48°44’48.06” n, 19°23’5.34” e; 625 m; 15°; e; e2 60%; e1 35%; e0 40%; 10; 8.11.2013 6. shrubland vegetation with moderately developed soil; 48°44’47.64” n, 19°23’5.04” e; 624 m; 0°; -; e2 50%; e1 25%; e0 30%; 7; 8.11.2013 7. sparse shrubland vegetation with moderately developed soil; 48°44’47.28” n, 19°23’4.38” e; 624 m; 0°; -; e2 20%; e1 40%; e0 20%; 6; 8.11.2013 8. sparse shrubland with moderately developed soil; 48°44’47.04” n, 19°23’3.84” e; 624 m; 0°; -; e2 35%; e1 5%; e0 35%; 6; 8.11.2013 9. tree cover with picea abies on coarse scree but developed soil; 48°44’47.34” n, 19°23’2.94” e; 629 m; 20°; ne; e3 85%; e1 5%; e0 13%; 5; 8.11.2013 10. bare coarse scree; 48°44’48.36” n, 19°23’4.92” e; 631 m; 20°; sw; e0 15%; 1; 8.11.2013 11. grassland on coarse scree but moderately developed soil; 48°44’51.96” n, 19°23’10.14” e; 657 m; 10°; e; e1 90%; e0 15%; 4; 8.11.2013 12. bare coarse scree; 48°44’51.24” n, 19°23’9.60” e; 653 m; 10°; s; e0 10%; 1; 8.11.2013 13. bare medium-sized scree on steep slope; 48°44’53.77” n, 19°23’9.81” e; 676 m; 50°; sw; e0 3%; 2; 8.11.2013 14. grassland on coarse scree but moderately developed soil; 48°44’50.10” n, 19°23’8.94” e; 643 m; 5°; s; e1 95%; e0 10%; 2; 8.11.2013 15. grassland on coarse scree but moderately developed soil; 48°44’49.74” n, 19°23’8.28” e; 640 m; 0°; -; e1 97%; e0 15%; 1; 8.11.2013 16. shady sparse grassland on medium-sized scree with slightly developed soil; 48°44’48.60” n, 19°23’7.20” e; 633 m; 5°; sw; e1 10%; e0 8%; 1; 8.11.2013 17. tree cover with picea abies on developed soil; 48°44’43.80” n, 19°23’5.34” e; 613 m; 7°; nw; e3 90%; e1 20%; e0 30%; 4; 8.11.2013 18. sparse tree cover with picea abies on developed soil; 48°44’42.42” n, 19°23’3.00” e; 595 m; 0°; -; e3 70%; e 1 5%; e0 12%; 3; 8.11.2013 19. sparse vegetation cover on �ne scree and slightly developed soil; 48°44’42.84” n, 19°23’2.76” e; 596 m; 0°; -; e1 15%; e0 7%; 3; 8.11.2013 20. shady and wet vegetation cover with moderately developed soil; 48°44’42.06” n, 19°23’3.18” e; 595 m; 0°; -; e3 65%; e2 8%; e1 35%; e0 17%; 6; 8.11.2013 staré hory – richtárová (r) 21. bare scree on steep slope; 48°49’40.20” n, 19°8’0.12” e; 580 m; 60°; nw; e0 25%; 2; 23.11.2013 22. bare scree on steep slope; 48°49’40.08” n, 19°7’59.70” e; 583 m; 60°; n; e0 5%; 1; 23.11.2013 23. wet grassland vegetation cover on skeletal soil; 48°49’39.84” n, 19°8’0.18” e; 581 m; 20°; ne; e1 60%; e0 30%; 3; 23.11.2013 b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 39 24. sparse vegetation cover on coarse scree; 48°49’39.72” n, 19°8’0.42” e; 582 m; 10°; w; e1 10%; e0 20%; 6; 23.11.2013 25. sparse shrubland vegetation cover with high portion of forest litter and slightly developed soil; 48°49’38.34” n, 19°7’59.58” e; 589 m; 15°; n; e2 25%; e1 7%; e0 35%; 5; 23.11.2013 26. �at surface with sparse herbaceous cover on skeletal soil; 48°49’38.88” n, 19°7’59.04” e; 588 m; 0°; -; e1 25%; e0 30%; 7; 23.11.2013 27. sparse moss cover on coarse scree; 48°49’38.94” n, 19°7’58.86” e; 591 m; 15°; e; e1 5%; e0 15%; 3; 23.11.2013 28. moss cover on steep slope with coarse scree; 48°49’38.46” n, 19°7’58.80” e; 592 m; 50°; e; e1 3%; e0 50%; 6; 23.11.2013 29. herbaceous and moss cover on coarse scree; 48°49’38.04” n, 19°7’58.32” e; 595 m; 30°; nw; e1 20%; e0 15%; 3; 23.11.2013 30. dense moss cover in a tree overgrowth with picea abies on skeletal soil; 48°49’36.90” n, 19°7’58.27” e; 601 m; 10°; nw; e3 75%; e1 30%; e0 85%; 10; 23.11.2013 31. overgrown coarse scree slope with pioneer tree species; 48°49’35.29” n, 19°7’57.40” e; 613 m; 30°; e1 25%; e0 65%; nw; 7; 23.11.2013 32. bare �ne scree in the center of the mine heap; 48°49’31.08” n, 19°7’53.58” e; 651 m; 5°; n; e0 6%; 2; 23.11.2013 33. sparse vegetation cover on �ne scree; 48°49’31.97” n, 19°7’52.40” e; 656 m; 7°; ne; e1 15%; e0 10%; 3; 23.11.2013 34. tree cover with picea abies on a steep slope with developed soil and dense moss cover; 48°49’29.46” n, 19°7’53.40” e; 663 m; 20°; ne; e3 99%; e1 2%; e0 55%; 6; 23.11.2013 35. dense grassland vegetation; 48°49’28.44” n, 19°7’54.30” e; 664 m; 0°; -; e1 95%; e0 10%; 3; 23.11.2013 36. dense moss cover on coarse scree; 48°49’26.99” n, 19°7’55.71” e; 669 m; 20°; ne; e0 40%; 7; 23.11.2013 špania dolina – maximilián (m) 37. bare coarse scree; 48°48’31.04” n, 19°8’8.61” e; 764 m; 45°; sw; e0 1%; 1; 19.12.2013 38. bare coarse scree; 48°48’31.74” n, 19°8’8.40” e; 773 m; 45°; w; e0 15%; 4; 19.12.2013 39. coarse scree with slightly developed soil; 48°48’32.04” n, 19°8’8.46” e; 775 m; 10°; s; e1 10%; e0 25%; 7; 19.12.2013 40. coarse scree with slightly developed soil; 48°48’32.64” n, 19°8’9.18” e; 784 m; 0°; -; e1 5%; e0 20%; 4; 19.12.2013 41. coarse scree with slightly developed soil; 48°48’31.74” n, 19°8’11.70” e; 777 m; 5°; sw; e0 15%; 3; 19.12.2013 42. bare coarse scree; 48°48’31.43” n, 19°8’13.26” e; 775 m; 5°; s; e0 1%; 1; 19.12.2013 43. moss cover on coarse scree with slightly developed soil; 48°48’31.50” n, 19°8’15.00” e; 776 m; 5°; s; e0 40%; 3; 19.12.2013 44. coarse scree with high portion of forest litter; 48°48’31.86” n, 19°8’12.96” e; 780 m; 5°; nw; e1 3%; e0 30%; 1; 19.12.2013 45. bare coarse scree; 48°48’31.20” n, 19°8’9.18” e; 766 m; 40°; s; e0 30%; 2; 19.12.2013 46. bare coarse scree; 48°48’31.86” 19°8’8.64” e; 774 m; 30°; n; e1 2%; e0 15%; 1; 19.12.2013 47. coarse scree with slightly developed soil; 48°48’32.10” n, 19°8’8.46” e; 775 m; 30°; n; e1 5%; e0 12%; 5; 19.12.2013 48. coarse scree with slightly developed soil and higher portion of forest litter; 48°48’33.12” n, 19°8’8.58” e; 789 m; 0°; -; e1 7%; e0 30% 2; 19.12.2013 49. coarse scree with slightly developed soil; 48°48’33.18” n, 19°8’8.40” e; 789 m; 0°; -; e1 3%; e0 10%; 4; 19.12.2013 50. coarse scree with slightly developed soil and high portion of forest litter; 48°48’33.30” n, 19°8’7.74” e; 788 m; 0°; -; e0 25%; 2; 19.12.2013 p av el š irk a, in gr id t ur is ov á, a nn a p et rá šo vá 40 51. vegetation cover with picea abies on coarse scree and moderately developed soil; 48°48’33.42” n, 19°8’7.62” e; 788 m; 0°; -; e3 20%; e2 40%; e1 15%; e0 30%; 6; 19.12.2013 52. coarse scree with moderately developed soil and high portion of forest litter; 48°48’33.48” n, 19°8’7.20” e; 787 m; 0°; -; e0 25%; 2; 19.12.2013 53. tree cover with picea abies and dense moss cover on developed soil; 48°48’34.20” n, 19°8’7.14” e; 794 m; 15°; w; e3 90%; e1 10%; e0 70%; 6; 19.12.2013 54. tree cover with picea abies and dense moss cover on developed soil; 48°48’34.02” n, 19°8’4.50” e; 780 m; 10°; s; e3 75%; e2 7%; e1 5%; e0 45%; 4; 19.12.2013 b ryophytes of c u-m ine heaps in the vicinity of b anská b ystrica (c entral s lovakia) 41 abstract �e knowledge of bryophytes growing on metal-contaminated sites is still insu�cient in slovakia. �is study deals with bryophyte �ora of three mine heaps (podlipa, richtárová and maximilián) with copper-rich substrate. a total of 54 relevés was made, in which a total of 45 bryophyte species (43 mosses and 2 liverworts) was recorded. species-richest mine heap was podlipa with 31 bryophyte species (29  mosses and 2 liverworts) and species-poorest was maximilián with 20 species of mosses. 11 species were mutual for all three mine heaps, while 9 species where present only on podlipa, 8 on richtárová and 4 on maximilián. �e most representative bryophytes, in terms of their occurrence and cover, are ceratodon purpureus, pohlia cruda, dicranum scoparium, hypnum cupressiforme, pleurozium schreberi, brachythecium salebrosum and plagiomnium a�ne. occurence of bryophytes on metal-contaminated sites was discussed. key words: bryophytes, mine heaps, ľubietová, staré hory, špania dolina, central slovakia received: [2016.04.16] accepted: [2016.09.30] mszaki z hałd kopalni miedzi w okolicach bańskiej bystrzycy (centralna słowacja) streszczenie na słowacji nadal niewystarczająca jest znajomość mszaków rosnących na terenach zanieczyszczonych metalami, takimi jak miedź. niniejsze badania dotyczą �ory mszaków trzech hałd kopalnianych (podlipa, richtárová i maximilián), o podłożu bogatym w ten metal. na analizowanym terenie wykonano 54 zdjęcia �tosocjologiczne, w których łącznie stwierdzono 45 gatunków mszaków (43 mchy i 2 wątrobowce). najbogatsza gatunkowo okazała się być hałda kopalniana z podlipa, na której odnotowano 31 gatunków mszaków (29 mchów i 2 wątrobowce), a najuboższa hałda maximilián z 20 gatunkami mchów. jedynie 11 gatunków było wspólnych dla wszystkich trzech hałd kopalnianych, natomiast tylko 9 gatunków występowało wyłącznie na hałdzie podlipa, 8 na richtárovej i 4 na maximilián. najczęstszymi mszakami, zarówno pod względem występowania, jak i pokrycia w płatach były: ceratodon purpureus, pohlia cruda, dicranum scoparium, hypnum cupressiforme, pleurozium schreberi, brachythecium salebrosum i plagiomnium a�ne. słowa kluczowe: mszaki, hałdy kopalniane, ľubietová, staré hory, spania dolina, centralna słowacja information on the authors pavel širka majored in ecology at the faculty of natural sciences, department of biology and ecology, matej bel university in banská bystrica. he is currently in 3rd year of ph.d. studies. his dissertation thesis deals with bryophyte assemblages on mine heaps in slovakia with di�erent mineral content of substrate in relation to chosen biotic and abiotic factors. ingrid turisová in her current scienti�c work she is focused on plant indication and bioaccumulation of heavy metals in a disturbed environment, botanical and ecological analysis of grassland habitats and their management. she collaborates in the studies of vegetation in the concept of ecological landscape integrity. anna petrášová until 2015 she worked as a lecturer at the faculty of natural sciences, department of biology and ecology, matej bel university in banská bystrica, where she was engaged mainly in the studies of bryophyte �ora and didactics of biology. she currently serves in a  managerial position at an elementary school, where in addition to managerial work she also works as a teacher. 152 annales universitatis paedagogicae cracoviensis studia naturae, 3: 152–161, 2018, issn 2543-8832 doi: 10.24917/25438832.3.12 arkadiusz gruca institute of biology, faculty of geography and biology, pedagogical university of cracow, cracow, poland, arkadiusz.gruca@up.krakow.pl a brief review of microbial induced corrosion research corrosion is a natural process of the gradual conversion of re�ned materials, such as metal or concrete, into a more chemically stable form, e.g., sulphide, nitrate, or oxide. corrosion closely corresponds with destruction of materials exposed to the environment (schweitzer, 2010). some microorganisms possess the ability to accelerate corrosion. �is process is called microbial induced corrosion (mic). mic is associated with formation of bacterial bio�lms. bio�lm is a bacterial community embedded in extracellular matrix formed by eps (extracellular polymeric substances) secreted by bacteria. products of bacterial metabolism are very corrosive to metal and concrete surfaces that the bio�lm is attached to, and so microbial induced corrosion is a signi�cant threat to metal and concrete surfaces (javed et al., 2015). pipelines, fuel tanks, ship hulls, sewage systems, and other elements exposed to freshwater, seawater, sewage, or soil are especially susceptible to mic (cayford et al., 2017; hunsucker et al., 2018; grengg et al., 2018). repairing damage caused by bacteria costs billions of dollars a year (koch et al., 2002). scientists recognise the threat posed by microorganisms and are conducting extensive research. mine study goals are the identi�cation of bacterial communities responsible for accelerated corrosion of materials, the explanation of the main microbial induced corrosion mechanisms and e�ective inhibitors of this kind of corrosion, and the creation of mic resistant materials. �e aim of this study is review the latest advances in microbial induced corrosion research, compare currently used biocorrosion prevention methods, and to discuss chemical and biological processes behind microbial induced corrosion. corrosion inducing bacteria corrosion is induced by wide range of bacteria. �e most prevalent of over 13 phyla related to biocorrosion in various environments are bacterioidetes krieg et al. 2012, a brief review of m icrobial induced corrosion research 153 proteobacteria stackebrandt et al., 1988 and firmicutes gibbons & murray 1978 (cayford et al., 2017; li et al., 2017a; hunsucker et al., 2018; li et al., 2018). overall, more than 20 classes of bacteria were proven to induce corrosion (li et al., 2017), the most abundant of them being deltaproteobacteria stackebrandt et al. 1988, clostridia rainey 2010 and gammaproteobacteria stackebrandt et al. 1988 (cayford et al., 2017; li et al., 2017a; hunsucker et al., 2018). on the genera level, the most common mic causing bacteria are desulfovibrio kluyver & van niel 1936, desulfobacter widdel 1981 and desulfotomaculum campbell & postgate 1965, all three belonging to sulphate reducing bacteria (srb) group (hamilton, 1985; jia et al., 2017; wan et al., 2018). sulphate reducing bacteria are considered to be the typical mic causing microorganisms, thanks to their ability to accelerate corrosion in anaerobic environments (videla, 1986; sherar et al., 2011; dec et al., 2016). studies have shown that srb, apart from corrosion acceleration, can also lead to corrosion inhibition. sulphides created by bacteria can form �lms on the surface. �in �lms work as corrosion inhibitors, while more bulky �lms can accelerate the corrosion rate (videla et al., 2005; xu et al., 2013). as srbs are strictly anaerobic, most of the research regarding corrosion e�ects of srbs is focused around anaerobic environments. in recent years, another group of corrosion inducing bacteria has gained a lot of researchers’ attention, that group being nitrate reducing bacteria (nrb). nrbs have proven to induce corrosion include gene bacillus cohn 1872, acidithiobacillus kelly & wood 2000 and alcaligenes castellani & chalmers 1919, (wang et al., 2014; liu et al., 2016; herisson et al., 2017). studies have shown that corrosion caused by nrbs can be more serious that that caused by srbs (wan et al., 2018). despite the intensive research, corrosion the mechanism of nrb still needs more investigation. mic as a topic of scientific research in recent years, microbial induced corrosion has been gaining more and more attention among scientists from a range of scienti�c �elds. due to the material destructing nature of mic, most of the research revolves around the development of corrosion resistant materials, corrosion inhibitors, and the recognition of mic mechanisms. mic has been proven as one of the main factors in concrete degradation. corrosion of wastewater networks poses a high risk to the environment and public health (world health organisation, 2000; li et al., 2017b). �e range of volatile organic compounds (vocs) produced as bacterial metabolites constitute considerable health and safety issues for sewage systems operators and community workers (alexander et al., 2013; gutierrez et al., 2014). despite intensive in situ and lab research, corrosion resistant concrete is still not available for wide usage. not one of the currently a rk ad iu sz g ru ca 154 used concrete mixtures can resist mic for their projected operating lifetime (goyns, alexander, 2014; herisson et al., 2014). experience has shown that physiochemical concrete parameters are very important for mic resistance (vincke et al., 2002; herisson et al., 2014). mixtures with high bacterial created acid neutralisation capacity and small pores were proven to be especially resistant to corrosion (gu et al., 2011; li et al., 2017b). antibacterial additives, such as zno powder, were also proven to be e�ective in slowing down mic (schultz et al., 2011). water transportation is another �eld in which mic causes considerable loses every year. ship hulls, and fuel and ballast tanks are especially endangered. seawater is a  perfect environment for bacteria, thanks to abundance of organic and mineral compounds necessary for bacteria to thrive, and a relatively stable temperature. intercontinental water transport highly contributes to the propagation of bacteria around the globe (souza et al., 2016). �e accumulation of bacterial bio�lm, responsible for corrosion on ship hulls causes increased drag, which leads to higher fuel consumption and increased exhaust emissions (swain, 2010). to negate this problem, biocides and anti-adhesion coatings are used (lee et al., 2012). because of high toxicity of biocides, new methods of protecting ship hulls are being developed. one of the new methods that show promise is grooming (hunsucker et al., 2018). grooming is based on brushing the surface attacked by bacteria and removing bio�lms, and other contaminations. �e groomed surface is smoother and thereby more resistant to bacterial adhesion. however, more research is required to re�ne grooming tools and procedures. studies have shown that biodiesel fuels can accelerate the corrosion of carbon steel fuel tanks in contact with marine microbes (bellige et al., 2015). cu-ni coatings used for protection of fuel tanks against corrosion were proven to be not e�ective against mic. bacterial sulphate reduction corresponding with fuel biodegradation can lead to rapid penetration of the protecting coating and the corrosion of external steel layers (lv et al., 2017). latest research shows that the type of fuel is a major factor in cu-ni coating corrosion (hunsucker et al., 2018). to �ght this, new generations of biofuels are being implemented (liang et al., 2017). �e environment of the oral cavity is a perfect incubator for bacteria (long, rack, 1998). because of that, large emphasis is given for development of mic resistant dental implants. �e most widely used material for implant production is titanium, known for its biocompatibility and corrosion resistance (navarro et al., 2008; diaz et al., 2018). �e corrosion resistance of titanium comes from its ability to passivate and create a 2-5 nm thick protective oxide layer on the implant surface. however, recent studies have shown that bacteria can accelerate titanium corrosion in the oral cavity environment (li et al., 2017). roughness of the implant surface also plays a major role in mic resistance, and because of that, a range of surface modi�cations are being a brief review of m icrobial induced corrosion research 155 extensively tested (souza et al., 2016). �e goal is to achieve high corrosion resistance without lowering biocompatibility of the implant. mic prevention methods because of high maintenance costs of elements a�ected by mic, a lot of emphasis is given to the development of e�ective anti-corrosion agents, coatings, and corrosion resistant materials. �e application of biocides, such as bronopol and innovative coatings containing antibacterial nanoparticles are being tested. bronopol (2-bromo 2-nitropropane-1.3-diol) is a well-known anti-microbial agent. it can form a protective layer on the surface of metal, thus protecting it against bacteria. studies have shown that bronopol can considerably reduce the corrosion rate of mild steel (narenkumar et al., 2018). however, according to sharma et al. (2017), high concentrations of bronopol can lead to an increase in the corrosion rate. because of that, high dosages of bronopol should be avoided. �e use of bioengineered silver nanoparticles (nps) is an innovative method of preventing mic (narenkumar et al., 2018). �anks to their high antibacterial potential, silver nanoparticles are very e�ective in stopping bio�lm development (sondi, salopek-sondi, 2004; kim et al., 2007; narenkumar et al., 2018). analyses have shown that silver nps can be absorbed by the metal surface and form a protective layer, which adds to their anticorrosive properties. unfortunately, silver nps have been proven as highly toxic and hazardous for the environment (hajipour et al., 2012; bondarenko et al., 2013; yuan et al., 2017). �e usage of many anti mic agents is very limited due to their high toxicity and destructing in�uence on the natural environment. because of that, eco-friendly alternative solutions are being developed. one of them is the usage of plant-based natural corrosion inhibitors (narenkumar et al., 2017; punniyakotti et al., 2017). many plants are well known for their antibacterial properties (raja, sethuraman, 2008; narenkumar et al., 2017; punniyakotti et al., 2017; aribo et al., 2017), and can be used to prevent corrosion. studies have shown that ginger (zingiber o�cinale rosc.) in concentrations as low as 20 ppm inhibits mic with over 80% e�ciency (narenkumar et al., 2017). despite high potential of natural inhibitors, more research is required to develop e�ective ways of implementing them in in situ conditions. mic mechanisms anaerobic bacterial metabolism can be divided into two types: fermentation and respiration (błaszczyk, 2010). with that classi�cation, anaerobic microbial induced corrosion can be divided into three main categories (xu, gu, 2011; gu, 2012; xu et al., 2013). a rk ad iu sz g ru ca 156 in standard conditions, organic carbon is a main source of nourishment for microbes. bacteria subjected to carbon starvation have been shown to accelerate carbon steel corrosion. with the lack of carbon, bacteria were using metallic iron as a source of electrons needed for the oxidation process (jia et al., 2017). metals that can be used as electron donors are more susceptible to biocorrosion (xu et al., 2016). in mic i, extracellular electrons realised in oxidation of iron are used by bacteria to reduce oxidants such as sulphate or nitrate in their cytoplasm. for this to happen, electrons must be transported through a cell wall, this is called extracellular electron transfer (eet). two main methods of eet are used by bacteria: mediated electron transfer (met) and direct electron transfer (det). �e addition of electron mediator into desulfovibrio vulgaris (hildenbor) culture medium accelerated corrosion (zhang et al., 2015). �is shows that electron transfer is a limiting factor in mic. a theory called biocatalytic cathodic sulphate reduction (bcsr) was proposed to describe the thermodynamics of microbial induced corrosion caused by srb (gu et al., 2009). in this theory, sulphate is the terminal electron acceptor, and iron oxidation occurs extracellulary, and sulphate reduction occurs in the srb cytoplasm. bcsr can be used as a base for computer modelling of mic caused by sulphate reducing bacteria. various factors (i.e., temperature, bio�lm aggressiveness, ph, [so42-]) in�uencing corrosion speed can be investigated through computer simulation (xu et al., 2016). biocatalytic cathodic nitrate reduction (bcnr) is a theory parallel to bcsr, and it can explain mic caused by nitrate reducing bacteria. nitrate reduction associated with extracellular iron oxidation can cause corrosion more severe than that linked to sulphate reduction (gu, 2012; xu et al., 2016). mic ii is caused by corrosive bacterial metabolites (i.e., organic acids and sulphides) released into bio�lm. in this type of mic, metabolites are used by bacteria to achieve redox balance (shuler, kargi, 2002). �e ph di�erence between bio�lm and surrounding liquid leads to the acidic corrosion of surface underneath the bio�lm (xu et al., 2016). it is still unknown if this process is deliberate and if bacteria secrete corrosive metabolites for the purpose of harvesting energy (li et al., 2018). type iii mic can be best described as the biodegradation of organic materials caused by microbes. in humid environments, microorganisms, such as fungi, excrete enzymes that digest organic matter transforming it into substances that can be absorbed into cells. �is kind of microbial induced corrosion can damage polymer insulations and lead to the failure of electrical systems (gu, 2003). conclusions �e state of the art knowledge of microbial induced corrosion was reviewed in this study. despite major advances in recent years, more research is still required to accua brief review of m icrobial induced corrosion research 157 rate the description of mic process in nature and the development of more e�ective biocorrosion inhibitors. computer simulation can help accelerate research speed and largely contribute towards new discoveries in mic studies. better understanding of mic mechanisms allowed for the development of corrosion resistant materials and new ways of �ghting corrosive microbes. scientists are on the right path, and rapid progress in microbial induced corrosion research is becoming more and more apparent. references alexander, m., bertron, a., de belie, n. 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(2015). electron mediators accelerate the microbiologically in�uenced corrosion of 304 stainless steel by the desulfovibrio vulgaris bio�lm. bioelectrochemistry, 101, 14–21. doi: 10.1016/j.bioelechem.2014.06.010 krótki przegląd badań nad biokorozją streszczenie korozja to ogół procesów prowadzących do niszczenia materiałów. jednym z  typów korozji jest korozja powodowana działaniem mikroorganizmów. tak zwana biokorozja w  znacznym stopniu przyczynia się do degradacji konstrukcji metalowych i betonowych. niektóre elementy tych konstrukcji, w szczególności te wystawione na działanie wody słodkiej, słonej, ścieków albo ziemi są szczególnie narażone na destrukcyjny wpływ mikrobów. korozja mikrobiologiczna w największym stopniu dotyka przemysłu na�owo-gazowego, transportu wodnego i  instalacji sanitarnych. niebagatelny problem stanowi także, powodowana przez bakterie znajdujące się w  jamie ustnej, korozja implantów dentystycznych. mimo, że mechanizmy a brief review of m icrobial induced corrosion research 161 powodujące biokorozję nie są dobrze znane, walka z tym zjawiskiem jest przedmiotem badań instytutów na całym świecie. ważnym zagadnieniem jest również projektowanie materiałów o zwiększonej odporności na biokorozję. celem tego artykułu jest podsumowanie dotychczasowego stanu wiedzy o zjawisku biokorozji, przybliżenie obecnie stosowanych metod jej zapobiegania, oraz omówienie procesów chemicznych i biologicznych stojących za korozją indukowaną przez mikroorganizmy. key words: bacteria, bio�lm, inhibitors, mechanism, microbial induced corrosion received: [2018.07.10] accepted: [2018.09.25] 72 annales universitatis paedagogicae cracoviensis studia naturae, 3 (suplement): 72–82, 2018, issn 2543-8832 doi:10.24917/25438832.3supp.10 ewa sosnówka-czajka*, iwona skomorucha department of poultry breeding, national research institute of animal production, kraków, poland, *ewa.sosnowka@izoo.krakow.pl selected blood parameters in organically raised hens fed with a purple coneflower supplemented diet** **carried out based on ministry of agriculture and rural development decision no. hor.re.027.5.2017 introduction organic farming requires continuous care for a high health status of birds to make poultry production pro�table and to obtain quality products, which is of direct interest to consumers. impaired immune function leads to deterioration in productivity, e.g., poorer laying performance, lower hatchability, or increased mortality (yunis et al., 2000; sivaraman et al., 2005). on the other hand, by stimulating the immune system through modi�ed nutrition, it is possible to reduce the negative e�ects of compromised immunity in poultry (truchliński et al., 2006; salim et al., 2018). environmental stress associated with the used rearing system may, in general, interfere with antibody production and cellular immune response, making poultry more vulnerable to vital infections (bartlett, smith, 2003). at present, organic farmers are focused on increasing their use of herbs and herbal mixtures, which have positive e�ects on the animals’ metabolism, health, productivity, feed conversion, and reproduction. a large number of biologically active therapeutic substances are found in purple cone�ower, including polysaccharides, glycoproteins, and alkaloids (dalby-brown et al., 2005). many of these substances are used to improve the body’s immune potential and to enhance macrophage activity (goel et al., 2002). allen (2003) reported that purple cone�ower extract can be used to increase resistance to coccidiosis in birds kept on litter. according to najafzadeh et al. (2011), purple cone�ower can be an e�ective means to prevent avian in�uenza in birds. �erefore, the aim of this study was to determine the e�ect of a dietary addition of dried purple cone�ower (echinacea purpurea (l.) moench) on selected blood parameters of chicks and hens of the native ‘greenleg partridge’ breed (z-11), maintained according to organic farming principles. 73 s elected blood param eters in organically raised hens fed w ith a purple coneflow er supplem ented diet material and methods animals and experimental design �e experiment was carried out on 252 ‘greenleg partridge’ hens (r-11) aged 20 weeks (wks) with 126 birds in each group with 6 subgroups per group. in each experimental subgroup, there was 1 rooster to 7 layers. hens were maintained on a certi�ed poultry farm according to organic farming principles. birds were divided into two groups according to diet: organic layer diet (group 1), and the same diet supplemented with certi�ed purple cone�ower (echinacea purpurea (l.) moench) from 20 wks of age at 10 g/ kg feed (group 2). layers were fed the certi�ed layer diet (17% protein and 11.1 mj me/ kg diet) following organic farming principles. �e diets contained organic components such as maize, wheat, soybean expeller, sun�ower cake, triticale, dehulled heat-treated lupin, peas, broad bean, soybean, maize gruel, dried lucerne, soybean oil, molasses, and monocalcium phosphate. �roughout the study, hens were fed ad libitum (optimal) and had constant access to water. hens were kept on litter (6 birds/m2 of �oor area) and were allowed access to grassy yards (4 m2/bird). �e yards were equipped with roofs and drinkers. �e microclimate conditions and the light programme were adjusted to the ages of the birds in keeping with pullet rearing and layer management standards. at 34 weeks of rearing, eggs were collected for experimental incubations (100 eggs from each group). eggs were incubated for 21 days under standard conditions. sample collection and laboratory analyses blood was collected from laying hens (from 15 birds per group – 2.5 ml from each hen) at 34 wks and from newly hatched chicks at one day of age (from 15 birds per group – 1 ml from each chick) to determine selected blood parameters. �e circulating immunoglobulin complex was determined by the procedure of lowry (lowry et al., 1951) modi�ed by ślebodziński et al. (1982), and igg was analysed by quantitation tests (chicken igg elisa kit, alpha diagnostic intl., inc.). �e results were read using a tecan spectra classic reader based on kcjunior so�ware (bio-tek instruments, inc.). blood for haematocrit determinations was centrifuged in haematocrit capillaries with an mpw-52 centrifuge, and the result was read from a reader placed on the centrifuge rotor. blood smears were stained with a may-grünwald-giemsa (mgg) protocol for calculation h:l. �e number of red and white blood cells was counted in a bürker chamber under a nikon ys 100 microscope. statistical analysis �e results were statistically analysed by one-way analysis of variance and estimated with duncan’s test. �e statistical calculations were made with statistica ver. 12 so�ware. ew a s os nó w ka -c za jk a, iw on a s ko m or uc ha 74 results appendix 1 – �gure 1 shows the levels of the circulating immunoglobulin complex in 35-week-old organic ‘greenleg partridge’ hens and in day-old chicks derived from these hens. �ere was only a tendency for higher immunoglobulin complex in birds fed the purple cone�ower diet. similar relationships were observed for igg, and the lack of statistically signi�cant di�erences results from a large scatter of data (appendix 1 – fig. 2). z-11 hens fed the purple cone�ower diet and chicks originating from them were characterised by the lowest h:l ratio at p ≤ 0.05 (appendix 1 – fig. 3). no signi�cant di�erences were observed between the experimental groups of z-11 hens and chicks in the level of leukocytes (appendix 1 – fig. 4), erythrocytes (appendix 1 – fig. 5) and haematocrit (appendix 1 – fig. 6). discussion �e body’s immunity depends on the maintenance of immune homeostasis which is associated with systemic homeostasis. �e e�ciency of the immune system is o�en determined from the level of immunoglobulins (sivaraman et al., 2005). according to wang et al. (2000), total ig level may be indicative of the potential e�ciency of the humoral system, whereas the igg level does not always re�ect the actual activity of the immune system. singh et al. (2006) concluded that circulatory immune complexes in broiler chickens average 2.46 g/dl and increase to 2.99 g/dl a�er stimulating the immune system with dietary vitamin e and selenium. in our study, the circulatory immune complexes tended to increase by about 15% in the blood serum of ‘greenleg partridge’ hens fed a purple cone�ower supplemented diet compared to the hens fed a standard organic diet. one of the indicators of chick quality is the body’s immunity, which allows the chicks to counteract the development of pathogens, thus having a positive e�ect on their survival during rearing (kogut, klasing, 2009; korver, 2012). in birds, maternal antibodies are transferred from layer’s serum to egg yolk and later to the yolk sac and blood circulatory system, which provides the embryos and newly hatched chicks with passive immunity (west et al., 2004; bencina et al., 2005). because the level of immunity in laying hens determines the immunity of the chicks, in our study, we found a similar relationship for the chicks from z-11 hens supplemented with purple cone�ower as in adult birds, namely, a tendency for the circulating immune complexes to increase by 9%. our study showed very large individual di�erences in the level of circulating immune complexes; therefore, they were not signi�cant, despite high differences between the groups. 75 serum igg level in chickens re�ects the body’s humoral immunity status (juul-madsen, sørensen, 2004). in our study, we found the level of immunoglobulins to vary widely between individual birds. �erefore, although between-group di�erences in igg concentration were high and reached 23%, they were not statistically signi�cant. similar correlations were also observed by wang et al. (2000). carlander et al. (2003) showed high individual di�erences in igy levels between egg yolks. according to najafzadeh et al. (2011), purple cone�ower can be e�ective in improving poultry immunity. yamada et al. (2011) found purple cone�ower extract to increase iga, igg, and igm levels. in our study, we did not �nd a conclusive e�ect of adding purple cone�ower to the feed of ‘greenleg partridge’ hens kept in accordance with the rules of organic farming. likewise, gurbuz et al. (2010) reported no unequivocal e�ect of purple cone�ower extract on the level of speci�c antibodies in 60-day-old hyline pullets. in contrast, chaves et al. (2007) and ma et al. (2009) showed the number of speci�c antibodies to increase following purple cone�ower supplementation. immune system e�ciency is determined, among others, from the level of lymphocytes (yurong et al., 2005). according to wang et al. (2000), lymphocytes play a very signi�cant role in the body’s immune response, and a change in the proportion of lymphocyte subclasses alters the immune function. yamada et al. (2011) reported purple cone�ower extract to increase the level of lymphocytes. jurkstiene et al. (2004) found that purple cone�ower supplement increased the leukocyte and lymphocyte counts. according to bany et al. (2003), purple cone�ower extract in�uences the level of granulocytes and lymphocytes. our study showed the e�ect of supplemental purple cone�ower on the level of blood heterophils and lymphocytes in z-11 hens. one of the parameters indicative of avian immunity is the heterophil to lymphocyte ratio (h:l). �e purple cone�ower supplement used in our study reduced the h:l ratio in layers, which may suggest that they were less stressed, more resistant, and showed better welfare compared to control hens. similarly, the h:l ratio was lower in chicks hatched from eggs of hens receiving purple cone�ower diets. during the production process, poultry is constantly exposed to stress factors, including those related to the rearing system. hangalapura et al. (2004a,b) hold that the cellular immune response becomes activated under chronic stress. stress in laying hens decreased the number of white blood cells (mashaly et al., 2004), but this can be counteracted through dietary modi�cation, e.g., the addition of herbal immunostimulants. goel et al. (2002) concluded that biologically active substances found in purple cone�ower can be used to improve the body’s immune potential and to increase macrophage activity. however, our study showed no e�ect of dietary purple cone�ower on selected blood count parameters of organically raised ‘greenleg partridge’ hens. in summary, the organically raised ‘greenleg partridge’ hens and chicks responded to the purple cone�ower supplement by changes in blood picture, which are impors elected blood param eters in organically raised hens fed w ith a purple coneflow er supplem ented diet ew a s os nó w ka -c za jk a, iw on a s ko m or uc ha 76 tant indicators of avian health and condition. as in z-11 chicks, purple cone�ower supplemented to the layer diet reduced the h:l ratio and induced a tendency for a higher level of the circulating immunoglobulin complex and igg in blood, which shows their better living comfort and higher immunity. high individual variation occurred for the circulating immunoglobulin complex and igg in the organically raised ‘greenleg partridge’ hens. references allen, p.c. 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(2001). e�ect of short–term thermal stress early in rearing on performance and physiological indicators of broiler chickens. annals of animal science, 1(2), 187–197. świerczewska, e., niemiec, j., noworyta-głowacka, j. (2003). a note on the e�ect of immunostimulation of laying hens on the lysosyme activity in egg white. animal science paper and reports, 21(1), 63–68. truchliński, j., krauze, m., cendrowska-pinkosz, m., modzelewska-banachiewicz, b. (2006a). in�uence of garlic, synthetic 1,2,4-triasole derivative and herbal preparation echinovit c on selected indices of turkey-hens non-speci�c immunity. polish journal of veterinary science, 9(1), 51–55. wang, y.w., field, c.j., sim, j.s. (2000). dietary polyunsaturated fatty acids alter lymphocyte subset proportion and proliferation, serum immunoglobulin g concentration, and immune tissue development in chicks. poultry science, 79, 1741–1748. doi: 10.1093/ps/79.12.1741 west, a.p., jr, herr a.b., bjorkman, p.j. (2004). �e chicken yolk sac igy receptor, a functional equivalent of the mammalian mhc-related fc receptor, is a phospholipase a2 receptor homolog. immunity, 20: 601–610. s elected blood param eters in organically raised hens fed w ith a purple coneflow er supplem ented diet ew a s os nó w ka -c za jk a, iw on a s ko m or uc ha 78 yamada, k., hung, p., park, t.k., park, p.j., lim, b.o. (2011). a comparison of the immunostimulatory e�ects of the medicinal herbs echinacea, ashwagandha and brahmi. journal of ethnopharmacology, 137(1), 231–235. doi: 10.1016/j.jep.2011.05.017. yunis, r., ben-david, a., heller, e.d., cahaner, a. (2000). immunocompetence and viability under commercial conditions of broiler groups di�ering in growth rate and in antibody response to escherichia coli vaccine. poultry science, 79, 810–816. yurong, y., ruiping, s., shimin, z., yibao, j. (2005). e�ect of probiotics on intestinal mucosal immunity and ultrastructure of cecal tonsils of chickens. archives of animal nutrition, 59(4), 237–246. abstract �e aim of the study was to determine the e�ect of dietary addition of dried purple cone�ower (echinacea purpurea (l.) moench) on selected blood parameters of chicks and hens of the native ‘greenleg partridge’ breed (z-11), maintained on a certi�ed poultry farm according to organic farming principles. birds were divided into two groups according to diet: organic layer diet (group 1), and the same diet supplemented with certi�ed purple cone�ower from 20 weeks of age at 10 g/kg feed (group 2). layers were fed the certi�ed layer diet (17% protein and 11.1 mj me/kg diet) following organic farming principles. experimental incubations were performed at 34 weeks of age. blood was collected from laying hens at 34 weeks and from newly hatched chicks at one day of age to determine selected blood parameters. �e organically raised ‘greenleg partridge’ hens and chicks responded to the purple cone�ower supplement by changes in their blood picture, which are important indicators of avian health and condition. as in z-11 chicks, purple cone�ower supplemented to the layer diet reduced the h:l ratio and induced a tendency for a higher level of the circulating immunoglobulin complex and igg in blood, which shows their better living comfort and higher immunity. dietary purple cone�ower was observed to have no e�ect on the level of haematocrit, erythrocytes, and leukocytes. high individual variation occurred for the circulating immunoglobulin complex and igg in the organically raised ‘greenleg partridge’ hens. key words: blood parameters, chicks, hens, immunity, organic production, purple cone�ower received: [2018.07.12] accepted: [2018.12.20] kształtowanie się wybranych parametrów krwi kur utrzymywanych zgodnie z założeniami rolnictwa ekologicznego i żywionych paszą z dodatkiem jeżówki purpurowej streszczenie celem badań była ocena wpływu dodatku do paszy suszonej jeżówki purpurowej (echinacea purpurea (l.) moench) na kształtowanie się wybranych parametrów krwi piskląt oraz kur rasy rodzimej ‘zielononóżka kuropatwiana’ (z-11) utrzymywanych zgodnie z założeniami rolnictwa ekologicznego na certy�kowanej fermie drobiu. ptaki przydzielono do dwóch grup zróżnicowanych pod względem diety: nioski żywiono ekologiczną mieszanką paszową dla kur nieśnych (grupa 1), a w grupie 2 zastosowano od 20 tygodnia życia dodatek do paszy certy�kowanej jeżówki purpurowej w ilości 10 g/kg paszy. nioski żywiono certy�kowaną mieszanką paszową dla kur nieśnych zgodnie z założeniami rolnictwa ekologicznego o zawartości białka 17% i energii metabolicznej wynoszącej 11,1 mj/kg paszy. w 34 tygodniu odchowu przeprowadzono lęgi doświadczalne. również w 34 tygodniu odchowu pobrano krew od kur nieśnych oraz w pierwszym dniu życia nowowyklutych piskląt w celu oznaczenia wybranych parametrów krwi. reakcją na podanie jeżówki purpurowej były zmiany w obrazie krwi kur oraz piskląt rasy ‘zielononóżka kuropatwiana’, utrzymywanych zgodnie z założeniami rolnictwa ekologicznego, będące ważnymi wskaźnikami zdrowotności i kondycji zwierząt. dodatek do paszy dla kur nieśnych jeżówki purpurowej wpłynął podobnie jak w przypadku piskląt z-11 na zawężenie stosunku h:l, a także na wystąpienie tendencji do wyższego poziomu kompleksu immunoglobulinowego oraz igg we krwi, co wskazuje na ich lepszy komfort bytowy i większą odporność. 79 nie wykazano natomiast wpływu jeżówki purpurowej w paszy na kształtowanie się poziomu hematokrytu, erytrocytów oraz leukocytów. stwierdzono występowanie bardzo dużej zmienności osobniczej w przypadku kompleksu immunoglobulinowego oraz igg u kur rasy ‘zielononóżka kuropatwiana’ w chowie ekologicznym. słowa kluczowe: parametry krwi, pisklęta, kury, odporność, produkcja ekologiczna, jeżówka purpurowa information on the authors ewa sosnówka-czajka https://orcid.org/0000-0003-3720-1685 she is an assistant professor at the department of poultry breeding in the national research institute of animal production in cracow. she is interested mainly in poultry welfare and the technology of poultry production. iwona skomorucha https://orcid.org/0000-0003-1241-7703 she is an assistant professor at the department of poultry breeding in the national research institute of animal production in cracow. she focuses also on poultry welfare and the technology of poultry production. s elected blood param eters in organically raised hens fed w ith a purple coneflow er supplem ented diet ew a s os nó w ka -c za jk a, iw on a s ko m or uc ha 80 appendix 1 fig. 1. �e level of the circulating immunoglobulin complex [g/dl]; the error bars denote the standard deviation (± sd) of the mean value; n = 15 fig. 2. �e level of igg [mg/ml]; the error bars denote the standard deviation (± sd) of the mean value; n = 15 81 fig. 3. heterophil to lymphocyte ratio – h:l; signi�cant di�erences between the treatments in groups in hens and chicks are indicated with di�erent letters (a, b – p < 0.05, duncan test); the error bars denote the standard deviation (± sd) of the mean value; n = 15 fig. 4. �e level of the leukocytes [thous/µl]; the error bars denote the standard deviation (± sd) of the mean value; n = 15 s elected blood param eters in organically raised hens fed w ith a purple coneflow er supplem ented diet ew a s os nó w ka -c za jk a, iw on a s ko m or uc ha 82 fig. 5. �e level of the erythrocytes [m/µl]; the error bars denote the standard deviation (± sd) of the mean value; n = 15 fig. 6. �e level of the haemoglobin [%]; the error bars denote the standard deviation (± sd) of the mean value; n = 15 39 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 39–46, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.5 vendula kuchařová1*, ondřej daněk1, miša škorič2, ivana veselá1, jaroslava tomenendálová1 1 department of physiology, university of veterinary and pharmaceutical sciences, brno, czechia, *kucharova.vendula@gmail.com 2department of pathological morphology and parasitology, university of veterinary and pharmaceutical sciences brno, czechia organ toxicity of diethylnitrosamine and capsaicin in mice – in vivo study introduction application of diethylnitrosamine (den) is commonly used in chemically induced animal models for hepatotoxicity and hepatocarcinogenesis. den belongs to a wide range of nitrosamines, substances well known for their general toxic properties. already in the 1930s, freund (1937) described the hepatotoxic e�ect of one of these substances, dimethylnitrosamine. application of den in di�erent doses and regimes was shown to induce histopathological changes in liver and kidneys a�ecting the biochemical parameters of hepatic injury (rezaie et al., 2013; paula-santos et al., 2014; cho et al., 2016; shaker et al., 2016). capsaicin (cap) is the main pungent principle in capsicum fruit, whose physical and chemical properties and biological e�ects have been already described and summarised more than 30 years ago (monsereenusorn et al., 1982). in more recent articles, various biological activities are described, such as thermogenic and weight reducing, analgesic, antioxidant, anti-in�ammatory, hypocholesterolemic and hypolipidemic, anti-diabetic, anti-ulcer, anti-apoptotic, and anti-cancer (basith et al., 2016; srinivasan, 2016). however, the last-mentioned e�ect is still controversial. some authors highlighted a possible dual e�ect of cap in mutagenicity and tumorigenicity (surh, lee, 1996; bode, dong, 2011). bley et al. (2012) highlight the source and method of extraction of cap and other capsaicinoid compounds which may have several impurities responsible for the potentially carcinogenicity e�ect. however, pure cap is a rather chemopreventive substance according to their study (bley et al., 2012). �e aim of our study was to assess the toxicity of den and cap in mice liver and kidney and to evaluate the chemoprotective e�ect of cap on den induced liver and kidney injury. 40 v en du la k uc ha řo vá , o nd ře j d an ěk , m iš a š ko rič , i va na v es el á, j ar os la va t om en en dá lo vá materials and methods chemicals diethylnitrosamine (den, sigma-aldrich) was dissolved in saline solution (0.9% nacl). capsaicin (cap, merck millipore) was �rst dissolved in ethanol to a stock solution, and this was further dissolved in saline to form a �nal solution containing 0.1 cap in 1 ml 1% ethanol solution (dosage 1.5 mg/kg body weight) or a �nal solution containing 0.05 mg cap in 1 ml 1% ethanol solution (dosage 0.75 mg/kg body weight). animals and experimental conditions 51 female icr mice at 4 weeks of age (13–16 g) were obtained from anlab s.r.o. (prague, czech republic). a�er 2 weeks of quarantine and acclimatisation, mice were randomly divided into �ve groups, housed in plastic cages with sawdust as bedding with a 12-h light/dark cycle, and temperature and humidity were monitored twice a day. drinking water and a standard mouse diet (biokron s.r.o., blučina, czech republic) was provided ad libitum throughout the experiment. all experimental procedures were conducted according to the czech animal welfare protection legal guidelines and eu directives. paper rolls were used to provide environmental enrichment. experimental design and sample collection a total of 51 female icr mice were randomly divided into �ve groups. mice from the �rst group (control, n = 11) received the ethanol solution (1%) intraperitoneally (ip) in weeks 3, 5, 7, 9, and 11. mice in the second group (cap, n = 10) received cap ip at a dose of 1.5 mg/kg body weight (bw) in weeks 3 and 5 and due to impaired tolerance the dose was halved in weeks 7, 9, and 11 (0.75 mg/kg bw). �ird group (den-cap, n = 10) den ip at a dose of 25 mg/ kg bw was administered in week 2 and from week 3 until the end of the experiment as in the second group (cap). mice in the fourth group (cap-den, n = 10) received cap ip at a dose of 1.5 mg/kg bw in week 1 and a week a�er den ip at a dose 25 mg/kg bw. in weeks 3, 5, 7, 9, and 11, ip saline was administered. mice in the last group (den, n = 10) received den ip at a dose of 25 mg/ kg bw in week 2 and in weeks 3, 5, 7, 9, and 11 saline ip. all procedures and applications were performed between 8 and 12 hours, and 24 hours a�er the last application all mice were euthanized using ether overdose and decapitation. �roughout the experiment, animals were observed daily to assess their general health, and the body weight was measured weekly. a�er euthanization, blood was collected from each mouse and clotting serum was obtained. complete necropsy was performed, and the livers and kidneys were weighed, examined macroscopically, �xed 41 o rgan toxicity of diethylnitrosam ine and capsaicin in m ice – in vivo study in 10% bu�ered formalin, and representative parts were embedded in para�n wax. biochemical parameters assessment and histological evaluation total serum protein (tp), albumin (alb), and activities of alanine aminotransferase (alt) and aspartate aminotransferase (ast) were assessed by a clinical chemistry analyser, abbott architect c4000 (abbott, abbott park, illinois, u.s.a.). tissue sections of 4 μm were processed and routinely stained with haematoxylins and eosin (h&e). slides were observed under light microscopy by di�erent experienced histopathologists in a blind fashion and results were compared. statistical analysis numerical data was statistically processed by medcalc for windows, version 15.11.0 (medcalc so�ware, ostend, belgium). anova and student t-tests were used. results �ere was no statistically signi�cant di�erence in body and organ weight between control and experimental groups (data not shown). levels of alt, ast activity, total protein, and albumin concentration were not statistically di�erent among control and experimental groups (tab. 1). tab. 1. results of biochemical parameters: tp – total serum protein, alb – albumin, alt – alanine aminotransferase, ast – aspartate aminotransferase; values shown as mean ± sd group tp [g/l] alb [g/l] alt [μkat/l] ast [μkat/l] control 54.48 ±2.03 38.92 ±1.76 0.37 ±0.13 2.43 ±0.72 cap 52.07 ±1.57 37.23 ±1.78 0.36 ±0.08 1.95 ±0.48 cap-den 54.57 ±3.21 38.33 ±2.69 0.35 ±0.12 2.43 ±0.76 den-cap 53.90 ±2.85 38.56 ±2.58 0.32 ±0.08 2.01 ±0.48 den 53.65 ±3.92 37.77 ±3.36 0.35 ±0.10 2.37 ±0.50 �e histopathological examination of the liver revealed multifocal lymphoplasmacytic reaction in parenchyma in the den treated group (fig. 1). cap used as both preventive and therapeutic agent caused a reduction in the number and extent of lesions. in the cap group, mitotic �gures were found suggesting xenobiotic-induced hepatotoxicity or regenerative changes (fig. 2). in the kidneys, den also revealed multifocal lymphoplasmacytic reactions (fig. 3) that have been mitigated by cap. moreover, histopathological observation of the kidney in the den group has revealed granular dystrophy of the renal tubules (fig. 4), which has not been presented in cap treated mice. in the control group, histological changes were observed neither in the liver nor in the kidneys. v en du la k uc ha řo vá , o nd ře j d an ěk , m iš a š ko rič , i va na v es el á, j ar os la va t om en en dá lo vá 42 fig. 1. lymphoplasmacytic reaction in liver parenchyma in den treated group. original magni�cation: a 200×, b 400×; h&e stain. (photo. kuchařová et al.) fig. 2. mitotic �gures in liver parenchyma in cap treated group. original magni�cation: 400×; h&e stain. (photo. kuchařová et al.) fig. 3. lymphoplasmacytic reaction in the renal cortex in den treated group. original magni�cation: a 100×, b 200×; h&e stain (photo. kuchařová et al.) 43 discussion intraperitoneal application of den to adult mice led to increased alt and ast activities during the �rst 24 or 48 hours, suggesting a physiologic reaction to the acute toxicity of this chemical (cho et al., 2016; hanna et al., 2016; shaker et al., 2016). when these parameters were assessed a�er a longer period a�er den application (20 or 40 weeks), the e�ect was not so prominent. sun et al. (2012) described increased alt activity a�er den-initiated and phenobarbital-promoted liver injury with no change in ast activity. however, healy et al. (2016) did not observe any change in alt activity a�er den application in female c57bl/6n mice. in our study, we did not �nd any den-induced changes in alt and ast activity, which was probably caused due to the late sampling a�er den application (11 weeks a�er den administration). capsaicin alone does not produce any changes in ast or alt activity, which is consistent with our results (hassan et al., 2012; bitencourt et al., 2015). in both studies, the e�ect of cap on carbon tetrachloride (ccl4)-induced hepatotoxicity was investigated with discordant results. hassan et al. (2012) found a signi�cant decrease of ccl4-induced elevation of these marker levels in rats, while bitencourt et al. (2015) observed further elevation of liver enzyme activities when cap was administered in mice. in more recent studies, the hepatoprotective e�ect of cap on alcohol or ‘high fat diet’ induced liver injury in mice was described (koneru et al., 2018; şekeroğlu et al., 2018). since we have not shown any e�ect of den on liver enzyme activity, we cannot assess the protective or harmful e�ects of cap. time-related den-induced histological changes were described in detail by paula-santos et al. (2014). �e duration of our experiment allowed the development of only early toxic lesions, and a longer time is needed for proliferative, dysplastic, or neoplastic lesions to development. mohammed et al. (2014) used fig. 4. granular dystrophy of the renal tubules in den treated group. original magni�cation: a 200×, b 400×; h&e stain (photo. kuchařová et al.) o rgan toxicity of diethylnitrosam ine and capsaicin in m ice – in vivo study v en du la k uc ha řo vá , o nd ře j d an ěk , m iš a š ko rič , i va na v es el á, j ar os la va t om en en dá lo vá 44 cap as a preventive and therapeutic agent against den-induced liver injury and hepatocellular carcinoma in rats with a prominent bene�cial e�ect of cap on liver histology, which is consistent with our results. �e renal toxicity of den was described by rezaie et al. (2013) with signs of acute tubular necrosis in histological sections. our results con�rm den-nephrotoxicity with the promising bene�cial e�ect of cap. conclusions in this study, a mild protective e�ect of cap on den-induced hepatotoxicity and nephrotoxicity was shown only in histopathological changes. �e toxicity of cap itself is questionable, and further studies should be performed to verify its chemopreventive potential. acknowledgement �is study was supported by grant iga vfu brno 114/2016/fvl. references basith, s., cui, m., hong, s., choi, s. (2016). harnessing the therapeutic potential of capsaicin and its analogues in pain and other diseases. molecules, 21(8), e966. doi: 10.3390/molecules21080966 bitencourt, s., stradiot, l., verhulst, s., �oen, l., mannaerts, i., van grunsven, l.a. 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(2016). in�uence of sex and developmental stage on acute hepatotoxic and in�ammatory responses to liver procarcinogens in the mouse. toxicology, 373, 30–40. doi: 10.1016/j.tox.2016.10.006 hassan, m.h., edfawy, m., mansour, a., hamed, a.a. (2012). antioxidant and antiapoptotic e�ects of capsaicin against carbon tetrachloride-induced hepatotoxicity in rats. toxicology and industrial health, 28(5), 428–438. doi: 10.1177/0748233711413801 healy, m.e., lahiri, s., hargett, s.r., chow, j.d., byrne, f.l., breen, d.s., kenwood, b.m., taddeo, e.p., lackner, c., caldwell, s.h., hoehn, k.l. (2016). dietary sugar intake increases liver tumor incidence in female mice. scienti�c reports, 6, 22292. doi: 10.1038/srep22292 koneru, m., sahu, b.d., mir, s.m., ravuri, h.g., kuncha, m., mahesh kumar, j., kilari, e.k., sistla, r. 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(2014). n-diethylnitrosamine mouse hepatotoxicity: time-related e�ects on histology and oxidative stress. experimental and toxicologic pathology, 66(9–10), 429–436. doi: 10.1016/j.etp.2014.07.002 rezaie, a., fazlara, a., haghi karamolah, m., shahriari, a., najaf zadeh, h., pashmforosh, m. (2013). e�ects of echinacea purpurea on hepatic and renal toxicity induced by diethylnitrosamine in rats. jundishapur journal of natural pharmaceutical products, 8(2), 60–64. doi: 10.5812/jjnpp.9686 şekeroğlu, v., aydın, b., atlı şekeroğlu, z., özdener kömpe, y. (2018). hepatoprotective e�ects of capsaicin and alpha-tocopherol on mitochondrial function in mice fed a high-fat diet. biomedicine & pharmacotherapy, 98, 821–825. doi: 10.1016/j.biopha.2018.01.026 shaker, m.e., ashamallah, s.a., el-mesery, m. (2016). �e novel c-met inhibitor capmatinib mitigates diethylnitrosamine acute liver injury in mice. toxicology letters, 261, 13–25. doi: 10.1016/j.toxlet.2016.08.015 srinivasan, k. (2016). biological activities of red pepper (capsicum annuum) and its pungent principle capsaicin: a review. critical reviews in food science and nutrition, 56(9), 1488–500. doi: 10.1080/10408398.2013.772090 sun, h., yu, l., wei, h., liu, g. (2012). a novel antihepatitis drug, bicyclol, prevents liver carcinogenesis in diethylnitrosamine-initiated and phenobarbital-promoted mice tumor model. journal of biomedicine and biotechnology, 2012, 584728. doi: 10.1155/2012/584728 surh, y.j., lee, s.s. (1996). capsaicin in hot chili pepper: carcinogen, co-carcinogen or anticarcinogen? food and chemical toxicology, 34(3), 313–316. doi: 10.1016/0278-6915(95)00108-5 abstract diethylnitrosamine (den) is proven to be toxic to kidneys and liver and to act as a potent carcinogen mainly in liver. capsaicin (cap) is an alkaloid produced by capsicum genus plants and is considered to be a protective agent against the toxicity and carcinogenicity of many substances including den. �e aim of this study was to assess the toxicity of den and cap in the livers and kidneys in mice. �e experiment started a�er two weeks of acclimatisation and was conducted according to the czech animal welfare protection legal guidelines. at the end of the experiment, the mice were euthanized and the toxicity of den and cap in the livers and kidneys were analysed. �e histopathological examination of the livers revealed multifocal lymphoplasmacytic reaction in parenchyma in the den treated group. cap used as both a preventive and therapeutic agent caused a reduction in the number and extent of lesions. in the cap group, mitotic �gures were found, indicating xenobiotic-induced hepatotoxicity or regenerative changes. in the kidneys, den also revealed multifocal lymphoplasmacytic reactions that have been mitigated by cap. moreover, histopathological observation of the kidneys in the den group revealed granular dystrophy of the renal tubules, which were not presented in cap treated mice. levels of alt, ast activity, total protein, and albumin concentrations were not statistically di�erent among control and experimental groups. in this study, the mild protective e�ect of cap on den-induced hepatotoxicity and nephrotoxicity was shown only in histopathological changes. �e toxicity of cap itself is questionable, and further studies should be performed to verify its chemopreventive potential. key words: capsaicin, diethylnitrosamine, hepatotoxicity, mice, nephrotoxicity received: [2018.06.07] accepted: [2018.11.21] o rgan toxicity of diethylnitrosam ine and capsaicin in m ice – in vivo study v en du la k uc ha řo vá , o nd ře j d an ěk , m iš a š ko rič , i va na v es el á, j ar os la va t om en en dá lo vá 46 toksyczność narządowa dietylo-nitrozaminy i kapsaicyny u myszy – badanie in vivo streszczenie udowodniono, że dietylo-nitrozamina (den) działa toksycznie na nerki i wątrobę oraz jako silny czynnik rakotwórczy, głównie w wątrobie. kapsaicyna (cap) jest alkaloidem wytwarzanym przez gatunki z rodzaju capsicum i jest uważana za czynnik ochronny przeciwko toksyczności oraz rakotwórczości wielu substancji, w tym również den. celem tego badania była ocena toksyczności den i cap w wątrobie oraz nerkach myszy. eksperyment rozpoczął się po dwóch tygodniach aklimatyzacji i został przeprowadzony zgodnie z czeskimi przepisami prawnymi dotyczącymi ochrony zwierząt. pod koniec doświadczenia myszy uśmiercano, następnie dokonano oceny toksyczności den i cap w wątrobie oraz nerkach. badanie histopatologiczne wątroby wykazało wieloogniskową reakcję limfoplazmatyczną w miąższu w grupie poddanej działaniu den. cap stosowany jako środek zapobiegawczy i leczniczy powodował zmniejszenie liczby oraz rozmiaru zmian. w grupie poddanej cap stwierdzono formy mitotyczne wskazujące na hepatotoksyczność wywołaną ksenobiotykami lub też na zmiany regeneracyjne. w nerkach poddanych den stwierdzono również wieloogniskową reakcję limfoplazmatyczną, która była łagodzona przez cap. ponadto obserwacja histopatologiczna nerki w grupie den ujawniła dystro�ę ziarnistą kanalików nerkowych, której nie przedstawiono u  myszy poddanych cap. poziomy aktywności alt, ast, stężenia białka i albuminy nie były statystycznie różne w grupach kontrolnych i eksperymentalnych. w badaniu tym łagodny ochronny wpływ cap na indukowaną przez den hepatotoksyczność i nefrotoksyczność wykazano tylko w zmianach histopatologicznych. toksyczność samej cap jest wątpliwa i należy przeprowadzić dalsze badania w celu zwery�kowania jej potencjału chemoprewencyjnego. słowa kluczowe: kapsaicyna, dietylo-nitrozoamina, hepatotoksyczność, myszy, nefrotoksyczność information on the authors vendula kuchařová she is a phd student and assistant at the department of physiology at the university of veterinary and pharmaceutical sciences brno. �e topic of her phd study is �e e�ect of capsaicin on tumour progression – in vivo model. ondřej daněk he studies in the master’s degree study programme at the faculty of veterinary medicine at the university of veterinary and pharmaceutical sciences brno. he is interested in cell biology and animal physiology. miša škorič he is an associate professors and leader of the discipline “pathological morphology” at the department of pathological morphology and parasitology at the university of veterinary and pharmaceutical sciences brno. he is a member of esvp (european society of veterinary pathologists), and his research activities include the mycobacterioses of animals, the detection of tissue defects in transplanted pulmonary valve (porcine model), the repair of bone defects, and colorectal anastomoses (rabbit). ivana veselá she is an assistant professor at the department of physiology at the university of veterinary and pharmaceutical sciences brno. she is interested mainly in cell biology and natural substances such as aeruginosin-865, resveratrol, and capsaicin in medicine. jaroslava tomenendálová she is an assistant professor and leader of the discipline “pathological physiology” at the department of physiology at the university of veterinary and pharmaceutical sciences brno. she also focuses on cell biology and natural products, predominantly in cancer prevention and therapy. 17 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 17–23, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.2 anna čuvalová1*, imrich strapáč2, lívia handrová1, vladimír kmeť1 1institute of animal physiology, centre of biosciences of the sas, soltesovej 4/6, 040 01 kosice, slovak republic, *cuvalova@saske.sk 2department of chemistry, biochemistry and biophysics, institute of pharmaceutical chemistry, university of veterinary medicine and pharmacy, komenskeho 73, 041 81 kosice, slovak republic antibiofilm activity of mushroom extracts against staphylococcus aureus f. j. rosen. introduction in recent years, a growing interest has developed in the mechanisms of the action of natural products, because they are a major source of chemical diversity and have provided important therapeutic agents for many bacterial diseases (payne et al., 2007). mushrooms have long been appreciated for their taste, �avour, desirable aroma, texture, and nutraceutical and medicinal attributes (strapáč et al., 2016). moreover, they are a renowned source of products with an array of bioactivities, from antibacterial to antiviral, cytotoxic, anti-in�ammatory, anti-feeding, antifungal or antioxidant and might be a valuable resource in the search of new bioactive extracts to inhibit bio�lm production (martín-rodríguez et al., 2014; alves et al., 2014). in this context, �avonoids and phenolic compounds have been revealed as potential inhibitors of bio�lm formation and the production of virulence factors in the pathogenic bacteria by interfering with quorum sensing mechanisms (nazzaro et al., 2013). �e main factors associated with bio�lm formation are the iron uptake system and adhesive matrix proteins. adhesion is favoured by the presence of virulence factors known as adhesins, which are grouped in a family known as the microbial surface components recognising adhesive matrix molecules (mscramm). major proteins adhesins in this group include �bronectin binding proteins a and b (fnbpa, fnbpb), bone sialoprotein binding protein (bbp), iron regulated surface determinants a and b (isda, isdb), and serine aspartate repeat gene proteins d and e (sdrd, sdre) (rasmussen et al., 2013; cucarella et al., 2001). staphylococcus aureus f. j. rosen also produces cytotoxins and hemolysins (𝛼, 𝛽, 𝛾 and δ), which possess the ability to form pores in host cells enabling lysis. in staphylococci, the expression of a series of toxins and virulence factors are controlled by the accessory gene regulator (agr) system (jarraud et al., 2002). a nn a č uv al ov á, im ric h s tr ap áč l ív ia h an dr ov á v la di m ír k m eť 18 in the present study, a water extract obtained from a sample of macrolepiota procera (scop.) singer, pleurotus ostreatus (jacq.) p. kumm., auricularia auricula-judae (bull.) quél., armillaria mellea (vahl) p. kumm. and laetiporus sulphureus (bull.) murrill was explored for its antibio�lm activity against staphylococcus aureus strains. material and methods bacterial strains �e following staphylococci strains were used in this study from our own laboratory: staphylococcus aureus no. 5 and s. aureus no. 51, isolated from ixodid ticks (acari); s. aureus no. 12, and s. aureus no. 14, isolated from ewe´s milk. all cultures were identi�ed by matrix-assisted laser desorption/ ionization (maldi) biotyper (bruker daltonik, leipzig, germany). all staphylococci strains were cultured at 37°c on blood agar (blood agar base no. 2, oxoid, basingstoke, united kingdom and with 5% de�brinated sheep blood). mushroom extracts �e preparation of mushrooms water extracts were determined as described previously (strapáč et al., 2016). for our analysis, we used 1 kg of freshly harvested fruiting bodies of macrolepiota procera, armillaria mellea and laetiporus sulphureus, collected in the autumn of 2014 in an area of dargov, bankov near košice and ižkovice, respectively, in the slovak republic. �e last two are commercially available mushrooms, pleurotus ostreatus and auricularia auricula-judae. water extracts were prepared by the extraction of 100 mg samples in 2 cm3 of water for 24 h with occasional vigorous stirring at 8°c in a refrigerator. �en, the extracts were �ltered and stored at 4°c. bio�lm production assay for the detection of bio�lm formation, the crystal violet method with nunc maxisorp plates were used (nunc, roskilde, denmark) by a previously published method (o´toole, 2011) with some modi�cations. overnight cultures of staphylococcus aureus were removed from each well and 3 times washed with saline, �xed with methanol, and stained with 0.1% crystal violet. �e bound dye was released with 33% acetic acid, and the optical density (od) at 570 nm was measured by using a synergy ht multi-mode microplate reader (biotek, winooski, vermont, usa). anti-bio�lm activity of mushroom extracts to detect the e�ects of mushrooms water extracts of macrolepiota procera, pleurotus ostreatus, auricularia auricula-judae, armillaria mellea, and laetiporus sulphureus on staphylococcal bio�lm formation, 10 µl of extracts were added to the each well and 19 a ntibiofilm activity of m ushroom extracts against staphylococcus aureus f. j. r osen. the plates were incubated 24 h at 37°c. �e bio�lm quanti�cation has been described above. wells containing medium were used as blank controls. �e percentage of the inhibition of biofilm formation was calculated according to the following equation: (1 – od590 of test / od590 of untreated control) × 100%. polymerase chain reaction �e presence of virulence genes of staphylococci was carried out by polymerase chain reaction (pcr). strains were screened for the following genes: α-hemolysin gene (hla) as described by (jarraud et al., 2002), serine-aspartate repeat proteins e gene (sdre) (sabat et al., 2006), iron-regulated surface determinants a gene (isda) (verkaik et al., 2010) and b gene (isdb) (waryah et al., 2016), bone sialoprotein – binding protein gene (bbp) (tristan et al., 2003), �bronectinbinding protein a gene (fnbpa) (booth et al., 2001), iron-siderophore transporter gene (sirb) (dale et al., 2004), and accessory gene regulator (agr ii) (shopsin et al., 2003). statistical analysis all assays were performed in eight replicates and the means as well as the standard deviations were calculated. a one-way analysis of variance (anova) and tukey’s test were used to compare data utilising statistica 9.0 so�ware (statso�, tulsa, oklahoma, usa). results in present study, genes hla and isda were found to occur in all staphylococci. �e presence of sdre gene was detected in three out of four strains. genes agrii, isdb, bbp and sirb were only detected in strains isolated from ixodid ticks. gene fnbpa was detected in staphylococcus aureus no. 51. �e bio�lm formation of s. aureus strains was reduced by all mushrooms extracts without a�ecting the bacterial growth. �e best results were observed for the armillaria mellea (70.87%), pleurotus ostreatus (67.00%), laetiporus sulphureus (64.14%) and auricularia auricula-judae (62.77%), while macrolepiota procera showed the lowest reduction of bio�lm formation (47.72%). �e extracts reduced bio�lm formation in the range of 47.72–70.87%, which means that the bio�lm was formed in the presence of extracts in the range of 29.13–52.28%. we showed that a more signi�cant anti-bio�lm e�ect of the extracts, except for m. procera, was of staphylococcus aureus isolated from ixodid ticks (82.00%) in comparison to s. aureus isolated from ewe’s milk (50.00%) (tab. 1). �e extract from m. procera had a similar e�ect on strains isolated from ixodid ticks (46.50%) and ewe’s milk (49.00%). a nn a č uv al ov á, im ric h s tr ap áč l ív ia h an dr ov á v la di m ír k m eť 20 tab. 1. �e e�ect of mushrooms extracts on bio�lm formation of staphylococcus aureus f. j. rosen. 5 isolated from ixodid ticks; signi�cant di�erences are indicated with asterisks (* – p < 0.05, ** – p < 0.01, *** – p < 0.001, tukey’s test) name of mushrooms mean value n = 8 standard deviation (±sd) control 0.216 0.100 armillaria mellea 0.037** 0.006 laetiporus sulphureus 0.042** 0.014 pleurotus ostreatus 0.025*** 0.009 auricularia auricula-judae 0.039** 0.010 macrolepiota procera 0.150 0.048 discussion we investigated the main factors associated with bio�lm formation. �e results of our study are in agreement with kateete et al. (2011) who reported that 100% isolates had hla gene and with verkaik et al. (2010) who reported that 100% isolates had isda gene. tristan et al. (2003) reported positivity rates of fnba and bbp among staphylococcus aureus isolates 28.00% and 22.00%. liu et al. (2015) showed that 68.10% of isolates contained the sdre gene. our study focused on the antibio�lm activity of mushrooms extracts. we have con�rmed here the great potential of mushrooms to produce antibio�lm compounds, and we showed good antibio�lm e�ects of aqueous extracts in terms of the reduction of bio�lm formation. similar observations have been made previously by others with di�erent bacteria (kostić et al., 2017), and showed the antibio�lm activity of armillaria mellea extract against pseudomonas aeruginosa. antibio�lm activity was associated with content of phenolic compounds and organic acids. another study that worked on the organic extracts of macrolepiota procera and laetiporus sulphureus showed antibio�lm activity against s. aureus (carvalho et al., 2015). li and dong (2010) reported the inhibition of escherichia coli t. escher. bio�lm formation (73.00%) by auricularia auricula-judae extract. �is is a pioneer study since, as far as we know, there are no reports on the antibio�lm activity by the mushroom extracts of pleurotus ostreatus, against s. aureus; nevertheless, other studies are required to elucidate the mechanism of action. conclusion extracts from mushrooms are a complex of di�erent chemical compounds. an identi�cation and understanding of the mechanisms of mushrooms extracts action will enable their further application to new innovative strategies for the control of microbial contamination and infection via the food chain. 21 acknowledgement �is study was supported by the slovak projects apvv 14-0274 and vega 2/0085/18. references alves, m.j., ferreira, i.c.f.r., lourenço, i., costa, e., martins, a., pintado, m. 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(2003). use of multiplex pcr to identify staphylococcus aureus adhesins involved in human hematogenous infections. journal of clinical microbiology, 41(9), 4465–4467. doi: 10.1128/jcm.41.9.4465-4467.2003 verkaik, n.j., boelens, h.a., de vogel, c.p., tavakol, m., bode, l.g., verbrugh, h.a., van belkum, a., van wamel, w.j. (2010). heterogeneity of the humoral immune response following staphylococcus aureus bacteremia. european journal of clinical microbiology & infectious diseases, 29(5), 509–518. doi: 10.1007/s10096-010-0888-0 waryah, c.b., gogoi-tiwari, j., wells, k., eto, k.y., masoumi, e., costantino, p., kotiw, m., mukkur, t. (2016). diversity of virulence factors associated with west australian methicillin-sensitive staphylococcus aureus isolates of human origin. biomed research international, 1–10. doi: 10.1155/2016/8651918 abstract mushrooms are a renowned source of products with an array of bioactivities, from antibacterial to antiviral, cytotoxic, anti-feeding, antifungal, or antioxidant and might be a valuable resource in the search of new bioactive extracts to inhibit bio�lm production. we demonstrate the e�ect of �ve mushroom water extracts, macrolepiota procera, pleurotus ostreatus, auricularia auricula-judae, armillaria mellea, and laetiporus sulphurous on bio�lm formation of four staphylococcus aureus strains isolated from ixodid ticks (acari) and ewe´s milk. �e pcr was used for the detection of virulence genes (hla, isda, b, bbp, sirb, fnbpa, sdre, agr ii). �e ability of bio�lm formation and anti-bio�lm activity of mushrooms extracts was assessed in a quantitative crystal violet assay. �e bio�lm formation of s. aureus strains was signi�cantly reduced by all mushrooms extracts (p < 0.001). we showed that a more signi�cant anti-bio�lm e�ect of the extracts was of staphylococcus aureus isolated from ixodid ticks in comparison to staphylococcus aureus isolated from ewe´s milk. in the present study, a. mellea, p. ostreatus, l. sulphurous, a. auricula-judae, and m. procera extracts inhibited bio�lm formation by 70.87%, 67.00%, 64.14%, 62.77% and 47.71%, respectively. �e results suggest that compounds in mushrooms extracts might be useful to control and handle detrimental infections caused by animal and human pathogens. key words: bio�lm, fungi, ixodid ticks (acari), milk, staphylococcus aureus received: [2018.05.30] accepted: [2018.11.12] 23 aktywność antybiofilmowa ekstraktów grzybowych przeciw staphylococcus aureus f. j. rosen. streszczenie grzyby są znanym źródłem produktów bioaktywnych – począwszy od środków przeciwbakteryjnych po przeciwwirusowe, cytotoksyczne, przeciwlękowe, przeciwgrzybicze lub przeciwutleniające. mogą być cennym źródłem nowych bioaktywnych ekstraktów poszukiwanych w celu zahamowania produkcji bio�lmu bakteryjnego. w pracy pokazano wpływ pięciu wodnych ekstraktów z macrolepiota procera, pleurotus ostreatus, auricularia auricula-judae, armillaria mellea i laetiporus sulphurous na tworzenie się bio�lmu czterech szczepów staphylococcus aureus, izolowanych z kleszczy (acari) i mleka owczego. do wykrywania genów wirulencji (hla, isda, b, bbp, sirb, fnbpa, sdre, agr ii) zastosowano metodę pcr. zdolność tworzenia bio�lmów i aktywność anty-bio�lmową ekstraktów grzybów oceniano w analizie ilościowej �oletem krystalicznym. tworzenie bio�lmu szczepów s. aureus było znacznie mniejsze w środowisku ekstraktów z grzybów (p < 0,001). wykazaliśmy, że bardziej wrażliwy na działanie anty-bio�lmowe ekstraktów grzybów był s. aureus wyizolowany z kleszczy niż wyizolowany z mleka owczego. w  niniejszych badaniach, a. mellea, p. ostreatus, l. sulphurous, a. auricula-judae i  m. procera, hamowały tworzenie się bio�lmu (o odpowiednio 70,87%, 67,00%, 64,14%, 62,77% i 47,71%). wyniki sugerują, że związki zawarte w wodnych ekstraktach z grzybów mogą być przydatne do kontrolowania i zwalczania szkodliwych infekcji powodowanych przez patogeny zwierzęce i ludzkie. słowa kluczowe: bio�lm, grzyby, kleszcze (acari), mleko, staphylococcus aureus information about authors anna čuvalová she is interested in anti-bio�lm activities of natural compounds using the static and dynamic bio�lm models with resistant staphylococci (mrsa and mrcons), escherichia coli (esbl and cefotaximases) and pseudomonas aeruginosa on various surfaces (plastics, catheters and food grade stainless sheet). imrich strapáč he is interested in bio�lm and anti-bio�lm activities of natural compounds using the static and dynamic bio�lm models with escherichia coli, and resistant staphylococci (mrsa and mrcons). �e area of his interest is genetic ecology and genes encoding factors of virulence, metabolism, and the spreading of these genes. he studies the resistance occurrence in animal, which could serve as a reservoir of antibiotic resistance in indicator bacteria. lívia handrová http://orcid.org/0000-0002-0985-1771 �e main area of her interest is genetic ecology and the spread of antibiotic resistance genes. she studies the resistance occurrence in small mammals, which could serve as a reservoir of antibiotic resistance (esbl, plasmid encoded chinolone resistance, carbapenemases) in indicator bacteria escherichia coli, pseudomonas aeruginosa and staphylococcus spp. vladimír kmeť http://orcid.org/0000-0002-8081-8579 he is interested in bio�lm and anti-bio�lm activities of natural compounds using the static and dynamic bio�lm models with escherichia coli, resistant staphylococci (mrsa and mrcons). �e area of his interest is genetic ecology and gene encoding factors of virulence, metabolism, and the spreading of these genes. he studies the resistance occurrence in animal, which could serve as a reservoir of antibiotic resistance in indicator bacteria. a ntibiofilm activity of m ushroom extracts against staphylococcus aureus f. j. r osen. 144 annales universitatis paedagogicae cracoviensis studia naturae, 1: 144–161, 2016, issn 2543-8832 peter repka department of biology and ecology, matej bel university, 974 01 banská bystrica, tajovského 40, slovakia, repka.peter@gmail.com management and environmental education in veľká fatra national park (comparison with other parks of visegrád group) introduction with regard to nature protection, e�ort is currently being made to promote four basic trends. �e �rst trend is the development of the biological aspects of nature protection through a better knowledge of species and their relationships in the ecological complex in which they are generated. �e second trend is the development of philosophical and ideological aspects of nature protection through new ecological ethics, which explores human relationship to the entire nonhuman world, to earth; from the moral point of view ecological ethics are considered a historical necessity at that stage and development (keller, 2005; kohák, 2006). �e third trend is to strengthen environmental legislative instruments, including the ones in relation to ensuring better information of the population and also building an open democratic society. �e fourth trend is practical and it deals with caring for ecosystems through active procedures related to maintainance, guidance, improvement or restoration of a favorable conservation status of habitats (vološčuk, 2005; kopcová, tuhárska, 2006; sabo et al., 2011). from the above mentioned, the main area of concern is the practical conservation of nature and paying special attention to the protected area (pa). protection methods are based and rely on exact and empirical experience and knowledge. �e aim of nature protection is to achieve biodiversity conservation and promotion of ecological stability in ecosystems with the acceptance of social and economic aspects as part of human society. �e main aim of the study is to evaluate the management of veľká fatra national park, and its comparison with the selected three analogous national parks (np) in the countries of the visegrád four, as well as to evaluate environmental consciousness and environmental edi�cation of students of grammar schools and secondary schools, not only within the region where np is located, but also outside it. �e study also aims to 145 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) evaluate the gained information about the condition of environmental consciousness and edi�cation in nps in individual countries, their reciprocal evaluation and comparison with veľká fatra np. based on the main objective the following partial aims were developed: 1 – to characterise the conditions of veľká fatra np and its current condition with regard to ecological, economic and social impact a�ecting its development; 2 – to evaluate and compare the management of veľká fatra np and analogous nps in visegrád four through the expert system ipam; 3 – to undertake a survey of environmental consciousness of students of grammar and secondary schools in the region veľká fatra np and outside that region, and to compare their evaluation with the environmental consciousness of students of grammar and secondary schools in the selected analogous nps of the visegrád four; 4 – to propose steps and procedures for strengthening and streamlining environmental education of veľká fatra np; 5 – to propose changes that are needed for the improvement and streamlining of the management of veľká fatra np from the point of view of its sustainable development. materials and methods investigated area �e research was focused on four np’s of visegrád four: veľká fatra np 48°59′ n, 19°05′ e (slovakia), bohemian switzerland np 50°53′ n, 14°23′22″ e (czech republic), bieszczady np 49°06′ n, 22°39′ e (poland) and bükk np 48°02′ n, 20°31′ e (hungary) (fig. 1). swot analysis, force �eld and problem analysis in the beginning of research, the method of swot (strengths, weaknesses, opportunities, threats) analysis was used to characterize the conditions in the veľká fatra np. �e study has begun in 2011 and it has been updated throughout the whole research. when the swot analysis was used for evaluating the intensity of mutual relations, a three scale score was kept. �e positive relationship was evaluated on a scale ranging from 1 to 3, and the negative correlation was evaluated on a scale ranging from -3 to 0. subsequently tables were drawn up where there were assessed strengths with opportunities, strengths with threats, weaknesses with opportunities and weaknesses with threats. a�er the aggregation of the results, the matrix interaction of individual components was made out, where based on the results, the most appropriate strategy was suggested, strategy which should be targeted by the management of veľká fatra np in order to achieve certain objectives. in order to assess more accurately, input data about veľká fatra np was evaluated a�er description of the weaknesses and threats by “force �eld analysis” and “problem analysis”. 146 p et er r ep ka methodology ipam a�er summarizing all the relevant information about the condition of the np, the methodology of the system integrated protected area management (ipam) was used to evaluate management. ipam so�ware it based on apache 2.0 web server, which is a widely used http server for the internet that runs on the linux operating system. pa managers and professionals access the system publicly from common web browsers. user identi�cation is performed on the portal ipam (jungmeier et al., 2005). �roughout assessing the management of pa’s via the system ipam the following operation was elected: create a user pro�le on page www.ipam.info → create a pro�le assessment of pa → self-assessment and control of individual areas of management of pa’s in the preparatory phase, the phase of the basic and detailed planning, implementation phase and the phase of connecting into the network → evaluation report → recommendations. procedure assumes knowledge of all areas of management of pa’s by the respondent (vološčuk, švajda, 2008). fig. 1. localization of the national parks selected for the study. 1. bohemian switzerland national park (�e czech republic), 2. bükk national park (hungary), 3. bieszczady national park (poland), 4.veľká fatra national park (slovakia) 147 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) in order to obtain input data, the exploratory method is used – standardised questionnaire or structured interview, in which respondents are being asked questions that have been made out by an ipam (121 questions intended to assess management areas). questions were evaluated in three categories – as completed, as started, as not started. self-assessment is completed by a report about development and standardised recommendation within the management. however, the most detailed management evaluated was the one applied in veľká fatra np. at the same time the recommendations for change within the management were adapted, especially since work was focused mainly on this np. for this reason the conditions in veľká fatra np were evaluated by several evaluation tools for the application management. in the bohemian switzerland np and bükk np recommendations within applied managements were developed for those areas to which ipam system in evaluating of management allotted high priority. recommendations within management of bieszczady np have been developed for all departments, since the ipam system did not assign a high priority for even one of them. survey methodology of environmental consciousness and education a survey of environmental awareness and education took place between students of grammar schools and secondary schools in the areas where the nps are located and outside the regions, over the years 2012–2014. tabular overview of states, cities and municipalities, schools and numbers of respondents for the individual states participating in our survey is given in table 1. �e percentages of respondents for individual states and their region of residence are expressed in table 2. �e ratio of respondents in each country of the region where the np was located and outside the np is almost balanced and represents 53% (695 respondents) and 47% (606 respondents). �e statistical population included 1301 respondents. a survey of environmental consciousness was realised by the method of interview. as chosen method in order to obtain the necessary information, a tool of exploratory research was used; such tool was proposed by us in the form of a non-standardised questionnaire. �e questionnaire consisted of 20 questions which were divided into three areas: p – knowledge about the park (8 questions), e – knowledge about environmental consciousness (5 questions) and o – general knowledge in the area of nature conservation (4 questions). �e �rst three questions were for identi�cation purpose; participants were asked to provide information about age and place of residence. �e questionnaire was distributed in four language versions. in the statistical analysis of the questions in the areas p, e, o, codes from -5 to 5 were assigned. code “-5” was assigned in case the respondents did not answer correctly. code “5” was assigned in case respondents presented correct answer and code “0” when they didn’t know how to answer. �en their average was calculated for each 148 p et er r ep ka tab. 1. general characteristics of the target groups of the study national park/ country �e city in the vicinity of the national park �e city further away from the national park name of school number of respondents bohemian switzerland np/ czech republic šluknov – secondary school of forestry and social vocational school 127 – nymburk middle school of nymburk 133 – kolín middle school of kolín – čáslav middle school and secondary technical school of education – kutná hora middle school of jiří orten bükki np/ hungary heves – catholic school of technology and economics of janos vak bottyán and middle school 99 – debrecen gabor bethlen economic second-ary school and vocational school 94 bieszczady np/ poland ustrzyki dolne – team of high schools of józef piłsudzki and lyceum 170 krosno – municipal team of school no. 4, ii lyceum of the constitution of may 3rd 30 – kraków ii lyceum of king jan iii sobieski 100 kraków x lyceum of the national educa-tion commission 96 great fatra np/ slovakia žilina – trade school of st. �omas aqui-nas 171 martin – middle school of william paulíny-tóth 98 – poprad secondary vocational school “svit“ 118 – lučenec middle school of božena slančíko-va-timrava 65 total 6 8 15 1301 tab. 2. division of respondents according to their region of residence residence czech republic hungary poland slovakia total village city village city village city village city in the vicinity of the national park 36 91 50 49 96 104 130 139 695 further away from the national park 60 73 26 68 77 119 65 118 606 total 96 164 76 117 173 223 195 257 1301 percentage 7 13 6 9 13 17 15 20 100 149 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) area individually. �e questions were processed in the statistical so�ware ibm spss statistics 19 and in ms excel 2007. at the beginning, groups were studied by homogeneity test of variance of several independent random choices (leven’s test). leven’s test results have con�rmed that the di�erent groups are not homogeneous. moreover, not all of the groups were the same size. �erefore, in other analyses, non-parametric tests had to be used. in the case of all tests, there is a signi�cant di�erence at p ≤ 0.05. a�er evaluating all the answers within the survey three null hypotheses were determined: h01 = in response to the questions concerning the following aspects: p – students awareness about the park, e – environmental consciousness of the students, o – general knowledge in the area of nature protection, there is no signi�cant di�erence between students from the city and the village; h02 = in response to the questions concerning the following aspects: p, e, o, there is no signi�cant di�erence between students from the region of the np and outside the np; h03 = in responses to the questions concerning the following aspects: p, e, o, there is no signi�cant di�erence between students from individual countries. based on h01, h02, h03 alternative hypotheses (h1, h2, h3) were speci�ed. hypothesis h01 and h02 were tested using the mann-whitney’s test and h03 hypothesis was tested using the kruskal-wallis test. additionally (individually for each country and collectively for all countries) statistical signi�cances of the level of students knowledge were compared, through the number of points obtained in the questions: “how many national parks are there in your country?” and “what is the logo of the analysed national park?” (kruskal-wallis test), “what is the logo of the analysed national park?” at individual states level (kruskal-wallis test and mann-whitney’s test), “what sources did you obtain the information about the national park from?” and “which attractions do you use most o�en when visiting the national park?” (friedman’s test), “would you like to engage in environmentally bene�cial activities organised by the management of the national park?”, “do you agree with the principles of conservation in the national park?” and also the question “will closing the national park as an institution have a negative impact on the region?”; a comparision has been made between all countries, as well as between individual countries (wilcoxon’s test); �nally, the order for the individual countries in the quantity of the most frequently presented responses to these questions was set. results swot analysis of veľká fatra national park a�er assigning assessment scales, the relationship between the various constituents was reached a�er summing up all the values into one �nal amount for each comparison. when comparing strengths with opportunities, 895 positive values were obtained. 150 p et er r ep ka when comparing strengths with threats, the negative value of -690 was reached. when comparing weaknesses with opportunities 239 positive values were obtained, and a�er comparing weaknesses with threats we got 754 positive values (fig.  2). a�er the summary of the �nal results, for individual components was designed the matrix of interactions of individual components, where based on the highest value of the strengths the most appropriate strategy was suggested which the administration of veľká fatra np should focus on and follow in order to achieve de�ned objectives. �e most appropriate recommendation was to use the strategy maxi-maxi. in working out the “force �eld” analysis, the swot analyses were based on de�ned weaknesses and threats. for each de�ned problem, the worst development of the issue was characterised, in case it is not possible within the management to reduce the forces which are deepening it. also a possible alternative was suggested as a solution for these de�ned problems. �e analysis showed that the most de�ned problems concerning the prevention of pollution in the pa, support the development of tourism and so� tourism. comparison of considered management through ipam system in surveyed national parks to view the di�erences between the various nps in the achievement of the performance, the �ve phases of management systems assessment ipam were monitored, and the graphical comparison was made (fig. 3). at all stages the best percentage score was achieved by bieszczady np. in the pre-phase the weakest rating was obtained by fig. 2. comparison of swot analysis components based on obtained points n um be r of p oi nt s 151 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) bohemian switzerland np. in the phase of basic planning and at the phase of connection in the network the worst rating was reached by the veľká fatra np. in the detailed planning phase and the implementation phase bükk np had the weakest percentage evaluated. in the pre-phase, in the detailed phase and in the implementation phase, veľká fatra np had the second best score. environmental consciousness and edi�cation (education) in the surveyed national parks comparing whether there is a signi�cant di�erence in knowledge between all students from city and village in the aspects: p, e, o together, in all countries were obtained the following results of mann-whitney’s test: in area p: z = -0.581; p = 0.561 > 0.05 → h01; in area e: z = -0.761; p = 0.447 > 0.05 → h01; in area o: z = -0.052; p = 0.958 > 0.05 → h01. based on these results in all three areas h01 was adopted. in the evaluation of the di�erences between students from the region of the nps and out of the region in all aspects p, e, o together, in all countries were obtained the following results of mann-whitney’s test: in area p: z = -1.303; p = 0.193 > 0.05 → h02; in area e: z = -0.764; p = 0.445 > 0.05 → h02; in area o: z = -2.914; p = 0.004 < 0.05 → h2. based on these results, the hypothesis h02 was rejected and adopted the alternative hyfig. 3. comparison of percentage of performance in the individual management phases in evaluation system ipam in all national parks; bs np – bohemian switzerland national park, bnp – bükki national park, bdnp – bieszczady national park, vf np – veľká fatra national park management phases pe rc en ta ge 152 p et er r ep ka pothesis h2. in aspect o students from the region of the nps introduced more answers which show that they understand the issue of nature conservation than the students from the region outside the nps in all surveyed countries. when comparing whether there is a di�erence in knowledge among all students in the aspects p, e, o together, between individual countries were obtained the following results of kruskal-wallis test: in area p: χ2 = 201.461, df = 3, p = 0.000 < 0.05 → h3; in area e: χ2 = 22.461, df = 3, p = 0.000 < 0.05 → h3; in area o: χ2 = 225.201, df = 3, p = 0.000 < 0.05 → h3. based on these results, the hypothesis h03 was rejected and alternative hypothesis h3 was adopted in all areas. di�erences in responses under the questions “how many national parks are in your country?” and “what is the logo of the analysed national park?” were statistically signi�cant (kruskal-wallis test for the �rst question: χ2 = 315.856, df = 3, p = 0.000 < 0.05, for the second question: χ2 = 320.197, df = 3, p = 0.000 < 0.05). under the �rst question the order was: poland, czech republic, slovakia, hungary and for the second question: czech republic, poland, hungary, slovakia. �e accuracy of the answers to the question “what is the logo of the analysed national park?” is a signi�cant di�erence between all surveyed countries (kruskal-wallis test: χ2 = 320.197, df = 3, p = 0.000 < 0.05). �e order was as follows: czech republic, poland, hungary and slovakia. when testing the di�erences between individual countries, the results were as follows: czech republic – poland (mann-whitney’s test: z = -1.657, p = 0.097 > 0.05; is not a signi�cant di�erence), hungary – poland (mann-whitney’s test: z = -9.294, p = 0.000 < 0.05; is signi�cant di�erence), hungary – slovakia (mann-whitney’s test: z = -2.302, p = 0.021 < 0.05; is signi�cant di�erence). �e order of the answers in the question “what sources did you obtain the information about the national park from?” was clear: school, internet, at home, media (tv, radio), publications (friedman’s test, χ2 = 722.559, df = 4, p = 0.000 < 0.05). �e order of the answers in the question “which attractions do you use most o�en when visiting the national park?” was clear: hiking trails, other (cultural monuments, ski resorts), spa (friedman’s test, χ2 = 1096.653, df = 4, p = 0.000 < 0.05). when testing the di�erence between the questions “would you like to engage in environmentally bene�cial activities organised by the management of the national park?”, “do you agree with the principles of conservation in the national park?” and “will closing the national park as an institution have a negative impact on the region?”, a clear di�erence was con�rmed (wilcoxon’s test: for the �rst and second question – z = -17.440, p = 0.000 < 0.05; for the second and third question – z = -5.658, p = 0.000 < 0.05). testing the di�erences between the �rst and the second question and between the second and the third question for individual countries, a clear di�erence in all countries was con�rmed (wilcoxon’s test: czech republic, for the �rst and second question – z = -7.160, p = 0.000 < 0.05, for the second and third question – z = -2.500, 153 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) p = 0.012 < 0.05; hungary, for the �rst and second question – z = -3.780, p = 0.000 < 0.05, for the second and third question – z = -3.130, p = 0.002 < 0.05; poland, for the �rst and second question – z = -10.818, p = 0.000 < 0.05, for the second and third question – z = -2.000, p = 0.046 < 0.05; slovakia, for the �rst and second question – z = -1.103, p = 0.000 < 0.05, for the second and third question – z = -3.545, p  = 0.000 < 0.05). regarding the number of responses which are indicator of su�cient knowledge of the respondents in activities bene�cial to the np among individual countries, they were in the following sequence: the �rst question – hungary, czech republic, slovakia and poland, the second question – slovakia, poland, czech republic, hungary, and the third question – poland, slovakia, czech republic and hungary. comparing the level of students knowledge of all the countries of the visegrád four, which was expressed by the number of points obtained in the questionnaire for individual areas p, e, o, students achieved the best results in the aspect e (50% of respondents reached less or more than 12.5 points), and the worst results in the aspect o (50% of respondents reached less or more than 7.5 points) (fig. 3). it also detected in aspect e the greatest dispersion of results in comparison to other aspects. �ere are respondents who have extremely low knowledge in comparison to other aspects, some of them achieve even negative values. discussion swot analysis based on the results of the swot analysis (fig. 2), maxi-maxi was selected as the best strategy, aiming to maximize the usage of the opportunities – its strengths and it focuses on their continuing strengthening and also on predicting the development of the environment in the np, either internal or external (lesáková, 2004; papula, 2005). �e strengths of veľká fatra np are in the size, condition and preservation of its area. �erefore, the individual measures should aim to support the protection of this area with regard to the development of economical tourism (považan et al., 2014). �rough summarizing and naming all the �ndings of veľká fatra np, it is possible to divide the negatives named by this analysis into: problems connected with anthropic e�ects, complications of vulnerable ecosystems, the disadvantages associated in the park territory with land owners and with other stakeholders. complications are due to the lack of environmental education and problems are connected with the absence of strategies and strategic documents (in various �elds) necessary for the management of the park. �e administration of veľká fatra np should therefore focus not only on the education and edi�cation of pupils and students, but also surrounding population and other stakeholders (powell et al., 2011; stern et al., 2012; weiland, morrison, 2013; stern et al., 2014). in addition to the activities that must be implemented by employees 154 p et er r ep ka of the administration of veľká fatra np, whenever possible, the landowners must be informed of the vulnerability of the ecosystems, as well as the care they require. �rough “problem analysis” the reasons why the negatives arised were speci�ed. on the other hand, objectives on how to resolve these issues were characterised. as key solution the education of all target groups was proposed, so as to achieve sustainable development of veľká fatra np: strengthening the competence of veľká fatra np as an extension of the area that the administration of the care program of the np will be able to manage; the implementation of measures aimed to the extension of competences of the care program of the np; strengthening the promotion of the park (repiský, švajda, 2012). �e recommendation of the ipam system for the di�erent phases of management in the national park �e most detailed management applied in the veľká fatra np was evaluated and also recommendations for changes within the management were adapted, especially since the presented work focused on this speci�c np. in the bohemian switzerland np and bükk np recommendations were worked out within applied managements for those areas to which the ipam system in evaluating management allotted high priority. recommendations under management of bieszczady np have been developed for all areas, since the ipam system did not assign a high priority to even one of them (fig. 3; tab. 3). tab. 3. recommendations of ipam system for the analysed national parks, concerning each phase of management basic planning phase – working out of the plan step by step. detailed planning phase – working out regional economic programs, speci�c planning, support for investment through appropriate incentive. implementation phase – recommends the creation of the custom module for internal control needs, focus on the preparation of new �nancing methods, – in the �eld of communication and participation iii of the reassessment forms of communication, – field of regional development protected areas aims to create and by sequel implementation of the regional economic program, – in the �eld of the �nancing and �nancial plan recommended to work out a  business plan of the organization, – in the �eld of impact appraisal and limitations it is necessary to work out overview of potential threats within the environment in the protected areas. networking phase – connection to the networking general, connection to the networking economic, connection to the networking social. 155 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) environmental consciousness and environmental edi�cation of students when comparing whether there is a signi�cant diference in knowledge between all students from city and village in areas: p – awareness of the park, e – environmental consciousness, o – common knowledge regarding the protection of nature, within all the states, there was not a statistically signi�cant di�erence. it is considered that the possibility of access to information of students living in the village and in the city is similar, because they have similar options concerning access to the internet, as well as to other media. while evaluating the di�erence between the students within the region where the np is located and outside that region, in areas p, e, o together, in all countries was found a signi�cant statistical di�erence only in area o. in this area students from the region of the np indicated more answers which show that they understand the topic of the protection of nature like the students from outside the np region. �erefore it was asserted that the nps as local centers of environmental education have an impact on the understanding of the need to protect nature, but their educational activity is insu�cient in areas p and e for all the groups of respondents, and in area o the nps should focus on strengthening the educational action for students in regions outside the np. strengthening of cooperation with schools was recommended, because the respondents answered the question “what sources did you obtain the information about the national park from?” in the following sequence: school, internet, home, media and publications. furthermore, managements of the nps should focus on updating websites and making them more attractive not only to the monitored target group, but also to others (haas et al., 2008; urban, 2009; urban, bitušík, 2015). �e signi�cant statistical di�erences were found in comparing knowledge between the students of all countries in areas p, e, o – together and between individual countries. while evaluating the environmental consciousness and the edi�cation of students, the weakest results were obtained by students from hungary and the best ones by students from poland (fig. 4). it was assumed, that this condition is a�ected by the fact that in poland, the teaching of environmental education is a part of training at middle and high schools. �ere is a curriculum which is closely and strictly de�ned and which students have to manage (guziak, 2012). within this subject the training focuses also on the knowledge of the region of residence (stern et al., 2012). in czech republic, the environmental education in middle and high schools is coordinated by ecological education coordinators; each school has such coordinators and this helps teachers to apply ecological education into each subject. it places great emphasis on the professionalisation of environmental education coordinators (mosely, reinke, 2005; švecová 2008). 156 p et er r ep ka in slovakia, environmental education is present in middle and high schools and it is a part of the curriculum, but it is up to the teachers to what extent they will abide to it. as a cross-cutting theme in slovakia the following problems are encountered at the moment: formalism, super�ciality, implementation of complexity, incompleteness and fragmentation of information and the lack of teacher quali�cation (kancír, suchá, 2012). in hungary there is a tuitional subject called “getting to know nature”, where students learn how to build a relationship with nature. �is subject is not required in the curriculum, but it is recommended and it is therefore up to the teachers, to what extent they will abide to it. within vocational subjects such as biology and ecology, especially in hungary, attention is not paid to this issue of environmental education. ultimately, it was concluded that teachers are the ones responsible for choosing textbooks and teaching methods. �e administration introduces only a list of suitable textbooks (szalatnyai, 2010). on the grounds of the results achieved slovakia, as well as in the hungarian conditions with regard to schools, it was suggested to build a system of environmental coordinators according to the czech model (repka et al., 2014). fig. 4. comparison of the level of students knowledge together for all the countries of the visegrád group, expressed as the amount of points obtained in surveys into areas: knowledge about the park – p (median: 9.5), general environmental students awareness – e (median: 12.5), and understanding the necessity of nature protection – o (median: 7.5); 1 – maximum, 2 – upper quartile, 3 – mediana, 4 – lower quartile, 5 – minimum, 6 – extremum 157 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) �e lack of knowledge possessed by the slovak respondents with regard to the question “how many national parks are there in your country?”, as well as the question “what is the logo of the analysed national park?”, justi�ed that the administration of veľká fatra np is not perceived by the target group in the same form as the other foreign nps administrations which were analysed. likewise, it could be said that within the research dealing with the comparison of np websites of slovakia, czech republic and poland, the level of np websites in slovakia was assessed as rather low (repiský, švajda, 2012). while testing the question “which attractions do you use most o�en when visiting the national park?”, according to research, it was found out that the top places are occupied by hiking trails, followed by other places (such as cultural monuments, ski resorts) and spa. it is necessary to operate seasonal tourist information centers and also to update the information for tourists on the websites of the np regarding weather, accessibility of tourist paths and means of transport (car, bus, train and parking, the availability of paved roads in the np etc.) on the main tourist paths (beregszászi, 1995). most responses, demonstrating that students possess awareness about environmental bene�cial activities organised by the administration of the np, and which are necessary for the protection of the administered area, were given by students from czech republic and hungary, and the least by students from slovakia and poland. �e di�erences in the public interest regarding volunteering in di�erent nps are explained by the fact that in 2014, in the bohemian switzerland np there were 18 voluntary work sessions carried out with involvement of 458 participants. in the bükk np voluntary work is carried out in several places on a particular day. in the veľká fatra np there is approximately one voluntary work session per year in one particular place. in bieszczady np, in 2014, 35 people volunteered to help the np, but in this case volunteering is limited because it is not a one-time and a one-day event, but volunteers will spend more days in activities bene�cial for the np (travel, food and accommodation are reimbursed). furthermore, volunteers have to be consenting adults, they have to have good physical condition etc. in czech republic and hungary the situation in this area is different also because the conditions are not set as strictly as in poland (repka et al., 2014). conclusions in veľká fatra np it is possible to divide negative problems to: disadvantages associated with anthropic e�ects, complications of vulnerable ecosystems, the disadvantages associated with land owners and with other stakeholders in the park. �e complications of education and environmental problems are associated with the absence of policies and strategic documents. for a better functioning of the organisation were suggested the strengthening of sta� of the administration of veľká fatra np – [1]. 158 p et er r ep ka for the assessment and subsequent comparison of applied management in veľká fatra np and analogous nps of visegrad four, it was concluded that bieszczady np was the best evaluated, through the assessment methodology ipam. �e appropriate application of management is also in�uenced by the fact that the bieszczady np has all the strategic documents and has legal personality. in addition to the bene�ts of legal personality, nps in poland are charging fees (hiking trails, caves) which is generating regular bene�ts – [2]. research has con�rmed a statistically signi�cant di�erence in the environmental consciousness of students in surveyed countries within the region where the np is located and outside that reagion. when comparing the results it was found that with regard to the environmental consciousness within the individual questions, students from poland and czech republic were better than the students from slovakia and hungary. �e system of environmental education in poland and in czech republic is well adjusted in schools. – [3]. in the area of environmental education and edi�cation it was recommended to strengthen the cooperation of the np vf with middle and high schools. it is necessary to understand that the np or any other type of pa does not represent the educational system in the country, but it is intended to be helpful in deepening and acquiring experiences of students in areas that school as an institution is not able to provide with quality to the students – [4]. in veľká fatra np was recommended the implementation of measures that will lead to the elimination of defects named within the applied methodology assessment of the management of the pas. �e shortcomings in the management of the np were caused by de�ciency of documents, as well as their inadequate preparation, such as: care program, zoning of the np, tourism strategy both as part of the school programs, as well as regional economic programs – [5]. �e analysis of the certain partial aims [1–5] in this study showed, that there are many opportunities to improve the management of veľká fatra np, especially with regard to its perception as an institution. references beregszászi, p. 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(2008). ohrozenie prírodných systémov a vývoj koncepcií ochrany prírody, integrovaný krajinno-ekologický manažment tatranského národného parku. in: monogra�cké štúdie o národných parkoch 7. zvolen: vydavateľstvo tu. [in slovak] weiland, i.s., morrison, j.a. (2013). �e integration of environmental education into two elementary preservice science methods courses: a content-based and a method-based approach. journal science of teacher education, 24(6), 1023–1407. doi: 10.1007/s10972-013-9336-1 abstract �e present study deals with the evaluation of the management of veľká fatra national park and its comparison with three other selected national parks of the visegrád four: bohemian switzerland national park (czech republic), bieszczady national park (poland) and bükk national park (hungary). �e information about veľká fatra national park was acquired through swot analysis. �e weaknesses and threats found following swot analysis were further evaluated by an analysis of the “force �eld” and “problem analysis”. �e management applied in individual parks was evaluated through the expert system integrated protected area management (ipam) with the aim to improve the present management. another part of the examination is the evaluation of the environmental edi�cation and of the consciousness of students of grammar and secondary schools, both in the region where the national park is located and outside that region. �e examination for this study was carried out between 2010–2014. a survey of the environmental consciousness and edi�cation has been implemented in all surveyed countries and the tool of exploratory method was a questionnaire, which was distributed to 1301 respondents. �e survey managed to con�rm the assumption that in some areas the level of environmental consciousness of the target group is very low. �is situation is justi�ed by the fact that schools do not have a precise de�nition in what form and to what extent attention should be paid to this �eld. another reason is the insu�cient cooperation between schools and national parks. key words: bieszczady national park, bohemian switzerland national park, bükk national park, environmental education, ipam, swot analysis, veľká fatra national park received: [2016.06.12] accepted: [2016.09.25] zarządzanie i edukacja ekologiczna w parku narodowym wielka fatra (porównanie z innymi parkami grupy wyszehradzkiej) streszczenie opracowanie dotyczy oceny zarządzania w parku narodowym (pn) wielka fatra oraz porównania jego jakości z wybranymi trzema pn czwórki wyszehradzkiej: pn czeska szwajcaria (republika czeska), bieszczadzkim pn (polska) oraz pn góry bukowe (węgry). informacje wstępne na temat pn wielka fatra uzyskano dzięki analizie swot. słabe strony zarządzania i zagrożenia, sprecyzowane dzięki analizie swot, zostały następnie ocenione analizą „pola siłowego” oraz analizą „problemową”. w celu poprawy sposobu zarządzania, obecne zarządzanie w poszczególnych pn zostało ocenione przez zintegrowany system zarządzania obszarami chronionymi (integrated protected area management – ipam). drugą częścią badań była ocena wpływu sąsiedztwa pn na edukację i świadomość środowiskową uczniów gimnazjów oraz szkół średnich w regionie, w  którym znajduje się pn, a  także poza tym regionem. badanie przeprowadzono w  latach 2010–2014 we wszystkich czterech analizowanych krajach. zastosowano technikę ankiety, w której narzędziem był kwestionariusz ankiety audytoryjnej, dystrybułowany wśród 1301 respondentów. wyniki badań potwierdziły założenie, że w  niektórych obszarach edukacji środowiskowej poziom świadomości populacji statystycznej był bardzo niski, zarówno w regionie pn, jak i poza tym regionem. jedną z przyczyn takiego stanu jest fakt, że programy szkolne nie de�niują precyzyjnie w jakiej formie i zakresie powinna być realizowana edukacja środowiskowa. innym powodem może być niedostateczna współpraca między szkołami i pn. 161 m anagem ent and environm ental education in v eľká fatra n ational p ark (com parison w ith other parks of v isegrád g roup) słowa kluczowe: bieszczadzki park narodowy, park narodowy czeska szwajcaria, park narodowy góry bukowe, edukacja środowiskowa, ipam, analiza swot, park narodowy wielka fatra information on the author peter repka his scienti�c interests focus on engagement in the management of protected areas, environmental education and consciousness. 191 annales universitatis paedagogicae cracoviensis studia naturae, 4: 191–194, 2019, issn 2543-8832 21st international symposium “parasitic and allergic arthropods – medical and sanitary significance” (may 4–6, 2019, janowiec near vistula river, poland) xxi międzynarodowe sympozjum „stawonogi pasożytnicze, alergogenne i jadowite – znaczenie medyczne i sanitarne” (4–6 maja 2019, janowiec nad wisłą, polska) on may 4–6, 2019, in janowiec near vistula river, the 21st international symposium “parasitic, allergenic and poisonous arthropods – medical and sanitary signi�cance” took place. as every year, the organisers of the symposium included the medical university of lublin and the foundation for the control of ticks and prevention in tick-borne diseases in lublin. �e aim of these cyclical symposia is to present current research conducted in domestic and foreign scienti�c units, on parasitic, allergenic and poisonous arthropods, especially ticks. in this year’s edition, symposium participants also discussed the problems of tick-borne diseases and the search for e�ective ways to prevent and monitor this type of threats. on the �rst day of this year’s congress (may 4), participants were registered and dinner was planned. �e o�cial opening of the 21st symposium by prof. alicja buczek, head of the department and faculty of biology and parasitology of the medical university of lublin, took place in the oblasówka guesthouse on may 5. further proceedings of this year’s symposium were also held there. w  dniach 4–6 maja 2019 roku w  janowcu nad wisłą, odbyło się xxi międzynarodowe sympozjum „stawonogi pasożytnicze, alergogenne i  jadowite – znaczenie medyczne i  sanitarne”. organizatorami sympozjum, jak co roku, były następujące jednostki: uniwersytet medyczny w lublinie oraz fundacja na rzecz zwalczania kleszczy i  pro�laktyki w  chorobach odkleszczowych w lublinie. celem tych cyklicznych sympozjów jest prezentacja aktualnych badań, prowadzonych w  krajowych i  zagranicznych jednostkach naukowych, dotyczących stawonogów pasożytniczych, alergogennych i  jadowitych, w  tym szczególnie kleszczy. w tegorocznej edycji uczestnicy sympozjum dyskutowali również nad problemami chorób odkleszczowych oraz nad poszukiwaniem skutecznych sposobów zapobiegania i monitorowania tego rodzaju zagrożeń. w pierwszym dniu tegorocznego zjazdu (4 maja) odbywała się rejestracja uczestników oraz została zaplanowana kolacja. o�cjalne otwarcie xxi sympozjum przez prof. dr hab. alicję buczek, kierownika katedry oraz zakładu biologii i  parazytologii uniwersytetu medycznego w lublinie, miało 192 r ep or ts �e �rst paper was presented by prof. michał stanko from the institute of parasitology of the slovak academy of sciences, who reported on the many years of research results in monitoring the presence of ticks in kosice. �en, you could listen to another interesting paper on the impact of the electromagnetic �eld on the meadow tick dermacentor reticulatus fabr., delivered by prof. branislav petko from the same scienti�c centre (fig. 1). a�er a short co�ee break, oral presentations continued, among others, by assoc. prof. slawomir pancewicz from the medical university of bialystok discussed the subject of tick-borne diseases in poland, especially lyme disease. as a  practitioner, he presented many facts about this disease, while dispelling the commonly circulating myths miejsce w  pensjonacie oblasówka w  dniu 5 maja. tam również odbywały się dalsze obrady tegorocznego sympozjum. pierwszy referat został zaprezentowany przez prof. dr hab. michała stanko z instytutu parazytologii słowackiej akademii nauk, który zreferował wieloletnie wyniki badań z  monitoringu występowania kleszczy w koszycach. następnie można było wysłuchać drugiego ciekawego referatu, dotyczącego wpływu pola elektromagnetycznego na kleszcza łąkowego dermacentor reticulatus fabr., wygłoszonego przez prof. dr hab. branislava petko z  tego samego ośrodka naukowego (ryc. 1). po krótkiej przerwie kawowej kontynuowano prezentacje ustne, m.in. prof. dr hab. sławomir pancewicz z  uniwersytetu medycznego w  białymstoku omawiał tematykę fig. 1. prof. branislav petko during lecture (photo. a. kocoń) ryc. 1. prof. dr hab. branislav petko w trakcie wykładu (fot. a. kocoń) 193 r eportsabout it. �e interesting subject of rickettsiosis dragged from travelling on the example of african tick fever was also discussed by phd magdalena tudrujek-zdunek from the medical university of lublin. while phd agnieszka pawełczyk from the university of warsaw presented the results of research related to the species diversity of borrelia spirochetes in ixodes ricinus l. ticks collected from humans in 2016–2018. on the same day (may 5) a�er dinner, subsequent papers were delivered, among others by phd students from the pedagogical university of krakow: msc anna kocoń presented preliminary results on ticks attacking domestic dogs and cats in southern poland, and msc natalia malejky-kłusek presented research on the repellent e�ect of plants and substances contained in them on selected beetles – pests of stored cereal grain. at the end of this paper session, phd grzegorz kania from the medical university of lublin discussed the importance of millipedes in nature, and phd elżbieta rożej-pabijan from the pedagogical university of krakow presented research on changes in the species composition of bees on variable-moist meadows (the molinion caeruleae relation w. koch 1926). a  poster session took place on the last day of the symposium (may 6). during this session, symposium participants could learn about interesting topics explored by phd students and other scientists from all over poland. for example, msc slawomir dudek from the silesian medical university illustrated the e�ect of plant extracts on borrelia burgdorferi spirochetes (johnson et al. emend. baranton et al.) under in vitro conchorób odkleszczowych w polsce, szczególnie boreliozy. jako lekarz praktyk, przedstawił wiele faktów na temat tej choroby, dementując jednocześnie powszechnie krążące o  niej mity. interesujący temat riketsjoz zawlekanych z podróży na przykładzie afrykańskiej gorączki odkleszczowej został również omówiony przez dr magdalenę tudrujek-zdunek z uniwersytetu medycznego w  lublinie. natomiast dr agnieszka pawełczyk z  uniwersytetu warszawskiego przedstawiła wyniki badań związanych z  różnorodnością gatunkową krętków borrelia u kleszczy ixodes ricinus l., zebranych z ludzi w latach 2016–2018. tego samego dnia (5 maja) po obiedzie, kolejne referaty wygłosili, m.in. doktoranci z uniwersytetu pedagogicznego w krakowie: mgr anna kocoń zaprezentowała wstępne wyniki na temat kleszczy atakujących psy i  koty domowe na terenie południowej polski oraz mgr natalia malejky-kłusek przedstawiła badania repelentnego wpływu roślin i  substancji w  nich zawartych na wybrane chrząszcze – szkodniki magazynowanego ziarna zbóż. na koniec tej sesji referatowej dr grzegorz kania z uniwersytetu medycznego w  lublinie omówił znaczenie krocionogów w  przyrodzie, a  dr elżbieta rożej-pabijan z  uniwersytetu pedagogicznego w  krakowie zaprezentowała badania zmian składu gatunkowego pszczół na zmienno-wilgotnych łąkach trzęślicowych (związek molinion caeruleae w.koch 1926). w  ostatnim dniu sympozjum (6 maja) odbyła się sesja posterowa. w  trakcie tej sesji uczestnicy sympozjum mogli zapoznać się z  ciekawymi tematami eksplorowanymi przez doktorantów i  innych naukowców z  całej polski. na przykład, mgr sławomir 194 r ep or ts dudek z  śląskiego uniwersytetu medycznego zilustrował wpływ ekstraktów roślinnych na krętki borrelia burgdorferi (johnson et al. emend. baranton et al.) w  warunkach in vitro, mgr aleksandra izdebska zaprezentowała temat repelentnego działania anetolu na kapturnika zbożowca rhyzoptera dominica f. (coleoptera: bostrichidae), mgr anna kocoń przedstawiła zagadnienia bezpieczeństwa człowieka podczas snu – czy na pewno jesteśmy wtedy bezpieczni i wolni od pasożytów?, a  mgr natalia malejky-kłusek zobrazowała tematykę wprowadzania naturalnych wrogów działających na szkodliwe stawonogi magazynowe. w  tegorocznym sympozjum uczestniczyło 30 osób, z  10 instytucji naukowych. w  trakcie całego zjazdu, uczestnicy tegorocznej edycji mieli możliwość przedstawienia swoich wyników badań i jak co roku utworzyły się nowe pomysły współpracy, nie tylko z jednostkami naukowymi w kraju, ale również za granicą. anna kocoń institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland; a_kocon@wp.pl ditions, msc aleksandra izdebska presented the topic of repetitive action of anethole on the rhyzoptera dominica f. (coleoptera: bostrichidae), msc anna kocoń presented the issue of human safety during sleep – are we sure that we are safe and free from parasites?, and msc natalia malejky-kłusek depicted the subject of introducing natural enemies a�ecting harmful warehouse arthropods. 30 people from 10 scienti�c institutions participated in this year’s symposium. during the entire conference, the participants of this year’s edition had the opportunity to present their research results and, as every year, new ideas for cooperation were created, not only with the scienti�c units in the country, but also abroad. 67 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 67–71, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.9 drahomíra sopková*, radoslava vlčková, zuzana andrejčáková, soňa gancarčíková department of anatomy, histology and physiology, institute of physiology, university of veterinary medicine and pharmacy, košice, slovak republic, *drahomira.sopkova@uvlf.sk diet supplementation with flaxseed stimulates gut metabolism in mice introduction �e close relationship between intestinal microbiota and health and diseases has aroused a great interest in the use of probiotics and prebiotics in the nutrition of humans and animals. �e main reason is the modulation of digestive processes via increased counts of bene�cial bacteria and their enzymatic activity (axling et al., 2012; borovská et al., 2013). one of the possibilities of increasing the e�ect of bene�cial bacteria is combining them with polyunsaturated fatty acids (pufas) that are an integral component of cellular membranes and are able to modify the adherence of bacteria to intestinal epithelium (borovská et al., 2013). under our conditions, one of the richest plant sources of ω-3 pufas is the �axseed (linum usitatissimum l.). of total fatty acids (fa) found in �axseed oil, 9% are saturated, 18% monounsaturated, and 73% polyunsaturated fa containing 50−61% of ω-3 pufa α-linolenic acid. at the same time, �axseed is a rich source of �bres and the soluble (slimy) �brous component is considered functional (borovská et al., 2013; martinez et al., 2013). in the digestive tract, it acts as a prebiotic and creates a suitable environment for bacteria bene�cial to health. �e aim of the study was to investigate the e�ects of the forti�cation of the diet with �axseed in a model experiment on mice with a focus on intestinal metabolism. material and methods plant material �e experiment was approved by the state veterinary and food administration of the slovak republic (no 1177/14-22) and carried out on 24 mice of balb/c line, 5 weeks old, allocated to two groups: control group (k-; n = 12) fed a standard diet for mice d ra ho m íra s op ko vá , r ad os la va v lč ko vá , z uz an a a nd re jč ák ov á, s oň a g an ca rč ík ov á 68 st–1 (altromin 1311; velaz; czechia) at a dose of 1.75 g/head/day; and the experimental group (mk; n = 12) fed diet supplemented with crushed �axseed variety flanders for the period of 35 days, at 5% concentration in feed. faeces of animals were collected and examined during the experiment and before euthanasia. �e animals were killed by cervical dislocation and caecum samples were collected for the determination of the concentration of organic acids using the method of capillary isotachophoresis (itp). statistical analysis �e results obtained were evaluated statistically using so�ware graphpad prism 3.0 for windows (graphpad so�ware, san diego, california, usa). �e dynamics of changes in individual parameters were evaluated by repeated measures anova and tukey post hoc tests and the di�erences between the groups by means of t-tests. �e signi�cance level was set to p < 0.05. fig. 1. level of organic acids in the caecum and faeces of groups k and mk; the level of organic acids in mice caecum a�er 35 days of supplementation of �axseed and the dynamics of concentration of acetoacetic, lactic, succinic, acetic, propionic, butyric, and valeric acids in mice faeces; k – group of mice fed exclusively the standard diet; mk – group of mice fed standard diet supplemented with crushed �axseed (5% added to the feed); mean values (mean ± sem) are presented in mmol.l-1; a,b,c,x,y,z di�erences between columns with the same superscript are signi�cant (a,x = p < 0.05; b,y = p < 0.01; c,z = p < 0.001); the superscripts a,b,c indicate intergroup di�erences while superscripts x,y,z indicate di�erences between groups d iet supplem entation w ith flaxseed stim ulates gut m etabolism in m ice 69 results supplementation of diet with �axseed (fas) caused a signi�cant increase in the level of organic acids in mice caecum (acetic acid, p < 0.01; propionic, butyric and valeric acids p < 0.001) in comparison with group k (fig. 1a). examination of faeces of �axseed-fed mice showed increased concentrations of lactic acid on days 7 and 28 (p < 0.01), acetic acid on days 14 (p < 0.05), 21 and 28 (p < 0.01) and butyric acid on days 14 (p < 0.001), 21 and 28 (p < 0.01) of supplementation in comparison with control group k (fig. 1b-d). �e most pronounced e�ect of the supplementation of �axseed on the level of all investigated organic acids was observed on day 28 of the feeding experiment (lactic, acetic, and butyric acids p < 0.01). on day 35 of the experiment, we observed a pronounced decrease in the concentration of acetic (p < 0.001), lactic (p < 0.01), and butyric (p < 0.05) acids in the forti�ed group. discussion a positive in�uence on microbiocenosis of the intestinal tract and thus also on the resistance of the macro-organism has been ascribed inter alia also to the prebiotic component of the additive, the �axseed (smith et al., 2006; hekmatdoost et al., 2008). polyunsaturated fatty acids found in �axseed are capable of a�ecting the adherence of bacteria by the modi�cation of the lipid composition of the intestinal wall or the bacterial cell wall. for example, (yu et al., 2014) reported the stimulation of growth and the adherence of lactobacilli in the digestive tract of mice and (nemcová et al., 2012) found similar results in pigs. �e ω-3 fatty acids reduce intestinal permeability by bacteria and the number of apoptotic cells in ileal mucosa (generoso et al., 2015). �e slimy �brous material in �axseed can serve as a speci�c growth substrate for bene�cial bacteria that slow down the production or absorption of toxic products of metabolism, and additionally, by their action, the �brous material decomposes to volatile fa that ful�l important protective role against infections and maintain the integrity of intestinal mucosa (smith et al., 2006; wong, jenkins, 2007). one of the mechanisms of the inhibition of pathogenic microorganisms by bene�cial micro�ora involves the production of antibacterial substances, such as organic acids. �e concentrations of these acids were signi�cantly increased in the caecum (acetoacetic, acetic, lactic, propionic, and butyric) and faeces (lactic, acetic, butyric) of experimental animals in our study. �e bene�cial bacteria are able to reduce the production of harmful nitrogenous compounds that cause damage to intestinal mucosa; and, in this way, they exert a positive e�ect on intestinal function, improve digestive processes, and subsequently increase the weight gain of animals (serban, 2014). adhesion d ra ho m íra s op ko vá , r ad os la va v lč ko vá , z uz an a a nd re jč ák ov á, s oň a g an ca rč ík ov á 70 of probiotics to intestinal mucosa is important for the promotion of health and the action of bacteria bene�cial to intestinal health can be positively a�ected by ω-3 pufas. �e 35-day forti�cation of mice diet with �axseed stimulated intestinal metabolism and fermentation activity of bene�cial autochtonous intestinal bacteria in mice caecum, which indicates its prospective use for pronounced improvement and protection of animal health. references axling, u., olsson, c., xu, j., fernandez, c., larsson, s., ström, ahrné, s., holm, c., molin, g., berger, k. (2012). green tea powder and lactobacillus plantarum a�ect gut microbiota, lipid metabolism and in�ammation in high-fat fed c57bl/6j mice. nutrition & metabolism, 9, 105. doi: 10.1186/1743-7075-9-105 borovská, d., nemcová, r., gancarčíková, s., koščová, j. (2013). �e synbiotic e�ect of lactobacilli and �axseed on selected intestinal micro�ora and organic acid levels in weaned piglets. microbiology, 2(10), 82–86. generoso de vasconcelos, s., rodrigues, n.m., trindale, l.m., paiva, n.c., cardoso, v.n., carneiro, c.m., ferreira, de matos, a.d., faria, a.m.c., maioli, t.u. (2015). dietary supplementation with omega-3 fatty acid attenuates 5-�uorouracil induced mucositis in mice. lipids in health and disease, 14(54), 1–10. doi: 10.1186/s12944-015-0052-z hekmatdoost, a., feizabadi, m.m., djazayery, a., mirsha�ey, a., eshraghian, m.r., yeganeh, s.m., sedaghat, r., jacobson, k. (2008). �e e�ect of dietary oils on caecal micro�ora in experimental colitis in mice. indian journal of gastroenterology, 27, 186–189 kiarie, e., nyachoti, c.m., slominski, b.a., blank, g. (2007). growth performance, gastrointestinal microbial activity and nutrient digestibility in early-weaned pigs fed diets containing �axseed and carbohydrase enzyme. journal of animal science, 85, 2982–2993. doi: 10.2527/jas.2006-481 martinez, f.a.c., balciunas, e.m., converti, a., cotter, p.d., oliveira, r.p.s. (2013). bacteriocin production by bi�dobacterium spp.: a review. biotechnology advances, 31, 482–488. doi: 10.1016/j. biotechadv. 2013.01.010 nemcová, r., borovská, d., koščová, j., gancarčíková, s., mudroňová, d., buleca, v., pistl, j. (2012). �e e�ect of supplementation of �ax-seed oil on interaction of lactobacillus plantarum – biocenol™ lp96 and escherichia coli o8:k88ab:h9 in the gut of germ-free piglets. research in veterinary science, 93, 39–41. doi: 10.1016/j.rvsc.2011.07.031 serban, d.e. (2014). gastro-intestinal cancers: in�uence of gut microbiota, probiotics and prebiotics. cancer letters, 345(2), 258–270. doi: 10.1016/j.canlet.2013.08.013 wong, j.m., jenkins, d.j. (2007). carbohydrate digestibility and metabolic e�ects. journal of nutrition, 137, 2539–2546. doi: 10.1093/jn/137.11.2539s yu, h-n., zhu, j., wen-sheng, pan, w-s., shen, s-r., shan, w-g., das, u.n. (2014). e�ects of �sh oil with a high content of n-3 polyunsaturated fatty acids on mouse gut microbiota. archives of medicinal research, 45(3), 195–202. doi: 10.1016/j.arcmed.2014.03.008 acknowledgement �e study was supported by the project vega no. 1/0476/16. abstract essential polyunsaturated fatty acids (pufas) in the feed may a�ect the gastrointestinal microbiota. �e present study investigated the e�ect of 35-day supplementation of mice diet with 5% concentration of high-ω-3 pufas in �axseed with a focus on the intestinal metabolism of mice. �e capillary isotachophod iet supplem entation w ith flaxseed stim ulates gut m etabolism in m ice 71 resis method was used for the assessment of the level of organic acids in the gut material and faeces. supplementation of �axseed increased the level of organic acids in the caecum (acetic, propionic, butyric, and valeric acids) and faeces (lactic, acetic, butyric acids). �e most signi�cant e�ect was observed on day 28 of �axseed supplementation. �e investigated additive had a stimulatory e�ect on the intestinal metabolism and fermentation activity of bene�cial bacteria. key words: �ax seed, intestine, mice, microbiota, organic acids received: [2018.05.30] accepted: [2018.12.11] suplementacja diety siemieniem lnianym stymuluje metabolizm jelit u myszy streszczenie niezbędne w  pożywieniu długołańcuchowe wielonienasycone kwasy tłuszczowe (ang. pufas) wpływają na mikro�orę jelitową. celem niniejszej pracy było zbadanie wpływu 35-cio dniowej suplementacji diety siemieniem lnianym o 5% stężeniu ω-3 pufas na metabolizm jelitowy myszy. przy użyciu techniki izotachoforezy kapilarnej (ang. citp) oznaczono stężenie kwasów organicznych w jelicie i kale. suplementacja siemieniem lnianym powodowała wzrost poziomu kwasów organicznych w  kątnicy (kwas octowy, kwas propionowy, kwas masłowy, kwas walerianowy) i kale (kwas mlekowy, kwas octowy, kwas masłowy). największy istotny wpływ obserwowano w 28 dniu suplementacji siemieniem lnianym. badany dodatek siemienia miał stymulujące działanie na metabolizm jelitowy i aktywność procesów fermentacyjnych prowadzonych przez pożyteczne bakterie jelitowe. słowa kluczowe: siemię lniane, jelita, myszy, mikro�ora, kwasy information on the authors drahomíra sopková her scienti�c �eld is aimed at lipid metabolism. radoslava vlčková her scienti�c �eld is focused on animal reproduction. soňa gancarčíková her research is focused on the metabolism of gut environment. zuzana andrejčáková her scienti�c �eld is aimed at animal physiology and biochemistry. 207 �e xxviii international conference entitled “�e importance of the bieszczady national park for scienti�c research and ecological education” was held in zatwarnica from 19th to 21st september. �e conference was organized by the bieszczady national park in cooperation with the połoniny national park (slovakia). �e honorary patronage over the conference was held by the secretary of state, chief nature conservator małgorzata joanna glińska, podkarpackie voivode ewa leniart and the marshal of the podkarpackie voivodship władysław ortyl. �e honorary guest of this year’s conference was prof. fedir hamor from the carpathian biosphere reserve (ukraine). �e conference was opened, on september 19th, by the director of the bieszczady national park, phd ryszard prędki, who warmly welcomed all participants of this year’s conference, especially guests from ukraine and slovakia. a�er the opening, invited guest prof. fedir hamor delivered a lecture “on some aspects of scienti�c activity and ecological education in the carpathian biosphere xxviii international conference “the importance of the bieszczady national park for scientific research and ecological education”, september 19–21, 2019, zatwarnica, poland xxviii międzynarodowa konferencja „znaczenie bieszczadzkiego parku narodowego dla badań naukowych i edukacji ekologicznej”, 19–21 wrzesień 2019, zatwarnica, polska xxviii międzynarodowa konferencja pt. „znaczenie bieszczadzkiego parku narodowego dla badań naukowych i edukacji ekologicznej” odbyła się w zatwarnicy w dniach 19–21 września 2019 roku. organizatorem konferencji był bieszczadzki park narodowy przy współudziale parku narodowego połoniny (słowacja). patronat honorowy nad konferencją objęli sekretarz stanu, główny konserwator przyrody małgorzata joanna glińska, wojewoda podkarpacki ewa leniart oraz marszałek województwa podkarpackiego władysław ortyl. gościem honorowym tegorocznej konferencji był prof. dr hab. fedir hamor z karpackiego rezerwatu biosfery (ukraina). 19 września br., otwarcia konferencji dokonał dyrektor bieszczadzkiego parku narodowego dr ryszard prędki, który serdecznie powitał wszystkich uczestników tegorocznej konferencji, zwłaszcza gości z ukrainy i słowacji. po otwarciu zaproszony gość prof. dr hab. fedir hamor wygłosił referat „o niektórych aspektach działalności naukowej i edukacji ekologicznej w annales universitatis paedagogicae cracoviensis studia naturae, 4: 207–210, 2019, issn 2543-8832 208 r ep or ts reserve (ukraine)” which met with great interest from conference participants. before the co�ee break, two lectures were presented by scientists from ukraine, among others, assoc. prof. oksana maryskevych from the institute of ecology of the carpathians nan of ukraine “prospects for the functioning of the new ukrainian bojkiwszczina national natural park”. in the second scienti�c session, the following papers were presented, among others: “a network of large protected areas on the northern slope of the eastern carpathians and prospects for international cooperation” – prof. platon tretiak from nan state museum of natural history of ukraine, “a synthetic presentation of information on research topics implemented in bdpn in the last decade” – phd eng. stanisław kucharzyk from the bieszczady national park, “diatoms (bacillariophyta) as indicators of the state of water environments – spring and head-streams of river sections of the rivers of the bieszczady national park. �e importance of shortand long-term studies” – assoc. prof. joanna żelazna-wieczorek from the university of lodz. in total, six papers were presented in this session. in the a�ernoon, the conference organisers invited all participants to the newly renovated ecological education field station in suche rzeki. �e grand opening was executed by the director of the bieszczady national park phd ryszard prędki and prof. bogdan zemanek from the jagiellonian university. �e park director thanked everyone who have contributed to the renovation of this facility and invited conference participants for refreshments. karpackim rezerwacie biosfery (ukraina)”, który spotkał się z dużym zainteresowanie ze strony uczestników konferencji. przed przerwą kawową referaty wygłosiło dwóch prelegentów, m.in. doc. dr oksana maryskevych z instytutu ekologii karpat nan ukrainy „perspektywy funkcjonowania nowego ukraińskiego przyrodniczego parku narodowego bojkiwszczina”. w drugiej sesji naukowej, swoje referaty wygłosili, m.in. prof. dr hab. platon tretiak państwowe muzeum przyrodnicze nan ukrainy „sieć dużych obszarów chronionych na północnym stoku karpat wschodnich a perspektywy współpracy międzynarodowej”, dr inż. stanisław kucharzyk bieszczadzki parku narodowy „syntetyczna prezentacja informacji o tematach badawczych realizowanych w bdpn w ostatnim dziesięcioleciu”, dr hab. joanna żelazna-wieczorek uniwersytet łódzki „okrzemki (bacillariophyta) jako wskaźniki stanu środowisk wodnych – źródeł i źródłowych odcinków rzek bieszczadzkiego parku narodowego. znaczenie badań krótkoi długoterminowych”. w tej sesji zaprezentowano w sumie sześć referatów. po południu, organizatorzy konferencji zaprosili wszystkich uczestników do nowo wyremontowanej terenowej stacji edukacji ekologicznej w suchych rzekach. uroczystego otwarcia dokonał dyrektor bieszczadzkiego parku narodowego dr ryszard prędki oraz prof. dr hab. bogdan zemanek z uniwersytetu jagiellońskiego. dyrektor parku serdecznie podziękował wszystkim osobom, które przyczyniły się do remontu niniejszego obiektu oraz zaprosił uczestników konferencji na niewielki poczęstunek. 209 r eportsin the next scienti�c session, which was continued in suche rzeki, the following papers were presented: “ecological education in the bieszczady national park” – phd grażyna holly from the bieszczady national park, “current activities in the �eld of environmental education implemented in the lkp lasy bieszczadzkie” – msc eng. mateusz świerczyński from cisna forest district together with msc eng. ewa wydrzyńska-scelina from baligród forest district, “degradation and renaturalisation of soils a�ected by hiking in the bieszczady national park” – prof. marek drewnik from the jagiellonian university in kraków, “recreation of water fauna in degraded streams: can nature cope alone?” – prof. krzysztof kukuła from the university of rzeszów, “new hieracium species (asteraceae) from bdpn” – prof. zbigniew szeląg from the pedagogical university of krakow and others. nine papers were presented in this session. a�er the paper session, there was time for a poster session. during this session, the results of the research were presented: “�e speci�c architecture of the bieszczady national park” – msc łukasz kielar cracow university of technology and “beaver’s activity and changes in the local �ora in the syhłowaciec stream valley (western bieszczady) – preliminary research results” – msc rita rakowska from the jagiellonian university in kraków. a�er returning to zatwarnica, the participants took part in the integrative meeting. �e next day, september 20th, in the early morning the conference participants went to slovakia to participate in a two-day �eld session. as part of it, they admired the educational qualities of the “great rawka-nova w kolejnej sesji naukowej, którą kontynuowano w suchych rzekach, referaty zaprezentowali: dr grażyna holly bieszczadzki park narodowy „edukacja ekologiczna w bieszczadzkim parku narodowym”, mgr inż. mateusz świerczyński nadleśnictwo cisna wraz z mgr inż. ewa wydrzyńska-scelina nadleśnictwo baligród „obecne działania w zakresie edukacji ekologicznej realizowane na terenie lkp lasy bieszczadzkie”, prof. dr hab. marek drewnik uniwersytet jagielloński w krakowie „degradacja i renaturyzacja gleb znajdujących się pod wpływem turystyki pieszej w bieszczadzkim parku narodowym”, prof. dr hab. krzysztof kukuła uniwersytet rzeszowski „odtwarzanie się fauny wodnej w potokach zdegradowanych: czy przyroda może poradzić sobie sama?”, prof. dr hab. zbigniew szeląg uniwersytet pedagogiczny w krakowie „nowe gatunki hieracium (asteraceae) z bdpn” i inni. w tej sesji zaprezentowano dziewięć referatów. po zakończeniu sesji referatowej nastąpił czas na sesję posterową. w jej trakcie wyniki badań zaprezentowali: mgr łukasz kielar politechnika krakowska „specy�czna architektura bieszczadzkiego parku narodowego” oraz mgr rita rakowska uniwersytet jagielloński w krakowie „działalność bobra a zmiany w lokalnej �orze w dolinie potoku syhłowaciec (bieszczady zachodznie) – wstępne wyniki badań”. po powrocie do zatwarnicy uczestnicy wzięli udział w wieczorze integracyjnym. następnego dnia, tj. 20 września br., uczestnicy konferencji w godzinach wczesno porannych wyjechali na słowację w celu uczestnictwa w dwudniowej sesji terenowej. w jej ramach podziwiali walory edukacyjne transgenicznej polsko-słowackiej ścieżki 210 r ep or ts sedlica” transgenic polish-slovak nature path and the “havesowa” reserve in the połoniny national park. �is year’s xxviii international conference gathered 77 representatives from three countries – poland, ukraine and slovakia. as in previous years, the conference integrated scientists from various countries and �elds of science. all the papers met with great interest from the conference participants. most importantly, the conference showed how important scienti�c research is, but also how nature education is conducted by the bieszczady national park and surrounding forest districts. more information on the conference can be found on the bieszczady national park website and in next year’s issue of the bieszczady yearbooks. przyrodniczej „wielka rawka-nova sedlica” oraz rezerwat „havesowa” w parku narodowym połoniny. tegoroczna xxviii międzynarodowa konferencja zgromadziła 77 przedstawicieli z trzech państw – polski, ukrainy i słowacji. tak jak w ubiegłych latach konferencja integrowała naukowców z różnych państw i dziedzin nauki. wszystkie referaty spotkały się z dużym zainteresowaniem od strony uczestników konferencji. co najważniejsze ukazała jak ważne są badania naukowe ale również edukacja przyrodnicza prowadzona przez park jak i okoliczne nadleśnictwa. więcej informacji na temat konferencji można znaleźć na stronie bieszczadzkiego parku narodowego oraz w przyszłorocznym wydaniu roczników bieszczadzkich. rita rakowska institute of botany, faculty of biology, jagiellonian university, gronostajowa 3 st. 30-387 kraków, rita.rakowska@doctoral.uj.edu.pl 7 annales universitatis paedagogicae cracoviensis studia naturae, 2: 7–26, 2017, issn 2543-8832 doi: 10.24917/25438832.2.1 jan przemysław kubik*, beata barabasz-krasny department of botany, institute of biology, pedagogical university of kraków, podchorążych 2, 30-084 kraków, poland, *shemgodd@gmail.com the succession of abandoned glades and its impact on the diversity of flora in beskid mały mountains (southern poland) introduction for hundreds of years, pasturage was the most optimal way of maintaining pasture plant communities. �e animal species bred on pastures in beskidy mountains, which included sheep, goats, cows, and oxen. in the 18th century, when shepherding was at its peak, tens of thousands of cattle were kept and bred in the beskidy mountains. �e primeval beech and �r forest provided feed for animals during summer and winter as well as wood for fuel and for construction of pens, huts, and houses. �e animals that grazed on the pastures naturally mowed the plants and fertilised the soil, and, at the same time, they provided a livelihood to the shepherds. �e shepherding industry that developed was based on products made from ewe’s milk, meat, wool, leather, and it constituted the backbone for the development of local economies and local communities. in the shepherding huts, butter and cheeses were produced – both so� cheeses such as ‘bundz’ and ‘bryndza’ as well as hard cheeses such as ‘oscypek’ (leszczyński, 1932; pawłowska, 1965; gołek et al., 2015). in the second half of 19th century, the shepherding industry began to decline. �e shepherds, who depended on primeval forest as a  source of feed for their animals, started to feel the e�ects of the industrial revolution. �e forest owners began to cut down trees for construction materials. spruce trees were planted in place of primeval forest tree species, which grew much faster but could not be used as feed for the animals (pawłowska, 1965). on top of that, more and more pastures were converted into farm �elds, and the farmers also began to set up their farms at higher altitudes in the mountains. shepherding experienced its greatest decline at the beginning of 20th century. it slightly rebounded a�er the first world war; however, as a  result of fast economic changes in the countryside, the era of extensive shepherding came to an end at that time (kufa, 2005; gołek et al., 2015). ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 8 poland economic transformation, which took place in 1980s and 1990s, took a  heavy toll on the polish farming industry as whole. due to drops in prices of agricultural products, the pro�tability of farming activities signi�cantly declined during that time. for example, in south-eastern poland, the sheep population declined by 86% between 1987 and 1996. in addition, during those years, production shi�ed from state-owned enterprises into the private sector (cach-czaja, 1998). at that time, many polish farmers reduced the pasture areas by either fallowing them or turning them into farm �elds to produce crops mostly for their own needs (ostromęcki, piechota, 1996). �e abandonment of pastures and the discontinuation of mowing and grazing by animals resulted in the gradual degradation of meadows and pastures, which began to accumulate the overgrown plant matter and dead plant remains (kornaś, 1990). additionally, such fallowed pastures experienced uncontrolled growth of pasture weeds as rumex alpinus, juncus sp., leading to reduction and, ultimately, complete disappearance of plants for grazing and valuable plant species (gołek et al., 2015). �e aim of this paper is to prove that cessation of grazing, mowing, and other usable treatments on mountain glades in the beskid mały mountain range results in the reduction of �oristic diversity of meadows and pasture areas. study area �e studied area includes the mountain massif of łamana skała (929 m.a.s.l.) – 49°45′49.0″n; 19°23′45.7″e and leskowiec (918 m.a.s.l.) – 49°47′19.0″n; 19°26′36.4″e, in beskid mały (southern poland). �e łamana skała group is a part of the beskid mały mountain range, positioned in its central part. it is located to the east from the kocierz group, ranging from the beskidek pass to the skawa river valley, and includes the leskowiec massif (truś, 2008). according to physiogeographic regionalisation by kondracki (2011), this area belongs to the beskidy zachodnie (western beskidy) macroregion, and the beskid mały mezoregion. according to geobotanical division of poland by w. szafer, the studied area belongs to zachodnio-karpacki (western carpathian) section, the beskidy region, and the śląsko-babiogórski sub-region (szafer, zarzycki, 1977). in terms of landscape, the beskid mały is a group of larger and smaller mountain ranges, valleys, as well as a  number of saddles and passes. it spans two altitudinal zones – the �ysch-plateau zone (pogórze �iszowe) (300–600 m.a.s.l.) and the mixed-forest zone (regiel dolny) (600–1150 m.a.s.l.). �e hillsides, especially on the northern side of the range, are quite steep (between 15° and 30°), while crests and peaks are rather �at. �e mountains are being shaped by various erosion processes, including �ysch landslides, wearing down by water, washout of farm �elds, erothe succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 9 sion attributable to changes in temperature, etc. �e area consists in 90% of hard sandstone and shale (łupki godulskie) of medium and lower levels, which formed during lower cretaceous epoch. numerous valleys are covered by brown earth (mostly clays), which, in combination with large number of springs, creeks, and rivers, is conducive to agriculture (ziemońska, 1973; matuszczyk, 1981). besides brown soils (which represent approx. 90% of soils), the beskid mały also has podsolic soils (approx. 5%), rusty soils, and aerosols. �e area also features alluvial soils, delluvial soils, luvisolic soils, pseudo-gley soils, and gley soils (forest management plan…2006/2015). according to division by m. hess (1965), the studied area spans two horizontal mountainous vegetation regions, corresponding to the following two altitudinal zones: the �ysch-plateau zone (250–600 m.a.s.l.), with an average annual temperature of +8°c, a total annual precipitation of 800 mm, and a vegetation period length of 220 days, and the mixed-forest zone (600–1100 m.a.s.l.), with an average annual temperature of +4°c, a total annual precipitation of 1400 mm, and a vegetation period length of 170 days. such areas experience frequent and very strong winds, especially in early spring and autumn, the intensity of which is increasing with altitude. methods studies were conducted from january to october 2015 and from may to july 2016. 54 squares (plots) with area of 25 m2 each (5×5 m) were randomly designated on southern slopes of leskowiec and łamana skała massif in the beskid mały. �e following information was recorded for each plot: the name of the township/glade, geographic coordinates, altitude in meters above sea level, and the date and time of conducting the �oristic survey. geographic coordinates for the plots were taken from a gps device and veri�ed with use of a tourist map of beskid mały in the scale of 1:50000 (2014). altitude in meters above sea level was read from the ‘altimeter’ application and veri�ed with the aforementioned tourist map (appendix 1 – tab. 1). to determine the botanic composition of each plot, the klapp (1965) estimation method was used. �is method was used to determine the percentage share of species in each plot with accuracy of about 1%. all the species that were identi�ed in the given plot were recorded in the following sequence: grasses, legumes, other herb species, as well as trees and shrubs. �e share of individual species within the group was estimated, starting with the most commonly occurring ones. �e species whose share was less than 1% were assigned with the ‘+’ sign. �e samples of species, which were not identi�ed directly in the �eld, were collected and then identi�ed in the laboratory of the department of botany, the institute of biology of the pedagogical university of kraków. during �eld work, particular attention was paid to di�erences between the ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 10 glades that were used and those which were unused. areas located directly beneath power lines as well as areas transformed by human activities or defaced by animals, e.g., by boars were not included in the research. bogs and waterlogged meadows and glades on which activities are conducted only partially, e.g., where self-planted trees or shrubs are cut down and removed, were also excluded from the research. �e �oristic data from all the analysed plots were entered in the turboveg database, and then they were subjected to hierarchical numerical classi�cation (gauch, 2012) based on the percentage coverage of species. classi�cation was performed with use of mulva-5 so�ware package (wildi, orlóci, 1996). similarities among plots were calculated according to van der maarel’s formula, and then the plots were grouped using the ward’s method (minimum variance clustering). �is method uses the variance analysis approach to estimate distance between cluster centres (dzwonko, 2007). in the obtained dendrogram, the groups of plots, which had the same quantitative coverage of species, were separated. for the individual groups of plots, weighted average values of ellenberg indicator values were calculated, based on percentage coverage by species, for light (l), moisture (f), soil ph (r), and nitrogen content in soil (n) (ellenberg et al., 1992). in addition, shannon-weaver general diversity indicators (h) (shannon-weaver, 1963), pielou uniformity indicators (j) (pielou, 1975), and dominance indicators (c) (simpson, 1949; shannon, weaver, 1963) were compared. for biodiversity indicators, a�er levene’s previous test, the statistical signi�cances between the groups were determined using the kruskal-wallis nonparametric test (for h and j) and the one-way anova parametric test (p < 0.05). plant names according to mirek et al. (2002). fig. 1. classi�cation of the studied plots based on the percentage coverage of species; a – pasture with nardus stricta; b – hay-meadow; c – unused plots with pteridium aquilinum; d – unused plots with shrubs the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 11 results hierarchical numerical classi�cation, which was performed on the basis of percentage species coverage in the plots, yielded the following four utility-�oristic groups: a  – pasture with nardus stricta, b – hay-meadow, c – unused plots with pteridium aquilinum, and d – unused plots with shrubs (fig. 1). �e �oristic characteristics and the quantitative share of species in the aforementioned groups are shown in tables 2–5 (appendix 1). �e �rst two groups distinguished on the study area were a – pasture with nardus stricta, and b – hay-meadow, which include the glades on which grazing, mowing and other usable treatments are still conducted. �ey have a higher average number of species in the plots – ranging from 21 to 22 (tab. 6). �ey are dominated by grasses such as festuca rubra and poa trivialis (appendix 1 – tab. 2–3). �e other two groups of plots (c and d) are unused glades. �e highest total number of species was recorded in unused plots with shrubs d – 69, and the lowest was recorded in unused plots with pteridium aquilinum c (fig. 2). fig. 2. total number of species in �oristic tables (appendix 1 – tab. 2–5) of the distinguished groups of plots 42 53 37 69 10 17 5 22 0 20 40 60 80 pasture with nardus stricta hay-meadow unused plots with pteridium aquilinum unused plots with shrubs number of plots number of species �e comparison of diversity, uniformity, and dominance indicators (tab. 6) has shown that the highest value of the (h) indicator was recorded in pasture with nardus stricta, and the lowest was recorded in unused plots with pteridium aquilinum. �e greatest uniformity was recorded in unused plots with shrubs and ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 12 pasture with nardus stricta, and the smallest in unused plots with pteridium aquilinum. �e highest dominance indicator was recorded in unused plots with pteridium aquilinum and in unused plots with shrubs, the lowest in multi-species plots of hay-meadow. tab. 6. average values of shannon-weaver general diversity indicators – h, uniformity – j and dominance – c, calculated for groups of plots distinguished on studied glades of beskid mały; in column for h and j no statistical signi�cance with kruskal-wallis test, p < 0.05; for c – a, b statistical signi�cance with tukey test, p < 0.05 group name of plots average number of species in plot ± sd biodiversity indicators (values and ranges) h j c a – pasture with nardus stricta 21 ±5.37 2.69 2.48–3.04 0.91 0.79–1.07 0.07ab 0.03–0.17 b – hay-meadow 22 ±2.43 2.68 2.32–3.02 0.87 0.75–1.01 0.05b 0.04–0.09 c – unused plots with pteridium aquilinum 18 ±3.27 2.46 2.09–2.81 0.86 0.74–0.97 0.11ab 0.06–0.14 d – unused plots with shrubs 18 ±3.94 2.62 2.07–2.86 0.91 0.78–0.99 0.11a 0.04–0.44 tab. 7. average values of ellenberg indicators (light – l, moisture – f, soil ph – r, nitrogen content in soil – n) calculated for groups of plots distinguished on studied glades of beskid mały group name of plots ellenberg indicators (values and ranges) l f r n a – pasture with nardus stricta 7.13 6.96–7.33 5.68 5.37–6.03 5.20 4.65–6.07 5.04 4.71–5.68 b – hay-meadow 7.10 6.81–7.41 5.50 5.25–5.88 5.90 5.33–6.52 5.68 4.90–6.43 c – unused plots with pteridium aquilinum 6.56 6.14–7.04 5.44 5.20–5.79 3.99 3.65–4.19 4.06 3.73–4.46 d – unused plots with shrubs 6.71 5.81–7.43 5.92 4.96–7.08 4.59 2.78–6.50 4.68 3.60–6.38 �e comparison of average values of ellenberg indicators (tab. 7) has shown that the greatest number of photophilous species occurred in plots of pastures with nardus stricta, and the smallest number of such species occurred in unused plots with pteridium aquilinum. in those plots, the values of indicators for light (l) were lowest. �e highest average indicator for moisture (f) was recorded in unused plots with shrubs, and the lowest was recorded in plots with pteridium aquilinum. �e highest average values of soil ph indicator (r) and the nitrogen content in soil (n) were recorded in hay-meadows, and the lowest values of those indicators were recorded in plots with pteridium aquilinum. the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 13 discussion pastures and glades in beskidy mountains were created arti�cially as a result of grazing, mowing, and other usable treatments conducted by the people who had been migrating to that area since the middle ages. initially, their activities were limited to cutting down forests in valleys and at the foothills of mountain ranges, to set up farm �elds. later periods, namely 14th and 15th centuries, witnessed the wave of vlach migrations that came from romania along the carpathian arc. �ey began to cut down forests at higher altitudes to make space for pastures, some of which were still used as recently as the beginning of the 20th century. extensive pasturage activities resulted in the emergence of diverse plant communities, which are categorised as semi-natural (pelc, 1958; pawłowska, 1965). �ey were dominated by domestic species that needed a  speci�c form of human activities, such as mowing or shepherding (gołębiowski, 1990; michalik, 1990). in the past, the areas covered by the botanical composition survey mostly included gladiolo-agrostietum (br.-bl. 1930) pawł. et wal. 1949 meadows at di�erent stages of their transition towards anthoxantho-agrostietum sillinger 1933 pastures mentioned in, among others, czech sources – hájek (2007) but not mentioned by matuszkiewicz (2007), as well as poor in nutrients hieracio-nardetum kornaś 1955 n.n. em. balcerk. 1984 swards. �e direction of the transition of those phytocoenoses depended on the manner of their usage, and the intensity of usage and maintenance activities conducted (zwolińska, 1960; kornaś, 1990; rozbrojová et al., 2010; gołek et al., 2015). �e botanical composition of one of the groups that was determined on the analysed area, referred to in this study as pasture with nardus stricta – a (appendix 1 – tab. 2), is similar to hieracio-nardetum kornaś 1955 n.n. em. balcerk. 1984 sward (matuszkiewicz, 2007), which had been said to occur in that area. �e phytocoenoses of that kind usually developed in places with extensive sheep pasturage and with insu�cient organic fertilisation, and this is con�rmed by relatively lower values of the soil nitrogen (n) indicator among other used plots (tab. 7). �e soils, on which such plant communities emerge, are barren and poor in nutrients. �ey give hay of very poor quality, and they very quickly grow over when unused. such swards serve the erosion-prevention function. nardus stricta is a grass with very thin leaf-blades, whose roots and shoots are very closely packed together at the base of the plant. in the past, pastures with nardus stricta were a very widespread plant community that developed on the areas with pasturage activities. currently, they are becoming increasingly hard to come by, and they occupy small surfaces. �ey can still be found next to shepherds huts, in sheep-pens, and on the edges of the glades (michalik, 1990; rozbrojová et al., 2010; gołek et al., 2015). �eir relatively high species richness shows in the dynamic changes occurring here as a result of the cessation of traditional management (tab. 6). ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 14 other species identi�ed in the plots of that type include potentilla anserina, festuca rubra, anthoxanthum odoratum, and carex rostrata (appendix 1 – tab. 2). �e most important plant community of the mountain glades of the mixed-forest zone in the western carpathians is still the gladiolo-agrostietum meadow (kornaś, 1967; kornaś, medwecka-kornaś, 1967). until recently, this plant community dominated on almost every mountain glade. currently, due to discontinuation of usage of the glades, the patches of typical gladiolo-agrostietum meadows are becoming increasingly rare. gladiolo-agrostietum is a  plant community of hay-meadows, which better serve as a  source of hay rather than a  pasture. it develops on glades that are moved once or twice a  year, where the grazing takes place only a�er mowing, and which are fertilised on regular basis by setting up sheep-pens or scattering the manure. �is plant community is characterised by a  large number of species, reaching more than 50 �owering plant species per 100 m2. �e dominating species include primarily grasses, including but not limited to agrostis capillaris, dactylis glomerata, arrhenatherum elatius, festuca rubra, and anthoxanthum odoratum. �is plant community also features perennial plants such as gladiolus imbricatus, lychnis �os-cuculi, heracleum sphondylium, achillea millefolium, leontodon hispidus, hypericum perforatum as well as alchemilla sp. (kotońska, 1991; pacyna, 2004; gołek et al., 2015). many of the aforementioned species were recorded in group b, which was referred to in this article as ‘used hay-meadow’ (appendix 1 – tab. 3). however, contrary to typical gladiolo-agrostietum meadow, this group did not have gladiolus imbricatus. it was found to contain numerous species of grasses for grazing and a group of legumes, proving that the analysed plots are useful from the agricultural standpoint. nevertheless, this group of plots shows certain symptoms of meadow degradation because of the presence of species such as anthoxanthum odoratum or nardus stricta. �e most probable hypothesis is that, as a  result of incorrect care, this type of meadows are gradually turning into anthoxantho-agrostietum pastures which, according to some authors, are becoming increasingly common throughout beskidy (gołek et al., 2015). �is plant community usually emerges as a result of discontinuation of usage or inadequate fertilisation of hay-meadows. although they are considered pasturable plant communities, only extensive farming activities are conducted on them. �e di�erence between typical used hay-meadows and anthoxantho-agrostietum pasture is in terms of its physiognomy, namely, its vegetation is much lower and less abundant. �is plant community is dominated by quite low and feeble grasses, such as agrostis capillaris, festuca rubra, and anthoxanthum odoratum. anthoxantho-agrostietum is gradually replacing gladiolo-agrostietum meadows to become a  dominating feature of many glades (gołek et al., 2015). �e plots which were included in the hay-meadow group (appendix 1 – tab. 3) are most likely the various stages of transition of gladiolo-agrostietum towards anthoxantho-agrostietum. the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 15 a�er discontinuation of grazing, mowing, and other usable treatments, mountain glades experience secondary succession consisting in, among others, the return of forests to the areas previously occupied by them (fig. 3). �e changes that ensue involve not only plants but the entire landscape (kornaś, 1990; kaźmierczakowa, poznańska, 1992; zarzycki, kaźmierczakowa, 2007; zarzycki, 2008; barabasz-krasny, 2010). initially, the overall number of species increases because the patches now contain plants from various communities – meadows and pastures as well as shrubs and trees (radkowski, barabasz-krasny, 2007; barabasz-krasny, 2010). an example of this in the current �oristic studies of the beskidy glades is ‘unused plots with shrubs’ – group d. �is group has the highest number of species in the �oristic table among all four distinguished groups and a high value of the general diversity (h) and dominance (c) indicators (fig. 2; tab. 6). when the photophilous species associated with open areas are completely eliminated, the general species diversity of such plots will also decline (zarzycki, kaźmierczakowa, 2007; barabasz-krasny, 2010), especially since some clearly dominant species are in plots of this group now, e.g., rubus sp., vaccinium myrtillus (appendix 1 – tab. 5). although unused plots with shrubs are characterised by large general diversity, the average number of species in the plot has been relatively low, which is evidence for the instability of �oristic composition and dynamically occurring species replacement processes. fig. 3. former hala rzycka – now completely covered with forest; october 2015 (photo. j. przemysław kubik) ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 16 similar processes were observed in plots characterised by a  presence of a  single dominating plant, namely unused plots with pteridium aquilinum (appendix 1 – tab. 4; fig. 4). in those plots, the meadow species still occur but in lower quantities, because they are overshadowed and dominated by pteridium aquilinum. �is clearly affects the general species diversity of such phytocoenoses (appendix 1 – tab. 4; tab. 6). in this case, photophilous plant species, which are typical to open areas, are eliminated even before the plots are overgrown by shrubs and trees, as con�rmed by the lowest light indicator value (l) for this group of plots (tab. 7). additionally, the acidi�cation of the soil and the reduction of the nitrogen content, which appear from unfavourable �ora changes as a result from the cessation of the use of meadows and pastures and the abandonment of care treatments, also promote the process of species elimination. conclusions �e results obtained through the conducted research con�rm the thesis that the cessation of grazing, mowing, and other usable treatments leads to the disappearance of abundant meadow phytocoenoses as well as the depletion and loss of their diversity. unused plots are characterised by lower average number of species. �e plants that grow on them usually include trees and shrubs, which supplant grasses and other spefig. 4. unused plot with pteridium aquilinum – plot no 50: zamczysko jaskinia lodowa i (photo. j. przemysław kubik) the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 17 cies typical for hay-meadows. cessation of grazing and other usable treatments results among other in reduction of the average number of species in plot, the increase of dominance indicators, and the reduction of ph and nitrogen content in soil. references barabasz-krasny, b. 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[in polish] the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 19 appendix 1 tab. 1. location of the studied areas number of plot in area name of glade geographical coordinates altitude (m.a.s.l.) used glades 1. kocoń 49°44ʹ01ʹʹn 19°24ʹ04ʹʹe 560 2. polana gałasie 49°44ʹ43ʹʹn 19°24ʹ00ʹʹe 670 3. gibasów groń 49°45ʹ11ʹʹn 19°22ʹ26ʹʹe 820 4. skolarówka 49°44ʹ31ʹʹn 19°20ʹ50ʹʹe 680 5. przełęcz płonna 49°44ʹ45ʹʹn 19°18ʹ43ʹʹe 700 6. zamczysko jaskinia lodowa 49°44ʹ56ʹʹn 19°18ʹ15ʹʹe 740 7. kocierz basie 49°45ʹ54ʹʹn 19°20ʹ56ʹʹe 640 8. kocierz kiczora 49°46ʹ03ʹʹn 19°19ʹ49ʹʹe 630 9. kocierz sołdrówka 49°45ʹ45ʹʹn 19°19ʹ29ʹʹe 530 10. kocierz mieszczaki 49°45ʹ44ʹʹn 19°18ʹ34ʹʹe 560 11. wysokie 49°45ʹ40ʹʹn 19°16ʹ45ʹʹe 540 12. groń jana pawła ii 49°47ʹ40ʹʹn 19°26ʹ48ʹʹe 850 13. podbucznik 49°46ʹ21ʹʹn 19°27ʹ06ʹʹe 670 14. targoszów pagórek 49°46ʹ09ʹʹn 19°27ʹ08ʹʹe 620 15. targoszów wieczorki 49°45ʹ54ʹʹn 19°27ʹ24ʹʹe 540 16. gronik 49°45ʹ31ʹʹn 19°27ʹ46ʹʹe 520 17. krzeszów harańczykówka 49°45ʹ50ʹʹn 19°28ʹ32ʹʹe 500 18. jaworzyna iii 49°46ʹ40ʹʹn 19°28ʹ00ʹʹe 650 19. jaworzyna ii 49°46ʹ42ʹʹn 19°27ʹ50ʹʹe 670 20. jaworzyna i 49°46ʹ45ʹʹn 19°27ʹ40ʹʹe 680 21. warmuzówka 49°48ʹ54ʹʹn 19°30ʹ43ʹʹe 420 22. jaszczurowa gancarzówka 49°47ʹ56ʹʹn 19°30ʹ20ʹʹe 420 23. tarnawa 49°46ʹ56ʹʹn 19°29ʹ10ʹʹe 490 24. pod makowską górą 49°47ʹ08ʹʹn 19°29ʹ03ʹʹe 560 25. polana 49°47ʹ54ʹʹn 19°28ʹ30ʹʹe 640 26. targoszów gajka 49°45ʹ33ʹʹn 19°26ʹ49ʹʹe 550 27. hala na potrójnej 49°46ʹ34ʹʹn 19°22ʹ07ʹʹe 840 unused glades 28. warmuzówka 49°48ʹ54ʹʹn 19°30ʹ43ʹʹe 420 29. jaszczurowa suszyce 49°48ʹ23ʹʹn 19°30ʹ39ʹʹe 400 30. jaśkowa arka 49°48ʹ07ʹʹn 19°28ʹ17ʹʹe 730 31. polana semikowa 49°47ʹ12ʹʹn 19°27ʹ04ʹʹe 800 32. pod jaworzyną 49°46ʹ42ʹʹn 19°27ʹ41ʹʹe 640 33. polana łazy 49°46ʹ17ʹʹn 19°28ʹ17ʹʹe 600 34. pod palusową górą 49°45ʹ58ʹʹn 19°28ʹ06ʹʹe 580 35. gronik 49°45ʹ31ʹʹn 19°27ʹ46ʹʹe 520 36. targoszów jurczakówka 49°45ʹ33ʹʹn 19°26ʹ49ʹʹe 550 ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 20 37. przełęcz midowicza 49°47ʹ35ʹʹn 19°26ʹ46ʹʹe 850 38. polana – leskowiec 49°47ʹ17ʹʹn 19°26ʹ43ʹʹe 890 39. gancarz 49°46ʹ37ʹʹn 19°26ʹ58ʹʹe 800 40. pagórek 49°46ʹ08ʹʹn 19°27ʹ06ʹʹe 610 41. targoszów wieczorki 49°45ʹ47ʹʹn 19°27ʹ26ʹʹe 520 42. targoszów ćwiękałówka 49°45ʹ35ʹʹn 19°26ʹ09ʹʹe 660 43. polana zaprzelina 49°46ʹ20ʹʹn 19°21ʹ14ʹʹe 760 44. kocoń 49°44ʹ04ʹʹn 19°24ʹ06ʹʹe 560 45. gałasie i 49°44ʹ37ʹʹn 19°24ʹ13ʹʹe 650 46. gałasie ii 49°44ʹ50ʹʹn 19°23ʹ46ʹʹe 720 47. polana – pietrasowa 49°44ʹ51ʹʹn 19°23ʹ19ʹʹe 800 48. gibasów groń 49°45ʹ08ʹʹn 19°22ʹ24ʹʹe 800 49. czarne działy 49°44ʹ58ʹʹn 19°21ʹ25ʹʹe 780 50. zamczysko jaskinia lodowa 49°44ʹ53ʹʹn 19°18ʹ23ʹʹe 740 51. ścieszków groń 49°44ʹ52ʹʹn 19°18ʹ04ʹʹe 760 52. kocierz walaszki 49°45ʹ56ʹʹn 19°20ʹ18ʹʹe 570 53. kocierz basie 49°45ʹ54ʹʹn 19°21ʹ10ʹʹe 620 54. słonków 49°45ʹ58ʹʹn 19°22ʹ04ʹʹe 760 tab. 2. floristic composition and species coverage [%] in plots of pasture with nardus stricta; + – species occurring in less than 1% successive number of plot 1 2 3 4 5 6 7 8 9 10 number of occurrence number of plot in area 7 5 27 4 3 12 18 2 26 20 group in dendrogram a number of species in plot 28 24 29 16 12 19 20 18 24 18 grass and sedges festuca rubra 15 40 15 15 40 15 15 15 15 40 10 poa trivialis 15 15 15 15 3 15 15 15 65 15 10 nardus stricta 15 3 15 15 15 3 3 15 3 . 9 phleum pratense + . 3 3 . 3 . . + 3 6 anthoxanthum odoratum 3 15 3 . . . 3 40 . . 5 dactylis glomerata . 3 3 . . . 3 . + 3 5 carex rostrata . . + 3 . . . . + . 3 legumes trifolium repens 3 3 3 3 3 3 3 3 3 3 10 vicia cracca + + + . . . . . . . 3 other herbaceous species hypericum perforatum 3 3 3 3 3 3 3 3 + 3 10 achillea millefolium 3 + + 3 3 3 + 3 + + 10 veronica chamaedrys 3 + + + 3 3 3 3 + + 10 rumex acetosa + 3 + 3 15 3 3 3 3 3 10 potentilla anserina + + + + 3 + + + 3 + 10 juncus articulatus 3 3 + + + . + 3 + + 9 ranunculus acris + . + + 3 3 + 3 + + 9 the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 21 leontodon hispidus 3 15 + + . . 3 . + + 7 plantago lanceolata + + + . . . 3 3 + + 7 stellaria graminea 3 + 3 + . 3 . . . + 6 lychnis �os-cuculi + . + . . . + 3 + + 6 campanula patula + + + . . + . . + . 5 leucanthemum vulgare + + + . . . + . + . 5 ajuga reptans . + + + . . . . . + 5 cruciata glabra . . . . . + 3 . + 3 4 alchemilla monticola . . . . . + + + + . 4 aegopodium podagraria + . . . . + . . + . 3 pteridium aquilinum . . . . + . + 3 . . 3 trees and shrubs rubus sp. (c) . 3 + . . + . . . . 3 species occurring sporadically grass and sedges: arrhenatherum elatius 27:+. legumes: vicia grandi�ora 7:3, 5:+; trifolium pratense 7:+, 27:3. other herbaceous species: carlina acaulis 7:+, 5:+; viola arvensis 7:+, 5:+; urtica dioica 7:+, 2:+; scabiosa sp. 5:+, 12:+; veronica o cinalis 27:+, 26:+; cirsium rivulare 2:+, 26:+; fragaria vesca 7:+; galium verum 7:+; rhinanthus minor 27:+, senecio integrifolius 27:+; dianthus deltoides 12:+. tab. 3. floristic composition and species coverage [%] in plots of hay-meadow; + – species occurring in less than 1% successive number of plot 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 number of occurrence number of plot in area 11 6 19 10 9 25 8 15 13 17 16 23 1 22 21 24 14 group in dendrogram b number of species in plot 22 18 22 22 28 22 23 23 22 22 21 22 23 22 27 20 22 grass and sedges festuca rubra 15 15 15 15 40 15 . 40 40 40 40 15 40 40 15 15 40 16 poa trivialis 15 15 15 15 15 15 15 . 3 15 15 15 15 15 15 15 15 16 dactylis glomerata 40 15 3 15 3 3 15 3 + . . 3 3 3 15 3 + 15 phleum pratense 15 3 . 15 3 65 + + . + 3 . 3 3 3 3 . 13 anthoxanthum odoratum . 3 3 3 + 3 15 3 3 15 3 3 15 . . + . 13 arrhenatherum elatius 15 15 3 . . . . 3 3 . 3 15 15 15 15 . . 10 holcus lanatus . . . + . . 40 15 . 15 15 15 . 3 3 3 + 10 nardus stricta . . 3 . . . . . . + . . . 3 3 3 3 6 lolium perenne . . . 3 . . . + . . . 3 . . . . . 3 legumes trifolium repens 15 . 3 3 15 15 15 15 3 + 15 3 3 3 3 3 + 16 t. pratense + . . + 3 + 3 15 3 3 . . 3 + 3 . . 11 vicia grandi�ora + 3 + + 3 . + . + + . . + . . . . 9 vicia cracca + . . . . . . + . + . . . . + . . 4 ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 22 other herbaceous species rumex acetosa + 3 3 3 + + + . 3 . + + 3 + + + + 15 achillea millefolium 3 3 + + + + . + + + + + . 3 + + + 15 hypericum perforatum + + + + + + + + + + + . . . + + + 14 ranunculus acris + + + + + . 3 + + 3 + + 3 . . . + 13 leontodon hispidus + + + . + + + 3 + . . + . 3 + + + 13 taraxacum o cinale . 3 . . + + + + + . . + + + 3 3 + 12 leucanthemum vulgare . . . + + + + + + + + + + + + . . 12 plantago lanceolata . 3 . . + . + 3 3 + + + 3 . 3 . 3 11 stellaria graminea . . + + + + + + . + + + + . . + . 11 veronica chamaedrys + 3 + + 3 + 3 + . . . . . . + . + 10 potentilla anserina + . + . + . + . . + . + + . + + + 10 campanula patula + . . . . + . + + . 3 . . + + + + 9 juncus articulatus . . . . + + . . + + + . 3 + + . + 9 alchemilla monticola + + . + + . + . . . + + 3 . . . . 8 aegopodium podagraria 3 + + + + . 3 . . . . . . . + . . 7 lychnis �os-cuculi . . . + . + . + . + + . . . . + + 7 senecio integrifolius . . . . . . . + + . . . . + + + + 6 euphorbia cyparissias + + 3 . . . . . . . . . . + . . . 4 fragaria vesca + . + . . . . . . . . . . . + . + 4 viola arvensis . . + . + + . . + . . . . . . . . 4 rhinanthus minor . . . . + . . + + + . . . . . . . 4 galium verum . . . . . . + . . . . + . + + . . 4 cruciata glabra . . 3 . . . . . . . + + . . + . . 4 urtica dioica 3 . . + + . . . . . . . . . . . . 3 pteridium aquilinum . . + . . . . . . . . . . . . + + 3 plantago major . . . + + . . . . . . + . . . . . 3 ajuga reptans . . . . . . + . + . . . . . . . + 3 cirsium rivulare . . . . . . . . . + + . + . . . . 3 trees and shrubs rubus sp. (c) . . . . . + + . . . . . . . . + . 3 species occurring sporadically grass and sedges: carex rostrata 9:+. legumes: trifolium dubium 16:+, 1:+. other herbaceous species: dactylorhiza majalis 17:+, 23:+; lamium maculatum 9:+; dianthus deltoides 25:+; digitalis purpurea 25:+; humulus lupulus (c) 1:3; bellis perennis 1:+; juncus conglomeratus 22:+; convolvulus arvensis 22:+. trees and shrubs: rosa canina (c) 22:+. the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 23 tab. 4. floristic composition and species coverage [%] in unused plots with pteridium aquilinum; + – species occurring in less than 1% successive number of plot 1 2 3 4 5 number of occurrence number of plot in area 52 45 50 43 41 group in dendrogram c number of species in plot 15 15 23 18 18 grass and sedges festuca rubra 15 + 3 3 3 5 poa trivialis + + + 3 3 5 agrostis capillaris 15 + + . . 3 phleum pratense + + 3 . . 3 nardus stricta 15 . . . 3 2 legumes vicia grandi�ora + + + . . 3 other herbaceous species pteridium aquilinum 15 90 90 90 90 5 hypericum perforatum + + 3 + + 5 stellaria graminea + + + + 3 5 potentilla anserina + . + + . 3 juncus articulatus + . + + . 3 rumex acetosa . . + + + 3 veronica chamaedrys . . + + + 3 trees and shrubs acer pseudoplatanus (a) . . + + + 3 rubus sp. (b) 15 3 + 15 15 5 species occurring sporadically grass and sedges: holcus lanatus 52:3, 45:3; anthoxanthum odoratum 52:+, 43:+; carex rostrata 45:+; lolium perenne 41:3; arrhenatherum elatius 41:+. legumes: vicia cracca 45:+, 50:+; trifolium pratense 50:+; t. repens 50:+. other herbaceous species: juncus conglomeratus 52:3, 43:3; senecio integrifolius 45:+, 43:+; galeopsis sp. 45:+, 41:+; leontodon hispidus 45:+, 41:+; achillea millefolium 50:+, 41:+; aegopodium podagraria 50:+, 41:+; ranunculus acris 50:+; scabiosa sp. 50:+; vaccinium myrtillus 43:15; veronica o cinalis 43:+; campanula patula 41:+; leucanthemum vulgare 41:+. trees and shrubs: betula pendula (a) 50:+, 43:15; picea abies (a) 50:+, 43:15. tab. 5. floristic composition and species coverage [%] in unused plots with shrubs; + – species occurring in less than 1% successive number of plot 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 number of occurrence number of plot in area 40 34 36 35 37 33 31 30 38 42 32 28 29 51 48 47 44 54 39 49 53 46 group in dendrogram d number of species in plot 21 28 23 18 10 17 17 15 15 22 22 17 14 15 17 17 22 21 18 18 14 21 grass and sedges festuca rubra 3 15 15 15 3 3 15 15 3 15 15 . 15 3 15 15 15 15 3 15 15 3 21 ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 24 poa trivialis 3 . 15 15 + 15 15 15 15 3 15 3 . + 15 15 3 . 3 40 . + 18 nardus stricta . . 3 3 3 3 15 15 15 15 15 3 . . + 3 3 15 3 3 3 15 18 holcus lanatus . + + 3 . . . . . . . 3 15 + 3 . + 3 . . 3 15 11 dactylis glomerata 3 . + + . . . . . . + 3 . . + 3 + . + . . . 9 phleum pratense 3 15 . 15 . . . . . . 3 . . . . + 3 + + . . . 8 agrostis capillaris . . . . . . . . . . . . . 3 15 3 15 15 . 15 15 + 8 arrhenatherum elatius + + + . . . . . . . 3 15 . + . . + . . . . . 7 anthoxanthum odoratum . . . . . + + . . . + . . . + . . . . . . + 5 carex rostrata . + . . . . . . . . + . . . . . . + . . . . 3 legumes vicia cracca . . . . . . . . . + . + . + . + 3 . . . . . 5 vicia grandi�ora . + . + . . . . . . . . . . . + + . . . . . 4 other herbaceous species hypericum perforatum + + + . + + . + . + 3 + . + + 3 3 3 + + + + 18 potentilla anserina + + + + + + + + + + + . + . . + + 3 + + . . 17 stellaria graminea 3 + 40 + . . + + . + . . . + + + . + . + 3 . 13 rumex alpinus + + + + + + + + + . . . . + 3 . . . . + . + 13 senecio integrifolius + + + . . . . . . . + . 3 . . . + + + . + + 10 juncus articulatus . + . . + . + . . + + . + . . . . + . + . + 9 achillea millefolium 3 + . . . . . . . + + + 3 . + . . . . + . . 8 veronica chamaedrys + + 3 . . . . . . + + . . . + . . + + . . . 8 leontodon hispidus + + . . . . . . . . + . 3 . . . + . + + . + 8 juncus conglomeratus . . 3 40 . 15 . + . + . . . . . . . . + + 3 . 8 vaccinium myrtillus . . . . . 3 90 65 90 . . . . . . 40 . 3 3 . . 40 8 ranunculus acris + + + + . . . . . + . . . . . + . . . . . . 6 veronica o cinalis . . . . + . + . . . + . . . . . . + + . . + 6 cirsium rivulare . + 3 + . + . . . . . + . . . . . . . . . . 5 aegopodium podagraria . + + . . . . . . . . + . + . . + . . . . . 5 galeopsis sp. . . . + . . + + . + . . . . + . . . . . . . 5 campanula patula + + . . . . . . . . . . . . . . . . . . + + 4 leucanthemum vulgare . + . . . . . . . + + + . . . . . . . . . . 4 urtica dioica . . + + . . . . . . . 3 . + . . . . . . . . 4 pteridium aquilinum . . . . . . . + + . . . . . . . + . . . + . 4 plantago lanceolata . + . . . . . . . + + . . . . . . . . . . . 3 equisetum sylvaticum . . + + . 15 . . . . . . . . . . . . . . . . 3 the succession of abandoned glades and its im pact on the diversity of flora in b eskid m ały m ountains (s outhern p oland) 25 trees and shrubs betula pendula (a) . . . . . 3 + 3 + 15 + + 15 40 . . 3 3 65 15 65 15 15 picea abies (a) . . . . . . . . 15 . . . . . . . . 15 40 15 . . 4 acer pseudoplatanus (a) . . . . . . . . + . . . . 90 3 . . . . + . . 4 rubus sp. (b) 65 40 + 3 90 15 3 15 3 15 40 15 + 3 + 3 15 15 15 65 3 15 22 frangula alnus (b) 3 40 + 40 65 3 . . 3 . . . + . . + + + + . + + 14 sorbus aucuparia (b) 3 . . . . + + . 3 . . . . . + + 3 + + + + + 12 crataegus sp. (b) 15 . 3 . . . . . . 3 . . . . 3 . . + . . . + 6 rosa canina (b) . . . . . . . . . 15 . 65 + . . . 3 . . . . . 4 species occurring sporadically grass and sedges: lolium perenne 51:+, 44:+; cynosurus cristatus 34:+. legumes: trifolium repens 42:+, 32:3; t. pratense 42:+. other herbaceous species: lychnis �os-cuculi 34:+, 42:+; dactylorhiza majalis 34:+, 47:+; alchemilla monticola 34:+, 46:+; galium verum 36:+, 28:3; polytrichum commune (d) 33:3, 38:+; scabiosa sp. 40:+; plantago major 34:+; briza media 36:+; veratrum lobelianum 31:3; ajuga reptans 31:+; digitalis purpurea 30:+; juncus e�usus 38:+; taraxacum o cinale 32:+; fragaria vesca 29:15; cruciata glabra 29:+; euphorbia cyparissias 29:+; gladiolus imbricatus 54:+; carlina acaulis 49:+. trees and shrubs: pinus sylvestris (a) 33:3, 28:+; quercus robur (a) 40:+; fagus sylvatica (a) 31:+; abies alba (a) 30:+; larix decidua (a) 46:+. ja n p rz em ys ła w k ub ik , b ea ta b ar ab as zk ra sn y 26 abstract �e in�uence of grazing, mowing, and other usable treatments on the �ora diversity of glades in the beskid mały in southern poland was investigated. �e �eld research was carried out between 2015 and 2016. flora analysis consisted mainly of comparing the botanical composition of glades abandoned for several decades with the botanical composition of glades currently used as pastures. on selected plots, botanical composition was determined using the klapp (1965) estimation method. all �oristic lists from the study plots were analysed by using hierarchical numerical classi�cation. based on the numerical classi�cation of plots on analysed glades in the beskid mały, four utility-�oristic groups were distinguished: pasture with nardus stricta a, hay-meadow b, unused plots with pteridium aquilinum c, and unused plots with shrubs d. �e results of studies con�rm the thesis that species composition is a  re�ection of management practices or lack thereof. it was demonstrated that the cessation of the grazing and mowing on the mountain glades of beskidy caused adverse changes in the structure of species composition and a reduction in �oristic diversity. initially, it causes an increase in the number of species in the sward, followed by elimination of the photophilous species, which lowers general species richness. key words: klapp estimation method, meadows and pastures, mountain pasturage, succession received: [2017.05.15] accepted: [2017.09.19] sukcesja nieużytkowanych polan i jej wpływ na zróżnicowanie flory w beskidzie małym (południowa polska) streszczenie zbadano wpływ zaprzestania działalności pasterskiej i  łąkarskiej oraz innych zabiegów użytkowych na zróżnicowanie �ory hal oraz polan w beskidzie małym w południowej polsce. badania w terenie wykonano w latach 2015–2016. analiza �orystyczna polegała głównie na porównaniu składu botanicznego polan od kilkudziesięciu lat odłogowanych, ze składem botanicznym polan aktualnie użytkowanych gospodarką pasterską. skład botaniczny określono na wybranych poletkach, przy pomocy metody szacunkowej klappa (1965). wszystkie spisy z analizowanych poletek poddano hierarchicznej klasy�kacji numerycznej. na podstawie klasy�kacji numerycznej poletek wyróżniono na badanych polanach beskidu małego cztery grupy �orystyczno-użytkowe: pastwisko z  nardus stricta a, łąka kośna użytkowana b, płaty nieużytkowane z  pteridium aquilinum c oraz płaty nieużytkowane zakrzaczone d. badania potwierdzają tezę, że skład gatunkowy jest odzwierciedleniem prowadzonych zabiegów gospodarczych lub ich braku. wykazano, że zaprzestanie gospodarki pasterskiej i kośnej polan górskich w beskidach skutkuje niekorzystnymi zmianami w strukturze gatunkowej runi oraz obniżeniem różnorodności �orystycznej. początkowo powoduje wzrost liczby gatunków w murawie, a następnie eliminację gatunków światłolubnych, co obniża ogólne bogactwo gatunkowe. słowa kluczowe: metoda szacowania klappa, łąki i pastwiska, pasterstwo górskie, sukcesja information on the authors jan przemysław kubik his scienti�c interests include secondary succession of mountain glades a�er cessation of agricultural activities. beata barabasz-krasny her main scienti�c interests include �oristics and phytosociology of non-forest plant communities with particular emphasis on the course of succession processes in the areas where agricultural activities were discontinued, the transformation of plant cover of thermophilic swards, and active protection of nonforest plant communities. 137 annales universitatis paedagogicae cracoviensis studia naturae, 2: 137–158, 2017, issn 2543-8832 doi: 10.24917/25438832.2.11 anna kocoń, magdalena nowak-chmura* department of invertebrate zoology and parasitology, institute of biology, cracow pedagogical university, cracow, podchorążych 2, 30-084 cracow, poland; *magdalena.nowak-chmura@up.krakow.pl skin ectoparasites of domestic animals small pets have long accompanied their owners and play a positive role and strengthen their owners’ psychological well-being and their physiological welfare, as their life companions. according to research by krassowska (2014), the majority of poles that have pets at home choose a dog (canis familiaris l.) (83%), fewer people decide on a cat (felis catus l.) (44%), the third place of the most popular animals accompanying poles at home are birds (4%), rodents (3%), and reptiles (1%). �e frequent phenomenon in accompanying animals is the infestation of their skin by insects or parasitic mites, causing onerous symptoms, such as discomfort, itching, or allergic reactions. due to the close contact of people with pets, we are also exposed to the attacks of dangerous arthropods, causing, e.g., pediculosis, sarcoptosis, otodectomy, and demodicosis and transmitting many dangerous zoonotic diseases, such as lyme disease tularaemia, rickettsiosis, ehrlichiosis, and others. skin ectoparasites that most o�en attack domestic animals are mites (e.g., demodex spp., sarcoptes spp., ticks) and insects such as �eas and lice (tab. 1). symptoms of attacks of dangerous ectoparasites may include skin itching, allergic reactions, skin redness, discomfort, the loss of hair, and skin discoloration. because of the rapid reproduction of parasites, immediate assistance is required for animals attacked by parasitic arthropods, primarily to protect the animals against parasite infestation on the whole body and against the destruction of the animal’s organism. �e prevention and protection against the attacks of skin ectoparasites is important, because, by protecting the animals we also protect ourselves against zoonotic diseases. review of the selected domestic animals and ectoparasites attacking them 1. dog (canis familiaris l.) �e dog is the most common domestic animal in polish homes (krassowska, 2014), and the oldest domesticated carnivore mammal. it probably derives from the grey a nn a k oc oń , m ag da le na n ow ak -c hm ur a 138 wolf (canis lupus l.). �e period of taming falls approx. 12 thousand years ago, when the portion of the grey wolf population was approaching human habitation. initially, man used the help of dogs to hunt and to guard the herds. as a result of more or less conscious selection and crossing, about 400 breeds and varieties of a domestic dog have been grown. �e breeds vary among themselves, with height, body weight, appearance, utility, and colour. �e excellent sense of smell, hearing, good olfactory and auditory memory, the ability of spatial orientation, intelligence, and the ease of attachment to people facilitate its education and training. demodex canis leydig – dog demodex (appendix 1 – fig. 1a), is a widespread animal parasite observed in nearly 80% of dogs. it occurs in the hair follicles and sebaceous glands, and sometimes also in the lymph nodes, internal organs, and blood. it tab. 1. skin ectoparasites of domestic animals no. parasite species host parasitic disease/carrier (vector) of the diseases 1. demodex canis canis familiaris demodicosis 2. sarcoptes scabiei var. canis c. familiaris sarcoptosis 3. ctenocephalides canis c. familiaris ktenocefalidosis/vector 4. linognathus setosus c. familiaris lice 5. cheyletiella yasguri c. familiaris cheyletiellosis 6. trichodectes canis canis familiaris, felis catus malofagosis 7. ixodes ricinus c. familiaris, f. catus vector 8. i. crenulatus c. familiaris, f. catus vector 9. i.hexagonus c. familiaris, f. catus vector 10. i. rugicollis c. familiaris, f. catus vector 11. dermacentor reticulatus c. familiaris, f. catus vector 12. otodectes cynotis c. familiaris, f. catus otodectomy 13. notoedres cati felis catus sarcoptosis 14. demodex cati f. catus demodicosis 15. cheyletiella blakei f. catus cheyletiellosis 16. ctenocephalides felis f. catus ktenocefalidosis 17. felicola subrostratus f. catus malofagosis 18. dubininia melopsittaci melopsittacus undulatus --------------------19. sideroferus lunula m. undulatus --------------------20. neopsittaconirmus gracilis m. undulatus malofagosis 21. trixacarus caviae cavia porcellus --------------------22. chirodiscoides caviae c. porcellus --------------------23. gliricola porcelli c. porcellus malofagosis 24. demodex cavinae c. porcellus demodicosis 25. d. criceti mesocricetus auratus demodicosis 26. d. aurati m. auratus demodicosis 27. d. �agellurus mus musculus demodicosis 28. d. musculi m. musculus demodicosis s kin ectoparasites of dom estic anim als 139 occurs naturally in small amounts in most dogs, and it usually does not cause clinical symptoms. occasionally, the mite can cause a disease called mange, and then lead to one of the most onerous skin problems encountered in veterinary practice (gauguere, 1994). �e infection with demodex already takes place in the �rst days of the life of puppies, most o�en during feeding, or being licked by the mother. in older dogs, the infection may take place through direct contact. human infection is very rare. factors a�ecting the mange in animals at a young age include the occurrence of metabolic disorders, malnutrition, systemic infections and cancers, diabetes, as well as a decreased resistance to natural factors, and stress. �e most common symptoms of the attack of d. canis are skin peeling, the appearance of purulent pimples coming out of the hair follicles, increased pigmentation, thinning of the hair, and red erythematous spots. skin lesions appear mainly around the head, muzzle, lips, ears, torso, and less frequently in the paw area, and if so, on the front paws. heavy and generalised mange may lead to the death of the animal (gauguere, 1994). sarcoptes scabiei l. var. canis – dog itching mites (appendix 1 – fig. 1b) causes a disease called sarcoptosis. mites attack many mammals, including humans and most livestock and domestic animals. dog scabies is very infectious among dogs. �e most vulnerable are dogs living in poor environmental conditions or staying in larger concentrations, e.g., in shelters or animal hotels with poor maintenance standards. mites can survive up to two weeks in the host, so the transmission does not require physical contact with other infected animals, because they are passively transmitted in close contact with the host. �is parasite spends all his life on the same host. dogs may pick up mites from the nearest environment. s. scabiei does not have external vectors (insects, worms, birds), which carry these mites, as is the case with many other ectoparasites. mites burrow tunnels under the skin. �eir saliva has strong digestive enzymes, which dissolve the skin tissues, and they do not suck blood. adult females lay their eggs in subcutaneous tunnels, which hatch within a few days. dog scabies will last only a few days without the host. damages caused by parasites may be signi�cant. �e tunnels cause skin irritation, which increases the allergic reactions to saliva and faeces excretion of the mites. pimples and lumps grow on the infected skin, the animal’s fur is brittle, and the skin becomes bald and hard, which results in the formation of skin folds. �e initial symptoms of the mite infection include redness and small balding areas near the lips, eyes, ankles or elbows, and on the periphery of the ears, where the skin is thin, which then spread to the rest of the body, most o�en covering the elbows, armpits, chest and abdomen of the animal (klockiewicz, 2003). dogs infected with s. scabiei su�er from severe itching, shaking their heads, o�en lick the infected parts of their body, vigorously scratch their head, ears, and they rub against objects (trees, furniture, etc.) which sometimes leads to self-mutilation. a nn a k oc oń , m ag da le na n ow ak -c hm ur a 140 untreated disease causes the destruction of the body and may lead to the death of the animal (klockiewicz, 2003). ctenocephalides canis curtis – dog �ea (appendix 1 – fig. 1c) is an external parasite, cosmopolitan, whose main hosts most o�en include dogs (c. familiaris), foxes (vulpes vulpes l.), and grey wolves (c. lupus). �ese �eas can also parasitize humans, cats, and other mammals (sandner, 1976). eggs laid by the female �eas are most o�en found in the animal’s litter. in neglected, sick dogs, the eggs can be laid directly on the skin. �is insect can transmit the plague, endemic dandelion, as well as be the intermediate host of �ea tapeworm (dipylidium caninum (l.) leuckart) and the vector of roundworm larvae diro�laria immitis leidy. infestation with �eas, called ktenocefalidosis, results in animal anxiety, itching caused by the irritating secretions introduced into small wounds along with saliva. strong itching o�en causes scratching and rubbing of the dogs against various objects, leading to secondary skin lesions, particularly in such areas of the body like the head, neck and tail (stefański, 1968). �is constant scratching and biting of the skin can cause the dogs skin to become red with in�ammation, and the dog can also lose hair. dog �eas can cause allergic dermatitis in dogs allergic to the saliva of these insects. in this case, the above-mentioned symptoms are more pronounced. �ese ectoparasites can also result in the animal’s anaemia, when �eas suck up a very large amount of blood in young, elderly, or sick individuals (patyk, 1978). fleas occur in a variety of environments, and they rapidly spread to another host by leaping, so it is important to take care of the hygiene of your animal and your own and to use specialist �ea�ghting preparations. linognathus setosus olters – dog louse (appendix 1 – fig. 1d) is a cosmopolitan ectoparasite, and it causes a disease called pediculosis. it infests the hairy scalp, ears, neck, back, and near the root of the tail, spreading to the whole body in the case of a severe infestation. nits, lice eggs, are �xed by the female with the so-called cement, at the base of the hair, usually visible to the naked eye with a bigger number of eggs. most of the time, the louse is a practically still animal (sadzikowski, gundłach, 2004). dog lice are not common in clean, healthy, well fed, and maintained accompanying dogs. however, it o�en attacks weakened, old, exhausted, and malnourished animals. attacked dogs are anxious, are itching, and o�en scratching, which leads to intense licking, rubbing, and biting of the infected areas. �e animal’s fur becomes rough and the skin is reddened, which can lead to dermatitis. infestation with lice may lead to anaemia (blagburn et al., 1981). unlike �eas, lice do not jump from one dog to another, but they can move from one host to another through direct contact with the infected dog, e.g., in the animal shelter, in parks, or in other places with a lot of dogs. due to the fact that lice cannot survive a long time outside the host, they cannot be found freely in the s kin ectoparasites of dom estic anim als 141 environment, which distinguishes them from, e.g., �eas and ticks. infected dogs can be quickly and easily cured with specialised insecticides. cheyletiella yasguri smiley – cosmopolitan mite (appendix 1 – fig. 1e), cheyletiella spp. is responsible for causing the skin disease called cheyletiellosis, which, due to its symptoms, is o�en called ‘walking dandru� ’. �e name ‘walking dandru� ’ comes from the dynamically moving mites in search of food between the exfoliated skin in which they move, thus making the skin move. �ese ectoparasites live on the skin surface and feed on keratin, the main structural skin protein, exfoliated skin, hair, and skin secretions. �ey periodically attach themselves to the skin of the host, which they pierce with mandibles and suck out the exudates coming out from the skin micro-damages (bronswijk, kreek, 1976). cheyletiella spp. are absolutely bound to their host, and they are not likely to survive outside their host in the environment. �ese mites can be transmitted between di�erent animal species, including humans, and in addition, some animals (mainly rabbits and cats). �e infection usually has a direct character by rubbing, licking the fur, sleeping or playing together. �erefore, all dogs can be the potential source of threat to each dog, but also other domestic animals (mainly cats and rabbits), as well as humans. �ese mites have been found on insects bigger than themselves with a parasitic lifestyle, e.g., on �eas, mice, and �ies, which is important for the spread of cheyletiella spp. among various mammals, including dogs. in very few cases, infections may also occur through indirect contact (shared blankets, toys, towels, etc.). �e risk of infection increases in large groups of dogs, among young animals, or sick ones with a weakened immune system, and among malnourished or poorly nourished individuals. dogs with long hair are more susceptible to infections. itching and dandru� are the most characteristic symptoms of the invasion. sick animals are restless, and they are scratching, licking, shaking, or rubbing against various objects incessantly, which may lead to skin damage, numerous wounds, or thinning hair. continuous peeling of the epidermis makes the animals look as if they were sprinkled with �our. dandru� is located in the area of the hindquarters, from where it extends to the back – along the spinal line – to the head. as a result of scratching, the mites spread to other parts of the body, sometimes covering the whole body. continuous scratching leads to skin eruptions, and lesions may be accompanied by severe seborrhoea and a very unpleasant odour. at an advanced stage, it may involve weight loss, sadness, apathy, fever, and systemic complications (saevik et al., 2004). trichodectes canis de geer – chewing lice, a cosmopolitan parasite of dogs and cats (appendix 1 – fig. 1f ). it dwells on the whole body, in the hair, mainly on the neck and head (stefański, 1968). t. canis is a well-known vector for dipylidium caninum. usually, it does not pose serious problems to the host; however, in severe infections, it can be very irritating. wild, sick, neglected dogs are o�en attacked by this ectoparasite. �is parasite mainly feeds on the �uids produced by the irritated skin. it causes stress a nn a k oc oń , m ag da le na n ow ak -c hm ur a 142 in the host, creates wounds on the surface of the skin, and it also leads to hair loss due to increased scratching, and biting the hair out (sadzikowski, gundłach, 2004). dogs in poland are also attacked by four tick species from the ixodidae family: ixodes ricinus l., i. crenulatus koch, i. hexagonus leach, i. rugicollis schulze et schlottke and one species from the amblyommidae family: dermacentor reticulatus fabricius. �ere have also been cases of bringing the rhipicephalus sanguineus latreille species to warsaw, a typical dog parasite, which does not occur in the polish fauna. moreover, probably also other tick species can attack domestic dogs and cats in poland (ixodes persulcatus schulze, haemaphysalis punctata canestrini & fanzago and h. concinna koch) (siuda et al., 2007). ticks thrive on the hosts body most o�en in places well supplied with blood vessels, on delicate skin, which are di�cult to access for the animals own hygiene: around the snout, behind the ears, on the back of the neck, the neck, on the back and groin, at the base of the tail or around the anus (siuda, 1993). ixodes ricinus l. – common tick (appendix 1 – fig. 2a) is the most common and widespread species of ticks in poland that attacks domestic dogs and cats (siuda et al., 2002, 2004; nowak-chmura, siuda, 2012). �is parasite is the vector and carrier of many pathogens, which are dangerous, due to the fact that they cause serious medical and veterinary consequences, the most important ones include, among others, the following: borrelia burgdorferi, b. garinii, b. afzelli, virus from the flaviviridae family, rickettsia slovaca, r. conorii, anaplasma phagocytophilum, coxiella burnetti, toxoplasma gondii, �eileria mutans, and others (nowak-chmura, siuda, 2012; nowak-chmura, 2013). narrow lanes, forest paths, tall grass, bushy scrubs, and deciduous and mixed forests are ideal walking locations for domestic dogs, and thus perfect conditions for i. ricinus to attack the host (siuda, 1993). i. hexagonus leach – hedgehog tick (appendix 1 – fig. 2b) are nest and burrow parasites, which can also occur in the near vicinity of dog kennels, in close proximity to human homes and settlements (hillyard, 1996; siuda, nowak, 2011). it is probably spread throughout poland (michalik et al., 2010; kilar, 2011; nowak-chmura, 2013). �e tick-borne encephalitis virus, borrelia burgdorferi, b. garinii, b. afzelii, rickettsia helvetica, anaplasma phagocytophilum are the main pathogens transmitted by this tick species (nowak-chmura, 2013). another tick species found in domestic dogs in poland is ixodes rugicollis (siuda et al., 2010). �e natural habitat of i. rugicollis usually includes burrows, mammal hiding places, low humidity habitats (nest and burrow parasite) (nowak-chmura, 2013). �e medical and veterinary signi�cance of i. rugicollis has not been fully investigated. i. crenulatus koch – is also a nest and burrow species, the parasite dwells in burrows of predatory mammals and rodents. it easily adapts to various habitats: lowlands, mountains, subalpine meadows, deserts (mihalca et al., 2012). �e tick was found in di�erent places throughout poland (siuda, 1993; kadulski, 2007; siuda et al., 2010). s kin ectoparasites of dom estic anim als 143 �e role of this species in the transmission of infectious diseases is poorly known. it is the carrier of coxiella burnetii and an important vector of borrelia burgdorferi s. in areas where i. ricinus is absent (estrada-peña et al., 1995). dermacentor reticulatus fabricius, meadow tick – the main habitat of this tick species includes wooded, bushy river valleys, streams, swampy mixed forests (szymański, 1986). �ere is a currently widespread coverage of this species in central europe and in neighbouring countries (dautel et al., 2006; nowak, 2011; petney et al., 2012). �e largest number of habitats of this tick can be found in the north of poland, in the areas of north-eastern and eastern poland (siuda, 1993; bogdaszewska, 2004; kadulski, izdebska, 2009; nowak-chmura, 2013). d. reticulatus is a signi�cant vector of tick-borne pathogens in europe, the greatest importance is related to the transmission of babesia canis protozoa, and this is the main etiologic agent of the ‘babesiosis canum’ disease. �is disease can lead to the dysfunctions of many of the host’s body organs. �e outbreaks of this disease have also been documented in poland (zygner et al., 2007, 2008; welc-falęciak et al., 2009). domestic dogs should be checked a�er each walk for the presence of ticks, especially around the ears, mouth, eyes, and the whole torso. proper hygiene should be maintained while removing the feeding ticks. 2. domestic cat (felis catus l.) �e domestic cat is the second most common animal accompanying polish families (krassowska, 2014), and currently the most popular pet in the world (driscoll et al., 2009). �is species probably originates from the nubian cat (felis silvestris ssp. lybica forster), and it was domesticated about 9.5 thousand years ago. for a few thousand years, about a hundred breeds of the domestic cat have been bred, varying in hair colour, length, and size. a large number of domestic cats live alone in towns and villages. previously, cats were used for killing mice and rats, which were destroying the farmers’ harvests before domestication of cats; in addition, they were taken hunting in order to scare the game (lasota-moskalewska, 2005). otodectes cynotis hering – scabies (appendix 1 – fig. 2c). it infests the external auditory duct of domestic cats, less o�en in dogs. in advanced cases, it can attack other areas of the body: neck, back, loin, and tail. �e infection occurs through direct contact with the sick individual. it is much more common in kittens than adult cats. �e microorganism can also be transferred to humans. considerable amounts of dark brown, waxy, dry secretions can be observed in the auricle, where these microorganisms accumulate. in the advanced state of the disease, the accumulated grease can clog the auditory canal, which leads to the situation where a cat cannot hear. an untreated invasion can damage the tympanic membrane. symptoms of scabies, the disease caused by this parasite, is a characteristic strong itching of the ears, during which the cat is shaking its head, scratching its ears and crooking its head, which is a nn a k oc oń , m ag da le na n ow ak -c hm ur a 144 due to the mechanical and chemical tissue irritation by the ectoparasites preying in them. infection is also accompanied by local allergic reactions, local in�ammation, and severe pruritus. constant itching forces the animal to continue scratching causing further irritation of the infected areas, thus leading to the aggravation of disease symptoms and mechanical spread of parasites to other body areas. skin lesions appearing on the skin take the form of localized balding and severely itchy follicular e�orescence (bowman, 2012). notoedres cati hering – feline scabies (appendix 1 – fig. 2d). �ese parasites are transmitted directly from a diseased individual to a healthy one; however, sometimes the physical contract is not required, because they can be easily collected in places shared with sick individuals. �ey resemble the sarcoptes scabiei mites both in terms of construction and behaviour. �e infection o�en covers the animals head and ears, and then it spreads to the neck, back, and other parts of the body. �e skin a�ected by the invasion of itchy mites is wrinkled, with hair loss, o�en with scabs on it. �e symptoms of the feline mite invasion include intense itching, vigorous and continuous scratching, and licking of the infected body parts. from time to time, the notoedres mites can infect humans, but are unable to complete their development on them (kotomski, 2000). demodex cati hirst – feline demodex (appendix 1 – fig. 2e), as in the case of demodex canis, it may occur as a natural �ora in healthy cats. skin lesions associated with the invasion of feline demodex tend to be localised on the head, neck, and eyelids, where they cause hair thinning and alopecia. it can also lead to seborrhoea, the formation of lumps, pustules, and skin in�ammation. mites can also be present in the auditory canals, causing the in�ammation of the middle ear (dembele, 2000). cheyletiella blakei smiley – small mites and ectoparasites of domestic cats. cheyletiellosis, the disease caused by this microorganism, can be transferred from cats to humans. cheyletiella yasguri occurring in dogs and c. blakei occurring in cats are absolutely not speci�c to typical hosts. �e presence of mites on the animal’s skin may be tolerated well by them, and excessive peeling of the skin is the only clinical symptom of infestation; however, other animals may develop itching with varying degrees of severity. �e symptoms of a cat being attacked by the mite are similar to the symptoms of domestic dogs being attacked by c. yasguri (appendix 1 – fig. 2f ) (fagasiński, 2000). ctenocephalides felis bouche – feline �ea (appendix 1 – fig. 2g). a cosmopolitan parasite, in addition to a domestic cat, it can also attack humans, domestic dogs and other domestic and wild animals. symptoms of cat infestation by �eas are the same as in the case of c. canis infestation of domestic dogs, i.e. severe itching that causes constant scratching and rubbing at various objects, which leads to skin lesions in areas of the body, such as the head, neck, and tail areas. �e animal’s skin becomes red, o�en balding (furmaga, 1985). s kin ectoparasites of dom estic anim als 145 felicola subrostratus nitzch in burmeister – a lice species, cosmopolitan, occurring all over the body in the hair (appendix 1 – fig. 2h). it is found in older or sick cats, especially in longhaired breeds. in case of invasion, the cat is constantly scratching, biting the areas with the itchy skin, rubbing, and is restless. �e longer dwelling time of the lice on the body and their increasing number causes the skin redness, numerous scratches on the skin, and the loss of hair by the animal (kotomski, 2000). domestic cats are attacked mainly by four tick species: ixodes ricinus, i. crenulatus, i. hexagonus and i. rugicollis (siuda et al., 2007; kocoń et al., 2017), which were described above for the domestic dog. 3. budgerigar (melopsittacus undulates shaw) budgerigar has been the most popular parrot occurring in breeds all over the world for over 200 years. �e main places where the parrots occur include steppes overgrown with various species of grass. it is a nomad species, if the food runs out in the place where it lives, it moves to another area. parrots are relatively easy to tame, especially because they are usually accompanied by people from birth. �ey �rst appeared in europe in 1840, in england, and for the next 20 years, they have spread in breeding in other parts of europe. �e essential food for budgerigars includes the properly selected grain mix, including millet, canary grass, senegal millet, japanese millet, oats, and a small amount of oil seeds. in 2012, 28 varieties of parrots were described (biel�eld, 1997). dubininia melopsittaci atyeo & gaud and sideroferus lunula robin & mégnin – acari, which do not normally threaten their hosts, look like mites. bigger invasions take place as a result of stress, poor nutrition, and inappropriate living conditions of the parrots, as well as due to the reduced animal immunity. under such conditions, these parasites reproduce in excess, which causes skin irritation for the birds, but they do not lead to feathers falling out. it is believed that d. melopsittaci (appendix 1 – fig. 3a) is more pathogenic than s. lunula (appendix 1 – fig 3b). s. lunula is usually located on large wing feathers and tail, while d. melopsittaci on smaller ones. �ese species are located in di�erent places and do not compete with each other. �ey spent all their lives dwelling among the host’s feathers. �ey move very well between the feathers, thanks to ‘sticking’ to the feathers, so they do not fall o� during the �ight. �ey feed mainly on the feathers with the glandular secretions, exfoliate skin, nd the feather residues. parasites pass from one bird to another during direct contact with the infected animal, especially during breeding, from adults to chicks. �e symptoms of invasion of these mites are scratching by the parrots and plucking feathers. changes in the appearance of the feathers are visible on the underside of the feather, where these parasites dwell, and the feathers appear greasy, stained, and devoid of natural glow. in budgerigars, apathy, anorexia, and the destruction of the body may occur during very large invasions. mite allergens can cause irritation of the skin and mucous membrane a nn a k oc oń , m ag da le na n ow ak -c hm ur a 146 in humans (atyeo, gaud, 1987). neopsittaconirmus gracilis guimaraes – lice, one of the feather-eating species (appendix 1 – fig. 3c). �is parasite feeds on the feather residues, exfoliated epidermis, and adult lice, as well as larvae, can actively collect blood. �e invasion is usually asymptomatic, as a result of a decrease in immunity, and bad conditions. mass infestation causes clinical symptoms, such as bird anxiety and weakness of the body. �e infection takes place through direct contact of birds, but sometimes they can get infected in the nest. �ey also nibble on the base of the feathers, causing bleeding damage to the skin, and scabs form in the lesion areas. �ey can cause in�ammation and allergic reactions on the skin, which o�en manifests as pruritus. additional skin damage occurs when birds are pecking the itchy spots. symptoms of feather-eaters feeding on the feathers (�ag loss, �ag holes) and eggs laid by them along the feather can be seen with the naked eye. however, the visible losses in the �ags do not always denote the presence of feather-eaters, and they can be the e�ect of mechanical damage. �ese lice are most o�en found on the back of the head and neck; however, during larger invasions, they cover the whole body (sychra, 2005). 4. guinea pig (cavia porcellus l.) cavy is a rodent species, commonly known as guinea pigs, which were domesticated by south american indians, who used them for many purposes, including culinary, ritual, and folk medicine. it came to europe as a hobby animal in the xvith and xixth centuries, and it was used as a laboratory animal. �e guinea pig is a herbivore, and it is well suited for hobby breeding. c. porcellus should not be kept alone, because it is a social animal, and it maintains close relations with the herd members. currently, there are many breeds and colour varieties of this rodent. �ese breeds are di�erent in terms of structure and length of hair (beck, 2014). trixacarus caviae fain, hovell & hyatt – mites that move directly from the infected animal to another one, and they rarely leave their host. in guinea pigs, they cause scratches, biting of the skin, hair loss, and in severe cases, seizures (appendix 1 – fig. 3d). with severe infection, they can be life threatening due to the lack of food, severe pain, and discomfort. �ey are not transferred to humans. in healthy animals, the mites can be dormant for several months or years, becoming a problem in pregnant, sick, stressed, and older guinea pigs (mederle, indre, 2009). chirodiscoides caviae hirst (appendix 1 – fig. 3e), mites that cause minor discomfort, even if they are present in large quantities, do not show signs of harmful e�ects on the rodent. sometimes, they are visible to the naked eye on the animal’s fur, attached to the hair. �ey do not cause skin irritation, in some cases, they can cause itching (d’ovidio, santoro, 2014). gliricola porcelli schrank (appendix 1 – fig. 3f ) are lice that most o�en infest the s kin ectoparasites of dom estic anim als 147 guinea pig. lice are visible to the naked eye in the rodent’s hair, and they feed on hair and exfoliated skin. because of their high species speci�city, they do not pose a threat to other animals or humans. �e infection occurs through direct contact with a sick animal. �e course of the disease may be asymptomatic. with more severe invasions, the animals experience pruritus, and they rub against objects or scratch the itchy areas, which leads to the formation of wounds. however, we usually observe only the deterioration in the quality of the coat, with its thinning and seborrhoea. with severe, long-lasting, and untreated invasions, the licheni�cation of the skin around the ears can be observed, as well as seizures (przyjałkowki, 1977). demodex caviae bacigalupo (appendix 1 – fig. 3g), demodex attacking guinea pigs. �e symptoms of the occurrence of d. caviae are the same as in the case of d. canis and d. cati described above (hawkins, bishop, 2012). 5. golden hamster (mesocricetus auratus waterhouse) it is the most well-know hamster species, which is a rodent, bred as a pet. �is species was �rst described in 1839 by the british zoologist, waterhouse. m. auratus did not stand out with special features, the interest in it was not high, and over time, it was observed less and less, until �nally it was considered to be the extinct species. in 1930, professor aharoni from the university of jerusalem, who conducted excavations in syria, found a female with young ones. �ree individuals from this family (2 females and a male) went to england, where zoologists created favourable conditions for them to live and breed, and the hamster population grew rapidly. �e �rst hamsters were imported to poland in 1950 from france. �e rodent feeds mainly on plant food (stromenger, 1995). demodex criceti nutting & rauch and d. aurati nutting – demodex are the most commonly ectoparasite recognised in hamsters. d. criceti and d. aurati initially do not cause any clinical symptoms on the skin, and they usually feed naturally on the animal. d. criceti, is a smaller mite (appendix 1 – fig. 4a) than d. aurati (appendix 1 – fig. 4b), while d. aurati is more pathogenic. �e symptoms of demodex invasion are rough coat, skin in�ammation, balding, mainly on the back. �e sick, old, and undernourished animals are attacked most o�en (karaer et al., 2009). 6. house mouse (mus musculus l.) �e house mouse is a species of small mammals, which is synanthropic and cosmopolitan. it probably derives from mice inhabiting prairies and semi-desert areas. currently, it can be found wherever humans live, and it is omnivorous. �anks to high reproduction and a lack of high breeding requirements, mice are willingly used as laboratory animals and are also grown at homes. it has very high adaptability capabilities (zientek, 2015). a nn a k oc oń , m ag da le na n ow ak -c hm ur a 148 demodex �agellurus bukva and d. musculi oudemans – demodex is most common in mice. d. �agellurus (appendix 1 – fig. 4c) occurs mainly in the czech republic. in poland, the species has been found in the northern and central parts of the country, but there is no detailed information about this parasite. d. �agellurus is one of the largest known mites in mice, and it is mainly found near the genital area. �ey spread easily between hosts. in some populations, they show a high degree of rodent infestation. no symptoms of the diseases were noted for these demodex on the animals’ skin. d. musculi is associated with the areas of the whole body and it has also been observed in wild populations of mice and laboratory mice (izdebska, rolbiecki, 2015). conclusions and recommendations external parasites are important factors in the health of domestic animals and humans. �ey cause skin damage and pathological reactions from the immune system. �ey also play a role as a source of many types of pathogenic agents and can have negative e�ects on the bonds that exist between humans and animals. other negative e�ects of parasitic infestation are skin lesions leading to secondary bacterial or fungal infections, as well as dermatitis. under the in�uence of contact with saliva of the ectoparasites, allergic reactions may occur. �e most important of which is the allergic �ea dermatitis (sadzikowski, gundłach, 2004). due to the transfer of pathogens by ectoparasites that cause diseases in many cases with more clinical relevance than the parasite infestation itself, caring for our animals is particularly important. when deciding on a pet, we should be aware of such dangers and carefully combat the dangerous ectoparasites to keep our pets in good health and �tness. organisms with an invasion of external parasites can be a source of infestation for their owners or other animals living in the same area, which can be a huge problem. �e direct infestation of the host by ectoparasites may not be limited only to the skin level, but it can also lead to anaemia in the case of arthropods that suck blood in large quantities. in the years 2012–2014, the presence of ectoparasites collected from domestic animals was studied in cooperation with veterinary clinics in the silesian and lesser poland voivodeships. skin and hair samples were collected from 77 infected hosts (62 from lesser poland, 15 from silesia). �e following species dominated among the mites: otodectes cynotis var. cati (85.6%), o. cynotis var. canis (4.4%), demodex canis (5.2%), cheyletiella yasguri (3.5%). �e presence of the following was found in smaller numbers: sarcoptes scabiei var. canis (0.8%), notoedres cati (0.2%), chirodiscoides caviae (0.1%). out of the external insects, the majority of rat lice were collected – polyplax spinulosa (92.9%), gliricola porcelli (3.1%), columbicola columbae (2.2%), ctenocephalides felis (1.3%), and ceratophyllus gallinae (0.4%) (pawełczyk et al., 2016). �e studies on demodecid mites found that the most common species attacking s kin ectoparasites of dom estic anim als 149 domestic dogs is demodex canis; moreover, two other species can be found in dogs: d. injai occurring in adult sick dogs with hypothyroidism and cushings syndrome, and d. cornei, which o�en causes seborrheic dermatitis. all of these species occur in poland. �ree species of demodex were also observed in cats: the most common d. cati, d. gatoi causing highly infectious lesions, described at homes, where a lot of cats live together, and d. felis. two species were found in the golden hamsters – d. criceti and d. aurati, and in mice – d. arvicolae found in the skin of the eyelids and around the eyes, and d. �agellurus (jańczak et al., 2017). in the years 2003–2007, 382 specimens of bred reptiles imported to poland were studied, belonging to the species: testudo sp., iguana sp., varanus sp., gongylophis sp., python sp., spalerosophis sp., and psammophis sp., from which the ticks were collected. �e most infected reptiles came from ghana (africa) and these were varanus exanthematicus boso and python regius shaw. during the entire study, 2104 tick specimens were collected of the amblyomma genus (detected in poland for the �rst time): amblyomma exornatum koch, a. �avomaculatum lucas, a. latum koch (most abundant), a. nuttalli dönitz, a. quadricavum schulze, a. transversale lucas, a. varanense supino, amblyomma spp. and hyalomma: h. aegyptium l. studies were also performed in terms for the presence of pathogens. in two ticks, ambylomma �avomaculatum, the presence of anaplasma phagocytophilum dumler et al. was detected (nowak, 2010). frequent weather changes, frequent journeys, frequent changes in the locations of the animals probably a�ect the existing epizootic situation of some external parasites and the pathogens they carry, and they can cause the spread of these organisms to new areas. �is may lead to an increase in the incidence of rare parasitic diseases. domestic animals, which accompany us on a daily basis, not only play a positive role, but also a negative one, because they are a perfect source for carrying numerous ectoparasites. we should pay special attention to keeping our animals healthy by protecting them against attacks by dangerous parasites. in the case of the ectoparasite invasion, we 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(2008). prevalence of babesia canis, borrelia afzelli and anaplasma phagocytophilum infection in hard ticks removed from dogs in warsaw (central poland). veterinary parasitology, 153, 139–142. doi: 10.1016 / j.vetpar.2008.01.036 a nn a k oc oń , m ag da le na n ow ak -c hm ur a 154 appendix 1 fig. 1. demodex canis – a (source: http://people.upei.ca/), sarcoptes scabiei var. canis – b (source: https:// www.vectorbase.org), ctenocephalides canis – c (source: http://entnemdept.u�.edu), lingonathus setosus – d (source: https://veteriankey.com), cheyletiella yasguri – e (source: http://www.swiatczarnegoteriera. republika.pl), trichodectes canis – f (source: http://atlas.sund.ku.dk) s kin ectoparasites of dom estic anim als 155 fig. 2. satiated ixodes ricinus female – a, female i. hexagonus – b (photos from the collection of the department of the invertebrate zoology and parasitology), otodectes cynotis – c (source: http://www. swiatczarnegoteriera.republika.pl/), notoedres cati – d (source: https://www.esccap.fr/), demodex cati – e (source: https://www.cliniciansbrief.com), cheyletiellosis in cats – f (source:http://vetbook.org), ctenocephalides felis – g (source: https://pl.pinterest.com), felicola subrostratus – h (source: http:// mites-and-parasites.org) a nn a k oc oń , m ag da le na n ow ak -c hm ur a 156 fig. 3. dubininia melopsittaci – a (source: http://www.federmilben.de), sideroferus lunula – b (source: http://www.federmilben.de), feather-easting on feather �ags – c (source: http://www.golebiepocztowe.eu), �e results of the occurence of trixacarus caviae – d (source: http://www.guinealynx. info), chirodiscoides caviae – e (source: https://www.esccap.fr), gliricola porcelli – f (source: https://www. esccap.fr), demodex cavinae – g (source: http://www.happy-knu�els.de) s kin ectoparasites of dom estic anim als 157 fig. 4. demodex criceti – a, d. aurati – b, d. �agellurus – c (source: https://www.researchgate.net) a nn a k oc oń , m ag da le na n ow ak -c hm ur a 158 ektopasożyty skórne zwierząt domowych streszczenie zwierzęta towarzyszące ludziom, w tym zwierzęta domowe odgrywają ważną rolę w otaczającym nas środowisku. jednak większość z nich może być narażona na kontakt z gatunkami pasożytniczych roztoczy i owadów, o  dużym znaczeniu medycznym i  weterynaryjnym, stanowiących zagrożenie dla zdrowia zarówno zwierząt, jak i ludzi. do najbardziej rozpowszechnionych ektopasożytów skórnych zwierząt należą: demodex canis, d. cati, d. criceti, d. aurati, d. caviae, otodectes cynotis, sarcoptes scabiei, cheyletiella yasguri, c. blakei, chirodiscoides caviae, ctenocephalides canis, c. felis, trichodectes canis, linognathus setosus, felicola subrostratus, notoedres cati, dubininia melopsittaci, sideroferus lunula, neopsittaconirmus gracilis, gliricola porcelli, trixacarus caviae, ixodes ricinus, i. hexagonus, i. crenulatus, i. rugicollis i  dermacentor reticulatus. różne gatunki pasożytów zewnętrznych, które mogą być również przenoszone na ludzi są odpowiedzialne za najczęstsze choroby pasożytnicze skóry zwierząt: sarkoptozę, otodektozę, ktenocefalidozę, wszawicę, wszołowicę, demodekozę i chejletielozę. ponadto kleszcze z rodziny ixodidae odgrywają znaczącą rolę w przenoszeniu chorób, do najważniejszych z nich należą: borelioza z lyme, babeszjoza, riketsjoza, erlichioza, tularemia. w  niniejszej pracy przedstawiono najczęściej występujące ektopasożyty skórne u zwierząt domowych takich jak: pies domowy (canis familiaris), kot domowy (felis catus), papużka falista (melopsittacus undulatus), kawia domowa (cavia porcellus ), chomik syryjski (mesocricetus auratus), mysz domowa (mus musculus). key words: demodex, ectoparasites, �eas, lice, mites, pets, ticks received on: [2017.09.17] accepted on: [2017.11.12] 55 annales universitatis paedagogicae cracoviensis studia naturae, 3: 55–69, 2018, issn 2543-8832 doi: 10.24917/25438832.3.4 iwona konieczna1*, grzegorz rut1, angelika kliszcz2 1institute of biology, pedagogical university of cracow, podchorążych 2 st. 30-084 kraków, poland, *i.konieczna08@gmail.com 2department of agrotechnology and agricultural ecology, university of agriculture in kraków, mickiewicza st. 21, 31-120 kraków, poland effects of copper and vanadium salts on morphology of carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) and wheat (triticum aestivum l.) plants introduction plants growing in natural conditions are constantly exposed to biotic and abiotic factors that cause stress reactions. among the stress factors, a reaction with other organisms can be mentioned from a biotic side, and the abiotic side includes temperature, water availability, solar radiation, salinity, and environmental pollution, among others, by heavy metals (kozłowska, 2007). in the ecotoxicological context, heavy metals are considered to be poisons, i.e. substances that have toxic properties on living organisms. toxicity is the ability of matter to cause physiological disturbances or the death of a living organism (wierzbicka, 2015). the intensive development of industry and means of transport contributes unquestionably to the increase of environmental pollution with heavy metals (luo et al., 2012). copper is a chemical element from the group of transition metals. it creates a large variety of compounds, does not react with water, and it darkens on the air and takes on a red or red-brown colour. natural sources of copper emissions include volcanic eruptions, the decomposition of organic matter, forest fires, and seawater aerosols. anthropogenic copper pollution results from industrial activity, including the metallurgical industry, power plants, waste incineration plant effluvium, mining waste dumps, copper mine tailings, sewage (including municipal sewage), agricultural inputs (fungicides), and road transport (wierzbicka, 2015). plants collect copper in a passive or active way associated with metabolic processes, usually in the form of divalent cu2+ ions. the amount of copper consumed by plants is proportional to its concentration iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 56 in the substrate (kabata-pendias, pendias, 1999). copper from anthropogenic sources is better absorbed than its natural forms found in soils (grupe, kuntze, 1988). to meet the physiological needs of plants, small concentrations of copper are sufficient (> 2 ppm/dry mass). regardless of the harmfulness of excess copper, it is also an indispensable element for the proper growth and development of plants. it participates in the basic processes of photosynthesis, respiration, the transformation of nitrogen and protein compounds, the transport of carbohydrates, the metabolism of cell membranes (affecting their permeability), the standardization of dna and rna formation processes, and indirectly participates in the immune mechanisms (kabata-pendias, pendias, 1999; guerrero, 2005). vanadium is an element that belongs to the group of heavy metals. it is generally available in nature, and it is also widely used in industry. it is used, for example, in the production of non-ferrous alloys, highly resistant non-carbon steels, and in the chemical, glass, ceramic, paint, and photographic industries. in the atmosphere, this element comes mainly from anthropogenic sources, such as the combustion of biolites (crude oil), whose emissions are estimated at above 80%. it naturally occurs as a component of marine aerosols and comes from volcanic emissions (urban et al., 2001). in the dissolved form of anions (vo43and vo3-), it is easily absorbed by plants. as its concentration and acidity of soil increase, it is intensively absorbed in the soil (kabata-pendias, pendias, 1999). the largest accumulation of vanadium occurs in leaves (280–2700 ppb), but it can also occur in fruits and seeds (0.5–60 ppb). the harmful effect of vanadium for plants can be observed at concentrations from 500 to 1.400 μg/l, which inhibit root development, overall chlorosis, and dwarfism of plants. the vanadium content in soils ranges from 10 to 220 ppm. the largest concentration of that element is found in rendzinas, and the lowest in organic soils. in soil, it is usually associated with iron oxides and clay minerals. leaching from the level of podzolic soil and concentrations in loamy and ferruginous layers determine its mobility. vanadium concentration is increasing in industrial areas – metal works, cement plants, coal fired power plants, and oil refineries (kabata-pendias, pendias, 1999). carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) belongs to the apiaceae family (former umbelliferae). it is a precious vegetable plant cultivated around the world. it was probably brought to europe about 600 years ago from afghanistan (burnie, 2005). its cultivation is common, and as many as 60 species of this genus are edible plants. in the wild state, an annual, biennial, or perennial plants occur. the proper conditions for cultivation are in airy and non-acidic soils with a permeable subsoil level. it should not be cultivated on wet soils with a high level of groundwater, loam, and poor structure. the edible root has a wide range of applications in the kitchen, because of the content of carbohydrates, proteins, carotene, volatile compounds, and pectins. the vitamins like b, c, e, h, k, pp, and 57 effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants mineral salts (e.g., calcium, iron, copper, or phosphorus) are also found in the roots (kunachowicz et al., 2017). wheat (triticum aestivum l.) is one of the most important cereal species in the world. like other grasses, it belongs to the poaceae family. it probably comes from south-west and central asia. due to its nutritional value, this species is valued all over the world. wheat grains contain 69% sugars, 12% protein compounds, and 2% fats. these three main groups of organic compounds account for a total of 83% of the dry mass of the grains. wheat is also a source of riboflavin, thiamine, niacin, carotene, tocopherol, and mineral salts (e.g., sodium, calcium, potassium, magnesium, phosphorus, sulphur, and iron). it occupies a leading position among arable crops – third place in grain production in the world (gwóźdź, 2017). among others, it is used in the food industry, in cosmetics, as well as a raw material for energy production (karcz, 2013). the aim of this study was to determine the influence of copper and vanadium salts with different molar concentrations on the germination and growth of carrot (daucus carota subsp. sativus (hoffm.) schübl. & g. martens) and winter wheat (triticum aestivum l.). material and methods experiments were carried out under laboratory and greenhouse conditions at the department of plant physiology, pedagogical university of cracow. carrot seeds and wheat grains were used for testing. morphometric analyses of carrot and wheat treated with copper and vanadium salts at the germination and growth stages were performed. the experiment was carried out in two series in 10 replications. the copper and vanadium salt solutions were prepared from copper (ii) sulphate (cuso4) and ammonium metavanadate (v) (h4no3v, azanium; oxido(dioxo)vanadium, according to iupac), at concentrations of 0.3 mm, 0.6 mm, 3 mm, and 6 mm. the control group was samples watered with distilled water. 50 carrot seeds and 50 wheat grains were placed in sterile petri dishes with copper and vanadium solutions, using tweezers. the seeds on petri dishes were kept for 9 days in the dark at room temperature. after this time, biometric analysis, the fresh and dry mass, and the percentage of water content of carrot and wheat seedlings were measured. a few seedlings with similar morphology were selected from the control group and put in pots. river sand, rinsed several times with running and distilled water, comprised the medium of seedlings in the pots. then, they were kept in a greenhouse. in the first week, plants were watered regularly 2–3 times a week with distilled water (10 ml each) and once a week with steiner medium (10 ml each). in the next four weeks, the plants were watered once a week, respectively with copper (cuso4) and vanadium (h4no3v) salts, with both concentrations of 0.6 mm and 3 mm (10 ml each) and iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 58 steiner medium (10 ml each). the control samples were watered twice a week with distilled water (10 ml each) and once a week with steiner medium (10 ml each). biometric analysis the biometric analysis of carrot and wheat seedlings was made with a ruler (with an accuracy of 0.1 cm). measurements of the length of underground and aboveground plant organs were carried out as well. fresh, dry mass and water content fresh mass fm of seedlings and plants was determined on an electronic balance (radwag wps 210c). dry mass dm was determined after oven-drying (wamed sup 100) plant materials at 110°c for 48 hours. on the basis of the obtained results, the percentage of water content wc (%) was determined according to the formula: % h2o = 100 – [(dm × 100) / fm]. the measurements were made on seedlings and on the underground and aboveground parts of carrot and wheat plants. statistical analysis the results presented in the paper are mean values from 10 independent replications with the standard deviation (± sd), calculated for each experimental variant. analyses of the significance of differences between objects were made using the one-way anova/manova parametric statistical test, single and multi-factorial, based on the duncan test, with p ≤ 0.05. calculations were made using statistica, v. 13.0 (statsoft, inc. 2017). results germination percentage copper and vanadium salts had a clear effect on daucus carota subsp. sativus seeds and triticum aestivum grain germination (fig. 1). the negative effect of copper ions on d. carota seeds germination in each of the concentrations of copper solution (cuso4) was observed. only carrot seeds treated with 0.6 mm copper solutions germinated in the same amount as in the control – 34%. the carrot seed germination was inhibited to 10% on 3 mm cuso4 solution. on petri dishes with 6 mm cuso4, a complete inhibition of germination was revealed. in contrast, vanadium salt solutions (h4no3v), had a positive effect on the germination of carrot seeds as compared to the control treatment. the germination of t. aestivum grains depended on the concentration and the type of used solutions. the copper and vanadium solutions used at the lowest concentration, i.e. 0.6 mm, slightly decreased the amount of germinated grains, compared to 59 fig. 1. germination of carrot seeds (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – (a and b) and winter wheat grains (triticum aestivum l.) – (c and d), treated with copper (cuso4) and vanadium (h4no3v) solutions at different concentrations (0.6 mm, 3 mm, and 6 mm) a b c d effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants the control. with the increasing of copper and vanadium concentrations (3 mm and 6 mm, respectively), inhibition of wheat germination was revealed. biometric analysis of seedlings the growth of carrot seedlings with increasing of copper and vanadium concentrations was significantly inhibited. in comparison to seedlings germinated on distilled water (control), the elongation growth of carrot seedlings was completely inhibited at the highest concentration of copper sulphate solution (6 mm cuso4). the root length of wheat seedlings was inhibited in each concentration of both the copper and vanadium salt solutions (fig. 2). the copper and vanadium solutions at concentrations of 6 mm caused the highest inhibition of root growth. the control wheat seedlings had significantly longer coleoptiles than those saturated with copper and vanadium salt solutions. the only exception was the seedlings germinated on 0.6 mm cuso4, when compared to the control group, no statistically significant differences in the coleoptiles length were found. the length of whole wheat seedlings, independent of the concentration and type of salt solutions, was shorter in relation to the seedlings germinated on distilled water. fresh and dry mass values and water content of seedlings the fresh mass values of carrot seedlings were significantly increased only when saturated with 0.6 mm cuso4. in other cases, no significant changes in the values of this iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 60 fig. 2. length of carrot seedlings (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – (a), roots – (b), coleoptiles – (c) and whole seedlings – (d) wheat (triticum aestivum l.), germinated on substrates with copper (cuso4) and vanadium (h4no3v) solutions, at various concentrations (0.6 mm, 3 mm and 6 mm); mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 parameter were noted as compared to the control group. the dry mass of carrot seedlings was not significantly different than seedlings germinated on salt solutions and distilled water (control). the percentage of water content only significantly increased in carrot seedlings treated with copper solutions at a 0.6 mm concentration. none of the other concentrations of copper and vanadium were found to significantly differ in the case of water content relative to control seedlings (tab. 1). the fresh mass of wheat seedlings germinated on 3 mm and 6 mm cuso4 and 6 mm h4no3v was significantly decreased, compared to the control. the dry mass values of wheat seedlings significantly increased both at concentrations of 3 mm and 6 mm copper and vanadium salt solutions. the water content of wheat seedlings decreased with the increasing of concentrations of salt solutions when compared to the control. differences in water content values were observed between seedlings treated the 3 mm and 6 mm cuso4, 6 mm h4no3v solutions and control (tab. 1). 61 tab. 1. fresh, dry mass and percentage of the water content of carrot seedlings (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – a and wheat grains (triticum aestivum l.) – b, treated with copper (cuso4) and vanadium (h4no3v) salt solutions at different concentrations (0.6 mm, 3 mm and 6 mm) pa ra m et er s control concentration of the solution [mm] cuso4 h4no3v 0.6 3 6 0.6 3 6 a b a b a b a b a b a b a b fm [g] 0.19 9.31 0.35* 8.69 0.11 5.44* 0.11 5.31* 0.16 6.64 0.12 6.14 0.13 5.59* dm [g] 0.06 1.47 0.05 1.60 0.04 1.77* 0.05 1.77* 0.03 1.62 0.04 1.74* 0.04 1.80* wc [%] 67 84 85* 82 60 68* 54 67* 83 76 67 72 71 68* fm – fresh mass [g], dm – dry mass [g], wc (%) – water content [%]; average values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 tab. 2. fresh mass values of carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – a and wheat (triticum aestivum l.) – b organs, watered with copper (cuso4) and vanadium (h4no3v) salt solutions, at various concentrations (0.6 mm, 3 mm and 6 mm) organ control concentration of solution [mm] cuso4 h4no3v 0.6 3 0.6 3 a b a b a b a b a b root 0.40 4.20 0.25* 4.54 0.24* 4.10 0.21* 4.65 0.20* 4.38 fourth leaf 0.25 1.86 0.23 1.82 0.22 1.68 0.22 1.58 0.18* 1.35 other leaves 0.22 1.69 0.19 1.44 0.16* 1.34 0.22 1.73 0.20 1.62 the whole plant 0.87 7.75 0.67* 7.80 0.62* 7.12 0.65* 7.96 0.57* 7.35 mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 biometric analysis of plant organs tab. 3. dry mass values of carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – a and wheat (triticum aestivum l.) organs – b, treated with copper (cuso4) and vanadium (h4no3v) salt solutions at various concentrations (0.6 mm, 3 mm, and 6 mm) organ control concentration of the solution [mm] cuso4 h4no3v 0.6 3 0.6 3 a b a b a b a b a b root 0.18 0.40 0.10 0.24* 0.03* 0.28* 0.07* 0.21* 0.02* 0.20* fourth leaf 0.08 0.25 0.08 0.22 0.01* 0.22 0.06 0.22 0.04* 0.18 other leaves 0.15 0.22 0.15 0.19 0.01* 0.16* 0.12 0.22 0.08* 0.20 the whole plant 0.41 0.87 0.33 0.66 0.04* 0.630 0.21* 0.65 0.19* 0.57* mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 62 the biometric analysis of carrot roots showed a slower growth trend both on 3 mm cuso4 and h4no3v solutions, relative to control conditions. the length of carrot leaves was not significantly different than copper and vanadium solutions and the control group. the copper and vanadium salts in none of the concentrations significantly affected changes in wheat root length. the growth of wheat leaves was stimulated by cuso4 solutions at concentrations of 0.6 mm and 3 mm, compared to the plant watered with vanadium solutions and distilled water (control) (fig. 3). fresh and dry mass and the percentage of water content in organs of plants fresh mass of carrot organs watered with copper and vanadium salt solutions clearly differed from the control sample (tab. 2). the fresh mass of root in each of the used solutions was decreased. for the fourth leaf, a significant mass reduction was observed with 3 mm h4no3v. compared to the control carrot organs, the fresh mass of other leaves decreased only in plants watered with 3 mm cuso4. generally, the fresh mass of the whole plant decreased under the influence copper and vanadium salts. the dry mass of carrot plants decreased in samples treated with solutions of the highest salts concentrations. the vanadium salts decreased the dry mass values in each concentration. for the root and the whole plant, significant differences in dry mass values were revealed. the water content increased for all carrot organs treated with 3 mm copper and vanadium solutions compared to the control group (tab. 3–4). the fresh mass of wheat organs treated with copper and vanadium solutions did not significantly differ relative to the control plants. the only change was noticed in the dry mass of wheat root. the dry mass of whole plants decreased in plants watered with 3 mm h4no3v. the percentage of water content did not differ from the control treatment (tab. 2–4). tab. 4. comparison of the water content in carrot organs (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) – a and wheat (triticum aestivum l.) – b, treated with copper (cuso4) and vanadium (h4no3v) salt solutions at various concentrations (0.6 mm, 3 mm and 6 mm) organ control concentration of the solution [mm] cuso4 h4no3v 0.6 3 0.6 3 a b a b a b a b a b root 54 90 59 95 88* 94 65 95 91* 95 fourth leaf 67 86 66 87 98* 87 71 86 78 87 other leaves 34 87 24 87 96* 88 43 87 62* 88 the whole plant 53 89 51 92 94* 91 68 92 92 92 mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 63 fig. 3. comparison of the length of roots and leaves of carrot (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) and wheat (triticum aestivum l.), treated with the copper (cuso4) and vanadium (h4no3v) salt solutions, at different concentrations (0.6 mm and 3 mm); mean values (n = 10) marked with an asterisk (*) differ significantly according to duncan test at p ≤ 0.05 a b discussion unlike other undesirable substances, heavy metals cannot be broken down only by biotransformation, due to complex physico-chemical and biological processes occurring in the soil. these processes affect their mobility and bioavailability in the soil – plant system (qishlaqi, moore, 2007). depending on the form, type, and concentration in the environment, heavy metals may have a positive or negative effect on organisms. the positive influence refers to those of elements that are essential in the metabolic processes (e.g., fe, mn, cu, zn, and mo), whereas toxic elements (i.e. as, hg, pb, cr, and cd) present in environment in trace amounts have a negative impact on the living organisms (gil et al., 2006). germination is a set of complex physiological processes regulated at the molecular, subcellular, and cellular level. the imbibition phase includes intensive water intake and increased respiration. in the catabolic phase, the hydrolytic decomposition of storage materials occurs, and the synthesis of cellular components begins in effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 64 the anabolic phase. the shoot that grows from the seed penetrates the seed coating, becoming a sprout and then a seedling (bewley, 1997). heavy metals at the early phase of the germination process affect the functioning of cellular organelles, including cell membranes, mitochondria, lysosomes, endoplasmic reticulum, and nucleus. they also affect enzymes involved in metabolism, detoxification, and the repair of cell damage (wang, shi, 2001). heavy metal ions interfere with dna and nuclear proteins and cause damage to genetic material. moreover, they induce conformational changes that may intrude the cell cycle modulation or cause carcinogenesis and apoptosis (beyersmann, hartwig, 2008). the experiments showed that, even during germination, the copper and vanadium ions significantly influenced the germination of d. carota subsp. sativus and t. aestivum l. an inhibition in the germination of seeds was observed with increasing concentration of metals (fig. 1). zandi et al. (2017), and możdżeń and rzepka (2016) also demonstrated similar symptoms of the negative impact of heavy metals on plant germination. the maximum capacity of each plant to absorb heavy metals depends on the gradient of their concentration and availability in the soil. plants imbibe heavy metals by diffusion or selective transport (peralta-videa et al., 2009). in the first stage, some of the metals are absorbed by the apical area of the plant’s roots, and some are absorbed by the entire root surface. usually, heavy metal ions are retained in the root system and only in very small quantities transported to the shoots (krzesłowska, 2011; hossain et al., 2012; barberon, geldner, 2014). the most common symptoms of excessive accumulation of heavy metals in plants are growth disorders and changes in the biomass of organs. in the root, there are bulges in the apical part, a reduction in diameter and number of transport vessels, browning, discoloration of cell structures, lipid disturbances, and ion destabilisation (rucińska-sobkowiak, pukacki, 2006; qureshi et al., 2007). as a consequence, the suppression of the entire plant growth occurs, and, in extreme cases, the plants die (ciećko et al., 2000). after analysing the results of these studies, both changes in the mass and the morphometric properties of carrot’s and wheat’s seedlings were observed (fig. 1–3, tab. 1–4). with the increase in the concentration of heavy metals, the inhibition of plant growth and their masses has been demonstrated. the largest differences at the highest concentrations of copper and vanadium solutions in relation to the control plants were noted. the toxic effects of copper on growth and dm in tobacco (nicotiana tabacum l.), mung beans (vigna radiata (l.) r. wilczek), and chinese tea (camellia sinensis (l.) kuntze) have also detected (gori et al., 1998; manivasagaperumal et al., 2011; dey et al., 2015). similarly, janas et al. (2010) found a significant influence of copper on growth, lipid peroxidation, and the accumulation of phenolic compounds in lentil seedlings (lens culinaris medic.). możdżeń et al. (2017) observed a reduction in the growth of one-year-old needles of scots pine (pinus sylvestris l.) in the warcino for65 est district (northwest poland). however, in numerous studies, copper has positively influenced the growth of rape (brassica napus l.) (purnhauser, gyulai, 1993), barley (hordeum vulgare l.) (wojnarowiez et al., 2002), and annual peppers (capsicum annuum l.) (joshi, kothari, 2007). plants have a network of defence strategies that allow them to avoid or tolerate stress factors, including heavy metals. the first line of defence is physical barriers, which include some morphological structures (thick skin) and biologically active tissues (hair and cell walls). for example, trichomes are used for the detoxification of heavy metals or the secretion of various secondary metabolites to protect against the adverse effects of metals. when heavy metals overcome biophysical barriers and metal ions get into tissues and cells, plants initiate the second line of defence, i.e. cellular defence mechanisms that eliminate or alleviate the negative effects of heavy metals (emamverdian et al., 2015). soil is one of the main reservoirs that accumulate significant amounts of harmful chemicals. slow but continuous pollution in the environment with heavy metals leads to serious threats to microorganisms, plants, and animals. the basic condition for limiting the uptake of heavy metals by plants is to provide them with optimal growth conditions. cultivated areas should be located away from busy roads or mills. farmers should care about maintaining a stable ph of soils (ph = 6.5–7), regular organic fertilisation, such as manure, compost or ‘green manures’ (luo et al., 2012). there have been many experiments on the impact of heavy metals on living organisms at various levels of their organisation. for example, for cadmium (cd) and zinc (zn) as suggested by current studies, the uptake of their ions by plants is genetically correlated, and these metals are taken by the same or different transporters and controlled by common regulators (oves et al., 2016). in order to restore the ecological balance of the environment and learn about the laws governing it, it is necessary to conduct further research on the mechanisms of interaction of heavy metals on various plant species. only comprehensive laboratory experiments at the level of whole plants, organs, cells, and molecules combined with field research will be able to reveal the positive and negative significance of various heavy metal ions in nature and indicate effective ways to eliminate their excess (tchounwou et al., 2014). conclusion in this experiment, the copper and vanadium ions reduced the number of germinated carrot seeds (daucus carota subsp. sativus) and winter wheat grains (triticum aestivum) with the increase in the concentration of their solutions. growth of carrot and wheat seedlings in copper and vanadium salt solutions was inhibited, compared to control seedlings. the fresh mass of carrot seedlings on petri dishes watered with 0.6 mm cuso4 was increased. the dry mass of wheat seedlings increased in 3 mm effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 66 and 6 mm copper and vanadium salts solutions. the percentage of water content in carrot seedlings was only higher in 0.6 mm cuso4. in the growth phase, the reduction in the root length of carrot was observed. as for wheat, the stimulation of leaf growth of the specimens watered with solutions of concentrations of 0.6 mm and 3 mm was assessed. in comparison to the control group, with the concentration of copper and vanadium ions, there has been a reduction in the fresh mass of carrot’s organs. the dry mass values of carrot and wheat plants decreased under the influence of copper and vanadium salts. copper and vanadium ions, in the highest concentrations, significantly influenced the water content of carrot organs. the salts used in the experiment, both in germination and growth phases, significantly affected the morphology of carrot and wheat plants. references barberon, m., geldner, n. (2014). radial transport of nutrients: the plant root as a polarized epithelium. plant physiology, 166, 528–537. doi: 10.1104/pp.114.246124 bewley, j.d. (1997). seed germination and dormancy. plant cell, 9, 1055–1066. doi: 10.1105/tpc.9.7.1055 burnie, g. (2005). botanica: the illustrated a-z of over 10,000 garden plants and how to cultivate them. germany, königswinter: ullmann publishing. ciećko, z., wyszkowski, m., żołnowski, a. (2000). działanie zanieczyszczenia gleby ołowiem i nawożenia wapniem na plonowanie i skład chemiczny kukurydzy. zeszyty problemowe postępów nauk rolniczych, 472, 129–136. [in polish] dey, s., mazumder, p.b., paul, s.b. (2015). copper-induced changes in growth and antioxidative mechanisms of tea plant (camellia sinensis (l.) o. kuntze). african journal of biotechnology, 14, 582–592. emamverdian, a., ding, y., mokhberdoran, f., xie, y. 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(2012). molecular mechanism of heavy metal toxicity and tolerance in plants: central role of glutathione in detoxification of reactive oxygen species 67 and methylglyoxal and in heavy metal chelation. journal of botany, 37. doi: 10.1155/2012/872875 iupac nomenclature, https://pubchem.ncbi.nlm.nih.gov janas, k.m., zielińska-tomaszewska, j., rybaczek, d., maszewski, j., posmyk, m.m., amarowicz, r., kosińska, a. (2010). the impact of copper ions on growth, lipid peroxidation, and phenolic compound accumulation and localization in lentil (lens culinaris medic.) seedlings. journal of plant physiology, 167(4), 270–276. doi: 10.1016/j.jplph.2009.09.016. joshi, a., kothari, s.l. (2007). high copper levels in the medium improves shoot bud differentiation and elongation from the cultured cotyledons of capsicum annuum l. plant cell tissue and organic culture, 88, 127–133. doi: 10.1007/s11240-006-9171-6 kabata-pendias, a., pendias, h. (1999). biogeochemia pierwiastków śladowych. warszawa: wydawnictwo naukowe pwn. [in polish] karcz, h., kantorek, m., grabowicz, m., wierzbicki, k. (2013). możliwość wykorzystania słomy jako źródła paliwowego w kotłach energetycznych. piece przemysłowe i kotły, 11–12, 8–15. [in polish] kozłowska, m. (2007). fizjologia roślin. od teorii do nauk stosowanych. warszawa: państwowe wydawnictwo rolnicze i leśne, pp. 466–468. [in polish] krzesłowska, m. (2011). the cell wall in plant cell response to trace metals: polysaccharide remodelling and its role in defense strategy. acta physiologiae plantarum, 33, 35–51. doi: 10.1007/s11738-010-0581-z kunachowicz, h., przygoda, b., nadolna, i., iwanow, k. (2017). tabele składu i wartości odżywczej żywności. warszawa: wydawnictwo lekarskie pzwl. [in polish] luo, ch., yang, r., wang, y., li, j., zhang, g., li, x. (2012). influence of agricultural practice on trace metals in soils and vegetation in the water conservation area along the east river (dongjiang river), south china. science of the total environment, 431, 26–32. doi: 10.1016/j.scitotenv.2012.05.027 manivasagaperumal, r., vijayarengan, p., balamurugan, s., thiyagarajan, g. (2011). effect of copper on growth, dry matter yield and nutrient content of vigna radiata (l.) wilczek. journal of phytology, 3, 53–62. możdżeń, k., rzepka, a. (2016). rola łupiny nasiennej podczas kiełkowania i wzrostu nasion bobu (vicia faba l.) w obecności siarczanu ołowiu. annales umcs sectio e agricultura, 71(4), 55–65. [in polish] możdżeń, k., wanic, t., rut, g. łaciak , t., rzepka, a. (2017). toxic effect of high copper content on the physiological processes in pinus sylvestris l. photosynthetica, 55(1), 193–200. doi: 10.1007/s11099016-0229-3 oves, m., saghir khan, m., huda qari, a., nadeen felemban, m., almeelbi, t. (2016). heavy metals: biological importance and detoxification strategies. journal of bioremediation and biodegradation, 7(2), 1–15. doi: 10.4172/2155-6199.1000334 peralta-videa, j.r., lopez, m.l., narayan, m., saupe, g., gardea-torresdey, j.l. (2009). the biochemistry of environmental heavy metal uptake by plants: implications for the food chain. the international journal of biochemistry and cell biology, 41, 1665–1677. doi: 10.1016/j.biocel.2009.03.005. purnhauser, l., gyulai, g. (1993). effect of copper on shoot and root regeneration in wheat, triticale, rape and tobacco tissue cultures. plant cell tissue and organic culture, 35, 131–139. doi: 10.1007/ bf00032962 qishlaqi, a., moore, f. (2007). statistical analysis of accumulation and sources of heavy metals occurrence in agricultural soils of khoshk river banks, shiraz, iran. american-eurasian journal of agriculture and environmental science, 2(5), 565–573. qureshi, m.i., abdin, m.z., qadir, s., iqbal, m. (2007). lead-induced oxidative stress and metabolic alterations in cassia angustifolia vahl. biologia plantarum, 51, 121–128. doi: 10.1007/s10535-0070024-x effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants iw on a k on ie cz na , g rz eg or z r ut , a ng el ik a k lis zc z 68 rucińska-sobkowiak, r., pukacki, p.m. (2006). antioxidative defense system in lupin roots exposed to increasing concentrations of lead. acta physiologiae plantarum, 28, 357–364. doi: 10.1007/s11738006-0032-z starck, r. (1984). uprawa roli i nawożenie roślin ogrodniczych. warszawa: państwowe wydawnictwo rolnicze i leśne. [in polish] tchounwou, p.b., yedjou, c.g., patlolla, a.k., sutton, d.j. (2014). heavy metals toxicity and the environment. national institiutes health, 1–30. doi: 10.1007/978-3-7643-8340-4_6 the plant list. a working list of all plant species. http://www.theplantlist.org/browse/a/poaceae/triticum/ wang, s., shi, x. (2001). molecular mechanisms of metal toxicity and carcinogenesis. molecular and cellular biochemistry, 222, 3–9. weber, r., hryńczuk, b. (2000). effect of leaf and soil contaminations on heavy metals content in spring wheat crops. nukleonika, 45(2), 137–140. wierzbicka, m. (2015). ekotoksykologia rośliny, gleby, metale. warszawa: wydawnictwo uniwersytetu warszawskiego, p. 17. [in polish] wojnarowiez, g., jacquard, c., devaux, p., sangwan r.s., clement, c. (2002). influence of copper sulfate on anther culture in barley (hordeum vulgare l.). plant science, 162, 843–847. doi: 10.1016/s01689452(02)00036-5 zandi p., możdżeń k., basu s.k., gonzález-valdivia n.a., cetzal-ix w. (2017). effect of different cadmium nitrate concentrations on sweet maize (zea mays l. cv. landmark). in: s.k. basu, p. zandi (eds.), environment at crossroads: challenges, dynamics, solutions. iran: haghshenass publication. abstract around the world, a major concern in researches on heavy metals is placed on their toxic effect on living organisms. the problem with heavy metals occurrence in the environment is not only associated with their toxicity, but also with their ability to accumulate inside living organisms. this study presents the effect of copper and vanadium ions on the germination and growth of carrot and wheat plants. the experiment was carried out in two independent series with ten repetitions each. the water solutions of copper (cuso4) and vanadium (h4no3v) salts with the concentrations of 0.3 mm, 0.6 mm, 3 mm, and 6 mm were used. the control groups were objects watered with distilled water. the conducted experiment showed that the copper and vanadium ions had negative effects on the germination and growth of tested plants. with the increasing concentration of heavy metal ions, an inhibition of seeds germination was observed. the length of carrot and wheat seedlings in each salt solution was inhibited, compared to the control group. during the growth phase, the stimulation of wheat leaves growth only in copper solutions with concentration 0.3 mm and 0.6 mm was observed. depending on the copper and vanadium ions concentrations, changes in the fresh and dry masses and the content of water were observed. key words: cereal, root crop, germination, fresh and dry mass, copper, vanadium received: [2018.02.09] accepted: [2018.10.15] wpływ jonów miedzi i wanadu na morfologię marchwi uprawnej (daucus carota l. subsp. sativus (hoffm.) schübl. & g. martens) oraz pszenicy (triticum aestivum l.) streszczenie na całym świecie prowadzone są badania nad toksycznym wpływem metali ciężkich na organizmy żywe. problem z ich występowaniem w środowisku wynika nie tylko z toksyczności, lecz także ze zdolności akumulowania w  organizmach żywych. w  pracy przedstawiono wyniki badań nad wpływem jonów miedzi 69 i  wanadu na kiełkowanie i  wzrost marchwi uprawnej oraz pszenicy zwyczajnej. doświadczenie przeprowadzono dla każdego gatunku w dwóch niezależnych seriach, po 10 powtórzeń. w badaniach stosowano wodne roztwory soli miedzi (cuso4) i wanadu (h4no3v), o stężeniach molowych: 0,3 mm; 0,6 mm; 3 mm i 6 mm. grupę kontrolną stanowiły obiekty podlewane wodą destylowaną. przeprowadzone eksperymenty wykazały negatywny wpływ jonów miedzi i wanadu na kiełkowanie oraz wzrost badanych roślin. wraz ze wzrostem koncentracji jonów metali ciężkich obserwowano zahamowanie zdolności kiełkowania nasion, a  siewki podlewane roztworami soli miedzi i  wanadu były krótsze, względem podlewanych wodą. w  fazie wzrostu, jedynie u pszenicy zaobserwowano stymulację przyrostu liści pod wpływem związków miedzi o stężeniach 0,3 mm i 0,6 mm. w zależności od koncentracji jonów miedzi i wanadu odnotowano istotne zmiany w wartościach świeżej i suchej masy oraz procentowej zawartości wody w siewkach. słowa kluczowe: roślina okopowa, zboże, zdolność kiełkowania, świeża i sucha masa, miedź, wanad information about authors iwona konieczna https://orcid.org/0000-0002-9538-8516 she is a student of biology. she is interested in allelopathic interaction between plants and effect of heavy metals on plant morphology and physiology. grzegorz rut https://orcid.org/0000-0001-6719-3060 his research topic concerns the course of basic physiological processes in plants (photosynthesis, respiration, chlorophyll fluorescence, transpiration), concentration chlorophyll, antocyanins, and reactions of plants, mainly mosses (polytrichum piliferum, mnium undulatum), ferns (platycerium bifurcatum) and vascular plants (oxalis acetosella, o. corniculata) on abiotic stress factors (excess and deficiency of solar radiation, hypoxia, anoxia, excess heavy metals, herbicides, salinity, changes in carbon dioxide concentrations, drought stress and low temperature). angelika kliszcz she is focusing on enhancing the understanding of the influence of different factors on soil structure and fertility. particularly, she is investigating the interaction of plants with the physical, chemical, and biological properties of the soil. she is also interested in organic methods of plant production and soil-enriching substances. effects of copper and vanadium salts on m orphology of carrot (d aucus carota l. subsp. sativus (h offm .) s chübl. & g . m artens) and w heat (triticum aestivum l.) plants 195 annales universitatis paedagogicae cracoviensis studia naturae, 4: 195–201, 2019, issn 2543-8832 from 1 to 7 of july, the 58th polish botanical society (pbs) congress under topic “botany without borders” took place. it had not only nationwide form but also an international scienti�c conference character. �e organisers of this important event for botanists were: kraków division of the polish botanical society, w. szafer institute of botany (polish academy of science), institute of botany of the jagiellonian university, institute of biology of the pedagogical university of krakow and botanical garden of the jagiellonian university. all scienti�c events of the congress occurred in the main building of the pedagogical university of krakow at podchorążych 2 st. �e lectures and poster sessions were delivered in 14 topic related sections: aerobiological, bryological, dendrological, plant physiology and biochemistry, geobotany and protection of the floras, history of botany, plant tissue cultures, lichenological, mycological, botanical gardens and arboreta, paleobotanical, pteridological, plant structure and development and vascular plants taxonomy. on the �rst day of the congress (july 1st), organisational meetings were held related 58th polish botanical society congress, “botany without borders” (july 1–7, 2019, kraków, poland) 58. zjazd polskiego towarzystwa botanicznego „botanika bez granic” (1–7 lipca 2019, kraków, polska) w dniach 1–7 lipca 2019 r. odbył się 58. zjazd polskiego towarzystwa botanicznego (ptb), pod hasłem „botanika bez granic”, mający tradycyjnie formę nie tylko ogólnopolskiej, ale i międzynarodowej konferencji naukowej. organizatorami tego ważnego dla botaników wydarzenia były: oddział krakowski ptb, instytut botaniki im. w. szafera polskiej akademii nauk, instytut botaniki uniwersytetu jagiellońskiego, instytut biologii uniwersytetu pedagogicznego w krakowie oraz ogród botaniczny uniwersytetu jagiellońskiego. wszystkie obrady tegorocznego zjazdu odbywały się w budynkach uniwersytetu pedagogicznego, przy ul. podchorążych 2 w  krakowie. wystąpienia ustne oraz doniesienia plakatowe prezentowano w  14 sekcjach tematycznych: aerobiologicznej, briologicznej, dendrologicznej, fizjologii i  biochemii roślin, geobotaniki i  ochrony szaty roślinnej, historii botaniki, kultur tkankowych roślin, lichenologicznej, mykologicznej, ogrodów botanicznych i arboretów, paleobotanicznej, pteridologicznej, struktury i rozwoju roślin oraz taksonomii roślin naczyniowych. pierwszego dnia zjazdu (1 lipca) odbyły się spotkania organizacyjne, związane 196 r ep or ts to the functioning of the society: a meeting of the presidium of the main council of the pbs, a meeting of the main board of pbs and the general assembly of delegates. as a result, a number of resolutions important for the society were adopted and the chairman and new presidium of the main board of the pbs were elected for the 2019–2022 term. in the late a�ernoon members of pbs had the opportunity to participate in two outdoor sessions: “herbarium murorum cracoviensis – plants in the architecture of cracow” and “sightseeing of the botanical garden of the jagiellonian university”. �e next day of the congress (july 2nd) the o�cial opening of the 58th polish botanical society congress took place, during z  funkcjonowaniem towarzystwa: posiedzenie prezydium zarządu głównego ptb, posiedzenie zarządu głównego ptb oraz walne zgromadzenie delegatów. w  efekcie podjęto szereg ważnych dla towarzystwa uchwał oraz wybrano przewodniczącego i  nowe prezydium zarządu głównego ptb na kadencję 2019–2022. natomiast późnym popołudniem miały miejsce dwie sesje plenerowe: „herbarium murorum cracoviensis – rośliny w architekturze krakowa” oraz „wizyta w  ogrodzie botanicznym uniwersytetu jagiellońskiego”. następnego dnia (2 lipca) nastąpiło uroczyste otwarcie 58. zjazdu ptb, podczas którego swoje wystąpienia wygłosili dotychczasowi przedstawiciele władz towarzystwa oraz fig. 1. speech by vice-rector of development of the pedagogical university of krakow – assoc. prof. robert stawarz, during the inauguration of the 58th congress of the polish botanical society (photo. d. węgiel) ryc. 1. przemowa prorektora ds. rozwoju uniwersytetu pedagogicznego w krakowie – dr hab. roberta stawarza, w trakcie inauguracji 58. zjazdu polskiego towarzystwa botanicznego (fot. d. węgiel) 197 r eports which the actual authorities of the society and invited guests presented their speeches (fig. 1). awards and medals for the most active botany scientists were also given (among others: medal of prof. władysław szafer, medal of prof. bolesław hryniewiecki (fig. 2) and medal of prof. zygmunt czubiński). next, the plenary sessions occurred. during them several lectures of local (fig. 3) and foreign researchers were presented which highlighted both the history and perspectives of global botany. particularly worth mentioning is the interdisciplinary and international character of this part of the congress. in the evening, a�er the dynamic discussion on the actual trends in the biology and evolution of the plant kingdom, it was time for a mozaproszeni goście (ryc. 1). nadano również odznaczenia i  medale naukowcom zasłużonym w  dziedzinie szeroko pojętej botaniki (m. in.: medal im. prof. władysława szafera, medal im. prof. bolesława hryniewieckiego (ryc. 2) oraz medal prof. zygmunta czubińskiego). w  dalszej kolejności odbyły się obrady sesji plenarnych, podczas których przedstawione zostały referaty krajowych (ryc. 3) oraz zagranicznych wykładowców dotyczące, m. in. porównania dorobku oraz perspektyw ogólnoświatowej botaniki. na szczególną uwagę zasługuje interdyscyplinarny oraz międzynarodowy charakter tej części zjazdu. wieczorem, po burzliwej wymianie poglądów na temat najnowszych trendów w  badaniach nad biologią i  ewolucją królefig. 2. �is year’s winners of the medal of professor bolesław hryniewiecki decorated for disseminating botanical knowledge (photo. d. węgiel) ryc. 2. tegoroczni laureaci medalu im. prof. bolesława hryniewieckiego odznaczeni za upowszechnianie wiedzy botanicznej (fot. d. węgiel) 198 r ep or ts ment of re�ection and calm during the organ concert at the church of st. francis of assisi in kraków. without doubts it was exactly the moment when we all could understand that botany is really a science “without borders” in which there should be a place for everyone who loves the beauty and the scienti�c significance of plants. on the third day of the congress (july 3rd), lectures and poster sessions in the individual pbs sections began. in addition, at the beginning of the meeting, a bene�t dedicated to the activities of prof. małgorzata latałowa took place. it summarised her contribution as an outstanding researcher to the current state of paleobotanical and phytogeographic knowledge. many hours of deliberations and discussions ended on that day with a gala stwa roślin, przyszedł czas na chwilę re�eksji i  wyciszenia podczas koncertu organowego w  bazylice ojców franciszkanów w  krakowie. niewątpliwie był to ten moment, w którym wszyscy mogli uzmysłowić sobie, że botanika jest nauką „bez granic“, w której powinno być miejsce dla każdego, kto umiłował sobie piękno oraz naukowe znaczenie roślin. trzeciego dnia zjazdu (3 lipca) rozpoczęły się obrady w sesjach tematycznych, referatowych i plakatowych, w poszczególnych sekcjach ptb. ponadto na początku tych obrad miał miejsce bene�s okolicznościowy, poświęcony działalności pani prof. dr hab. małgorzaty latałowej, podsumowujący wkład tej wybitnej badaczki w  obecny stan wiedzy paleobotanicznej i  �togeogra�cznej. wielogodzinne obrady i  dyskusje zakończofig. 3. plenary session: professor martin kukwa (university of gdańsk): lichenology – where we were, where we are, where we are going (photo. d. węgiel) ryc. 3. sesja plenarna: prof. dr hab. martin kukwa (uniwersytet gdański): lichenologia – gdzie byliśmy, gdzie jesteśmy, dokąd zmierzamy (fot. d. węgiel) 199 r eportsdinner, accompanied by the sounds of music in the kraków opera building. on the fourth day of the conference (july 4th), in the thematic sections the lectures and poster sessions were continued. due to the very large number of presentations submitted, the meeting time was fully utilised to present the latest achievements and the concept of botanical studies. it was easy to see how far specialisation in individual branches of botany had come. it is important to emphasise the high substantive level and innovation of many of the scienti�c presentations and reports presented during the congress. when the o�cial scienti�c parts came to an end �eld excursions begun. �ey were an opportunity for both the tightening of the cooperation between di�erent institutions and to highlight the natural value of selected areas of southern poland. in the program of excursions a special importance was given to the presentation of natural characteristics and nature–human interactions in these ecosystems, which play the most important role in the discussion on the evolution of interdisciplinary methods of nature conservation and innovative use of its resources. on friday (july 5th), one-day �eld sessions took place: in the babiogórski national park, niepołomicka forest, as well as in the miechowska and ponidzie uplands. on saturday (july 6th), one-day trips to the gorczański national park, the ojców national park and the olkusz ore-bearing region were organised. �e sessions proposed by the organisers were undoubtedly an opportunity for direct contact with nature of primary environments, as well as those in which human activity overlaps with ne zostały tego dnia uroczystą kolacją, przy dźwiękach muzyki w  gmachu opery krakowskiej. w  czwartym dniu konferencji (4 lipca) kontynuowane były obrady sesji referatowych oraz plakatowych w  sekcjach tematycznych. ze względu na bardzo dużą liczbę zgłoszonych wystąpień czas obrad wykorzystany został w  pełni na przedstawienie najnowszych osiągnięć oraz koncepcji badań botanicznych. łatwo było zauważyć to, jak daleko zaszła specjalizacja w poszczególnych gałęziach botaniki. należy w  tym miejscu podkreślić wysoki poziom merytoryczny oraz innowacyjność wielu z przedstawionych podczas zjazdu wystąpień i doniesień naukowych. po zakończeniu wszystkich obrad w sekcjach, przyszedł czas na sesje terenowe, które stały się okazją, zarówno do pogłębienia współpracy pomiędzy przedstawicielami różnych ośrodków naukowych, jak i  do zapoznania się z  cechami przyrodniczymi wybranych rejonów polski południowej. przygotowując program sesji terenowych położono szczególny nacisk na zaprezentowanie walorów przyrodniczych oraz interakcji przyroda–człowiek tych ekosystemów, które zajmują ważne miejsce w dyskusji nad dalszą ewolucją metod interdyscyplinarnej ochrony przyrody i  innowacyjnego wykorzystania jej zasobów. w  piątek (5 lipca), odbyły się jednodniowe sesje terenowe: w  babiogórskim parku narodowym, puszczy niepołomickiej oraz na wyżynie miechowskiej i  ponidziu. w  sobotę (6 lipca) miały miejsce również jednodniowe sesje w  gorczańskim parku narodowym, ojcowskim parku narodowym oraz w  olkuskim okręgu rudnym. 200 r ep or ts the intrinsic rhythms of ecosystems. �e diversity of these areas in terms of topography and ecology gave the opportunity to present a wide spectrum of determinants of active and passive protection of natural resources in a gradient of anthropopressure and social expectations. parallel to the above-mentioned sessions, a three-day �eld session was also held (from friday to sunday, july 5–7th). it covered the topic of vegetation of the bieszczady mountains, sanocko-turczańskie mountains and the przemyśl foothills. participation in this event enjoyed particular interest among participants of the congress. in total, 450 participants attended the pbs congress, 200 oral presentations were lectured and 240 posters were delivered. 10 guests from abroad: czech, belgium, slovakia, switzerland, ukraine and great britain participated in the congress. �e 58th polish botanical society congress was undoubtedly an occasion to discover the actual trends in the biological and biotechnological plant science, and understand the problems of the conservation of the �ora of southern poland. �e polish botanical society is one of the oldest and biggest scienti�c associations in poland. it realises several goals in the �eld of the promotion of specialised research in the plant biology, ecology, taxonomy and phytogeography, the integration of the scienti�c society and the popularisation of botany in the society. �e cyclical organisation of meetings and conferences, such as the pbs congress, undoubtedly contributes to the achievement of the above goals. �e organisers of the congress prepared and electronically published the book of abstracts of oral presentations zaproponowane przez organizatorów sesje były niewątpliwie okazją do bezpośredniego kontaktu z  przyrodą środowisk pierwotnych, jak również tych, w których działalność człowieka nakłada się na samoistne rytmy ekosystemów. zróżnicowanie tych obszarów pod względem topogra�cznym oraz ekologicznym dało możliwość przedstawienia szerokiego spektrum uwarunkowań ochrony czynnej oraz biernej zasobów przyrodniczych w gradiencie antropopresji i oczekiwań społecznych. równolegle do powyższych sesji odbyła się także trzydniowa sesja terenowa (od piątku do niedzieli, 5–7 lipca). obejmowała ona tematykę szaty roślinnej bieszczadów, gór sanocko–turczańskich i  pogórza przemyskiego. udział w  tym wydarzeniu cieszył się szczególnym zainteresowaniem wśród uczestników zjazdu. łącznie w  58. zjeździe ptb brało udział 450 botaników, wygłoszonych zostało 200 referatów i  przedstawiono 240 plakatów. ponadto, w  zjeździe uczestniczyło 10 gości z  zagranicy: czech, belgii, słowacji, szwajcarii, ukrainy i  wielkiej brytanii. zjazd ten stał się okazją do zapoznania się z aktualnymi trendami w naukach dotyczących biologii i biotechnologii roślin, jak również do zrozumienia problemów ochrony szaty roślinnej polski południowej. polskie towarzystwo botaniczne jest jednym z  najstarszych i  najliczniejszych towarzystw naukowych w  polsce, realizującym cele w  zakresie promowania specjalistycznych badań nad biologią, ekologią, systematyką i  geogra�ą roślin, integrowania środowiska botaników oraz upowszechniania wiedzy botanicznej w  społeczeństwie. cykliczne organizowanie spotkań i  konferencji, takich jak zjazd ptb, niewąt201 r eports łukasz m. kołodziejczyk department of animal physiology and toxicology, institute of biology, pedagogical university of krakow; lukas.bios@wp.pl pliwie przyczynia się do osiągnięcia powyższych celów. na potrzeby tegorocznego zjazdu przygotowano i  opublikowano elektronicznie „streszczenia referatów i  plakatów” (dostępne pod adresem: https://hdl.handle. net/20.500.12333/254) oraz „przewodnik sesji terenowych” (dostępny pod adresem: https://hdl.handle.net/20.500.12333/255). z  wielką niecierpliwością oczekujemy kolejnych, równie owocnych zjazdów polskiego towarzystwa botanicznego! and posters (available at: https://hdl.handle. net/20.500.12333/254) and the guidebook for the excursions (available at: https://hdl. handle.net/20.500.12333/255). we look forward to the next, equally fruitful meetings of the polish botanical society! 98 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 98–105, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.13 thiep vo van1,2*, łukasz j. binkowski1, robert stawarz1 1institute of biology, pedagogical university of cracow, podchorazych 2, 30-084 kraków, poland 2faculty of agriculture, forestry and fisheries, quang binh university, 312 ly �uong kiet str, dong hoi, quang binh, vietnam, *thiep.vo-van@up.krakow.pl the concentration of mercury in organs of whipfin silver biddy (gerres filamentosus cuv.) and flathead grey mullet (mugil cephalus l.) in coastal central vietnam introduction vietnam is a part of south east asia bordered by the ocean on the west east and the south, with china to the north and cambodia and laos to the west. �e coastline stretches over 3.260 km, with an exclusive economic zone (eez) of over 1 million km2 where is a habitat of a vast array of aquatic species (us department of state, 1983; pham, masahide, 2007; teh et al., 2014). currently, with the increase in population, urbanization, industrialization, and agricultural practices, pollution with heavy metals in an aquatic ecosystem may occur (gupta et al., 2009). mercury (hg) is one of the most toxic metals in the aquatic ecosystems, which originates both from natural sources and human activities (luciana et al., 2005; seyed et al., 2013). hg cannot be degraded; it is deposited in the aquatic sediments and can be bioaccumulated and biomagni�ed via the food chain, and �nally assimilated by human consumers, which results in health risks (grimanis et al., 1978; adams et al., 1992; ermosele et al., 1995; smith et al., 1996; zweig et al., 1999; agah et al., 2009; malik et al., 2010). it is well known that �sh play an important role in the human diet. fish is not only a source of proteins and healthy fats, but it is also a unique source of essential nutrients, including long-chain omega-3 fatty acids, iodine, vitamin d, and calcium (fda, 2006; kruzikova et al., 2013; vicarova et al., 2015). however, it can represent a dangerous source of some heavy metals, especially hg (kruzikova et al., 2013). according to stankovic et al. (2014), microbes, fungi, plants, animals, and humans are used as bioindicators of heavy metals (including hg) originating from the air, water, sediment, soil, and the food web. �erefore, �sh could be a good and e�ective indicator of these elements in the aquatic environment. fish represent a speci�c level 99 the concentration of m ercury in organs of w hipfin silver biddy (g erres filam entosus c uv.) and flathead grey m ullet (m ugil cephalus l.) in coastal central v ietnam of the trophic pyramid and links hg to the ecosystem by bioaccumulation and biomagni�cation (stankovic et al., 2014; łuczyńska et al., 2016) in this study, two �sh species were collected: whip�n silver biddy and flathead grey mullet. �ese species are abundant and an easily accessible resources for artisanal �shing communities and are popular on vietnam �sh markets. �ere is some published literature on hg content of these two species in the world (legorburu et al., 1988; meng-hsien chen, 2002; yilmaz, 2003; 2005; chouba et al., 2007; sih-wei huang et al., 2008; frías-espericueta et al., 2016; türkmen et al., 2016; ruelas-inzunza et al., 2017; delgado-alvarez et al., 2017; dung et al., 2018), but the data for vietnam is scarce. �erefore, the aim of this study was to determine the e�ect of �sh species on hg concentrations in the selected organs (muscle, liver, and gills) of whip�n silver biddy and flathead grey mullet. �e data obtained were used to access the level of risk associated with consumption two these �sh species in vietnam. moreover, the study also evaluates di�erences between the content of hg in organs of the same �sh. material and methods during july, august, and september 2017, �sh samples were obtained from local �shermen and the �sh market of coastal vietnam of nghe an, ha tinh, quang binh, quang tri and hue. two �sh species were collected as whip�n silver biddy – gerres �lamentosus cuv. (n = 28) and flathead grey mullet – mugil cephalus l. (n = 48). �e muscle tissue from the dorsal area, the liver, and gill tissue were collected, placed in labelled polypropylene bags and stored at -20°c until analysis. total mercury concentrations in samples were determined by cold vapour atomic absorption spectrometry (nic, ma-2; limit of quanti�cation was 0.2 ng per sample). data was presented in µg g-1 wet weight. �e one-way analysis of variance anova and duncan’s test was used to test signi�cant interspeci�c di�erences in the content of mercury both between species and the organs of the same species. statistical signi�cance was declared when the p value was equal to or less than 0.05. results and discussion �e highest hg concentrations in flathead grey mullet were found in the liver, followed by muscle and gills (0.195, 0.097 and 0.046 µg g-1 w.w., respectively – tab.1). th ie p v o v an , ł uk as z j. b in ko w sk i, r ob er t s ta w ar z 100 tab. 1. mercury concentrations (µg g-1 wet weight) in gills, liver, and muscle of flathead grey mullet (n = 48) tissue mean ±sd min max gills 0.046 0.016 0.015 0.075 liver 0.195 0.101 0.063 0.494 muscle 0.097 0.037 0.027 0.184 note: sd – standard deviation, min – minimum, max – maximum �ere are several studies of hg concentrations of flathead grey mullet available in the literature. mostly, the concentrations reported are lower than the mean value found in this study (sankar et al., 2006; dural et al., 2007; ruelas-inzunza et al., 2008; sih-wei huang et al., 2008; squadrone et al., 2013; ruelas-inzunza et al., 2017). however, there were also some reported hg concentration higher than the mean we obtained (chouba et al., 2007; frías-espericueta et al., 2016) (tab. 2). tab. 2. mean total hg concentrations (µg g-1 wet weight) in flathead grey mullet mean hg concentration location references gills liver muscle 0.046 0.195 0.097 central vietnam �is study 0.050 0.032 california us state ruelas-inzunza et al. (2017) 0.503 0.036 northwestern mexico frías-espericueta et al. (2016) < 0.025 mediterranean sea squadrone et al. (2013) 0.016 northwestern mexico ruelas-inzunza et al. (2008) 0.044 0.110 tainan, taiwan sih-wei huang et al. (2008) 0.025 mediterranean sea dural et al. (2007) 0.242 0.098 tunisian lagoon (winter) chouba et al. (2007) 0.235 0.084 tunisian lagoon (spring) chouba et al. (2007) 0.231 0.056 tunisian lagoon (summer) chouba et al. (2007) 0.247 0.102 tunisian lagoon (autumn) chouba et al. (2007) 0.040 calicut, india sankar et al. (2006) tab. 3. mercury concentrations (µg g-1 wet weight) in gills, liver, and muscle of whip�n silver biddy (n = 28) tissue mean ±sd min max gills 0.077 0.040 0.036 0.220 liver 0.245 0.187 0.078 0.643 muscle 0.460 0.200 0.233 1.095 note: sd – standard deviation, min – minimum, max – maximum concentrations of hg in gills, liver, and the muscle of whip�n silver biddy di�ered between themselves (tab. 3). �e highest mean was noted in muscles (0.460 µg g-1 w.w), followed by the liver and gills (0.245 and 0.077 µg g-1 w.w, respectively). in this study, hg accumulation in muscles and the liver was higher than previously reported 101 by meng-hsien chen (2002), sih-wei huang et al. (2008), and dung le quang et al. (2018). according to meng-hsien chen (2002), hg concentrations in muscles and livers of whip�n silver biddy were both 0.025 µg g-1 w.w..in muscles, sih-wei huang (2008) recorded 0.45 µg g-1 w.w., and dung le quang (2018) recorded 0.358 µg g-1 w.w. �ere were signi�cant di�erences in mercury contents in muscles and gills between both species in this study (p<0.05). however, there was no di�erence in the concentration of hg in livers between flathead grey mullet and whip�n silver biddy (tab. 4). tab. 4. di�erences in hg concentration (µg g-1 wet weight) between the organs of both species tissue species mean statistical results (p) ±sd min max gills whip�n silver biddy 0.077 0.000 0.040 0.036 0.220 flathead grey mullet 0.046 0.016 0.015 0.075 liver whip�n silver biddy 0.245 0.132 0.200 0.233 1.095 flathead grey mullet 0.195 0.037 0.027 0.184 muscle whip�n silver biddy 0.460 0.000 0.187 0.078 0.643 flathead grey mullet 0.097 0.101 0.063 0.494 note: sd – standard deviation, min – minimum, max – maximum �e mean hg concentrations determined in this study for flathead grey mullet are one order of magnitude lower than the maximum permissible threshold (0.5 μg/g w.w. = 2 μg/g d.w.; fao-who, 2003; ministry of health of vietnam, 2007), which suggests wide limits of safety for the consumption of these �sh species. it is not the same in the case of whip�n silver biddy, because the hg concentration (0.460 µg g-1 w.w) in the muscle of this species almost reached the maximum permissible level. �e result of this study agree with the views of türkmen et al. (2011; 2016) and azevedo et al. (2012), which describe that, in most �sh species, the liver is the main storage place for metals. however, this is not a general rule in the case of hg. we detected higher content of hg in muscle than in liver in whip�n silver biddy, as did coelho et al. (2010) and polack-juszczak (2015). additionally, waltham et al. (2013) and diop and amara (2016) found no di�erences between hg concentrations in livers and muscles of some �sh species. �e di�erence in hg concentration in organs of �sh between the two species may be explained by several factors, such as resident time, trophic transfer, growth rate, prey type, and dietary quality. all of them can a�ect the hg bioaccumulation in �sh communities (hall et al., 1997; marugo-negrete et al., 2008). among others factors, feeding habits have been recognised as a prime reason for hg contamination (hall et al., 1997). the concentration of m ercury in organs of w hipfin silver biddy (g erres filam entosus c uv.) and flathead grey m ullet (m ugil cephalus l.) in coastal central v ietnam th ie p v o v an , ł uk as z j. b in ko w sk i, r ob er t s ta w ar z 102 conclusions statistically signi�cant di�erences in mean hg levels were observed between two �sh species investigated (whip�n silver biddy – gerres �lamentosus cuv. and flathead grey mullet – mugil cephalus l.) and their tissues, (except for their livers, there is no statistical di�erence between the two observed species). �ese results supply information on hg contents in tissues of the species examined in coast central vietnam and indirectly indicate hg levels in the marine environment. �ese results can be used to understand the chemical quality of �sh and to evaluate the possible risk associated with their consumption. references adams, w.j., kimerle, r.a., barnett, j.w. 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(2003). levels of heavy metals (fe, cu, ni, cr, pb and zn) in tissue of mugil cephalus and trachurus mediteraneus from iskenderun bay, turkey. environmental research, 92, 277–281. doi: 10.1016/s0013-9351(02)00082-8 yilmaz, a.b. (2005). comparison of heavy metal levels of grey mullet (mugil cephalus l.) and sea bream (sparus aurata l.) caught in iskendrun bay (turkey). turkish journal of veterinary and animal sciences, 29, 257–262. zweig, r.d., morton, j.d., stewart, m.m. (1999). source water quality for aquaculture: a guide for assessment. washington dc: �e world bank, 23–24. abstract �e concentration of mercury (hg) in �sh species has direct consequences on the health of humans and the ecosystem. �us, in this paper, the accumulation of hg in gills, livers, and muscles of two �sh species (whip105 �n silver biddy – gerres �lamentosus and flathead grey mullet – mugil cephalus) were measured by cold vapour atomic absorption spectrometry. �e �sh specimens were collected from local markets and direct �shing with the help of �shermen over the period from july to september 2017 in coastal vietnam. di�erences in the total hg were found both between two species and organs. �e concentration of hg in all organs investigated of whip�n silver biddy was higher than of flathead grey mullet (p < 0.05). �e content of hg in the muscles of whip�n silver biddy was higher than in the livers and gills (p < 0.05), 0.460, 0.245, 0.077 µg g-1 w.w., respectively. livers of flathead grey mullet had more hg accumulated than did the muscles and gills (0.195, 0.097, 0.046 µg g-1 w.w., respectively). �e results revealed that hg concentrations in flathead grey mullet did not exceed food �sh safety limits established for human consumption, while the concentration of this toxic element in the muscles of whipin silver biddy almost reached the maximum permissible level. keywords: coastal vietnam, gerres �lamentosus, mugil cephalus, mercury received: [2018.05.28] accepted: [2018.11.20] stężenia rtęci w organach pobranych od gerres filamentosus (cuv.) i mugil cephalus (l.) z wietnamu streszczenie akumulacja rtęci (hg) w rybach ma (duże, choć pośrednie) bezpośrednie znaczenie dla zdrowia ludzi i ekosystemu. dlatego w tym projekcie zbadano stężenia hg w skrzelach, wątrobie i mięśniach dwóch gatunków ryb (gerres �lamentosus i mugil cephalus). badania przeprowadzono z wykorzystaniem techniki atomowej spektrometrii absorpcyjnej z  przystawką zimnych par. osobniki ryb zostały (kupione) pozyskane na lokalnych targach lub bezpośrednio od rybaków w okresie między lipcem a wrześniem 2017 r. w wietnamie. różnice w stężeniach rtęci były obserwowane zarówno między badanymi gatunkami, jak i tkankami (materiałami). stężenia hg wykryte we wszystkich materiałach pobranych od gerres �lamentosus były wyższe niż w materiałach pobranych od mugil cephalus (p < 0,05). stężenia hg w mięśniach gerres �lamentosus były wyższe niż w wątrobie i skrzelach (p < 0,05), odpowiednio 0,460, 0,245 i 0,077 µg g-1 m.m. (mokrej masy). wątroba mugil cephalus zakumulowała wyższe stężenia hg niż mięśnie i skrzela (odpowiednio 0,195; 0,097 i  0,046 µg g-1 m.m. wyniki wskazują, że stężenia rtęci u  mugil cephalus (badanych gatunków) nie przekraczają norm ustalonych dla żywności do spożycia przez ludzi. jednak poziom hg w  mięśniach gerras �lamentosus jest zbliżony do tej wartości, co stanowi pewne zagrożenie dla potencjalnych konsunentów. słowa kluczowe: wietnam, gerres �lamentosus, mugil cephalus, rtęć information on the authors �iep vo van he is mostly interested in �sh biology and �sh toxicology. łukasz j. binkowski https://orcid.org/0000-0001-7271-2371 he is specialised in the ecotoxicology and biomonitoring of metals in the environment and animals. robert stawarz https://orcid.org/0000-0002-0495-1730 his main scienti�c interests are dedicated to toxicology and physiology. the concentration of m ercury in organs of w hipfin silver biddy (g erres filam entosus c uv.) and flathead grey m ullet (m ugil cephalus l.) in coastal central v ietnam 211 annales universitatis paedagogicae cracoviensis studia naturae, 4: 211–214, 2019, issn 2543-8832 a�er a four-year break, the department of biology and ecology at the faculty of natural sciences of the matej bel university in banská bystrica organised another, this time jubilee´ conference, 10th central european dipterological conference. of course, ten conferences in a row are not such an unusual event. however, it is unique that these conferences already have a fairly digni�ed tradition lasting half a century. �eir predecessors were seminars of czech and slovak dipterologists, which were initiated in 1969 by assist. prof. juraj čepelák – at the time the head of the department of zoology at the agricultural university of nitra. at this department, the important research centre for dipterous insects in europe was created by him. a ��y-year journey – from the �rst meeting of czechoslovak dipterologists to today’s international conference – has not always been easy. it is admirable that the continuity of these meetings maintained. over time in these seminars that were organised every 2–3 years, alternately on the czech and slovak side, were participated colleagues from poland and hungary, and therefore these events were transformed into international conferences in 1993. 10th central european dipterological conference in slovakia, kežmarské žľaby, september, 23–25, 2019 x środkowoeuropejska konferencja dipterologiczna na słowacji, kežmarské žľaby, 23–25 września 2019 po czteroletniej przerwie, katedra biologii i  ekologii wydziału nauk przyrodniczych uniwersytetu macieja bela w  bańskiej bystrzycy zorganizowała kolejną, tym razem jubileuszową x środkowoeuropejską konferencję dipterologiczną. oczywiście dziesięć konferencji z  rzędu nie jest aż tak niezwykłym wydarzeniem. jednak wyjątkowe jest to, że konferencje te mają już dość godną, trwającą pół wieku tradycję. ich poprzednikami były seminaria dipterologów czeskich i słowackich, których założycielem w 1969 r. był doc. juraj čepelák, ówczesny kierownik wydziału zoologii uniwersytetu rolniczego w  nitrze. w  ośrodku tym stworzył on ważne w  europie centrum badań nad owadami dwuskrzydłowymi. pięćdziesięcioletnia podróż – od pierwszego spotkania czechosłowackich dipterologów do dzisiejszej międzynarodowej konferencji, nie zawsze była prosta. godne podziwu jest to, że utrzymała się ciągłość tych spotkań. z  biegiem czasu w  seminariach, które były organizowane co 2–3 lata, na przemian po stronie czeskiej i  słowackiej, uczestniczyli koledzy z  polski oraz węgier, a  zatem wydarzenia te od 1993 212 r ep or ts �e last, 10th jubilee conference took place at the crocus recreation center in kežmarské žľaby (slovakia) under the patronage of the dean of the faculty of natural sciences of the matej bel university – assist. prof. jarmila kmeťová and the mayor of the high tatras – ing. ján mokoš. �is year’s conference was attended by 52 dipterologists from 12 countries – so far the most in the history of these meetings. in addition to slovakia and the czech republic, participants included experts from hungary, poland, ukraine, croatia, serbia, bulgaria, lithuania, great britain, germany and russia (fig. 1). during the 3 days (september 23–25), a  wide range of dipterological topics were presented and discussed, ranging from phylogeny and molecular taxonomy, to new results of studies on dipterous insects in di�erent parts of the world, their ecology and even palaeontology and paleoecology (fig. 2). roku zostały przekształcone w  konferencje międzynarodowe. ostatnia, x jubileuszowa konferencja odbyła się w  ośrodku rekreacyjnym crocus w kežmarské žľaby (słowacja), a patronowali jej dziekan wydziału nauk przyrodniczych uniwersytetu macieja bela dr hab. jarmila kmeťová oraz burmistrz tatr wysokich inż. ján mokoš. w  tegorocznej konferencji wzięło udział 52 dipterologów z 12 krajów – jak dotąd najwięcej w  historii tych spotkań. oprócz słowacji i  czech, uczestnikami byli eksperci z  węgier, polski, ukrainy, chorwacji, serbii, bułgarii, litwy, wielkiej brytanii, niemiec oraz rosji (ryc. 1). w  ciągu 3 dni (23–25 września) obrad omówiono i przedyskutowano szeroki zakres tematów dipterologicznych, począwszy od �logenezy i taksonomii molekularnej, po nowe wyniki badań nad dwuskrzydłymi w różnych częściach świata, ich ekologii, a  nawet paleontologii i  paleoekologii (ryc. 2). tradycyjfig. 1. participants of the 10th central european dipterological conference (september 23–25, 2019) – kežmarské žľaby, high tatras, slovakia (photo l. hamerlík) ryc. 1. uczestnicy x środkowoeuropejskiej konferencji dipterologicznej (23–25 września 2019 r.) – kežmarské žľaby, tatry wysokie, słowacja (fot. l. hamerlík) 213 r eports a traditional part of the conference was a competition for the best student presentation. as usual, the international evaluation committee was strict but fair. prizes funded in the competition by the mayor of the high tatras and the company kvant were awarded to valentin dorić (croatia) and andreas laug (germany). because science is a ‘living man’, there was also a gala dinner at the ždiarsky house museum. it was additionally enriched by the ždiar wedding show, in which selected conference participants took part in traditional costumes. certainly, this event contributed to the friendly and informal atmosphere of this part of the conference. at the end of the conference, for those interested, a trip to the green lake valley (dolina zeleného plesa) was organised, which was successful, despite the fact that the weather was not perfect. ną już częścią konferencji był konkurs na najlepszą prezentację studencką. jak zwykle międzynarodowa komisja oceniająca była surowa, ale sprawiedliwa. nagrody ufundowane w konkursie przez burmistrza tatr wysokich i �rmę kvant zostały przyznane dla valentina dorić (chorwacja) oraz andreasa laug (niemcy). ponieważ nauka jest ‘żywym człowiekiem’, odbyła się również uroczysta kolacja w muzeum żdiarskiego domu. wzbogacona została ona dodatkowo pokazem żdarskiego ślubu, w którym brali udział w tradycyjnych strojach wybrani uczestnicy konferencji. z  pewnością wydarzenie to przyczyniło się do przyjaznej i  nieformalnej atmosfery tej części konferencji. na zakończenie tegorocznej edycji konferencji, dla zainteresowanych, zorganizowano wycieczkę do doliny zielonego jeziora (dolina zeleného plesa), która fig. 2. prof. tadeusz zatwarnicki (poland) during his presentation (photo p. bitušík) ryc. 2. prof. tadeusz zatwarnicki (polska) w trakcie swojego wykładu (fot. p. bitušík) 214 r ep or ts at the end of this valley, mostly hidden in clouds, the extremely peculiar and photogenic spectacle was observed. it is not appropriate to boast, but according to the opinions of participants, it seems that this year’s conference has ended positively, both in terms of organisation and science, thus met the expectations. a�er a two-year break, it was again an opportunity for european dipterologists to share their knowledge and experience, renew old and make new contacts and spend pleasant moments together. we look forward to the next such scienti�c meetings. peter bitušík, tímea chamutiová department of biology and ecology, uniwersity of matej bel, tajovského 40, 974 01 banská bystrica, slovakia; peter.bitusik@umb.sk zakończyła się sukcesem, mimo że pogoda nie była idealna, a  wyjątkowo osobliwy i  fotogeniczny spektakl znajdujący się na końcu doliny, był w większości ukryty w chmurach. nie wypada się chwalić, ale zgodnie z opiniami uczestników wydaje się, że tegoroczna konferencja zakończyła się pozytywnie, zarówno pod względem organizacyjnym, jak i  naukowym, spełniając tym samym stawiane oczekiwania. po dwóch latach przerwy, była to ponowna okazja dla europejskich dipterologów, aby podzielić się swoją wiedzą i  doświadczeniem, odnowić stare i  nawiązać nowe kontakty oraz spędzić razem miłe chwile. z niecierpliwością oczekujemy kolejnych tego rodzaju spotkań naukowych. 125 annales universitatis paedagogicae cracoviensis studia naturae, 3: 125–134, 2018, issn 2543-8832 doi: 10.24917/25438832.3.9 iwona ałtyn, magdalena twarużek* department of physiology and toxicology, institute of experimental biology, faculty of natural sciences, kazimierz wielki university, chodkiewicza 30 st., 85-064 bydgoszcz, poland, *twarmag@ukw.edu.pl; https://orcid.org/0000-0002-8655-0067, https://orcid.org/0000-0002-2568-1178 heavy metal and mould contamination of herbal medicinal products – an overview the medicinal plant consists of many active chemicals, such as mucilage, polyphenols, polysaccharides, etc., which may modify the effects of the active principles. therefore, it is believed that some natural remedies may have toxic effects on the body or act as an agonist or antagonist of the active substance (dar, 2013). that is why toxicity tests for the pre-assessment of the efficacy of plant extracts are so important (hewawasam, 2016). herbal plants are exposed to different types of pollution, from chemical to microbiological, which, as mentioned above, can have a negative impact on the human body. there are numerous causes of chemical and microbiological pollution. one of them may be associated with the usage of human excreta, animal manures, and sewage as fertilisers. the world health organization’s (who) guidelines of good agricultural and collection practices (gacp) for medicinal plants prohibit the use of human manure as fertiliser. moreover, they insist that animal manure should be thoroughly composted (who, 2003). another cause of chemical pollution may be found in composted sewage, as well as in chemicals used in households and industrial chemicals, as the vast majority of them are found in analysed herbs and herbal materials. the listed potential sources of herbal contamination may lead to the appearance of unwanted chemicals, not just during the preparation process, but also in its final product. according to the world health organization’s research, the level of pollution present in medicinal plants may change in different stages of its manufacture, such as post-harvest processing (e.g., drying), herbal preparation – extraction process, as well as in the finished herbal products (who, 2007). therefore, the manufacture process of herbal preparation or its final pharmaceutical product should be controlled at each stage of production by gacp for medical plants and by good manufacturing practices (gmp) for herbal medicines. iw on a a łty n, m ag da le na t w ar uż ek 126 the purpose of this review is to show the mycological and chemical contamination of medicinal plants, as well as to indicate several important challenges related to the effective monitornig of their safety. contamination by heavy metals the rapid growth of industry and agriculture in recent times has caused a serious problem of contamination of air, soil, and water with heavy metals (orisakwe et al., 2012). heavy metals are generally defined as a collection of metalloids with anatomic density greater than 6 g/cm3. moreover, some of them (cadmium cd, chromium cr, copper cu, mercury hg, nickel ni, lead pb and zinc zn) possess toxic properties (awodele et al., 2013). in the sciences of biology and medicine, heavy metals are defined as elements used in the industry and simultaneously characterised by the toxicity to humans or the environment. the widespread environmental dispersion of heavy metals enter into the food chains through the epidemic pollution of air, water, and soil during cultivation (husain et al., 1995). the use of herbal plant contaminated by heavy metals for the production of herbal medicines can trigger chronic accumulation of metals in the human organs (ajasa et al., 2004; ang et al., 2006; arzani et al., 2007). bioaccumulation of heavy metals in the body can cause both short-term and long-term health damages. it can cause abdominal pain, disease of the foetus, which may lead to abortion and/or preterm labour, as well as mental retardation to children, while adults may suffer from hypertension, fatigue, as well as impaired kidney and brain damage (hifsa et al., 2009). prolonged consumption of products contaminated with heavy metals may also lead to skin eruptions, intestinal ulcers, and different types of cancer (shad et al., 2008). due to the high health risks, the who has developed limit guidelines for the maximum values of these compounds in herbal medicine. according to the who, remedies should be checked for the presence of different contaminants such as heavy/toxic metals, fungi, and microorganisms (who, 1998; 2007). several studies have tested herbal medicine preparations for heavy metal contamination. filipiak-szok et al. (2015) determined the highest concentration of lead pb (9.27 µg), cadmium cd (0.36 µg) and arsenic as (1.25 µg/g), in dietary supplements. for ni 0.16–14.21 µg/g dry mass (d.m.), while for ba 0.49–11.45 µg/tablet and 11.64 µg/g d.m. the lowest concentration of all analysed heavy metals was found for antimony sb. for plants it was at the level of 0.003 µg/g d.m. and 0.10 µg/g d.m. for dietary supplements. chuang et al. (2000) and singh and garg (1997), analysed the chinese root of panax ginseng c.a. meyer. the results showed the level of pb as – 0.16 ± 0.11 µg/g, cd as – 0.06 ± 0.04 µg/g and as as – 0.05 ± 0.03 µg/g; 4.62µg/g barium ba and 223 µg/g sb. h eavy m etal and m ould contam ination of herbal m edicinal products – an overview 127 olowoyo et al. (2012) showed that the maximum concentration of zinc zn in the roots of datura stramonium l. was recorded at value of 90.65 ± 1.22 μg/g. the concentration of cu from both plant parts ranged from 3.84 ± 0.33 μg/g – 14.05 ± 0.02 μg/g. contamination by nickel ni from the plant parts were in the range from 4.36 ± 0.25 μg/g to 15.78 ± 0.14 μg/g. pb concentration in all the plant parts ranged from 0.51 ± 0.01 μg/g to 2.13 ± 0.02 μg/g, while cr levels ranged from 5.65 ± 0.05 μg/g to 18.31 ± 0.01 μg/g. başgel et al. (2006) determined the high concentration of calcium ca from 965 mg/kg (rosae caninae fructus) to 17.740 mg/kg (tilia × vulgaris h. and urtica dioica l.). magnesium mg in the seven herbs and their infusions was in the range of 1643–3778 mg/kg and 610–2078 mg/kg, respectively. the iron fe content of the herbs and their infusions was in the range of 224–502.7 mg/kg and 4.90–107.4 mg/kg, respectively. the content of al in the herbs varied between 87 mg/kg (t. × vulgaris) and 596 mg/kg (u. dioica). manganese mn in the herbs varied in a wide range of 23–244 mg/kg (r. c. fructus); whereas, its concentration in the infusions varied between 4.30 mg/kg (foeniculum vulgare mill.) and 49.1 mg/kg (r. c. fructus). the highest concentration of cu was in the matricaria chammomilla l. infusion as 6.75 mg/ kg and the lowest value was determined in the cassia anqustifolia vahl infusion as 2.45 mg/kg. sr in the herbs was found between the range of 17.5 mg/kg (salvia officinalis l.) and 174 mg/kg (u. dioica); whereas, in the infusions, it varied between 2.45 mg/kg (s. officinalis) and 43 mg/kg (u. dioica). caldas (2004) showed that none of the 38 analysed samples of cynara cardunculus var. scolymus (l.) fiori, solanum melongena l. or paullinia cupana kunth contained detectable levels of cadmium cd (< 0.2 mg/g), mercury hg (< 0.01 mg/g), or lead pb (< 2.0 mg/g). cadmium concentrations varied from < 0.20 to 0.74 mg/g. centella asiatica (l.) urban. the levels of hg varied from < 0.01 to 0.09 mg/g with ginkgo biloba l. the levels of lead in the samples varied from < 2.0 to 1480 mg/g with aesculus hippocastanum l. having the highest concentrations. ting et al. (2013) research on chinese herbal medicine showed that out of the 6 metals (mn, pb, cu, cd, fe, zn), the highest concentration which reached the level of 1.394–18.545 mg/l belonged to mn. cd had the lowest level which was identified as 0.105–0.314 mg/l. other metals were all < 3 mg/l. naga raju et al. (2006) showed that ocimum sanctum l. were contaminated by ni (24.1 ± 4.7 µg/g). the same high level of ni 33.7 ± 50 µg/g was shown in the results of a study conducted by gowrishankar et al. (2010). the o. sanctum was an object of research conducted by professor kumar and his team. in their work, the results showed very high concentration of al at the 8249 µg/g (devi et al., 2008). harris et al. (2011) study showed that all samples of chinese herbal medicine were contained by at least one heavy metal. 34.4% of samples had detectable levels of all five metals. the highest concentration of cr and ar, were iw on a a łty n, m ag da le na t w ar uż ek 128 21 ppm and 20 ppm, respectively (egan et al., 2007; lendinez et al., 2001; saper et al., 2004; usda, 2009; us fda, 2007; us gao, 2010). fungal contamination moulds comprise a large group of around 100 thousand species, and they are widely distributed in nature. moreover, they are one of the most widespread environmental pollutants. the reason for this phenomenon is connected with their high ability to multiply on various raw materials, under favourable conditions (ahmad et al., 2014). therefore, it is estimated that approx. 25% of them have moulds (hussein et al., 2001). contamination by such fungi as aspergillus spp., penicillium spp., fusarium spp. and alternaria spp. is especially dangerous due to the production of toxic secondary metabolites – mycotoxins, which can cause both acute and chronic toxicities or even death (fao, 2001). herbal medicinal products preparations have been analysed for fungal contamination in a number of studies. efuntoye (1996) showed that samples were contaminated by the colony of aspergillus spp., pencillium spp., fusarium spp., and rhizopus spp. moreover, mucor spp., aspergillus spp. was isolated in all the samples of studied herbs, while the species aspergillus niger tiegh and a. flavus link were the most prevalent. the genus fusarium spp. was found in six of the plants studies. colonies of trichoderma viride pers. were isolated from azadirachta indica a. juss., plumbago zeylanica l. and jatropha curcas l., while colonies of cladosporium bantianum (sacc.) borelli and alternaria humieola oudem. were found in xylopia aethiopica (dunal) a.rich. and mangifera indica l., respectively. penicillium spp. were found in samples, with frequency at a level of 87.5% and makes up 18.5% of the total fungi. tournas et al. (2006) showed that 100% of the siberian, 56% of the chinese and 48% of the american ginseng root samples were contaminated with fungi. the highest contamination level (4.3×105 cfu/g) was observed in the locally grown american ginseng root, while the lowest (< 100 cfu/g) in ginseng extract. alternaria alternata (fr.) keissl. was found at levels as high as 4.0×104 cfu/g, while aspergillus spp. and cladosporium spp. were found at levels ranging between < 100 and 1.0×104 cfu/g. eurotium chevalieri l. was present in 11%, whereas penicillium spp., rhizopus spp. and yeasts were found in 33% of the tested samples. penicillium spp. levels were between < 100 and 1.0×103 cfu/g. the lowest level (< 100 – 4.0×102 cfu/g) of aspergillus niger was present in 22% of the samples. additionally, raman et al. (2004) also examined the dietary supplements including ginseng and the results showed that samples were contaminated with fungi, but the study did not quantify the fungal contaminants. zhang and zhang (2002) also isolated moulds from american ginseng seeds. according to the results of their work, researched samples were contaminated by high levels h eavy m etal and m ould contam ination of herbal m edicinal products – an overview 129 of fusarium spp., alternaria spp., penicillium spp., rhizopus spp. and low levels of aspergillus spp. rajeshwari and raveesha (2016) revealed that 6 of the herbal drugs’ raw materials were highly contaminated: withania somnifera (l.) dunal (100%) followed by curcuma angustifolia roxb. (92%), centella asiatica (l.) urban (88.6%), acorus calamus l. (88%), tinospora cordifloia (willd.) hook. f. & thomson (86%), and myristica fragrans houtt. (82%). therefore, researchers were able to identify and isolate 41 fungal species belonging to 16 genera. the most predominant were aspergillus spp. and penicillum spp., while the a. niger was the most frequently occurring fungi. roy and chourasia (1990) analysed traditional herbal drugs from india. the results showed that aspergillus spp., fusarium spp., penicillium spp. and trichoderma spp. were the common moulds isolated from most of the samples. aspergillus spp. were isolated from almost all samples with a frequency at 66.2% on strychnos nux-vomica l. seeds. the fusarium spp. was at the level of 11.6 ± 6.5 in the 3 samples of ichnocarpus frutescens (l.) w.t.aiton. hitokoto et al. (1978), while analysing herbal drugs’ samples, discovered that the most predominant fungi were aspergillus spp. and penicillium spp. mucor spp., rhizopus spp., cladosporium spp., and aureobasidium spp. were found in a few samples. the species of penicillium spp. predominated from powdered japanese peony roots – paeoniae radix pulverata (2.579 colonies/g) and powdered coptis – coptidis rhizoma pulveratum (9.500 colonies/g). aspergillus spp. with the highest frequency was found in powdered coptis (3.438/g), powdered scutellaria roots – scutellariae radix pulverata (2.241/g), powdered japanese peony roots (1.523/g), and powdered cininidium – cinidii rhizoma pulveratum (1.133/g). the species a. niger was the most frequently encountered group in the drugs, accounting for 24.6% of the total isolates. the aspergillus glaucus (l.) link and a. flavus link were the next most prevalent species with 9.3% and 7.8% of the total isolates, respectively. on the other hand, udagawa et al. (1976) showed that a. niger, a. glaucus and a. flavus were the most prevalent species in herbal drugs. matsushima et al. (1958) showed that aspergillus awamori nakaz., a. glaucus, a. mangini thom & raper, a. niger, a. ochraceus wihelm, penicillium frequentans west., p. variabile sopp., o.j., and rhizopus spp. were predominant. chourasia (1995) examined the herbal drugs from the indian pharmacopoeia. the maximum level of fungi was found in the fruits of piper longum l., p. nigrum l. and elettaria cardamomum (l.) maton. the most frequently isolated fungi were aspergillus spp. and fusarium spp., while the lowest frequently isolated ones were alternaria spp., emericella spp., mucor spp., penicillium spp., and chaetomium spp. rizzo et al. (2004) conducted a study to assess the presence of fungi in dried medicinal herbs. 27% of all samples were contaminated with aspergillus flavi section, iw on a a łty n, m ag da le na t w ar uż ek 130 25% circumdati section (aspergillus alliaceus thom & church, a. ochraceus and a. sclerotiorum g.a. huber) and 16% with fusarium spp. the most frequent contaminants were a. nigri. ahmad et al. (2014) studies showed that 90% of medicinal plant samples were contaminated with moulds and 70% of them exceeded the permissible limits determined by the united states pharmacopeia (2005). mould contamination in withania coagulans (stocks) dunal (3.4×1011 cfu g-1), papaver somniferum l. (6.1×1012 cfu g-1), olea europaea l. (1.08×105 cfu g-1), stevia rebaudiana (bertoni) bertoni (7.1×105 cfu g-1), ocimum basilicum l. (2.4×105 cfu g-1), aloe vera (l.) burm. f. (3.5×103 cfu g-1), opuntia monacantha haw. (5.8×104), glycyrrhiza gabra l. (2.2×105 cfu g-1), and cymbopogon citratus (dc.) stapf (4.6×103 cfu g-1) were observed less often but still in a significant quantity. the most frequently isolated species were aspergillus niger, a. flavus, a. parasiticus, speare, a. terreus thorn, penicillium verrucosum dierckx, p. citrinum thom. c, fusarium sp., rhizopus sp., and alternaria alternata. among them, aspergillus niger were found in 50% of the samples, followed by a. flavus reaching 43%. those results are in agreement with findings of abou-arab et al. (1999) and abou donia (2008), which showed that a. niger and a. flavus were the most dominant moulds in the collected samples of medicinal herbs in egypt. bugno et al. (2006) results showed that, in cymbopogon citratus (dc. ex nees) stapf, the level of fungal pollution was of 3.98×105 cfu/g-1. additionally, research conducted by jahani et al. (2013) on the subject of fungal contamination in barberry showed high levels of aspergillus spp. and penicillium spp. filipiak-szok et al. (2016) examination of plants and dietary supplements showed that the sum of moulds and yeast were lower than 69 cfu/g-1, with the most frequently species cladosporium spp. rawat et al. (2014) examined 40 samples of 8 different medicinal plants: saraca indica l., terminalia arjuna (roxb.) wight & arn., withania somnifera (l.) dunal, bacopa monnieri (l.) wettst., evolvulus alsinoides (linn.) linn., zingiber officinale roscoe. 92.5% of all analysed samples were found to be contaminated by more than one fungal species. those findings revealed that the highest frequency of contamination in samples belongs to aspergillus niger (38.56%) then mucor spp. (32.05%), rihzopus spp. (12.82%), a. flavus (12.82%), a. nidulans (2.56%) and myceliophthora spp. (1.28%). conclusions nowadays, due to the stressful working mode and lack of time for healthy meals, consumers are increasingly relying on medicinal plants as medications for health and ailments, or even dietary supplements. the presented results of multiple research studies indicate that plants for medical use should be carefully stored and evaluated for their possible contamination. long-term studies on heavy metal and fungal contaminants h eavy m etal and m ould contam ination of herbal m edicinal products – an overview 131 show how important monitoring is in determining the safe levels of pollutant concentration in medicinal plants and their products. each level of contamination that humans are exposed to by the intake of medicinal plants needs to be considered individually for every type of medicinal plant. therefore, to protect consumers against the contaminant health hazards, storage conditions of medicinal plants should be improved. moreover, good agricultural practice should be implemented to lower the presence of moulds on the medicinal plants. due to the wide range of different factors threatening a plant’s health, it is very important to continuously exert regular mycological evaluations. the presented research results not only allow one to determine the degree of potential threat but also, what is even more important, to raise the awareness about the importance of improvement in the processing techniques, i.e. harvesting, drying, transport, and storage. therefore, it can be stated that medicinal herb products for human consumption should be monitored for contaminants regardless of whether they are cultivated or collected from the wild. references abou-arab, a.a.k., kawther, m.s., el tantawy, m.e., badeaa, r.i., khayria, n. 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(2002). fungal detection of american ginseng seeds from beijing and northeast area in china. zhongguo zhongyao zazhi, 27(9), 658–661. przegląd zanieczyszczeń metalami ciężkimi i pleśniami roślin leczniczych streszczenie rośliny zielarskie są powszechnie wykorzystywane jako surowce w przemyśle farmaceutycznym. od czasów starożytnych ziół używa się w leczeniu i zapobieganiu chorób. jednakże w dzisiejszych czasach mogą one nie spełniać wymagań dotyczących jakości, bezpieczeństwa i  skuteczności. większość produktów ziołowych nie jest testowana, a ich efekty działania są słabo monitorowane. konsekwencją tego jest niewystarczająca wiedza na temat przebiegu ich działania, skutków ubocznych, przeciwwskazań oraz interakcji z występującymi na rynku produktami farmaceutycznymi oraz żywnością. przyczyną negatywnego wpływu surowców ziołowych na organizm może być ich zanieczyszczenie przez różne grzyby pleśniowe, powstające podczas zbioru, przetwarzania, przechowywania, a także dystrybucji. zanieczyszczenie surowców zielarskich może być również spowodowane przez różnorodne metale ciężkie, które występują w wielu aspektach współczesnego życia. celem niniejszej pracy jest przegląd informacji na temat stanu mykologicznego i chemicznego roślin leczniczych, a także wskazanie kilku ważnych wyzwań związanych z efektywnym monitorowaniem ich bezpieczeństwa. key words: contamination, heavy metals, medicinal plants, mould received: [2018.02.23] accepted: [2018.10.16] 83 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 83–89, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.11 jaromír vašíček1,2*, andrej baláži1, vladimír parkányi1, miroslav bauer1,3 1institute for farm animal genetics and reproduction, research institute for animal production nitra, nafc, lužianky, slovak republic, *jaromir.vasicek@gmail.com 2department of biochemistry and biotechnology, faculty of biotechnology and food science, slovak university of agriculture, nitra, slovak republic 3department of botany and genetics, faculty of natural sciences, constantine the philosopher university, nitra, slovak republic different macs sorting strategies for the enrichment of lin(cd34+ cd45-) hematopoietic progenitor cells: preliminary study introduction magnetic-activated cell sorting (macs) has become a standard method for cell separation in many di�erent �elds. numerous publications have demonstrated its use, from lab bench to the clinic, both small to large scale, from abundant cells to rare cells with complex phenotypes, and from human and mouse cells to many other species (bosio et al., 2009). �ere are various magnetic cell separation systems currently available. �ey di�er principally in two features: the composition and size of the magnetic particles used for cell labelling (miltenyi et al., 1990; ugelstad et al., 1998) and the mode (positive isolation – enrichment or negative isolation – depletion) of magnetic separation. at present, animal stem cells are widely used in the biomedical research. since rabbit is genetically very close to human, rabbit hematopoietic stem/progenitor cells (hsc/hpc) could be a precious animal model cell line for the study of human hematopoietic disorders. however, among the published studies, there is none available that describes either this type of rabbit cells or their isolation itself. cryopreservation of hsc/hpc for human therapeutic use in hematopoietic stem cell banks has been carried out for more than 30 years. �ree types of hsc/hpc harvests from bone marrow, mobilized peripheral blood, and umbilical cord blood (watt et al., 2007) are now routinely obtained by the macs enrichment of cd34 positive cells. �e same principle could be used for the isolation and cryopreservation of rabbit hsc/hpc. however, currently, there is no substantial data on the isolation and/or enrichment of rabbit hematopoietic stem cells, or on the detection of rabbit cd34+ cells. in the few published studies, mainly anti-human cd34 antibodies (vašíček et al., 2018) were ja ro m ír v aš íč ek , a nd re j b al áž i, v la di m ír p ar ká ny i, m iro sl av b au er 84 used, due to the unavailability of speci�c anti-rabbit cd34 antibodies in the market. on the other hand, hsc/hpc are de�ned as lineage negative cells that do not express lineage commitment markers typical for mature hematopoietic cells, including t and b cells, monocytes/macrophages, granulocytes, erythrocytes, and their committed precursors (gallacher et al., 2000). �us, mature cells expressing lineage commitment markers can be easily depleted from the heterogeneous mixture of hematopoietic cells (cheng et al., 2008) in order to enrich hsc/hpc. �erefore, the aim of this preliminary study was to isolate rabbit hsc/hpc via removing mature hematopoietic cells that express cd45 (common leukocyte antigen) from the rabbit peripheral blood and bone marrow. material and methods isolation of cells from rabbit blood and bone marrow mononuclear cells from rabbit peripheral blood (pbmcs) and bone marrow (bmmcs) were isolated from 3 young (5 months-old does) and clinically health rabbits of the nzw line as described previously (vašíček et al., 2016; 2018). indirect immunomagnetic sorting of cd45+ pbmcs and bmmcs brie�y, mononuclear cells were aliquoted into prepared tubes and incubated with anti-rabbit cd45 monoclonal antibody (l12/201, mouse igg1; biorad, uk) for 15 minutes on ice. a�er washing, samples were incubated with anti-mouse igg1 microbeads (miltenyi biotec, germany) according to the producer’s manual. �ree di�erent types of sorting strategy available by automacs pro separator (miltenyi biotec, germany) were used in the experiments: 1) depletes – depletion in sensitive mode i for the removal of cells with normal antigen expression, if purity is the highest priority or for the removal of cells with low antigen expression; 2) depl025 – depletion in sensitive mode iii for the removal of cells with low antigen expression, special program for very sensitive depletion; and, 3) posselds – positive selection in sensitive mode, double-column program for the isolation of cells with frequencies lower than 5% and low antigen expression. control samples were analysed for cd45 expression but not sorted. flow cytometry a�er macs sorting, the apc-conjugated labelling check reagent (lcr; miltenyi biotec, germany), which binds to the microbeads, was used to control the sorting e�ciency. moreover, control samples (unsorted cells) and both macs fractions (negative and positive) were incubated with another clone of anti-rabbit cd45 (isc18a, mouse igg2a; wsu, usa) and appropriate secondary anti-mouse igg2a-fitc antibody (ebioscience, austria). 7-aad (ebioscience, austria) was used in order to ex85 d ifferent m a c s sorting strategies for the enrichm ent of lin (c d 34 + c d 45 -) hem atopoietic progenitor cells: prelim inary study clude debris and dead cells from the analysis. labelled cells before and a�er sorting were evaluated using a facscalibur �ow cytometer (bd biosciences, usa). at least 50.000 events (cells) were analysed in each sample. double positive cells (cd45+lcr+) in macs fractions served as an indicator of the sorting (depletion) e�ciency in negative fractions. qpcr analysis of cd34 expression total rna isolation and cdna synthesis from the control (unsorted) samples and both cd45and cd45+ fractions as well as quantitative polymerase chain reaction (qpcr) were performed according to vašíček et al. (2017). since rabbit cd34 has predicted two transcript variants, we used universal primers that comprise both known transcripts (vašíček et al., 2017). results flow cytometry results are summarised in table 1. according to the obtained data, isolated rabbit pbmcs showed more than 98% positivity for cd45 antigen. on the other hand, only 70% of fresh rabbit bmmcs were positive for cd45. double staining (cd45+lcr+) of separated cells revealed the best depletion e�ciency in the negative fractions that were sorted using depl025 mode for both pbmcs and bmmcs (about 10 and 3%, respectively). tab. 1. sorting e�ciency of di�erent macs strategies in terms of removing cd45+ cells from the sorted samples assessed by �ow cytometry as the percentage of cd45+lcr+ cells [%] sample depletes depl025 posselds c o n (c d 45 + ) po s (c d 45 + l c r + ) n eg (c d 45 + l c r + ) c o n (c d 45 + ) po s (c d 45 + l c r + ) n eg (c d 45 + l c r + ) c o n (c d 45 + ) po s (c d 45 + l c r + ) n eg (c d 45 + l c r + ) pbmcs 98.8 93.4 37.4 99.1 95.5 10.0 98.8 95.4 44.6 bmmcs 67.3 94.4 22.4 65.7 92.6 2.7 75.8 98.2 32.1 note: con – control (unsorted) fresh sample analysed for the initial expression of cd45, pos – positive fraction, neg – negative fraction, lcr – labelling check reagent qpcr analyses were used to evaluate the relative expression of cd34 in the control and sorted samples. �e highest cd34 expression within the negative fractions were found in the pbmcs sample sorted using depl025 mode in comparison to the control and positive fractions or, in general, within the used sorting strategies (fig. 1). on ja ro m ír v aš íč ek , a nd re j b al áž i, v la di m ír p ar ká ny i, m iro sl av b au er 86 the other hand, bmmcs sample sorted by posselds mode revealed the highest cd34 expression in negative fraction in comparison to the control and positive fractions or, in general, within the used sorting strategies (fig. 2). fig. 1. relative expression of cd34 in control (unsorted) and sorted pbmcs normalised to b2m (beta-2-microglobulin) fig. 2. relative expression of cd34 in control (unsorted) and sorted bmmcs normalised to b2m (beta-2-microglobulin) 0 1 2 3 4 5 control depletes positive depletes negative depletes control depl025 positive depl025 negative depl025 control posselds positive posselds negative posselds r el at iv e ex pr es si on u ni ts macs sorting strategy cd34 0 1 2 control depletes positive depletes negative depletes control depl025 positive depl025 negative depl025 control posselds positive posselds negative posselds r el at iv e ex pr es si on u ni ts macs sorting strategy cd34 87 discussion we chose three di�erent macs sorting strategies for the negative or positive selection of cd45+ cells in order to obtain the highest sorting e�ciency or the maximum removal of mature hematopoietic cells (cd45+) from the sample. according to the producer’s manual, these strategies di�ered mainly in the sample loading rate and the number of used magnetic columns: 1) depletes – one column mode with loading rate 1 ml/min; 2) depl025 – one column mode with loading rate 0.25 ml/min; and, 3) posselds – two column mode with loading rate 1 ml/min for each column. �us, the sorting e�ciency of the used modes could di�er according to the sample loaded as well as the level of desired antigen expression. in this preliminary study, we assessed the sorting or depletion e�ciency of macs strategies used as the percentage of cd45+lcr+ cells. �e best removal of cd45+ cells in both pbmcs and bmmcs samples was obtained using depl025 mode (tab. 1). so, we might assume that negative fractions (cd45-) obtained by this mode can be enriched for cd34+ cells. �is �nding strongly agreed with qpcr analysis of rabbit pbmcs sorted by this mode, since the highest relative expression of cd34 was noticed in the negative fraction in comparison to other negative fractions or the control and positive fraction within the sample (fig. 1). however, the highest relative expression of cd34 in bmmcs was observed in the negative fraction of the sample sorted by a posselds mode (fig. 2) in comparison to other negative fractions or the control and positive fractions within the sample. although there are presently no any published studies focusing on the enrichment of rabbit lin(cd34+cd45-) cells, this technique based on the depletion of cells with lineage commitment markers is widely used for the puri�cation of mouse (ema et al., 2006) or human (cheng et al., 2008) hematopoietic stem and progenitor cells. conclusions �is preliminary study demonstrated the possibility to obtain a cell population from rabbit pbmcs or bmmcs that is enriched for cd34+ cells. �e best macs sorting strategy according to the relative expression of cd34 antigen in enriched (negative fractions) were depl025 for rabbit pbmcs or posselds for bmmcs. as far as we know, this is the �rst published study focusing on the enrichment of rabbit hsc/hpc (cd34+) cells using the removal of cd45+ cells. however, further experiments are required in order to prove these preliminary results. acknowledgement �is work was supported by the grants: apvv-14-0348 and vega 1/0160/18. d ifferent m a c s sorting strategies for the enrichm ent of lin (c d 34 + c d 45 -) hem atopoietic progenitor cells: prelim inary study ja ro m ír v aš íč ek , a nd re j b al áž i, v la di m ír p ar ká ny i, m iro sl av b au er 88 references bosio, a., huppert, v., donath, s., hennemann, p., malchow, m., heinlein, u.a.o. (2009). isolation and enrichment of stem cells. advances in biochemical engineering/biotechnology, 114, 23–72. doi: 10.1007/10_2008_38 cheng, p., corzo, c.a., luetteke, n., yu, b., nagaraj, s., bui, m.m., ortiz, m., nacken, w., sorg, c., vogl, t., roth, j., gabrilovich, d.i. (2008). inhibition of dendritic cell di�erentiation and accumulation of myeloid-derived suppressor cells in cancer is regulated by s100a9 protein. journal of experimental medicine, 205(10), 2235–2249. doi: 10.1084/jem.20080132 ema, h., morita, y., yamazaki, s., matsubara, a., seita, j., tadokoro, y., kondo, h., takano, h., nakauchi, h. (2006). adult mouse hematopoietic stem cells: puri�cation and single-cell assays. nature protocols, 1(6), 2979–2987. doi: 10.1038/nprot.2006.447 gallacher, l., murdoch, b., wu, d.m., karanu, f.n., keeney, m., bhatia, m. (2000). isolation and characterization of human cd34-linand cd34+linhematopoietic stem cells using cell surface markers ac133 and cd7. blood, 95, 2813–2820. miltenyi, s., müller, w., weichel, w., radbruch, a. (1990). high gradient magnetic cell separation with macs. cytometry, 11, 231–238. doi: 10.1002/cyto.990110203 ugelstad, j., prestvik, w., stenstad, p., kilaas, l., kvalheim, g. (1998). selective cell separation with monosized magnetizable polymer beads. in: w. andrä, h. nowak (eds.), magnetism in medicine – a handbook. weinheim: wiley, 473–490. doi: 10.1002/masy.19880170113 vašíček, j., bauer, m., baláži, a., chrenek, p. (2017). expression of cd34 in di�erent passages of rabbit endothelial progenitor cells. animal physiology 2017 – proceedings of international conference, košice: sav ústav fyziológie hospodárskych zvierat, 81. vašíček, j., kováč, m., baláži, a., bauer, m., chrenek, p. (2016). phenotypic analysis of rabbit mesenchymal stem cells using �ow cytometry and rt-pcr. slovak journal of animal science, 49(4), 160. vašíček, j., shehata, m., schnabl, s., hilgarth, m., hubmann, r., jäger, u., bauer, m., chrenek, p. (2018). critical assessment of the e�ciency of cd34 and cd133 antibodies for enrichment of rabbit hematopoietic stem cells. biotechnology progress. [in press]. doi:10.1002/btpr.2659 watt, s.m., austin, e., armitage, s. (2007). cryopreservation of hematopoietic stem/progenitor cells for �erapeutic use. in: j.g. day, g.n. stacey (eds.), methods in molecular biology: cryopreservation and freeze-drying protocols. totowa, nj: humana press inc., 237–259. doi: 10.1007/978-1-59745362-2_17 abstract magnetic-activated cell sorting (macs) has become a standard method for the isolation of hematopoietic stem/progenitor cells (hsc/hpc) in human or mouse models using cd34 antibodies. however, at present, there is no useable cd34 antibody that could be successfully used for the selection of rabbit hsc/hpc. �erefore, the aim of this preliminary study was to remove all mature cells (cd45+) from the heterogeneous mixture of rabbit peripheral blood and bone marrow mononuclear cells (pbmcs and bmmcs) in order to enrich these cell populations for the cd34+ cells. brie�y, cells were incubated with a cd45 antibody and proper magnetic microbeads. �ree di�erent macs sorting strategies were used in the experiment that di�ered mainly in the sample loading rate and the number of used magnetic columns. control (unsorted) and sorted cells were assessed for the sorting e�ciency (% of double positive cells for cd45 and labelling check reagent – lcr) by �ow cytometry and for the relative expression of cd34 antigen by qpcr. according to �ow cytometry, depl025 mode showed the best sorting e�ciency in terms of the lowest percentages of cd45+lcr+ cells for rabbit pbmcs as well as bmmcs. qpcr analysis con�rmed this mode as the best in terms of the relative cd34 expression for rabbit pbmcs. however, a higher relative expression of cd34 in bmmcs was obtained by another mode – posselds. in conclusion, this study demonstrates a possible 89 enrichment of rabbit (cd34+) hsc/hpc by the magnetic depletion of mature hematopoietic (cd45+) cells. key words: cd34, cd45, �ow cytometry, hematopoietic stem cells, macs, qpcr, rabbit received: [2018.05.30] accepted: [2018.11.22] różne strategie sortowania macs do wzbogacania hematopoetycznych komórek progenitorowych lin(cd34+ cd45-): badania wstępne streszczenie w  modelu ludzkim lub mysim, sortowanie komórek aktywowane magnetycznie (macs) stało się standardową metodą izolacji hematopoetycznych komórek macierzystych/progenitorowych (hsc/hpc) z użyciem przeciwciał cd34. obecnie jednak nie ma użytecznego przeciwciała cd34, które mogłoby być z powodzeniem zastosowane do selekcji króliczego hsc/hpc. dlatego celem tego wstępnego badania było usunięcie wszystkich dojrzałych komórek (cd45+) z heterogennej mieszaniny krwi obwodowej królika i jednojądrzastych komórek szpiku kostnego (pbmc i bmmc), aby wzbogacić ich populacje dla komórek cd34+. komórki inkubowano z  przeciwciałem cd45 i  odpowiednimi mikrokulkami magnetycznymi. w eksperymencie zastosowano trzy różne strategie sortowania macs, które różniły się głównie szybkością ładowania próbki i liczbą użytych kolumn magnetycznych. kontrolne (nieposortowane) i  posortowane komórki oceniano pod względem wydajności sortowania (% podwójnie dodatnich komórek dla cd45 i odczynnika do sprawdzania znakowania – lcr), za pomocą cytometrii przepływowej i względnej ekspresji antygenu cd34 przez qpcr. zgodnie z cytometrią przepływową tryb depl025 wykazał najlepszą skuteczność sortowania pod względem najniższego odsetka komórek cd45+ lcr+ dla pbmc królika, jak również bmmc. analiza qpcr potwierdziła ten tryb jako najlepszy do oceny względnej ekspresji cd34 dla pbmc królików. najwyższą względną ekspresję cd34 w bmmc uzyskano w innym trybie – posselds. podsumowując, niniejsze badanie demonstruje możliwe wzbogacenie hsc/hpc królika (cd34+) przez magnetyczne zubożenie dojrzałych komórek krwiotwórczych (cd45+). słowa kluczowe: cd34, cd45, cytometria przepływowa, hematopoetyczne komórki macierzyste, macs, qpcr, królik information on the authors jaromír vašíček https://orcid.org/0000-0003-4144-8584 expert in the �eld of the farm animal reproduction and the preservation of animal genetic resources. his specializations are the following: 1st – the study of the farm animal male generative cells, 2nd – research on farm animal somatic stem cells, and 3rd – the application of the biotechnological methods for the conservation of animal genetic resources via stem cell banking. andrej baláži expert in the �eld of animal cell isolation, magnetic cell separation, �ow cytometry, cell culture, and animal reproduction. vladimír parkányi senior researcher with long-time experiences in the �eld of cytogenetics of farm animal somatic and generative cells with skills in the �eld of animal cell magnetic separation, pcr techniques, and elisa. miroslav bauer expert in molecular biology experienced in genetic markers of farm animals, rdna and gene cloning, pcr, rt-qpcr, dna sequencing and bioinformatics. d ifferent m a c s sorting strategies for the enrichm ent of lin (c d 34 + c d 45 -) hem atopoietic progenitor cells: prelim inary study 24 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 24–32, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.3 lívia handrová*, anna čuvalová, vladimír kmeť institute of animal physiology, centre of biosciences of the sas, soltesovej 4/6, 040 01 kosice, slovak republic,*handrova@saske.sk the relationship between biofilm formation, genes of virulence and iron metabolism in escherichia coli introduction urinary tract infections (utis) are the most prevalent infectious diseases, and very problematic worldwide (navidinia et al., 2018). uropathogenic escherichia coli t. escher. (upec), which can colonise successfully in the urinary tract, is the primary etiologic agents associated with uti (peerayeh et al., 2018). avian pathogenic e. coli (apec) cause septicemia, polyserositis, aerosacculitis and other mainly extraintestinal diseases in chickens and other avian species. apecs are found in the intestinal microbiota of healthy birds, and most of the diseases associated with them are secondary to environmental and host predisposing factors (dho-moulin, fairbrother, 1999). �e common presence of a set of virulence-associated genes among as well as similar disease patterns and phylogenetic background indicate a genetic relationship between apec and upec isolates (kaper et al., 2004; moulin-schouleur et al., 2006; ron, 2006). �e success of e. coli in colonising such a wide range of hosts and environments is basically due to a noticeable ductility in exploiting the available resources. it is becoming increasingly clear that bio�lms have an enormous impact on medicine (mah, o’toole, 2001; wang et al., 2017), since 65% of human microbial infections involve bio�lms (labbate et al., 2004). microbial bio�lm formation is now recognised as a principle virulence factor in many localised chronic infections (hyun koo et al., 2017), and their role in infecting the biological devices among hospitalised patients is a universally accepted fact (vasudevan, 2014). in addition, recent experimental evidence indicates a role of bio�lm formation in acute infections (hannan et al., 2012; kumagai et al., 2011). understanding bio�lm formation to �nd e�ective ways to prevent bio�lms is important for combating disease. �e primary aim of this study was to detect e. coli strains with a bio�lm formation from animals and the detection of apec virulence genes presence in these strains in compared to the ability of production a bio�lm. 25 the relationship betw een biofilm form ation, genes of virulence and iron m etabolism in escherichia coli material and methods escherichia coli strains were isolated from broilers rectal swab coming from farms of eastern slovakia. samples were resuscitated overnight at 37°c in bu�ered peptone water (oxoid, basingstoke, uk) and subcultured on mac conkey agar (oxoid) and uriselect agar (bio-rad laboratories, hercules, ca, usa) again overnight at 37°c. �e colonies were isolated, identi�ed, and con�rmed as e. coli by commercial identi�cation microsystem enterotest24 (erbalachema brno, czech republic) and by using the maldi-tof ms biotyper (bruker daltonics, bremen, germany). nineteen strains were selected for further testing. bio�lm formation �e ability of bio�lm formation was assessed in a quantitative assay using a microtiter-plate test (nunc, roskilde, denmark). strains grown on bhi agar and colonies were re-suspended in bhi broth (oxoid, uk) to reach the 0.5 suspension mcfarland’s standard, and volumes of 200 μl of these cell suspensions were transferred to wells of the microplate. a�er incubation (24 h/37°c), adherent cells were washed three times using a saline solution and stained with a 0.1% crystal violet solution (mikrochem, pezinok, slovakia) for 15 min. a�erwards, excess stain was rinsed o� by �lling the wells with sterile distilled water. �e adhering dye was dissolved with 30% acetic acid for 15 minutes and the optical density was measured at 570 nm in synergy ht multi-mode microplate reader (biotek, usa) (čuvalová, 2018). we divided isolates of e. coli into four classes based on stepanovic et al. 2007. for classi�cation, we used average optical density (od) value and cut-o� value (odc) (de�ned as three standard deviations (sd) above the mean od of the negative control). �e �nal od value of a tested strain was expressed as the average od value of the strain reduced by the odc value. for interpretation of the results, strains were divided into the following categories: od ≤ odc = non-bio�lm producer; odc < od ≤ 2 x odc = weak bio�lm producer; and, 2 x odc < od ≤ 4 x odc = moderate and 4 x odc < od = strong bio�lm producers. detection of genes by pcr screening of e. coli isolates for apec virulence genes were carried out by polymerase chain reactions with the ampli�cation of the following: the receptor for aerobactin – iuta (johnson, stell, 2000); colicin v – cvac (johnson, stell, 2000); increased serum survival – iss (foley et al., 2000); temperature sensitive haemagglutinin – tsh (dozois et al., 2000); p �mbrial adhesion – papc (le bouguénec et al., 1992); capsular polysialic acid virulence factor – kps (johnson, stell, 2000); iron-regulated gene a homologue adhesion – iha (johnson et al., 2000) and genes of iron metabolism – putative iron transport gene – sita (rodrigues-siek et al., 2005); iron-related genes – gene which lí vi a h an dr ov á, a nn a č uv al ov á, v la di m ír k m eť 26 mediates ferric iron uptake feob (rodrigues-siek et al., 2005), encodes an iron-responsive element and putative sideropohore receptor gene – irea (russo et al., 2001) and iron repressible gene associated with yersiniabactin synthesis – irp2 (schubert et al., 1998), yersiniabactin receptor for ferric yersiniabactin uptake – fyua (schubert et al., 1998), and the catecholate siderophore receptor gene – iron (johnson, stell, 2000), and primers are listed in table 1. tab. 1. primers used for detection of virulence genes and genes of iron metabolism gene primer sequences (5’–3’) annealing [°c] size [bp] iuta f: ggctggacatgggaactggr: cgtcgggaacgggtagaatcg 63 300 cvac f: cacacacaaacgggagctgttr: cacacacaaacgggagctgtt 63 680 iss f: atcacataggattctgccgr: acaaaaagttctatcgcttcc 61 700 tsh f: ggtggtgcactggagtggr: agtccagcgtgatagtgg 55 620 papc f: gacggctgtactgcagggtgtggcgr: atatcctttctgcagggatgcaata 61 328 kps f: gcgcatttgctgatcgttgr: catccagacgataagcatgagca 63 272 iha f: ctggcggaggctctgagatca r: tccttaagctcccgcggctga 60 827 sita f: agggggcacaactgattctcgr: taccgggccgttttctgtgc 59 608 feob f: aattggcgtgcatgaagataactgr: agctggcgacctgatagaacaatg 59 470 irea f: tggtcttcagctatatggr: atctatgattgtgttggt 55 415 irp2 f: aaggattcgcgtgacr: tcgtcgggcagcgtttcttct 59 287 fyua f: tgattaaccccgcgacgggaar: cgcagtaggcacgatgttgta 55 880 iron f:aagtcaaagcaggggttgcccgr: gacgccgacattaagacgcag 60 655 results �e interpretation of obtained results requires a de�nition of the cut-o� value that separates bio�lm producing from non-bio�lm-producing strains. we divided isolates based upon the previously calculated od values, which was a modi�cation of method and classi�cation described by stepanović et al. (2007): very weak 12/19 (63.0% of strains), weak 2/19 (10.5%), moderate 2/19 (10.5%) and strong 3/19 (16.0%) bio�lm producers. �e occurrences of 13 detected genes are presented in �gure 1. among 19 e. coli isolates, all isolates contained the feob gene, 16 isolates contained the sita 27 gene, 13 isolates contained the iss and iron genes, 12 isolates contained the iuta gene, 11 isolates contained the fyua gene, 6 isolates contained the papc and irea genes, 5 isolates contained the cvac and tsh genes, 4 isolates contained the irp2 gene, and 2 isolates contained the kps gene. for better comparison of our results, we created two groups of strains (fig. 2). �e �rst of the two groups represented very weak bio�lm producers and the second group represented weak, moderate, and strong formers. representation genes of virulence were high in isolates from the �rst group – from seven genes were six highly, only papc was low. genes of iron metabolism were di�erent. genes sita, fyua, and irea were higher in the second group, and feob, irp2 and iron were higher in the �rst group. discussion genes coding adhesins, toxins, or iron acquisition systems have been described to be of particular importance during the pathogenesis of septicemia (gyles, 1994; babai et al., 1997; terlizzi et al., 2017; robinson et al., 2018), and iron acquisition is a requirement for upec survival in an environment that is as iron-limited as the urinary tract (skaar, 2010). isolated e. coli strains were investigated for the presence of thirteen virulence genes that are associated with colibacillosis and iron metabolism. two genes, fyua and irp2, coding proteins involved in iron acquisition, were described in yersinia sp., and this iron acquisition determinant has been found in human septicemic and enteroaggregative e. coli isolates (karch et al., 1999; pelludat et al., 1998; schubert et al., 1998). �e sequences of the irp2 and fyua genes in e. coli are alfig. 1. distribution of genes virulence in escherichia coli strains: iuta (12/19), cvac (5/19), iss (13/19), tsh (5/19), papc (6/19), kps (2/19), iha (1/19), sita (16/19), feob (19/19), irp-2 (4/19), fyua (11/19), iron (13/19) and irea (6/19) 0 4 8 12 16 20 iuta cvac iss tsh papc kps iha sita feob irp2 fyua iron irea d is tr ib ut io n of g en es v ir ul en ce escherichia coli strains the relationship betw een biofilm form ation, genes of virulence and iron m etabolism in escherichia coli lí vi a h an dr ov á, a nn a č uv al ov á, v la di m ír k m eť 28 most identical to those of yersinia spp. (germon et al., 2005) and have been described in apec isolates by gophna et al. (2001), subedi et al. (2018). from nineteen isolates in our study, we detected irp-2 gene in four and fyua in eleven strains. to increase survival and resistance, e. coli strains also form bio�lm, but published data is variable, depending on the strain origin, di�erent types of surfaces, culture medium, and the methodology used for quantifying bio�lm. in our study, the presence of genes of virulence was low in second group – better bio�lm formers and only papc was higher. to compare with another study (naves et al., 2008; pavlickova et al., 2017), papc was also determined in a strain with better forming bio�lm, and tsh, which was similar to our results, was detected in weak bio�lm formers. iuta was represented in both groups of strains with weak and strong production of bio�lm. genes of iron metabolism shows that sita, fyua, and irea were represented higher in the second group (weak, moderate and strong). naves et al. (2008) recorded the presence of fyua in all strong bio�lm producers, but iron, unlike our study, was higher in low bio�lm producers. �e literature contains only a few papers correlating the virulence factors investigated in this study with the ability of pathogenic e. coli to form bio�lms in vitro. further studies involving larger numbers of clinical strains are needed to corroborate our data concerning the interaction between bio�lm formation and virulence factors. conclusion bio�lms are of particular interest in the poultry industry and public health, because these �lms can harbour pathogenic microorganisms. in this study, escherichia coli strains were identi�ed and analysed for the presence of genes of iron metabolism, fig. 2. presence of detected genes expressed as a percentage; divided in two groups: �rst group = �rst column of data (very weak producers) and second group = second column of data (weak, moderate and strong bio�lm producers) 0 20 40 60 80 100 iuta cvac iss tsh papc kps iha sita feob irp2 fyua iron irea [% ] escherichia coli genes very weak producers (1. group) better producers (2,3,4 group) 29 virulence-associated genes, and bio�lm-forming abilities. all 19 e. coli strains evaluated were able to form bio�lms, with the majority exhibiting very weak bio�lm-forming potential. �e prevalence of the virulence-related genes was higher in low bio�lm producers, where the presence of the siderophore-related genes was variable, but no signi�cant di�erences were observed between strong and weak bio�lm producers. results provide a basis for the further study of the pathogenesis of apec and its abilities of formation bio�lms. acknowledgement �is study was supported by the slovak project vega 2/0085/18. references babai, r., blum-oehler, g., stern, b.e., hacker, j., ron, e.z. 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(2016). �e irp2 and fyua genes in high pathogenicity islands are involved in the pathogenesis of infections caused by avian pathogenic escherichia coli (apec). polish journal of veterinary sciences, 19(1), 21–29. doi: 10.1515/pjvs-2016-0004 vasudevan, r. (2014). bio�lms: microbial cities of scienti�c signi�cance. journal of microbiology experimental, 1–14. doi: 10.15406/jmen.2014.01.00014 wang, y., jayan, g., patwardhan, d., phillips, k.s. (2017). antimicrobial and anti-bio�lm medical devices: public health and regulatory science challenges. in: z. zhang, v. wagner (eds.), antimicrobial coatings and modi�cations on medical devices. springer, cham. doi: 10.1007/978-3-319-57494-3_2 abstract escherichia coli is known as one of the bacterial species with the widest adaptability to a variety of niches either within organisms or outside in environment. most strains of e. coli are of low virulence and associated with opportunistic infections, whereas others are highly virulent. �e success of e. coli in colonising such a wide range of hosts and environments is basically due to a noticeable ductility in exploiting the available resources. it is becoming increasingly clear that bio�lms have an enormous impact on medicine, because 65% of animal and human bacterial infections involve bio�lms. in the present study, we isolated strains of e. coli from animals. 19 interesting isolates were selected and tested by pcr ampli�cation to virulence – iuta, cvac, iss, tsh, papc, kps, iha and iron metabolism genes – sita, feob, irp2, fyua, iron, and irea. �e ability of bio�lm formation was assessed in a quantitative assay using microtiter-plate tests. bacterial strains were grown on bhi. we divided isolates of e. coli into four classes: very weak (63.0%), weak (10.5%), moderate (10.5%), and strong (16.0%) bio�lm producers. representation genes of virulence were high in isolates from very weak bio�lm producers – from 7 genes were 6 highly and only papc (p �mbrial adhesin) was low. genes of iron metabolism were di�erent. genes – sita, fyua, and irea in strong isolates producing bio�lm and feob, irp2, and iron in weak producers were most represented. �e results show a possible relation between presence virulence factors and low bio�lm formation. key words: bio�lm, virulence genes, iron metabolisms genes received: [2018.05.30] accepted: [2018.12.29] the relationship betw een biofilm form ation, genes of virulence and iron m etabolism in escherichia coli lí vi a h an dr ov á, a nn a č uv al ov á, v la di m ír k m eť 32 związek pomiędzy tworzeniem się biofilmu, genami wirulencji i metabolizmem żelaza u escherichia coli streszczenie escherichia coli znana jest jako jeden z gatunków bakterii o najszerszej zdolności adaptacji do różnych nisz w organizmach lub w środowisku zewnętrznym. większość szczepów e. coli ma niską wirulencję i wiąże się z infekcjami oportunistycznymi, podczas gdy pozostałe szczepy są wysoce wirulentne. sukces e. coli w kolonizowaniu tak szerokiego zakresu żywicieli i środowisk wynika przede wszystkim z zauważalnej ciągliwości w wykorzystywaniu dostępnych zasobów. staje się jasne, że bio�lmy mają ogromny wpływ na medycynę, ponieważ 65% zakażeń bakteryjnych zwierząt i ludzi dotyczy bio�lmów. w obecnych badaniach izolowano szczepy e. coli ze zwierząt. wybrano 19 interesujących izolatów i testowano je przez ampli�kację pcr pod względem wirulencji – geny metabolizmu iuta, cvac, iss, tsh, papc, kps, iha oraz żelaza – sita, feob, irp2, fyua, iron, irea. zdolność tworzenia bio�lmu oceniano w teście ilościowym, stosując test płytki mikrotitracyjnej. szczepy bakteryjne hodowano na bhi. izolaty e. coli podzielono na cztery klasy producentów bio�lmu: bardzo słabe (63,0%), słabe (10,5%), umiarkowane (10,5%) i silne (16,0%). geny reprezentacyjne wirulencji były w większości izolowane od bardzo słabych producentów bio�lmu – z 7 genów było 6 wysoko wirulentnych; tylko papc (adhezyna �mbrialna) była niska. geny metabolizmu żelaza były różne pod względem wirulencji. najbardziej reprezentowane były geny – sita, fyua, irea w silnych izolatach produkujących bio�lm oraz feob, irp2, iron u słabych producentów. wyniki pokazują możliwą zależność pomiędzy obecnym czynnikiem zjadliwości, a niską formacją bio�lmu. słowa kluczowe: bio�lm, geny wirulencji, geny metabolizmu żelaza information on the authors lívia handrová http://orcid.org/0000-0002-0985-1771 �e main area of her interest is genetic ecology and the spread of antibiotic resistance genes. she studies the resistance occurrence in small mammals, which could serve as a reservoir of antibiotic resistance (esbl, plasmid encoded chinolone resistance, carbapenemases) in indicator bacteria escherichia coli, pseudomonas aeruginosa and staphylococcus spp. anna čuvalová she is interested in anti-bio�lm activities of natural compounds using the static and dynamic bio�lm models with resistant staphylococci (mrsa and mrcons), escherichia coli (esbl and cefotaximases) and pseudomonas aeruginosa on various surfaces (plastics, catheters and food grade stainless sheet). vladimír kmeť http://orcid.org/0000-0002-8081-8579 he is interested in bio�lm and anti-bio�lm activities of natural compounds using the static and dynamic bio�lm models with escherichia coli, resistant staphylococci (mrsa and mrcons). �e area of his interest is genetic ecology and gene encoding factors of virulence, metabolism, and the spreading of these genes. he studies the resistance occurrence in animal, which could serve as a reservoir of antibiotic resistance in indicator bacteria. 81 annales universitatis paedagogicae cracoviensis studia naturae, 4: 81–90, 2019, issn 2543-8832 doi: 10.24917/25438832.4.4 angelika kliszcz*, joanna puła university of agriculture in krakow, department of agroecology and plant production, mickiewicza 21 ave, 30-120 kraków, poland; *angelika.kliszcz@student.urk.edu.pl assessment of earthworms activity based on eaten biomass from selected catch crops introduction in order to preserve the homeostasis of the soil environment and increase its fertility, the presence and activity of soil mesofauna, especially earthworms, is an essential factor (family lumbricidae ra�nesque-schmaltz, 1815). by feeding on plant and animal residues, as well as microorganisms, earthworms form resources of soil organic matter that are permanently associated with the soil’s mineral phase (wu et al., 2018). a network of corridors formed by individual ecological groups of earthworms contributes to the regulation of water-air relations in soil. according to bouché (1972), three main ecological groups of earthworms can be distinguished in the soil pro�le: anecic, endogeic and epigeic. for agroecosystems, the �rst two play a key role. anecic species (e.g. lumbricus terrestris l.) live deep in the soil pro�le (up to several meters) and form corridors with a further to the vertical slope, which communicate earthworms with a substantial food source, i.e. with plant debris le� on the ground (e.g. in the form of mulch). however, their basic component of the diet is the mineral part of the soil, which they eat while drilling corridors. �eir activity contributes to better water in�ltration and the creation of ‘fertile’ corridors on the walls of which reside bacteria in the organic matrix le� by earthworms (coprolytes, body’s excrement). �ey are also weed seed vectors, which enriches the soil seed bank located at deeper levels. endogeic species from the second ecological group intensify their activity in the arable soil layer, near plant roots (up to 30 cm deep) and create galleries with a horizontal slope, contributing to the formation of proper humus reservoir in the plant rhizosphere. also these species are vectors of microorganisms and seeds in the inhabited area (clause et al., 2017). �ey are also species that prefer a large proportion of organic remains in addition to the mineral part of the soil. a ng el ik a k lis zc z, j oa nn a p uł a 82 �e presence and activity of earthworms in the cultivated �eld can be limited by the intensi�cation of cultivation treatments (briones, schmidt, 2017) and the use of herbicides (kostecka, 1999; pelosi et al., 2014). �e introduction of plant biomass into the �eld, for example in the form of catch crops, is an additional source of food and can contribute to an increase in the earthworm population. �e presence and activity of earthworms increases the fertility of the soil habitat, which creates favourable conditions for the growth and development of crop plants. each animal selects, per se, the most rich in content – optimal food that it needs. for earthworms these are: plant and animal debris, living and dead soil organisms (bacteria, fungi, protozoa, algae, nematodes, amoebas) (curry, schmidt, 2007), as well as excrements of various living organisms, minerals, ions in the free state in soil solution. penetrating the soil pro�les of almost every square meter of soil on the earth’s surface, earthworms, when drilling corridors and taking food, decide what and in what quantity will be collected and processed by them. however, this fascinating mechanism of food preferences in earthworms is not yet fully understood. �e �rst work on the trophic behaviourism of earthworms appeared in ancient times (li et al., 2010), and the deliberations were continued with considerable publicity by charles darwin, who devoted the last 30 years of his life to studying the life and functions of earthworms in the process of forming soil organic matter. when food is consumed by the earthworm, a decisive role is played by a part of the neural ganglion, which is stimulated by chemical receptors located in the prostomium (above-mouth lobe) and from receptors on the entire body surface of the earthworm. �ese receptors provide the earthworm with information about the environment in which it is currently located. generally, the trophic mechanism in the earthworm consists in collecting through the mouth, then swallowed pieces pass through pharynx, the esophagus and enter the crop, in which they are temporarily kept and mixed with a concentrated suspension of calcium carbonate produced by calciferous gland’s secretory cells localised at the end of the esophagus. next, the food goes to a heavily muscled stomach (gizzard), then passes into the intestine, from where it is excreted in the form of casts through the anus. despite the absorption of nutrients by the body of the earthworm, droppings are a valuable and rich component of soil matrix and remain compact for a long time. it is interesting that despite high concentrations of phenolic compounds in plant material and their adverse e�ect on the precipitation of proteins in living organisms, plant biomass remains the main substrate for earthworms. �e research of liebeke et al. (2015) shows that drillodefensins (surface active lipophilic ions 259.1013 da, which m/z are consistent with a molecular formula of c 12h19o4s -) are produced in the earthworm body, which are produced in the foregut section of earthworms and help them digest phenolic-rich residues plant. a ssessm ent of earthw orm s activity based on eaten biom ass from selected catch crops 83 one way to assess the trophic activity of earthworms is to quantify their ability to eat food per unit of time. �e aim of the study was to evaluate the possibility of processing food (mixed with soil catch crop residues from white mustard (sinapis alba l.), buckwheat (fagopyrum esculentum moench) and tansy phacelia (phacelia tanacetifolia benth.) and crop biomass which was spring triticale (×triticosecale wittm. ex a.camus) by earthworms of the species lumbricus terrestris l. regard to control object (soil from the �eld). an interesting further question concerned examining whether any of the plants can be preferred (more willingly taken) by earthworms, thus may supposedly contribute to an increase in their numbers in the �eld. material and methods study material �e model organism in the experiment were individuals of lumbricus terrestris, a species from the anecic group, which are known for drawing plant organic matter from the soil surface into their corridors reaching 2 meters deep into the soil (bogdanowicz et al., 2004), and that while burrowing corridors, they also pass a signi�cant amount of the mineral part of the soil through the gastrointestinal tract (rouse, 2016). assessment of food intake by earthworms �e assessment of food intake by earthworms was performed in laboratory conditions in experiment with petri dishes. earthworms were purchased from a commercial supplier (ekagro) and kept for 4 hours in the dark (15°c) in a container with wet tissue paper to empty their digestive tract. �en 1 l. terrestris individual (average weight 4.99 g) was placed in one petri dish (∅ 11 cm), in seven replications, and was incubated in a vegetation chamber (darkness, 18°c) in a completely randomised system. �e food material (20 g per petri dish) was airdried fragmented plant biomass (7% w/w) sieved through a ∅ 1 mm sieve and mixed with soil sterilised at 105°c which was sieved through a ∅ 2 mm sieve and brought to �eld humidity (approx. 35%). �e amount of food eaten by each individual was assessed a�er 12 h, 24 h and 63 h with an accuracy of 0.0001 g. before measuring each dish with food (7 replicates), the earthworm and wet paper were removed, then excrements and coprolites le� on the petri dish during the animal’s activity were wiped with paper, and then the together with food was weighed (later subtract the weight of each dish). all results were corrected for values resulting from natural weight loss of food recorded simultaneously for objects without earthworms. a ng el ik a k lis zc z, j oa nn a p uł a 84 statistical analysis �e results regarding the food intake of earthworms were analysed based on anova with repeatable measurements and a grouping factor (food material) or one-way anova (c and n content; ratio of food intake) and subjected to the tukey hsd test at α = 0.05. results and discussion �e dynamics and amount of food intake at 12-hour intervals (fig. 1 a–b) indicates that this geophages species prefer soil as food. however, among the plant biomass supplements, earthworms most o�en took white mustard (sinapis alba l.) (0.80 g a�er the �rst 12 hours). �ese food tendencies of earthworms remained until the end of the experiment, and this is all the more surprising because plants of the brassicaceae burnett family have thioglycosides of volatile mustard oils in their tissues, which a�er being subjected to mechanical grinding and under the in�uence of the enzyme myrosinase, may decompose in white mustard to toxic and irritating so� tissues p-hydroxybenzyl isothiocyanate (sawicka, kotiuk, 2007). tab. 1. nitrogen (n) and carbon (c) content in the biomass of analysed plants and in soil object n [%] c [%] c/n ratio phacelia tanacetifolia benth. 1.81 ±0.018 b 40.04 ±0.004 c 22.1 sinapis alba l. 1.94 ±0.032 a 42.18 ±0.170 a 21.8 fagopyrum esculentum moench 1.57 ±0.040 c 40.27 ±0.083 c 25.7 ×triticosecale wittm. ex a.camus (spring) 1.83 ±0.021 b 41.68 ±0.077 b 22.8 soil (control) 0.08 ±0.003 d 0.64 ±0.023 d 8.54 mean values ±sd, n = 3; di�erent letters next to values indicate di�erent homogenous group, hsd tukey test, α = 0.05 �e third measurement, a�er more than 3-times a further exposure (63rd hour of the experiment), revealed even greater discrepancies in soil uptake (15.13 g) and soil with the addition of plant biomass (1.37 g on average). at 63rd hour of the study, there was a disruption in the metabolism of earthworms in the form of a secreted white substance in a facility with tansy phacelia (phacelia tanacetifolia benth.), which resulted in a lower uptake of eaten food than assessed a�er 24 hours and this also became the basis for the termination of the experiment. �e dynamics of food intake showed acceleration of consumption for some objects and exposure times, and sometimes earthworms took food constantly (fig. 1). for example, tansy phacelia are characterised by increased intake dynama ssessm ent of earthw orm s activity based on eaten biom ass from selected catch crops 85 fig. 1. accumulated mass of food eaten by earthworms lumbricus terrestris l. (a) a�er 12 hours, (b) a�er 24 hours, (c) a�er 63 hours, (n = 7); di�erent letters next to values indicate di�erent homogenous group inside the one object with a time, hsd tukey test, α = 0.05; ** for ×triticosecale p-value = 0.360579; object: 1 – phacelia tanacetifolia benth., 2 – fagopyrum esculentum moench, 3 – sinapis alba l., 4 – ×triticosecale wittm. ex a.camus, 5 – soil (control) a ng el ik a k lis zc z, j oa nn a p uł a 86 ics between 12 and 24 hours of exposure (190%), just as triticale between 12 and 24 hours (131%) and 24–63 hours (138%). white mustard and buckwheat were taken changeless at both time intervals, with more dynamic (though less quantitatively) uptake of buckwheat (83 and 57%, respectively) and balanced uptake dynamics for white mustard (57 and 55%, respectively). earthworms, as a biological component that increases soil fertility, are particularly valuable in soil and plant cultivation systems, in which it is not possible to increase this fertility by introducing fertilisers and soil conditioners into the system. �is is the case in the organic farming system. non-use of plant protection products and a larger amount of organic matter going back to the �eld mean that this system has a greater biodiversity, quantity and biomass of earthworms, although the plough tillage is a decisive factor (bilalis et al., 2009; munro et al., 2002). in this context, the conducted research may allow the identi�cation of species-speci�c trophic behaviour of earthworms (food processing capability, intake dynamics) and estimation of the impact of the earthworm population and their trophic behaviourism on the positive (or negative) e�ect on the growth and development of plants in organic crops. it should be noted, that the practice of leaving plant residues in the form of mulch on the soil surface has been known in agriculture for a long time and it is used now, especially in organic farming. although, according to jodaugiene et al. (2010) the largest amount and biomass of earthworms is concentrated under grass mulches (av. 185 number per m-2 and 42.5 g×m2 respectively), the selection of plants was not accidental in this experiment, because tansy phacelia, white mustard and buckwheat belongs to the plants widely used in organic farming, at least because of their phytosanitary properties in relation to commercial crops (majchrzak et al., 2005). plants used in the food material used in the experiment were subjected to biochemical analysis for nitrogen and carbon content (tab. 1). �e data show that white mustard, which the earthworms took the most, also had the highest c/n ratio (25.7), while in spring triticale, whose intake was the most dynamic during the whole experiment, this ratio was at the level of 22.8. a good estimator of the quantitative and qualitative food intake by earthworms is the ratio of the weight of food taken (per day) to the average body weight of the earthworm – f24/awe (tab. 2). based on the obtained results, it was observed that the activity of earthworms increased more than threefold in objects with an addition of catch crop biomass in relation to spring triticale. a ssessm ent of earthw orm s activity based on eaten biom ass from selected catch crops 87 tab. 2. comparison of the ratio of food intake from various plant species during the 24 hours to the average body weight of the earthworm (lumbricus terrestris l.); awe – average weight of the earthworm, awft – average weight of food taken in total a�er 24 hours, f24/awe – ratio of food intake per 24 hours to the average weight of the earthworm plant species awe [g] awft [g] f24/awe phacelia tanacetifolia benth. 5.00 ±1.022 0.70 ±0.365 0.14 b fagopyrum esculentum moench 4.93 ±1.555 1.19 ±0.545 0.24 b sinapis alba l. 4.86 ±1.614 1.31 ±0.348 0.27 b ×triticosecale wittm. ex a.camus (spring) 5.21 ±1.197 0.37 ±0.340 0.07 b soil (control) 4.93 ±0.965 6.93 ±1.982 1.41 a mean value for plants 5.00 ±1.375 0.89 ±0.557 0.18 average means ±sd, n = 7; di�erent letters next to values indicate di�erent homogenous group, hsd tukey test, α = 0.05 it is estimated that in maize cultivation in temperate climate, 100 individuals of l. terrestris species per 1 m2 process on average 840 kg of litter per year (bohlen et al., 1997). in this experiment, a new indicator, the decomposition rate, was used, which tells how many kilograms of food used (mixture of soil and plant biomass) pass through the digestive tract of 100 individuals per year, in this case l. terrestris (fig. 2). it can be useful in estimating the contribution of individual soil fauna species to the transformation processes of the organic substance of the agroecosystem in a time interval. estimation through this indicator gives the opportunity to assess the impact of a particular species, taking into account its synecology (population ecology), living in a given ecosystem and the ability to compare the e�ectiveness of organisms (including earthworms) in various crops. however, it is advisable to collect the organisms before calculating the decomposition index in order to estimate their real number in the studied area. �e most e�ective crops in terms of processing of plant biomass by earthworms of the species l. terrestris (assessment based on the calculated decomposition rate) would be white mustard and buckwheat (the highest values in �gure 2). on the other hand, in quantitative terms, the biomass of cereal plants represented by spring triticale was almost three times less processed compared to other tested plants. to correctly assess the impact of the earthworm population on the plant biomass cycle in the agroecosystem, it should be taken into account that earthworms as burrowers require more space for their activities, so that the spatial stress factor does not inhibit them from activity. �erefore, �eld experiments in the mesocosm system are the most commonly used in this type of research. on the other hand, laboratory experiments are short and re�ect the food preferences of individuals quite well. a ng el ik a k lis zc z, j oa nn a p uł a 88 conclusion �e conducted analyses show that lumbricus terrestris shows food preferences in relation to crop biomass, as the eurybiont of many habitats in temperate climate. during the 63 hour experiment, soil (15.1 g) constituted the most food material collected by earthworms, and among the plant components – the one with the addition of sinapis alba (2.03 g). however, the object with the addition of ×triticosecale spring was characterised by the highest consumption dynamics (the average of 135%). food material with the addition of tansy phacelia was taken up the fastest in the �rst 24 hours (190%), but later it fell sharply until changes in animal metabolism were recorded. �e ratio of food intake per day to the average body weight of one earthworm exceeded the unity threshold (1.41) only in the case of the soil object. in other cases (non-cereal plants) it oscillated around 0.22 and only in triticale reached three times lower (0.07). �e most favorable decomposition rate was recorded for white mustard, whose estimated amount of food eaten by earthworms (with a local population of 100 individuals) would be approximately 48 kg/m2/year in the �eld. �e assessment of the possibility of food processing by earthworms, as well as the determination of their population in the �eld, can be very helpful in the organic farming system, where the biological component plays a key role in increasing soil fertility. acknowledgment �e research was �nanced from the funds of the ministry of science and higher education as a part of a subsidy (bm no. 2119). con�ict of interest �e authors declare no con�ict of interest related to this article. fig. 2. estimated average mass of food eaten by 100 individuals (lumbricus terrestris l.) based on the results of a laboratory experiment; 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[in polish] a ng el ik a k lis zc z, j oa nn a p uł a 90 wu, y., shaaban, m., peng, q., zhou, a., hu, r. (2018). impacts of earthworm activity on the fate of straw carbon in soil: a microcosm experiment. environmental science and pollution research, 25, 11054–11062. doi: 10.1007/s11356-018-1397-4 abstract �e trophic activity of soil mesofauna, especially earthworms (the lumbricidae family), is a key element in increasing the fertility of agroecosystems. �e food strategies that earthworms use as part of the trophic networks in soil, and especially their food preferences, are still unknown. much is known about what is the food substrate of earthworms, but the food preferences of individual species, as well as the possibilities and dynamics of food processing are not fully understood. �e aim of the experiment was to observe the amount and dynamics of food uptake by the earthworms of the species lumbricus terrestris l., which is a common species of soil oligochaeta in agricultural areas, as well as to propose a new decomposition rate measuring the strength of the earthworm population and its contribution to the mechanism of processing plant organic matter. key words: agroecosystem, lumbricidae, cover crops received: [2019.05.07] accepted: [2019.10.11] ocena aktywności dżdżownic na podstawie pobrania biomasy z wybranych międzyplonów streszczenie aktywność tro�czna mezofauny glebowej, zwłaszcza dżdżownic (rodzina lumbricidae), stanowi kluczowy element w podnoszeniu żyzności agroekosystemów. strategie pokarmowe jakie stosują dżdżownice będące częścią sieci tro�cznych w glebie, a zwłaszcza ich preferencje pokarmowe są wciąż niepoznane. wiele wiadomo na temat tego co jest substratem pokarmowym dżdżownic, to jednak nie do końca poznano preferencje pokarmowe poszczególnych gatunków, a także możliwości i dynamikę przerobienia pokarmu. celem eksperymentu było zaobserwowanie ilości oraz dynamiki poboru pokarmu przez dżdżownice z  gatunku lumbricus terrestris l., będącej powszechnie występującym gatunkiem skąposzczetów glebowych na obszarach użytkowanych rolniczo, a także zaproponowanie nowego wskaźnika dekompozycji, mierzącego siłę i wkład populacji dżdżownic w proces przerabiania roślinnej materii organicznej. słowa kluczowe: agroecosystem, lumbricidae, międzyplony information on the authors angelika kliszcz https://orcid.org/0000-0002-1270-4414 she is focusing on enhancing the understanding of the in�uence of di�erent factors on soil structure and fertility. particularly, she is investigating the interaction of plants with the physical, chemical, and biological properties of the soil. she is also interested in earthworm ecology and mesofauna function in agroecosystems. joanna puła https://orcid.org/0000-0002-3672-5690 her research is connected with agrotechnology in plant cultivation and plant ecology. presently, she is interested in the use of the biomass of plants and other organic fertiliser like biochar in agriculture. 1 annales universitatis paedagogicae cracoviensis studia naturae, 8: xx–xx, 2023 issn 2543-8832 doi: 10.24917/25438832.8.x agnieszka tatoj 1 *, aleksandra smagieł 1 , himani yadav 2 , peiman zandi 3 1 department of botany, institute of biology and earth science, pedagogical university of krakow, 30-084 kraków poland; *agnieszka.tatoj@doktorant.up.krakow.pl 2 department of botany, chattrra marg university of delhi, new delhi-110007, india 3 international faculty of applied technology, yibin university, yibin 644000, china plants supporting the treatment of lyme disease introduction lyme disease (= tick-borne spirochete, lyme disease; latin borreliosis, morbus lyme) is a chronic multi-system disease caused by spirochaetes (order spirochaetes) of the species borrelia burgdorferi s.l. (in a broad sense of this taxon) (logiudice et al., 2003; stanek, reiter, 2011). transmitted by ticks of the genus ixodes latreille, from the tick family ixoidae. b. burgdorferi is a complex taxon, divided into a number of species that have pathogenic properties and operate in different regions of the world (tab. 1). a larva ambylomma tick containing spirochete-like cells was discovered in dominican amber by the american biologist george poinar in 2014 (nuwer, 2014). the age of this specimen was estimated at 15–20 million years. poinar also suspected that the first humans, who appeared on earth about 200,000 years ago, suffered from ailments caused by the bites of these invertebrates. however, he did not compare this fossil genetic material with currently known forms of lyme disease, as this would have destroyed his precious specimen. poinar (2014) noted that the route of infection probably took place in a similar way as at present – during food intake by a tick burrowing into the host's body. the earliest documented case of lyme disease dates back to 5,300 years ago and was discovered in an ice mummy. the genus name of the borrelia bacteria was given from the name of the researcher who discovered it in 1905 – amandee borrela (koperwas, 2013). arvid afzelius, a swedish dermatologist, was the first to associate the occurrence of erythema migrans with a tick bite. in 1909, he undertook to study this relationship. he was followed by others – charles garin and antoine 2 bujadoux, who in 1922 described the symptoms of neuroborreliosis. in 1941 and 1944, alfred bannwarth linked the neurological syndrome of erythema migrans with tick bites (nuwer, 2014). tab. 1. transmission of selected species of borrelia burgdorferi s.l. (according to sinski, welc-faleciak, 2012 – updated) species of b. burgdorferi s.l. vector tick species animal reservoir occurrence area borrelia burgdorferi johnson et al. 1984 emend. baranton et al. 1992 s.str. ixodes pacificus, i. ricinus, i. scapularis birds, rodents europe, north america b. afzelii manuela marin canica et al. 1994 i. persulcatus, i. ricinus rodents, other small mammals europe, asia b. garinii baranton et al. 1992 i. persulcatus, i. ricinus birds, rodents europe a, asia b. lusitaniae le fleche et al. 1997 i. ricinus rodents europe, north america, north africa b. spielmanii richter et al. 2006 i. ricinus rodents europe b. valaisiana wang et al. 1997 i. ricinus, i. columnae birds europe asia b. bavariensis margos et al. 2013 i. ricinus rodents europe b. bissettii = b. bissettiae gupta 2019 i. ricinus, i. scapularis, i. pacificus birds, rodents europe, asia, north america b. lusitaniae le fleche et al. 1997 i. ricinus lizards europe, north africa quite late, lyme disease began to be associated with non-specific symptoms. the year 1975 was a breakthrough because in that year lyme disease was diagnosed in the city of lyme (hence the common name of this disease). at that time, 12 cases of arthritis in children were described and associated with skin lesions that occurred after a tick bite (fraser et al., 1997; long, cohn, 2018). since then, the number of patients has been steadily increasing (magnarelli 2009; koperwas, 2013; stone et al., 2017). based on the most recent data from 2010–2018, approximately 476,000 people are diagnosed and treated with lyme disease in the united states each year (centers for disease control and prevention). in 1982, wilhelm burgdorfer isolated borrelia spirochetes, which were named after him – b. burgdorferi; it is one of the most common borrelia species in north america (tab. 1). thanks to this, he also proved the relationship between tick bites and lyme disease (fraser et al., 1997). infection occurs when an infected tick burrows into the skin to collect food – human blood, and releases saliva or the contents of the midgut, in other words, “vomit”. spirochetes are present in the digestive tract of the tick, after multiplication they pass into the salivary glands and then penetrate the skin, later migrating to distant human systems and organs, causing disease symptoms (sobieszczańska et al., 1998; cuningham 2005). 3 fig. 1. the main ways of spread of borrelia burgdorferi s.l. (according to matysiak, 2018 – changed) the bacterial dissemination cycle can be divided into several stages. ticks acquire the disease by biting into and coming into contact with bacteria from their host. the bacteria are then transferred by the tick into the blood or body fluids of the next host, which is a warmblooded animal (mammal). in the third stage, the bacteria live in the cells of the mammal, and in the fourth, the infected cells die and release the bacteria into the blood, where the tick can pick them up again (dumler, bakken, 1998; brochocka et al., 2018; matysiak, 2018). ticks have four stages of development – egg, larva, nymph and adult (fig. 1). lyme disease is a disease in which the symptoms are non-specific (ambiguous). the presence of erythema migrans on the skin is characteristic, but it does not appear in all infected people. it is estimated that erythema migrans occur in about 50–80% of patients. erythema is a circular skin rash with a pale rim between the centre of the rash and the outer circle. erythema migrans, as the name suggests, does not have to appear at the site of the bite, but it can wander all over the body (cameron et al., 2014). it also happens that patients do not experience any symptoms at first, and the untreated disease takes various forms. the most common symptoms of lyme disease include fatigue, headaches, migraines, dizziness, fever, 4 muscle and joint pain, general weakness, memory disorders, numbness of the limbs, muscle spasms, chills, sweats, visual disturbances, hair loss and the above-mentioned erythema migrans. less common symptoms include difficulty breathing, sinusitis, diarrhoea, nausea, constipation, enlargement of the spleen and liver, weight loss, cystitis, impotence, libido disorders, psoriatic lesions, lymphocytic infiltrates on the skin, etc. lyme disease also imitates mental disorders, it can cause severe anxiety attacks, combined with panic attacks (matysiak, 2018). in the case of lyme disease, patients reach for plant therapies when the disease has become chronic (late phase) and suffers from complications after antibiotic terapy, in the hope of improving their health (cianciara, juszczyk, 2012; adrion et al., 2015; aucott, 2015; middelveen et al., 2018; adams, 2022). however, according to the recommendations of scientific societies, such as the polish society of epidemiologists and doctors of infectious diseases (polskie towarzystwo epidemiologów i lekarzy chorób zakaźnych), antibiotics should be used in the treatment of lyme disease and neuroborreliosis, which, depending on the form of the disease, can be administered orally or intravenously. antibiotic treatment is not replaced by other therapies. taking any preparations (whether they are herbs or supplements) that have not undergone clinical trials carries the risk of poisoning or allergies. this is only supportive treatment. there is no conclusive evidence for the effectiveness of treating lyme disease other than antibiotics, as it is a bacterial infection. taking herbs or supplements often mobilizes patients to a more healthy lifestyle, which in turn brings an improvement in well-being. this way of taking care of health after antibiotic therapy is considered by specialists to be appropriate and less harmful.therefore, more and more different herbal preparations are commercially available for to support the treatment cycle and completed antibiotic therapy of lyme disease. the main aim of this study is to make a list of plants, contained in herbal preparations and dietary supplements used to support lyme disease therapy, available for sale without a prescription in poland. analysis methods the research consisted in analysing the composition of herbal preparations supporting the treatment of lyme disease (supplements and herbal medicines without a prescription). materials for the inventory were obtained from stationary in pharmacies and herbal shops in krakow (southern poland) and through a review of commercial offers of online herbal shops. 5 in total, 77 preparations available for sale were collected for analysis: stationary 29, online 48. among the preparations, capsules and herbs for brewing prevailed. other forms of preparations were less common (fig. 2). the plant species obtained based on the above-mentioned analysis are summarised in a table, with the latin name of the species and family affiliation, origin, type of herbal raw material and trade names of preparations in which they were recorded. fig. 2. the forms of the analysed group of herbal medicines and supplements available in trade the origin of species was assumed mainly according to podbielkowski, studnikwójcikowska (2003), while the names of raw materials were taken from product packaging and the internet atlas of vascular plants of poland (https://atlas-roslin.pl/roslinylecznicze.htm), with additions from various other internet sources. a list of species whose raw materials are most often used in the treatment and supplementation of lyme disease, the most numerously represented families and types of herbal raw materials used in the analysed group of preparations was also made. nomenclature and belonging to families of european species and acclimatised in europe were taken from the internet atlas of vascular plants of poland (www.atlas-roslin.pl) and the internet atlas of mushrooms (www.grzyby.pl), and exotic species from world flora online (https://wfoplantlist.org/plant-list/). the systematic arrangement of the families and their nomenclature was used according to reveal (2007). 6 results the conducted analysis showed that 77 preparations used in the treatment and supplementation of lyme disease contain herbal raw materials of 101 plant species and one fungus (tab. 2 – appendix 1). the most common species in the studied group of preparations include reynoutria japonica, dipsacus sylvestris, uncaria tomentosa, cistus ×incanus, and andrographis paniculata (fig. 3). fig. 3. the comparison of the most common species in the study group of herbal medicines and supplements among the recorded species, most are native to areas of asia, europe and africa (fig. 4). fewer species are native to the americas (a total of 18 have been recorded). some species are native to eurasia – e.g. chelidonium majus l., angelica archangelica l., and others occur virtually everywhere and can be considered cosmopolitan, e.g. trametes versicolor (l.) lloyd, equisetum arvense l., or polygonum aviculare l. one species of unknown origin was also recorded – citrus bergamia risso (risso) & poit. 7 fig. 4. the comparison of the number of species found in the study group of herbal medicines and supplements due to their origin in total, these species belong to 53 families (tab. 3). the most numerous families are lamiaceae and rosaceae. most families (as many as 34) are represented by one species. tab. 3. the comparison of the share of medicinal species recorded in the analysed group of preparations within families (fungus, horsetails, gymnosperms, angiosperms: monocotyledonous, dicotyledonous); the most represented families are distinguished in grey no. name of the family number of species no. name of the family number of species 1. polyporaceae fr. ex corda 1 28. rosaceae juss. 8 2. equisetaceae michx. ex dc 1 29. hydrangeaceae dumort. 1 3. pinaceae spreng. ex f. rudolphi 2 30. crassulaceae j. st.-hil. 1 4. asparagaceae juss. 1 31. saxifragaceae juss. 1 5. amaryllidaceae j. st.-hil. 2 32. fabaceae lindl. 3 6. smilacaceae vent. 1 33. rutaceae juss. 4 7. zingiberaceae martinov 4 34. geraniaceae juss. 1 8. bromeliaceae juss. 1 35. polygalaceae hoffmanns. & link 1 9. poaceae barnhart 5 36. combretaceae r. br. 3 10. annonaceae juss. 1 37. onagraceae juss. 1 11. menispermaceae juss. 2 38. myrtaceae juss. 4 12. berberidaceae juss. 1 39. zygophyllaceae r. br. 1 13. papaveraceae juss. 1 40. araliaceae juss. 1 14. lauraceae juss. 1789 1 41. apiaceae lindl 4 15. piperaceae giseke 1 42. vitaceae juss. 1 16. juglandaceae dc. 1 43. santalaceae r. br. 1 17. urticaceae juss. 1 44. oleaceae hoffmanns. & link 1 18. polygonaceae juss. 2 45. apocynaceae juss. 1 19. theaceae mirb. ex ker gawl. 1 46. rubiaceae juss. 2 20. cistaceae juss. 2 47. adoxaceae e. mey. 1 8 21. moringaceae martinov. 1 48. dipsacaceae juss. 1 22. caricaceae dumort. 1 49. solanaceae juss. 2 23. cucurbitaceae juss. 1 50. acanthaceae juss. 1 24. brassicaceae burnett 3 51. pedaliaceae r. br. 1 25. malvaceae juss. 1 52. lamiaceae martinov 9 26. phyllanthaceae martinov. 1 53. asteraceae bercht. & j. presl 4 27. ericaceae juss. 2 among the herbal raw materials, fruits, roots, herbs, oils, leaves and rhizomes are usually used in the analysed group of preparations, while fruiting bodies, tubers, and others. are used the least often (fig. 5). fig. 5. the comparison of the number of occurrences of various types of herbal raw materials in the analysed preparations discussion phytotherapy supporting the treatment of lyme disease early diagnosis of lyme disease increases the chance of its complete recovery (hinckley et al., 2014). when it is not possible to get a diagnosis early enough, the disease begins to pass into various forms that are difficult to treat and very debilitating for the body. therefore, in later supportive therapies, herbal medicine is often used to help the body weakened by antibiotics (buhner, 2005, 2015a, b; cianciara, juszczyk, 2012; smoleńska et al., 2016; feng 9 et al., 2020). herbal regimens to support the treatment of lyme disease include various protocols. below are three examples of herbal protocols to support the treatment of lyme disease. the buhner protocol is a herbal supportive therapy known all over the world, while the protocols of oruba and różański are known mainly in poland. when describing these three selected herbal protocols, it should be noted that there are no clinically proven studies of their effectiveness so far. stephen harrod buhner is one of the most famous american herbalists. he is the author of many books on herbal medicine and lyme disease (e.g. buhner, 2005, 2015a, b). according to him, herbal therapy is a very good substitute for synthetic drugs used in the treatment and elimination of the effects of lyme disease (however, this is not clinically proven). the buhner procedure is divided into basic protocol and extended protocol. the basic protocol consists of three herbs: reynoutria japonica, uncaria tomentosa, and andrographis paniculata. the extended protocol covers five medicinal species: stephania tetrandra, eleutherococcus senticosus, astragalus mongholicus, dipsacus sylvestris, smilax officinalis. in his books, buhner describes exact instructions for taking and combining herbs with each other in order to obtain the best therapeutic effects (buhner, 2015a, b). jan oruba, polish herbalist and author of the protocol supporting the treatment of lyme disease with liposomal herbs (liposomes are very small, water-lipid structures in the shape of a sphere/vesicle; discovered in the 1960s by the haematologist and biophysicist alec bangham). they are carriers of active compounds from herbs or oils and are the safest form of delivering these substances to the body. according to oruba, there is no one specific herb that treats lyme disease, as lyme disease can present differently in everyone and involve multiple systems. his supporting therapy consists of using of liposomal herbs, selected according to the dominant symptoms, characteristic of lyme disease, babesiosis, bartonella, mycoplasma. according to oruba, our immune system is able to prioritise itself and choose the dominant infection that is the greatest threat to it at a given moment (oruba, 2018). henryk różański is a long-term polish scholarship holder of the swiss büchner foundation (1993-2000), the author of many publications in the field of botany, phytopharmacology, phytotoxicology, and pharmaceutical biology (e.g. różański, h., petryja, 2021, 2022). according to różański, herbs supporting the treatment of lyme disease are divided into basic ones (uncaria tomentosa, dipsacus sylvestris), immunising (mud extracts, aloe sp., nigella sativa l. oil, artemisia absinthium l., echinacea purpurea (l.) moench root and herb, e. pallida (nutt.) nutt. and e. angustifolia dc.), anti-inflammatory (harpagophytum procumbens root, melampyrum sp. herb, rhinanthus alectorolophus herb 10 (scop.) pollich and r. serotinus (schönh.) oborný), odontites verna herb (bellardi) dumort., angelica archangelica root combined with filipendula ulmaria flower, buds or bark populus alba l., p. nigra l. or p. ×canescens (aiton) sm., p. tremula l. and the bark of salix alba l. or s. purpurea l.), antibacterial/disinfectant (bark, twigs, buds and leaves of padus avium mill., herb and root of sanguisorba officinalis and s. minor scop., rhizome of curcuma longa l. and flower and herb of tanacetum vulgare l.). they should be used in the right order to get the best phytotherapy results. herbal preparations commercially available during the collection of materials for this study, most preparations were obtained via the internet and in herbal shops. however, in most pharmacies, pharmacists recommended only dried herbs from cistus sp. for brewing, and this was usually the end of the commercial offer of preparations supplementing the treatment of lyme disease. many species are traditionally used in this type of supporting therapy (tokarski, denys, 2018), although their effects are not fully confirmed (tab. 4; fig. 6 – appendix 2). tab. 4. plants supporting the lyme disease treatment cycle and completed antibiotic therapy no. latin name common name 1. andrographis paniculata creat, green, chiretta 2. astragalus mongholicus mongolian milkvetch 3. cistus creticus pink rock-rose, hoary rock-rose 4. c. ×incanus hoary rock-rose 6. dipsacus silvestris wild teasel, fuller’s teasel 7. eleutherococcus senticosus devil’s bush, siberian ginseng, eleuthero 12. reynoutria japonica japanese knotweed, asian knotweed 13. rhodiola rosea golden root, rose root, roseroot 14. scutellaria baicalensis baikal skullcap, chinese skullcap 15. smilax officinalis sarsaparilla plant 16. stephania tetrandra, s. cepharantha fen fang ji 17. uncaria tomentosa, u. rhynchophylla cat’s claw, uña de gato, fish hook vine you can buy both ready-made preparations in various forms, from various raw materials (fig. 2, 5), as well as books with recipes for individual forms of lyme disease and other tick-borne diseases. in some shops, it is also possible to order mixtures created on the spot for specific symptoms of this disease. in the analysed 77 preparations, as many as 101 plants and one species of fungus were recorded (tab. 2 – appendix 1). most of the tested preparations contain species described in the buhner protocol (buhner, 2005, 2015a, b) and plants with a general effect, supporting the 11 proper functioning of the body. the most common plants are reynoutria japonica, dipsacus sylvestris, cistus ×incanus oraz uncaria tomentosa (fig.3.). the first two species belong to the ingredients obtained in europe easily from the wild, which certainly affects the lower price and better quality of their raw materials. in addition, these plants have a proven effect against lyme spirochetes (in cell line studies), and this determines the effectiveness of preparations based on them. for example, ethanol extracts from reynoutria japonica show strong activity against the growing b. burgdorferi as well as its non-growing stationary phase (feng et al., 2020). the main active ingredient of r. japonica is the polyphenol – resveratrol, which, according to goc and rath (2016), has a bacteriostatic and bactericidal effect against borrelia sp. spirochetes. however, it is less effective against rounded forms and has no significant effect against biofilm. another active ingredient here is emodin (6-methyl-1,3,8trihydroxyanthraquinone), which has documented activity against b. burgdorferi stationary phase cells (feng et al., 2015). r. japonica eliminates the symptoms of this disease in most people (buhner, 2015a, b), and has the closest effect to the cytokine dynamics caused by lyme bacteria. in herbal medicine, it is most often found as an ingredient in capsules, but also as a powdered root, dried leaves, dried rhizomes and in the form of tinctures. it grows naturally in japan, taiwan, china and korea (tab. 2 – appendix 1). it was brought to europe, where it spread quickly and is now considered an invasive plant (chmura et al., 2013). liebold et al. (2011) showed that ethyl acetate and dichloromethane extracts from the roots of dipsacus sylvestris, of lipophilic nature, have an inhibitory effect on b. burgdorferi, while the hydroethanolic extract does not inhibit the growth of spirochetes. according to saar-reisma et al. (2022), extracts from d. sylvestris leaves also show activity against b. burgdorferi. however, due to their cytotoxicity, the authors indicated the need to isolate the most active and less cytotoxic fractions. they also found that the highest cytotoxicity came from polyphenols. the iridoid-glycoside fraction, containing two main bioactive substances – sylvestrosides iii and iv, has reduced cytotoxicity and an effective effect against borrelia in the stationary phase. cistus ×incanus, on the other hand, has anti-inflammatory effects, inhibits the growth of lyme spirochetes, and its regular consumption helps the body produce a smell that is repellent to ticks (buhner, 2015a, b). it is found in europe and asia minor (tab. 2 – appendix 1). other species of the genus cistus, e.g. c. creticus l., also have an inhibitory effect on the proliferation of borrelia (hutschenreuther et al., 2010). two other species – uncaria tomentosa and cryptolepis sanguinolenta are foreign plants in europe, whose herbal raw material is imported. u. tomentosa is the strongest immunostimulating herb used in lyme 12 disease and its co-infections. it increases the amount of cd57 leukocytes, which are crucial in the body’s fight against lyme disease. it soothes muscle and joint pain and inflammation. according to feng et al. (2020), u. tomentosa shows strong activity against non-growing b. burgdorferi stationary phase cells and weaker activity against growing ones. it naturally occurs in central and south america and the caribbean (tab. 2 – appendix 1). in herbal medicine, it is mainly used in tablets, powders, infusions and tinctures. native to africa, cryptolepis sanguinolenta has anti-inflammatory, antibacterial, antifungal, antiamoeba and antimalarial properties. includes, e.g. the alkaloid cryptolepine, with bactericidal and bacteriostatic properties (paulo, gomes 1995; osafo et al., 2017). cryptolepin causes morphological changes and cell lysis as well as dna intercalation and inhibition of poisomerase ii. it shows strong activity against b. burgdorferi and is also highly effective against even its aggregated forms (feng et al., 2020). despite the proven effectiveness, this species was found in only two of the studied preparations (tab. 2 – appendix 1). other plant species, showing according to feng et al. (2020) strong activity against b. burgdorferi, are also present in a few analysed preparations: juglans nigra in one, artemisia annua in three, and scutellaria baicalensis in five. most species belong to the rosaceae and lamiaceae families (tab. 3). these are families rich in common species and well-documented in european native flora (mederska, 2021; mederska, mederski, 2021). on the other hand, it seems very interesting that in the analysed group of preparations, there are also quite often species from families that are not very representative or completely exotic in central europe flora, e.g. myrtaceae, rutaceae, zingiberaceae, combretaceae. lyme disease is still poorly understood, as are the properties of many plant species. not all plants supporting the treatment cycle and completed antibiotic therapy have a confirmed effect in this respect, as mentioned earlier. producers of preparations willingly use plant materials used in traditional far eastern medicine (fig. 4), although they are not sure about their effectiveness in this disease. the addition of raw materials known in the far east also greatly affects the commercial attractiveness of the products offered, because people prefer natural traditional herbal medicine (anczyk, 2021). a somewhat similar explanation may apply to a very large group of species native or cultivated in europe, which have been recorded here, e.g. garlic, onion, pine, horsetail, currant, cabbage, apple tree, rose, elderberry, hawthorn, etc. (tab. 2 – appendix 1 ). many of them have a general-strengthening effect, and some simply improve the functioning of the immune system (van wyk, wink, 2017; bigoś, 2019). these premises are already recognised by producers of herbal preparations as a sufficient reason for their use in lyme disease 13 supplements. for example, thompson et al. (2023) analysed 18 herbs commonly used by patients to treat lyme disease symptoms, seven of which had proven in vitro activity against b. burgdorferi, thirteen of which had proven in vitro antimicrobial activity, and 15 had documented anti-inflammatory properties. therefore, when buying herbal preparations that relieve the symptoms of lyme disease, it is worth checking their composition for the presence of species with proven therapeutic properties (at least in cell line studies), such as herbs from the basic or extended buhner protocol. many dishonest manufacturers, taking advantage of the poor knowledge of the average citizen about this disease, will unfortunately continue to offer supplements with poor healing properties. therefore, in addition to prophylaxis (nowak-chmura, siuda, 2012; sinski, welc-faleciak, 2012; nieto et al., 2018; hussain, 2021), the effectiveness of lyme disease detection and thorough examination of the course of the disease itself (hinckley et al., 2014), it is also worth promoting the knowledge already existing in this area about its herbal supplementation (feng et al., 2017; feng et al., 2020). however, it should always be remembered that so far the only effective and clinically proven method of treating lyme disease is antibiotic therapy. conclusions summing up the whole analysis, the following conclusions can be drawn: [1] herbal preparations used in lyme disease supplementation usually have the form of capsules, herbs for infusion or extract, and the most commonly used raw materials are fruit, root, herb, oil, leaf; [2] the species most commonly used in the preparations are reynoutria japonica, dipsacus sylvestris, cistus ×incanus and uncaria tomentosa – other species from the buhner protocol are also quite common (most preparations contained at least one of the ingredients from this group); [3] many plants contained in the preparations have an effect that generally supports the proper functioning of the body or targets a specific form of this disease; [4] most species belong to the rosaceae and lamiaceae families; representatives of other families appear quite often: myrtaceae, rutaceae, zingiberaceae, combretaceae; [5] the most of species used in lyme disease supplementation come from asia, europe and africa. alphabetical list of polish trade names of 77 analysed preparations: andrographis – magiczny ogród, andrographis 10% ekstrakt, andrographis paniculata nalewka – nanga, andrographis paniculata ziele cięte – nanga, astragalus root – swanson, bolleriocaps – herbapol, bolleriofix – herbapol, borelfix – herbarium św. franciszka, borelia med – berg life, borelio herbs – inwent herbs, borelio mieszanka ziołowa, boreliol, borelioza max, borelisspro, borelix forte – produkty bonifraterskie, borellvit, 14 borelyma – herbal monasterium, borrelia protect extra, borreliosis tea – everest ayuverda, cat’s claw koci pazur – swanson, cryptolepis – magiczny ogród, czystek – sekrety zielnika, czystek altermedica laboratories, czystek fix – zielnik apteczny, czystek kreteński – magiczny ogród, czystek – natura wita, eko czystek liść – natura wita, eleuthero ginseng royal jelly, kłącze rdestowca japońskiego – flos, koci pazur (vilcacora) – rozdrobniona kora, korzeń szczeci – flos, liposomalny rdest japoński, lyme protector, maść arnikowa elissa, maść szałwiowa elissa, mielona ashwagandha, nanobab – b&m, nanobarto – b&m, nanoborrel – b&m, nanomyko – b&m, now foods andrographis extract, now foods black walnut hulls, now foods cat’s claw koci pazur, oeparol – adamed, olejek z drzewa herbacianego – herbapol, optiborelia, probio borelio, rdest japoński – b&m research, rdestowiec – magiczny ogród, rdestowiec 500+ nanga suplements, rdestowiec japoński kłącze cięte – nanga, rdestowiec japoński kłącze – herbapol, rdestowiec japoński kłącze mielone – nanga, rdestowiec japoński korzeń mielony – plantago, rdestowiec japoński liść, rdestowiec japoński liść – nanga, rdestowiec japoński susz – astron, rdestowiec japoński ziele suszone – dary podlasia, resweratrol – medica herbs, rhodiola – magiczny ogród, sarsaparilla 400, sarsaparilla root – swanson, stephania – magiczny ogród, stop borelia herbatka ziołowa, szczeć – altermedica laboratories, szczeć pospolita kapsułki – futunatura, szczeć pospolita kapsułki – medica herbs, szczeć pospolita korzeń – natura wita, tarczyca bajkalska – magiczny ogród, traganek – magiczny ogród, yango borrelin, yango czystek, yango szczeć, zimnotłoczony olej z nasion wiesiołka – gal, żeń-szeń syberyjski – korzeń cięty, żeń-szeń syberyjski – magiczny ogród, żeń-szeń syberyjski 400. conflict of interest the authors declare no conflict of interest related to this article. references adams, w. 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(2017). medical plants of the world. publisher: cabi. 520 pp. world flora online (https://wfoplantlist.org/plant-list/) (access 2020-07-20) 18 appendix 1 tab. 2. list of all taxa (fungi, horsetails, gymnosperms and angiosperms: monocotyledonous, dicotyledonous) used as support in the treatment of lyme disease and improving immunity no. family, latin name of a plant/fungus origin herbal raw material polish trade names of medicinal preparations polyporaceae fr. ex corda 1. trametes versicolor (l.) lloyd cosmopolitan fungus trametes lyme protector equisetaceae michx. ex dc. 2. equisetum arvense l. cosmopolitan equiseti herba probio borelio pinaceae spreng. ex f. rudolphi 3. cedrus atlantica (endl.) g. manetti ex carrière north africa oleum cedri nanobarto – b&m 4. pinus sylvestris l. europe, northern zone of asia folium et gemma pini borelfix – herbarium św. franciszka; borelioza max asparagaceae juss. 5. asparagus officinalis l. mediterranean area, western siberia rhizoma asparagi borelisspro amaryllidaceae j. st.-hil. 6. allium cepa l. west asia allium cepa folium borelisspro 7. a. sativum l. central asia allii sativi bulbus borelisspro; borelioza max; optiborelia; probio borelio; yango borrelin smilacaceae vent. 8. smilax officinalis kunth central america and northwest part of south america smilax officinalis radix/radix sarsaparillae borelio mieszanka ziołowa; borelix forte – produkty bonifraterskie; borrelia protect extra; sarsaparilla 400; sarsaparilla root – swanson zingiberaceae martinov 9. curcuma longa l. india, malaysia curcumae longae rhizoma borreliosis tea – everest ayuverda; nanobab – b&m; probio borelio 10. c. zedoaria (christm.) roscoe south-east asia rhizoma zedoariae nanobab – b&m 11. hedychium spicatum buch.-ham. ex sm. china, himalayas, burma, ethiopia, thailand hedychium spicatum rhizoma nanoborrel – b&m http://www.wikidata.org/entity/q536945 http://www.wikidata.org/entity/q458004 19 12. zingiber officinale rosc. malaysia, india zingiberis rhizoma probio borelio bromeliaceae juss. 13. ananas comosus (l.) merr brazil ananas comosus fructus borelisspro poaceae barnhart (gramineae) juss. 14. cymbopogon citratus stapf india cymbopogon citratus folium borelfix – herbarium św. franciszka 15. c. flexuosus (nees ex steud.) w.watson india, sri lanka, burma, and thailand oleum cymbopogon flexuosus nanobab – b&m 16. c. martinii (roxb.) w.watson india and indochina oleum cymbopogoni martinii nanobarto – b&m 17. c. winterianus jowitt ex bor western malesia oleum cytronellae indicum nanoborrel – b&m 18. triticum vulgare l. cosmopolitan triticum vulgaris germen borelisspro annonaceae juss. 19. cananga odorata (lam.) hook.f. & thom. south-east asia oleum cananga odorata nanobab – b&m menispermaceae juss. 20. stephania tetrandra s.more china and taiwan radix stephaniae tetrandrine stephania – magiczny ogród 21. tinospora cordifolia (willd.) hook. f. & thom. indian subcontinent tinospora cordifolia radix and stipula borreliosis tea – everest ayuverda berberidaceae juss. 22. berberis thunbergii dc. japan and eastern asia radix berberidis borelioza max papaveraceae juss. 23. chelidonium majus l. eurasia chelidonii herba borelioza max lauraceae juss. 1789 24. cinnamomum verum j.presl sri lanka cinnamomi cortex nanoborrel – b&m 20 25. litsea cubeba pers. southern china oleum litsea cubeba nanoborrel – b&m piperaceae giseke 26. piper nigrum l. malabar coast (india) fructus piperis nigri borellvit; probio borelio juglandaceae dc. 27. juglans nigra l. north america pericarpium juglandis now foods black walnut hulls urticaceae juss. 28. urtica dioica l. eurasia, north africa, north and south america herba utricae dioicae czystek – sekrety zielnika polygonaceae juss. 29. polygonum aviculare l. cosmopolitan polygoni avicularis herba probio borelio 30. reynoutria japonica houtt. south-east asia polygoni cuspidati rhizoma et radix bolleriocaps – herbapol; borelia med – berg life; borelio herbs – inwent herbs; borelio mieszanka ziołowa; boreliol; borelioza max; borelisspro; borelix forte – produkty bonifraterskie; borellvit; borelyma – herbal monasterium; borrelia protect extra; kłącze rdestowca japońskiego – flos; liposomalny rdest japoński; optiborelia; rdest japoński – b&m research; rdestowiec – magiczny ogród; rdestowiec 500+ nanga suplements; rdestowiec japoński kłącze cięte – nanga; rdestowiec japoński kłącze – herbapol; rdestowiec japoński kłącze mielone – nanga; rdestowiec japoński korzeń mielony – plantago; rdestowiec japoński liść; rdestowiec japoński liść – nanga; rdestowiec japoński susz – astron; rdestowiec japoński ziele suszone – dary podlasia; yango borrelin theaceae mirb. ex ker gawl. 31. camellia sinensis (l.) kuntze south-east asia folium theae borelisspro; lyme protector cistaceae juss. 32. cistus creticus l. north africa, west asia, south and east europe cistus creticus herba borelfix – herbarium św. franciszka; borrelia protect extra; czystek kreteński – magiczny ogród; lyme protector; yango borrelin; yango czystek 33. c. ×incanus l. hybrids cistus folium/ herba/ oleum bolleriocaps – herbapol; bolleriofix – herbapol; borelia med – berg life; borelio herbs – inwent herbs; borelio mieszanka ziołowa; borelisspro, borelix forte – produkty bonifraterskie; borelyma – herbal monasterium; czystek – sekrety zielnika; czystek altermedica 21 laboratories; czystek fix – zielnik apteczny; czystek – natura wita; eko czystek liść – natura wita; optiborelia; probio borelio; stop borelia herbatka ziołowa moringaceae martinov. 34. moringa oleifera lam. indian subcontinent moringa herba probio borelio caricaceae dumort. 35. carica papaya l. south america, south regions of north america fructus caricae borelisspro cucurbitaceae juss. 36. cucumis sativus l. india, south china cucumis sativus fructus borelisspro brassicaceae burnett 37. brassica oleracea l. var. capitata (l.) duchesne europe, cultivar brassicae oleraceae folium borelisspro 38. b. oleracea l. var. italica plenck europe, cultivar brassica oleracea inflorescentia borelisspro 39. isatis tinctoria l. southeast europe, west asia isatidis radix nanomyko – b&m malvaceae juss. 40. hibiscus syriacus l. korea, south-central, southeast china, india flos hibisci syriacus bolleriofix – herbapol; czystek – sekrety zielnika phyllanthaceae martinov. 41. phyllanthus emblica l. south asia emblica officinalis fructus borreliosis tea – everest ayuverda ericaceae juss. 42. gaultheria procumbens l. northeastern north america oleum gaultheriae nanoborrel – b&m 43. vaccinium myrtillus l. asia, europe, north america fructus vaccinii borelisspro rosaceae juss. 44. armeniaca vulgaris lam. central asia armeniacae fructus borelisspro 45. aronia melanocarpa (michx.) elliott eastern north america aroniae fructus bolleriofix – herbapol 46. cerasus avium (l.) moench europe, western asia, asia minor pruni aviae fructus borelisspro 47. crataegus sp. temperate zone of the crataegi fructus probio borelio 22 northern hemisphere 48. filipendula ulmaria (l.) maxim. asia, northern and central europe filipendulae ulmariae herba borelioza max 49. fragaria ×ananasa duchesne hybrids fructus fragariae ananssae borelisspro 50. malus sylvestris (l.) mill. europe fructus malus sylvestris borelisspro 51. rosa canina l. europe, north africa, asia minor fructus rosae bolleriofix – herbapol; czystek – sekrety zielnika hydrangeaceae dumort. 52. hydrangea febrifuga (lour.) y.de smet & c.granados asia? radix dichroae nanobab – b&m crassulaceae j. st.-hil. 53. rhodiola rosea l. north america, europe, asia rhodiolae roseae rhizoma cum radicibus rhodiola – magiczny ogród saxifragaceae juss. 54. ribes nigrum l. europe, asia fructus ribis nigri bolleriofix – herbapol; borelisspro fabaceae lindl. (leguminosae juss., papilionaceae giseke) 55. astragalus mongholicus bunge temperate asia radix astragali borellvit; lyme protector; traganek – magiczny ogród; astragalus root – swanson 56. glycyrrhiza glabra l. south-east europe, asia minor liquiritiae radix borreliosis tea – everest ayuverda 57. pterocarpus marsupium roxb. india, nepal, sri lanka pterocarpus marsupium herba borreliosis tea – everest ayuverda rutaceae juss. 58. citrus bergamia risso (risso) & poit. unknown oleum citrus bergamia nanomyko – b&m 59. c. ×paradisi macfad. hybrids citrus paradisi fructus borelisspro 60. c. ×sinensis (l.) osbeck hybrids fructus citrus sinensis borelisspro 61. phellodendron amurense rupr. eastern asia cortex phellodendri nanobab – b&m; nanomyko – b&m http://www.wikidata.org/entity/q729917 http://www.wikidata.org/entity/q36706980 http://www.wikidata.org/entity/q36706980 http://www.wikidata.org/entity/q23649298 23 geraniaceae juss. 62. pelargonium sidoides dc. south africa pelargonii radix borelix forte – produkty bonifraterskie polygalaceae hoffmanns. & link 63. polygala tenuifolia willd. asia radix polygalae nanobarto – b&m combretaceae r. br. 64. terminalia arjuna (roxb. ex dc.) wight & arn. indian subcontinent terminaliae arjunae cortex borreliosis tea – everest ayuverda 65. t. bellirica (gaertn.) roxb. south and south-west asia terminalia bellirica fructus borreliosis tea – everest ayuverda 66. t. chebula retz. south asia fructus chebulae borreliosis tea – everest ayuverda onagraceae juss. (oenotheraceae c. c. robin) 67. oenothera biennis l. north america oleum oenotherae oeparol – adamed; zimnotłoczony olej z nasion wiesiołka – gal myrtaceae juss. 68. eucalyptus globulus labill. south-eastern australia eucalypti folium nanomyko – b&m 69. melaleuca alternifolia cheel australia melaleucae aetheroleum nanomyko – b&m; olejek z drzewa herbacianego herbapol 70. m. viridiflora sol. ex gaertn. northern australia, new guinea oleum melaleuca viridiflora nanobab – b&m; nanoborrel – b&m 71. syzygium aromaticum (l.) merr. & perry moluccas (indonesia) oleum caryophylli nanobarto – b&m zygophyllaceae r. br. 72. tribulus terrestris l. southern eurasia and north africa tribulus terrestris herba et fructus borreliosis tea – everest ayuverda araliaceae juss. 73. eleutherococcus senticosus maxim. east asia, china, japan, russia eleutherococci radix borelio herbs – inwent herbs; borelix forte – produkty bonifraterskie; borellvit; borrelia protect extra; eleuthero ginseng royal jelly; probio borelio; żeń-szeń syberyjski – korzeń cięty; żeń-szeń syberyjski – magiczny ogród; żeń-szeń syberyjski 400 apiaceae lindl. (umbelliferae juss.) 74. angelica archangelica l. eurasia angelicae archangelicae radix probio borelio 75. a. dahurica (hoffm.) benth. & hook.f. ex franch. & sav. siberia, russia far east, mongolia, northeastern china, japan, radix angelicae dahuricae nanomyko – b&m 24 korea, taiwan 76. centella asiatica (l.) urb. africa, asia, australia, islands in the western pacific ocean centellae asiaticae herba borreliosis tea – everest ayuverda 77. daucus carota l. eurasia, south africa daucus carota radix borelisspro vitaceae juss. 78. vitis vinifera l. mediterranean region, central europe, southwestern asia fructus vitis veniferae, vitis vinifera semen borelisspro; yango borrelin santalaceae r. br. 79. santalum album l. indian subcontinent oleum santali borreliosis tea – everest ayuverda oleaceae hoffmanns. & link 80. olea europaea l. mediterranean basin (europe, asia, africa) olea europaea fructus borelisspro apocynaceae juss. 81. cryptolepis sanguinolenta (lindl.) schltr., c. dubia (burm.f.) m.r.almeida africa; south, southeast asia radix cryptolepis sanguinolentae cryptolepis – magiczny ogród; nanobab – b&m rubiaceae juss. 82. galium aparine l. cosmopolitan herba galii aparinis borelioza max 83. uncaria tomentosa dc. south and central america, caribbean uncariae tomentosae cortex borelia med – berg life; bolleriocaps – herbapol; borelio herbs – inwent herbs; borelio mieszanka ziołowa; boreliol; borelioza max; borelix forte – produkty bonifraterskie; borellvit; borelyma – herbal monasterium; borrelia protect extra; cat’s claw koci pazur – swanson; koci pazur (vilcacora) – rozdrobniona kora; now foods cat’s claw koci pazur; optiborelia; probio borelio; yango borrelin adoxaceae e. mey. 84. sambucus nigra l. europe flos sambuci probio borelio dipsacaceae juss. 85. dipsacus sylvestris huds. europe, west asia, south africa dipsacus fullonum radix bolleriofix – herbapol; borelfix – herbarium św. franciszka; borelia med -berg life; borelio herbs – inwent herbs; borelio mieszanka ziołowa; borelix forte – produkty bonifraterskie; borellvit; optiborelia; probio borelio stop borelia herbatka ziołowa; szczeć – altermedica laboratories; szczeć 25 pospolita korzeń – natura wita; bolleriocaps – herbapol; borelioza max; borelisspro; borelyma – herbal monasterium; korzeń szczeci – flos; szczeć pospolita kapsułki – futunatura; szczeć pospolita kapsułki – medica herbs; yango szczeć; yango borrelin solanaceae juss. 86. lycopersicon esculentum mill. peru, ecuador (south america) lycopersicon esculentum fructus borelisspro 87. withania somnifera (l.) dunal africa, asia radix et fructus withaniae borelix forte – produkty bonifraterskie; borelyma – herbal monasterium; borreliosis tea – everest ayuverda; mielona ashwagandha acanthaceae juss. 88. andrographis paniculata (burm.f.) wall. india, sri lanka herba andrographitis andrographis – magiczny ogród; andrographis 10% ekstrakt; andrographis paniculata nalewka – nanga; andrographis paniculata ziele cięte – nanga; bolleriocaps – herbapol; borelia med – berg life; borelio mieszanka ziołowa; boreliol; borelix forte – produkty bonifraterskie; borelyma – herbal monasterium; borrelia protect extra; borreliosis tea – everest ayuverda; lyme protector; now foods andrographis extract; optiborelia pedaliaceae r. br. 89. harpagophytum procumbens dc. ex meisn. southern africa harpagophyti radix probio borelio lamiaceae martinov (labiatae juss.) 90. betonica officinalis l. europe, western asia, northern africa betonicae herba borelfix – herbarium św. franciszka 91. mentha ×citrata ehrh. hybrids menthae citratae folium nanomyko – b&m; czystek – sekrety zielnika 92. ocimum tenuiflorum l. indian subcontinent ocimum sanctum folium borreliosis tea – everest ayuverda 93. origanum vulgare l. mediterranean basin, siberia, himalayas origani herba borelisspro; nanoborrel – b&m probio borelio; yango borrelin 94. rosmarinus officinalis l. mediterranean basin rosmarinus officinalis folium borelfix – herbarium św. franciszka 95. salvia officinalis l. mediterranean basin, asia minor, syria salviae folium maść szałwiowa elissa 96. s. sclarea l. northern mediterranean basin, north africa, central asia oleum salviae sclarea nanobarto – b&m 97. satureja montana l. southern europe, the satureja montana nanoborrel – b&m 26 mediterranean, africa oleum 98. scutellaria baicalensis georgi china, korea, mongolia, russian far east, siberia radix scutellariae baicalensis bolleriocaps – herbapol; borelio mieszanka ziołowa; nanoborrel – b&m optiborelia; tarczyca bajkalskamagiczny ogród asteraceae bercht. & j. presl (compositae giseke) 99. arnica montana l. europe arnicae flos maść arnikowa elissa 100. artemisia annua l. southeastern europe, western asia artemisiae annuae herba borelioza max; lyme protector; nanobarto – b&m 101. solidago virgaurea l. s.str. eurasia, north africa herba solidaginis borelioza max 102. stevia rebaudiana bertoni brazil, paraguay steviae rebaudianae folium nanoborrel – b&m 27 appendix 2 fig. 6. selected plants traditionally used in the supporting treatment of lyme disease: andrographis paniculata – a, astragalus mongholicus – b, cistus creticus – c, c. ×incanus – d, dipsacus silvestris – e, eleutherococcus senticosus – f, gaultheria procumbens – g, harpagophytum procumbens – h, rhodiola rosea – i, juglans nigra – j, reynoutria japonica – k, scutellaria baicalensis – l, stephania tetrandra – m, uncaria tomentosa – n, u. rhynchophylla – o (source: public domen – https://pl.wikipedia.org/wiki/wikimedia_commons) 28 rośliny wykorzystywane w leczeniu boreliozy streszczenie borelioza z lyme jest chorobą bakteryjną przenoszoną przez kleszcze zakażone bakteriami z grupy borrelia burgdorferi. posiada nieswoiste objawy, takie jak: bóle głowy, gorączka, bóle mięśni, czy stawów, dlatego bywa często mylona z innymi chorobami. w jej przebiegu wyróżnia się 3 stadia: wczesne (rumień wędrujący), wczesne rozsiane (dolegliwości układu kostno-stawowego, ośrodkowego nerwowego) oraz późne (dolegliwości mięśniowo-szkieletowe, arytmia, jednostronne porażenie twarzy, stany zapalne mózgu, rdzenia kręgowego, ból e głowy itd.). leczenie polega na antybiotykoterapii. po zakończeniu leczenia, pacjenci mogą zastosować protokoły ziołolecznicze, które wspomagają organizm i mogą poprawić samopoczucie. celem głównym tego opracowania było sporządzenie wykazu roślin wykorzystywanych we wspomaganiu terapii boreliozy po zakończonej antybiotykoterapii, w oparciu o preparaty ziołowe i suplementy diety występujące w sprzedaży bez recepty na terenie polski. analizie poddanych zostało 29 preparatów dostępnych stacjonarnie oraz 48 preparatów roślinnych dostępnych przez internet (łącznie 77 leków i suplementów bez recepty). w składzie preparatów odnotowano 101 roślin i jeden gatunek grzyba. wiele roślin wykorzystywanych w tego rodzaju fitoterapii wspomagającej, nie wykazuje bezpośredniego działania przeciwko krętkom boreliozy i posiada działanie ogólnie wspierające prawidłowe funkcjonowanie organizmu. niektóre natomiast mają udowodnione właściwości bakteriobójcze (u większości nie zostało udowodnione bezpośrednie działanie w walce z boreliozą), wirusobójcze, wspierające prawidłowe działanie układu nerwowego oraz dostarczające wiele witamin i składników mineralnych potrzebnych dla zdrowia. słowa kluczowe: borrelia burgdorferi, fitoterapia wspomagająca, rośliny lecznicze, suplementacja ziołowa received: [2023.06.08] accepted: [2023.08.18] 103 annales universitatis paedagogicae cracoviensis studia naturae, 3: 103–122, 2018, issn 2543-8832 doi: 10.24917/25438832.3.8 erika remešicová1, peter andráš2*, radmila kučerová3 1nábrežie armádneho generála l. svobodu 5, 812 49 bratislava 1 bratislava, slovakia 2faculty of natural sciences, matej bel university, tajovského 40, 974 01 banská bystrica, slovakia, *peter.andras@umb.sk 3technical university ostrava, ostrava – poruba, czechia environmental characteristics of the mining area affected by sulphide minerals and acidification (banská štiavnica, slovakia) introduction the presence of sulphides in heaps and tailing ponds connected with the oxidation and subsequent acidification represents risk for the environment. iron sulphates and h2so4 are formed during reaction of the sulphides, air, and water. fe2+ may be consequently oxidized and produce more acids. bacteria have an important role in this process that significantly accelerates the oxidation (ledin, pedersen, 1996). the oxidation of the sulphides is a natural process, although naturally taking place much slower in geological time periods (akcil, koldas, 2006; moreno, neretnieks, 2006; lottermoser, 2007; jennings et al., 2008). acidification removes mould, ca, and mg from the soils, mobilizes metals, and reduces the sorption capacity of the upper horizon causing its gradual destruction. water entering the mining areas can enhance the oxidation process and pose a significant risk of acid mine water (amd) formation. the metals released from amd both to the surface water and groundwater are dangerous, particularly because of bioaccumulation, through which they are further spread into the food chain (salomons, 1995; thornton, 1996; bell et al., 2001; marqués et al., 2001; šottník, 2005). some of the mining waste pond sites may seem harmless, without producing amd and become a source of contamination in few years or decades after the termination of mining operations and revegetation (younger, wolkersdorfer, 2004). the aim of this study was to examine the environmental characteristics of the mining area affected by sulphide minerals and acidification (banská štiavnica, slovakia). er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 104 study area, materials and methods banská štiavnica is situated in the southern part of central slovakia and belongs to the protected landscape area štiavnické vrchy mts. in the past, banská štiavnica belonged to the most important agand pbzndeposits of europe. areas of interest in this study are the tailing pond sedem žien and nearby hydroquartzite quarry šobov. the deposit is situated in neovolcanic rocks (diorite, andesite, rhyolite, and younger basalts). the dominant vein minerals are quartz, carbonates, sphalerite, galena, pyrite, chalcopyrite, hematite, gold, argentite, and stephanite (koděra, 1963). the tailing pond was intentionally built according to the project of the mining company banský projekt košice (križáni, andráš, 2008). the pond is 44 m deep with the volume of 2.5 million cubic meters (masarovičová et al., 2007). the area of the tailing pond is approx. 22 ha, and the dam of the pond consists of mine waste rocks and local soils. the pond is filled with mud from the flotation treatment plant of galena-sphalerite ore (masarovičová et al., 2007). the tailing pond was in operation from 1963 until 1994. during the reclamation, the plain of the pond was covered with 2–3 fig. 1. localisation of the sampling points. s1 – 3: soil samples; r1 – 3: rock samples; w1 – 3: water samples (maps source: google earth) environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 105 meters of waste rocks coming from galena-sphalerite mine and partially of waste from the šobov quarry on which was spread a layer of dam soils (masarovičová et al., 2007). sampling sites for soil material were as follows: sample 1 s1 – soil forming the second terrace of the pond’s dam; s2 – soil from the fourth terrace of the pond’s dam; and, s3 – soil from the surface of the top plane of the tailings pond. rock samples were selected as follows: r1 – the rock material from the lower part of the šobov quarry; r2 – rock material from the upper parts of the šobov quarry; and, r3 – pb-zn ore. the analysed soils were sampled from the upper layer (20 cm) of the soil. at the sampling points of the water, ph, conductivity (eh), dissolved oxygen and total dissolved solids of the samples were measured. the šobov quarry is situated in the southwest direction from the pond. the deposit is formed by secondary quartz with dispersed pyrite. the tailing pond, sedem žien, and the šobov quarry fulfil the technical and legislative requirements for categorisation in the group of permanent environmental burdens (masarovičová et al., 2007; sazp.sk, 2015). three sampling sites for each type of sample are presented in figure 1. the following water sampling sites were selected: water sample 1 (w1) – red coloured drainage percolating sedem žien tailing pond; water sample 2 (w2) – transparent drainage percolating tailing pond; and water sample 3 (w3) – accumulated drainage from hydroquartzite quarry šobov. the mineralogical composition of the pulverised soils and rocks samples was determined by x-ray diffraction analysis (xrd) using a philips x’pert pro multipurpose x-ray diffractometer, and the mineral morphology of the samples was studied by scanning electron microscope fei xl 30. the concentrations of macro elements (ca, mg, na, k) and metals (fe, al, pb, zn, mn, co, cr, ni, cu) were measured in rock, soil, and water samples. the soils and rocks samples were moistened with 5 ml of hf to dissolve the silicates and with 1.5 ml of hclo4 to oxidize of the organic matter. the resulting compound was dried for 2 days at 150°c to dry salts. after the addition of 3.75 ml of 37% hcl and 1.25 ml of 69% hno3 the samples were evaporated for 1 hour and mixed with distilled water. analyses of metals and macro elements were realised by atomic absorption spectrometry using a fast sequential atomic adsorption spectrometer varian aa240 fs. the measurement of anions in the water was carried out with use of ion chromatography. measuring f-, cland sulphates (so4)2was performed by a dionex ics 1000 ion chromatography system. the rinse ph (soil reaction) was measured in a suspension of 5 g soil sample in 25 ml of distilled water. a similar procedure was chosen for the determination of paste ph, which was measured in a suspension of 5 g soil in 25 ml of 1 m kcl after 1 hour of mixing in a magnetic stirrer. er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 106 determination of isotopes of selected elements (carbon, oxygen, deuterium, sulphur, nitrogen) is crucial in identifying the origin of the elements. δ13c represents the ratio between carbon isotopes 13c/12c, δ18o represents the ratio between 18o/16o, δ34s is 34s/32s ratio, and δd is the ratio of deuterium to light hydrogen 2h/1h (mook, 2001; šgúdš, 2012). isotope measurements of sulphur (δ 14s) were carried out in all samples. the measurement of deuterium isotopes (δ d or δ 2h) and oxygen (δ 18o) was made in water samples, and carbon isotopes (δ 13c) measurements were made in rock and soil samples. in order to measure the isotopes of sulphur and carbon, solid samples were weighted in tin capsules and mixed with a catalyst of vanadium pentoxide (v2o5). for the sulphur isotopes measurements, precipitation of barium sulphate (baso4) from the water samples was needed. the ph value of the samples was adapted to a value of 1 by the application of hcl. after adding 4 g of nacl, the samples were boiled for 5 min, and consequently bacl2 was added to the solution until a sufficient amount of baso4 was precipitated. water samples measured for deuterium and oxide isotopes were filtered before analysis through a 0.2 µm membrane. deuterium and oxygen were measured by a laser absorption spectroscope laser water isotope analyser oa-icos dlt-100 from los gatos research, and isotopes c and n were measured by a mass spectrometry analyser flash ht plus elemental analyser connected to the instrument delta v advantage isotope ratio mass spectrometer from thermo scientific. isotopes are reported using the conventional δ notation relative to canyon diablo troilite (v-cdt) for 34s, vienna standard mean ocean water (v-smow) for d and 18o, and vienna pee dee belemnite (vpdb) for 13c (ďurza, 2007). tab. 1. quantitative representation of the minerals in studied rock samples sample ang anh ank ant cal clc ccp fsp gn ms po py qtz toz sp r1 + + ++ ++++ + r2 + * + ++ ++ +++ ++ r3 + + + + + +++ ++ +++ +++ ++++ explanatory notes: from + mild representation, to ++++ dominant representation, * traces of the mineral; ang – anglesite, anh – anhydride, ank – ankerite, cal – calcite, clc – clinochlore, ccp – chalcopyrite, fsp – feldspar, gn – galena, ms – muscovite, po – pyrrhotite, py – pyrite, qtz – quartz, toz – topaz, sp – sphalerite soil samples were analysed for the availability of metals and macro elements by single chemical extraction adapted from tipping et al. (2003). the extraction of soil samples with grain size ≤ 2 mm was performed with 0.43 m hno3. triplicates of each soil at a ratio of 2 g air dried soil to 20 ml of extractant were mixed by end-over-end shaking for 2 hours and filtered through a 0.2 µm membrane. samples were then analysed using aas varian aa240 fs equipment. environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 107 results mineralogical characteristics of the rock samples xrd and sem analyses of the rock samples’ mineralogical compositions showed that silica with sulphide minerals predominate in the samples. sample r3 represents sphalerite-galena ore. xrd analysis confirmed the presence of the following sulphide minerals in the next samples: r1 – pyrrhotite, chalcopyrite, and sphalerite; in r2 – pyrite; and, in r3 – pyrite, pyrrhotite, chalcopyrite, galena, and sphalerite (fig. 2; tab. 1). sem analysis enabled us to distinguish the following in sample r1: quartz – sio2, fine-grained pyrite – fes2 in the quartz matrix, galena – pbs, sphalerite – zns, anatase – tio2 and zircon – zrsio4 (appendix 1 – fig. i). sem analysis identified the following in sample r2: quartz, anatase, topaz, anhydride, and pyrite (fes2; appendix 1 – fig. ii). in rock sample r3 the following were identified: quartz, ankerite, clinochlore, dolomite, pyrite, galena, sphalerite, and chalcopyrite (appendix 1 – fig. iii). mineralogical characteristics of the soil samples xrd analysis of soil samples confirmed the presence of the following minerals: quartz, albite, biotite, calcite, clinochlore, illite, ilmenite, jarosite, kaolinite, muscovite, and orthoclas (fig. 3; tab. 2). fig. 2. the mineralogical composition of rock samples determined by xrd; explanatory notes: ang – anglesite, anh – anhydride, ank – ankerite, cal – calcite, clc – clinochlore, ccp – chalcopyrite, fsp – feldspar, gn – galena, ms – muscovite, po – pyrrhotite, py – pyrite, qtz – quartz, toz – topaz, sp – sphalerite er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 108 tab. 2. quantitative representation of the minerals in studied soil samples sample ab bt cal clc ill ilm jrs kln ms or qtz s1 ++ + ++ + ++ +++ s2 + + ++ + + ++ +++ s3 ++ + + + + ++ ++ +++ explanatory notes: from + mild representation, to ++++ dominant representation, * traces of the mineral; abbreviation of minerals as in figure 3 tab. 3. aas analysis of rock and soil samples sample cao mgo na2o k2o al2o3 fe2o3 mn pb co cr cu ni zn [mg.kg-1] r1 0.23 0.13 0.08 0.35 1.61 4.67 0.04 82.48 4.96 531 9.71 173 137 r2 0.27 0.10 0.12 4.65 22.47 75.82 0.01 <d.l. 14.85 315 19.34 78.19 34.74 r3 2.97 42.57 0.12 0.24 52.52 108 13.58 95594 43.03 173 2834 45.37 218767 s1 2.2 6.82 4.72 38.66 102 14.12 0.22 1584 <d.l. 83.06 233 10.67 383 s2 2.48 6.83 2.62 45.44 81.56 29.3 1.18 787 <d.l. 98.1 233 8.21 549 s3 21.21 9.46 6.08 25.06 127 42.87 1.48 797 8.36 120 93.99 18.48 862 sem analysis identified the following minerals in the soil samples: sample s1 – quartz, muscovite, jarosite, biotite, and cerium phosphate (appendix 1 – fig. iv); sample s2 – quartz, biotite, mica, chlorite, orthoclase, anglesite, and iron oxide (apfig. 3. the mineralogical composition of soil samples determined by xrd. explanatory notes: ab – albite, bt – biotite, cal – calcite, clc – clinochlore, ill – illite, ilm – ilmenite, jrs – jarosite, kln – kaolinite, ms – muscovite, or – orthoclase, qtz – quartz environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 109 pendix 1 – fig. v); sample s3 – muscovite, quartz, illite, plumbojarosite, apatite, and ilmenite (appendix 1 – fig. vi). heavy metals contamination the chemical analysis of the samples indicates that the investigated area is contaminated by heavy metals, which correspond to the metals occurring in rocks and ores from the study area (tab. 3). the quartzite samples (r1 and r2) contain an increased amount of fe2o3 due to the presence of pyrite in the samples. sample r3 represents pb-zn ore. samples contain a high amount of fe2o3 (108 g.kg-1), which influences the course of oxidation. soils from the 2nd and 4th terrace of the pond dam (s1 and s2) show a strongly acidic soil reaction (tab. 4). tab. 4. rinse and paste ph sample phh2o phkcl s1 2.28 2.07 s2 3.25 2.81 s3 7.26 6.95 water samples contain high concentrations of metals. the red coloured tailing drainage water (w1) contains a large amount of fe, mn, and zn, and the second drainage water percolating the tailing pond (w2) contains high concentrations of pb (3773 µg.l-1). w3 (from šobov quarry) is rich in fe (311 mg.l-1) and al (157 mg.l-1) (tab. 5). sample w3 is the only sample among analysed drainages that were highly acidic (ph 2.17), and it had the highest values of conductivity and total dissolved solids (tab. 6). high concentrations of sulphates (966-2599 mg.l-1, tab. 7) were found in water samples. tab. 5. aas analysis of water samples sample ca mg na k al fe mn pb co cr cu ni zn [mg.l-1] [µg.l-1] w1 297 59.11 16.01 34.85 0.30 9.63 4.37 73 <d.l. <d.l. 140 <d.l. 1720 w2 461 64.82 14.32 49.3 0.48 0.75 0 3773 <d.l. 0.3 160 <d.l. 400 w3 79.54 98.35 2.57 2.26 157 311 10.74 126 270 170 890 160 1690 tab. 6. ph, electroconductivity, dissolved oxygen and total dissolved solids values measured at the sampling points sample ph ec [µs/cm] do [mg.kg-1] tds [mg.kg-1] w1 7.60 2093 6.26 1049 w2 8.81 2870 10.22 1434 w3 2.17 6457 6.20 3226 explanatory notes: ec – electroconductivity. do – dissolved oxygen. tds – total dissolved solids er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 110 tab. 7. ic analysis of anions in water samples sample fcl(so4)2[mg.l-1] w1 1.08 52.76 966 w2 1.54 39.79 1666 w3 1.56 52.92 2599 percentages of macroand microelements in soils are shown in table 8. tab. 8. percentages of available elements in soils sample ph cao mgo na2o k2o al2o3 fe2o3 mn pb co cr cu ni zn [%] s1 2.28 13.11 5.38 0.3 0.11 0.69 39.27 11.3 2.37   – 1.93 27.23 5.19 57.34 s2 3.25 10.75 1.19 0.11 0.04 0.63 2.74 6.02 2.31  – 1.83 9.11 8.58 8.29 s3 7.26 99.89 9.92 0.22 1.17 2.17 4.82 52.74 56.85 55.55 3.22 33.69 11.09 75.06 the isotopic composition of the drainage water samples shows the meteoric origin of the waters. δd values fluctuate in the range from -42.73 to -74.78‰, and values δ18o vary from -6.11 to -10.17‰. the sulphur isotope composition δ34s in samples w1, w2, and w3 is 3.37, 4.6 and -2.19, while the isotopic compositions of sulphur in the solid samples vary from δ34s -1.48 to 4.48‰. isotopes 13c (-4.25 to -27.73‰) represent isotopic composition characteristic for carbonates (probably of biogenic origin) (tab. 9–10; fig. 4). tab 9. δ d, δ 18o and δ 34s isotopes in water samples sample δ d (‰vs. v-smow) δ 18o (‰vs. v-smow) δ 34s (‰vs.v-cdt) w1 -74.40 -10.17 4.60 w2 -74.78 -10.35 3.37 w3 -42.73 -6.10 -2.19 tab. 10. 34s and 13c isotopes in soil and rock samples sample δ 34s (‰ vs. v-cdt) δ 13c (‰ vs. vpdb) r1 <d.l. -27.73 r2 -1.48 -25.07 r3 4.48 -4.25 s1 1.37 -25.53 s2 -0.62 -24.54 s3 4.15 -24.65 environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 111 discussion the study confirmed the significant effect of the mining activity on the observed area. this phenomenon is conditioned by the geological structure of the area, including the mining activity, which facilitated the weathering of rocks and the subsequent release of hazardous elements into the environment. the soils of the tailing´s dam show very acidic reactions, while the soil from the pond surface has a neutral reaction, which is probably due to application of a limestone layer during reclamation. low ph is a cause of the depletion of ca and mg in soil samples s1 and s2. mobility of the metals is affected by many aspects. factors that can affect the activity of metals and other elements are, e.g., the ph and eh of the soil, moisture, temperature, soil type, the content of metals in the soil and their binding, the content of humus components in soil, the presence of clay minerals, cation exchange capacity, or the presence of ca and mg (čurlík et al., 2015). tipping et al. (2003) assumed that hno3 extraction is one of the methods providing the best measures of geochemical active metals. the high ca and mg contents are typical for alkaline soil reactions, and, on the contrary, the absence of these cations usually indicates acid soil reactions. the ca/ mg ratio in the sorption complex significantly affects the rate of the entering of heavy metals into plants. in soil samples s1 and s2, the ca/mg ratio is <1 (tab. 3), which is characteristic for the presence of acidic rocks (mccarten, 1992), whereas s3 sample shows ca/mg = 2.24 caused by landfill of caco3. extraction of the ca from the soils was higher than the extraction of mg. calcium is usually characterized by a higher level of availability than magnesium (čurlík et al., 2015). despite the neutral ph, soil fig. 4. overlap of wml – world meteoric water line and lml – local meteoric water line, with values measured for water samples er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 112 sample s3 showed the highest percentage of availability for all analysed metals, except fe. the ph dependence on the level of the extractability of metals from soils can be seen when comparing samples s1 and s2, where the metals of s1 with ph 2.82 were more accessible than the metals in sample s2 with ph 3.25. all investigated soils exceeded pb, zn, cu, and cr content limits for agricultural land under the act no. 220 coll. on the conservation … (2004). although the drainage waters percolating the tailing pond sedem žien have different appearances, they have similar composition and comparable ph. differences were observed in concentrations of fe, zn, and pb and sulphates. the most significant difference between water samples is higher fe concentration in sample w1 in comparison to sample w2, causing the red coloration. the drainage water from šobov quarry (w3) has a low ph value (2.17) and high al, fe, and sulphates content. iron concentrations in water samples w1 (9.63 mg.l-1) and w3 (311 mg.l-1) highly exceed the limit concentrations of fe (4.0 mg.l-1) for water resulting from the extraction of ores by government regulation 269/2010 coll., which stipulates criteria for achieving good water balance (tab. 3). water sample w2 is characterised by high pb content (> 0.5 mg.l-1, which is current slovak limit concentration for waters resulting from the extraction of the ores government regulation 269/2010 coll.). differences in ph values between water percolating the tailing and quarry may be caused by more rapid weathering of disulphides than monosulphides, which could explain a slower oxidation of galena – pbs and sphalerite – zns in the tailing pond in comparison to pyrite – fes2 dispersed in the quartz. another reason for the differences in ph may be the presence of acidophilic sulphide-oxidizing bacteria (acidithiobacillus ferrooxidans (temple & colmer) kelly & wood), which was previously found in the drainage water from šobov quarry (šlauková, bella, 2006; bella et al., 2010). the limiting factor for the oxidation is the transformation of fe2+ to fe3+, which is very slow in abiotic conditions with ph above 5 (šottník, 2005). interpretation of the geochemistry of stable isotopes (d, o, c, n, s) is complicated because of the fact that the minerals created by different processes may have the same isotopic composition and minerals created by same process; however, under different conditions, they may have significantly different isotopic compositions. therefore, waters which percolate rocks and soils affected by the composition of ore minerals reflect all of the mentioned aspects. variations in the isotopic composition of the natural material are the result of isotope exchange reactions and kinetic isotope effects (hladíková, 1988; rollinson, 1988). sulphur in natural materials can have various sources, and its isotopic composition may reflect different processes that material overcame. in the tailing pond sedem žien, the main sources of sulphur are hydrothermal sulphides of mined polymetallic (pb-zn) ore and dispersed pyrite from the nearby šobov quarry. environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 113 it is known that δ34s values in the surrounding of banská štiavnica are relatively homogeneous, and close to zero. such values indicate the sulphur origin from sedimentary rocks (sakai et al., 1982; chaussidon et al., 1989). the negative δ 34s value in sample w3 (-2.19) is typical for the sulphur originating from the sulphides (newman et al., 1991). the isotopic composition of sulphur in the solid samples (δ34s -1.48 to 4.48‰) mainly reflects the isotopic balance of the sulphides of the main pb-zn hydrothermal mineralization in the ore field of banská štiavnica as well as the isotopic characteristics of sulphur in pyrite dispersed in the hydroquartsite mined in the vicinity of the tailing pond. the homogeneity of the composition indicates a magmatic origin of ores and a meteoric origin of the waters (kantor, 1979; burian et al., 1985). δ34s values of drainage waters of the tailings pond correspond to the range of values for sulphur originating in central europe from atmospheric precipitation (+3‰ to 5‰; newman et al., 1991). regarding the oxidation of sulphides, many studies on the effects of the isotopic composition have been done. the results of studies of the effects of oxidation in the presence of phototrophic bacteria diverge, but the oxidation in the presence of chemotrophic bacteria shows clearer results. kaplan-rittenberg (1964) demonstrated the effect of oxidation in the presence of bacteria thiobacillus concretivorus, as did kelly & harrison, when changes reached the range of -18‰ (šgúdš, 2010). isotopes 18o compose, in comparison with 16o, just a small part of oxygen, and their ratio helps to provide information about origins of the waters (hoefs, 1987). the dependence between δ18o and δd ‰ values in the water indicates formation in the conditions of isotopic equilibrium. the isotopic composition of the δd values in the drainage water (δd 42.73 to -74.78‰) and of δ18o (-6.11 to -10.17‰) corresponds, according to taylor (1974), sheppard (1981), and rollinson (1988), to meteoric water. conclusion the studied area is contaminated by heavy metals (mainly by fe, pb, zn, and cu) and acidified. from the mineralogical point of view, the rock material is rich in quartz and sulphide minerals, which has an important influence on the chemical composition of soils and waters. the isotope study proved the meteoric origin of the waters, and sulphide origin of the sulphur in the waters. references act no. 220 coll. on the conservation and use of agricultural land (2004). zákon č. 220/2004 z. z. o ochrane a využívaní poľnohospodárskej pôdy a o zmene zákona č. 245/2003 z. z. o integrovanej prevencii a kontrole znečisťovania životného prostredia a o zmene a doplnení niektorých zákonov. 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(1974). the application of oxygen and hydrogen isotope studies to problems of hydrothermal alteration and ore deposition. economic geology, 843–883. doi: 10.2113/gsecongeo.69.6.843 thornton, i. (1996). impacts of mining on the environment; some local. regional and global issues. applied geochemistry, 11, 355–361. doi: 10.1016/0883-2927(95)00064-x tipping, e., rieuwerts, j., pan, g., ashmore, m.r., lofts, s., hill, m.t.r., farago m., thornton, i. (2003). the solid-solution partitioning of heavy metals (cu, zn, cd, pb) in upland soils of england and wales. environmental pollution, 125(2), 213–225. doi: 10.1016/s0269-7491(03)00058-7 younger, p.l., wolkersdorfer, ch. (2004). mining impacts on the fresh water environment: technical and managerial guidelines for catchment scale management. mine water and the environment, 23(1), 2–80. doi: 10.1007/s10230-004-0028-0 er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 116 appendix 1 fig. i. scanning electron micrographs showing mineral phases of the sample r1; a: qtz – quartz (sio2) and ant – anatase (tio2) phases with finely dispersed gn – galena (pbs); b: crystals of anatase; c: sp – sphalerite (zns) surrounded by quartz; d: other formations of anatase environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 117 fig. ii. scanning electron micrographs of sample r2; a, d: qtz – quartz matrix with dispersed toz – topaz and anh – anhydride needles; b, c : inclusions of py – pyrite in the quartz, ant – anatase and anhydride with si and al impurities; e, f: penetration of the quartz, topaz, anatase and anhydride er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 118 fig. iii. scanning electron micrographs of sample r3; a: py – pyrite and ccp – chalcopyrite inclusions surrounded by gn – galena; b: chalcopyrite, galena and sp – sphalerite; c: galena, sphalerite and chalcopyrite inclusions among clc – clinochlore and dol – dolomite; d: small inclusions of galena and chalcopyrite in the in the sphalerite matrix; e: sphalerite and galena surrounded by qtz – quartz and intersected by dolomite veins. environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 119 fig. iv. scanning electron micrographs of the sample s1; bt – biotite; qtz – quartz; ms – muscovite; jrs – jarosite; cerium phosphate fig. v. scanning electron micrographs of the sample s2; a: brt – barite, qtz – quartz, jrs – jarosite, ill – illite; b: , qtz – quartz, jrs – jarosite, ill – illite; c: jrs – jarosite, or – orthoclace, clc – clinochlore er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 120 fig. vi. scanning electron micrographs of the sample s3; qtz – quartz, ms – muscovite, jrs+pb – jarosite with pb impurities, ilm – ilmenite, or-orthoclase environm ental characteristics of the m ining area affected by sulphide m inerals and acidification (b anská š tiavnica, s lovakia) 121 abstract the area of sedem žien tailing pond and the nearby šobov hydroquartzite quarry affected by mining activity were investigated by geochemical and mineralogical methods to determine the contaminating chemical compounds and study their availability. degradation of the hydrothermal base mineralisation (galena, sphalerite, pyrite, pyrrhotite, and chalcopyrite) and of fine-grained pyrite oxidation, which forms impregnations in hydroquartzite, produce acid mine drainage (amd). the area is acidified and the components (soil, rock, water) are contaminated mainly by pb, zn, and fe. the tailing pond dam forming soils have an acidic ph of 2.28–3.25, whereas the soil on the tailing pond surface is close neutral (ph 7.26). the leaching availability of the metals from the soil is up to 75%. the amd from the hydroquartzite quarry is, in comparison with those percolating the tailing pond sediments, very acidic (ph 2.71) and contains high concentration of metals (fe 311 mg.l-1, zn 1690 µg.l-1, cu 890 µg.l-1, pb 126 µg.l-1). key words: heavy metals, contamination, acidification, availability received: [2018.07.11] accepted: [2018.10.25] charakterystyka środowiskowa obszaru górniczego skażonego minerałami siarczkowymi i zakwaszającymi (banská štiavnica, słowacja) streszczenie za pomocą metod geochemicznych i mineralogicznych, zbadano obszar osadnika sedem žien i pobliskiego kamieniołomu hydro-kwarcytu šobov, dotkniętych działalnością górniczą, w celu określenia związków chemicznych zanieczyszczających te tereny i zbadania ich dostępności. degradacja hydrotermalna oparta na mineralizacji (galena, sfaleryt, piryt, pirotyn i chalkopiryt) i utlenianiu drobnoziarnistego pirytu, której formy impregnują hydro-kwarcyt, produkuje kwaśne odcieki z  kopalń (amd). obszar jest zakwaszony, a  składniki lokalne (gleba, skała, woda) są zanieczyszczone, głównie przez pb, zn i  fe. zanieczyszczenia zapory osadnika tworzą gleby kwaśne (ph 2,28 3,25), natomiast zanieczyszczona gleba na powierzchni osadnika jest zbliżona do odczynu obojętnego (ph 7,26). dostępność ługowania metali z  gleby wynosi tu aż do 75%. amd z kamieniołomu hydro-kwarcytu, jest porównywalna z tymi przenikającymi, bardzo kwaśnymi (ph 2,71), zanieczyszczeniami sedymentacyjnymi osadnika i zawiera wysokie stężenia metali (fe 311 mg.l-1, zn 1690 µg.l-1, cu 890 µg.l-1, pb 126 µg.l-1). słowa kluczowe: metale ciężkie, zanieczyszczenia, zakwaszenie, dostępność information on the authors erika remešicová she studied environmental engineering during her master studies, and the field of her doctoral studies was environmental protection in the industry. during phd. studies, she dedicated her research to mining landscape in slovakia and to the possibilities of the decontamination of the acidic mine waters. currently, she works at water research institute in slovakia at the department of groundwater assessment. peter andráš https://orcid.org/0000-0002-5366-4041 he is a professor at the department of environment (matej bel university, banská bystrica). he is a geochemist, mineralogist, and environmentalist. he has worked for a long time as a research worker in the field of ore mineralogy, mineral deposits (mineralogy, isotope study, study of the ore origin etc.), and the impact of the mining activities on the environment. he gives lectures at several home universities and also abroad. he was a principal investigator in 7 grants and/or a member of the research team in 24 grant projects. he is an author of 11 monographs, 13 chapters in monographs, numerous current contents articles, and articles registered in wos and scopus databases. er ik a r em eš ic ov á, p et er a nd rá š, r ad m ila k uč er ov á 122 radmila kučerová she is the associate professor at the department of environmental engineering (faculty of mining and geology, všb – technical university of ostrava). she is mainly concerned with biotechnologies. she is the author of 3 monographs, articles registered in wos and scopus databases, and numerous articles presented at domestic and foreign conferences. she was a principal investigator in 3 grants and a member of the research team in 19 grant projects. 90 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 90–97, 2018, issn 2543-8832 doi: 10.24917/25438832.3supp.12 francesco vizzarri1*, marisa palazzo m1, donato casamassima1, carlo corino2, sara chiapparini2, lubomir ondruska3, nikola knizatova4, martin massanyi4, filip tirpak4, peter massanyi4 1department of agricultural and environmental science, university of bari aldo moro, bari, italy, *francesco.vizzarri@uniba.it 2department of health, animal science and food safety, university of milano, milano, italy 3national agricultural and food centre, nitra, slovak republic 4department of animal physiology, slovak university of agriculture, nitra, slovak republic dietary supplementation with algae and polyphenols in male rabbits: effects on semen quality traits introduction in animal production, poor reproductive performance currently a�ects livestock productivity. several e�orts toward overcoming this challenge (poor reproductive performance) have culminated in identifying oxidative stress as the main reason because animals’ productivity is impaired either directly or indirectly by di�erent unfavourable conditions (rahal et al., 2014). oxidative stress has been widely reported as the mechanism behind many pathological development and disease conditions, including reproductive ine�ciency (sikiru et al., 2018). in rabbit production, this aspect is a very important parameter for reproductive performance evaluation, because it determines pro�tability as well as products yield, and it is one of the major factor determining enterprise productivity and production objectives. in male organisms, pro-oxidant condition a�ects both seminal qualities and reproductive functions of the spermatozoa. high levels of reactive oxygen substances are also reported to induce oxidative damage of dna in the sperm plasma membrane, mitochondrion, and nuclear genome. �e danger in this damage is not associated with poor reproductive performance alone, but also with cancer development and inherited infertility in the o�-springs (aitken, krausz, 2001). in recent years, many studies have been focused on natural substances that can a�ect the health of animals and challenge the new animal welfare prospective. �e present research is based on the claim that exogenous supplementation of antioxidants is a proven tool of reducing oxidative stress associated with reproduc91 d ietary supplem entation w ith algae and polyphenols in m ale rabbits: effects on sem en quality traits tive performance and the need to discover di�erent sources of antioxidants capable of improving reproductive activities. a mix of di�erent extracts was used as a dietary supplement, and it consists exclusively of natural products. its main components are polyphenols from terrestrial and marine origins and plant polysaccharides. �e e�ect of this supplement on reproduction has not been reviewed in the past, and that is a reason why we decided to test its e�ect on the reproductive potential of male rabbits. �e aim of the present study is to determine e�ects of natural mix extracts during a 120-day in vivo experiment on selected reproductive traits of male rabbits. material and methods animals and experimental design �e trial lasted 120 days and was conducted in the animal production research centre in nitra, slovak republic, on 14 adult new zealand white rabbit bucks. all experimental procedures and the management of animals were conducted in accordance with european community guidelines n. 86/609/eec regarding the protection of animals for experimental purposes. �e tested animals (aged 15 ± 3 months) were divided into three homogeneous groups, and the body weights were recorded at the beginning and the end of experiment: the control (con; n = 5) was fed with commercial feed, the �rst experimental group (t1; n = 4) received a 0.3% feed additive mix, and the second experimental group (t2; n = 5) received a 0.6% feed additive mix. �e mix extract supplement, containing mainly polyphenols from algae and chestnut tannin extracts, was analysed using hplc-dad according to russo et al. (2017) and the following natural compounds were determined as the most prominent: neochlorogenic acid, elaigic acid, syringic acid, cynaroside, and rutin. �e natural extract was produced and provided by lombarda trading srl (casale belvedere, cremona, italy). �e ingredients and chemical composition of diets are reported in table 1. �e semen samples were collected on day 0 (basal), and days 30, 60, 90, and 120 of the feeding period with the help of an arti�cial vagina. �e obtained semen samples were diluted with physiological solution in the ratio 1:5. a�er processing, the samples were incubated at the temperature of 37°c and were analysed immediately in triplicate. each of prepared samples was evaluated using a computer assisted semen analyzer (casa) system – sperm vision (minitub, tiefenbach, germany) equipped with a microscope (olympus bx 51, japan) to assess the spermatozoa motility (massanyi et al., 2008). each sample was placed into makler counting chamber (depth 10 μm, se�–medical instruments, germany). using the rabbit speci�c set up, the following parameters were evaluated: spermatozoa concentration (conc, 106/ml), total motile spermatozoa (%, motility > 5 μm/s), and progressive motile spermatozoa (%, motility > 20 μm/s). fr an ce sc o v iz za rr i, m ar is a p al az zo m , d on at o c as am as si m a, c ar lo c or in o, s ar a c hi ap pa rin i, lu bo m ir o nd ru sk a, n ik ol a k ni za to va , m ar tin m as sa ny i, fi lip t irp ak , p et er m as sa ny i 92 tab. 1. ingredients and chemical composition of the diets [g/kg] ingredients experimental diet con t1 t2 maize 282 281 280 alfalfa hay 305 305 305 sun�ower meal 135 135 135 palm seed oil 8 8 8 soybean oil 7 7 7 wheat 80 80 80 cane molasses 20 20 20 carob bean meal 90 90 90 oat 53 53 53 calcium carbonate 7 7 7 sodium chloride 3 3 3 dicalcium phosphate 2 2 2 methionine (99%) 2.5 2.5 2.5 lysine (78.5%) 1.6 1.6 1.6 choline (75%) 1.4 1.4 1.4 vitamin and mineral premix* 2.5 2.5 2.5 experimental supplement** 0.00 3 6 chemical composition*** crude protein 184.0 183.6 183.5 ether extract 35.7 35.5 35.5 crude �bre 187.0 186.8 187.0 ash 86.0 85.7 85.8 nitrogen free extract 507.0 507.1 506.9 ndf 302.1 301.5 301.7 adf 195.8 195.4 195.3 adl 39.9 39.5 39.5 notes: *supplied per kg diet: 13.500 i.u. vitamin a (trans-retinyl acetate); 800 i.u. vitamin d3 (cholecalciferol); 35 mg vitamin e (α-tocopherol min 91%), 35 mg copper (cupric sulphate pentahydrate), 150 mg aminoside sulphate; ** quantities of plant extract, t1 – experimental group fed 0.3% of natural mix supplement and t2 – experimental group fed 0.6% of natural mix supplement; *** analyses determined in triplicate �e superoxide dismutase levels (sod), which catalyses the dismutation of superoxide radical reaction in hydrogen peroxide and molecular oxygen, together with glutathione peroxidase (gpx) determination, were determined using a commercial colorimetric kit-assay provided by randox (randox laboratories ltd., united kingdom). sod activity was expressed in units per milligram of protein [u/mg], and gpx activity was expressed in units per gram of protein [u/g]. �e ferric reducing antioxidant power (frap) test, developed by benzie and strain (1996), measures the antioxidant capacity of plasma and is used to assess the ability to reduce the ferric iron complex in an acidic environment. one unit frap is expressed in mmol/ml and d ietary supplem entation w ith algae and polyphenols in m ale rabbits: effects on sem en quality traits 93 indicates the number of moles of ferric ion (fe3+) reduced to ferrous ion (fe2+) from one mol of tested antioxidants. statistical analysis obtained data was statistically analysed with the help of the pc program excel and a  commercially available statistics package sas 8.0 (sas institute inc., usa) using student’s t-test and sche�e’s test. statistical signi�cance was indicated by p values of less than 0.05, 0.01, and 0.001. tab. 2. semen characteristics (x ± sd) of control (con, n = 5) and experimental (t1, n = 4; t2, n = 5) buck rabbits items dietary treatment1 con t1 t2 p-value2 concentration [10 6 × ml-1] basal 0.656±0.270 0.695±0.428 0.728±0.532 n.s. 30d 0.638±0.280 0.635±0.602 0.607±0.490 n.s. 60d 0.647±0.1754 0.699±0.345 0.871±0.387 n.s. 90d 0.640±0.245 0.538±0.307 0.586±0.461 n.s. 120d 0.663±0.251 0.685±0.123 0.609±0.213 n.s. motility [%] basal 79.640±2.240 81.630±2.760 82.959±4.040 n.s. 30d 84.760±2.694 85.840±9.272 77.700±12.740 n.s. 60d 85.090±7.526 87.210±8.245 89.380±8.023 n.s. 90d 82.840±4.511 87.630±4.567 77.330±12.710 n.s. 120d 92.100±4.209 88.670±5.257 85.680±7.959 n.s. progressive motility [%] basal 66.440±3.130 66.530±13.690 62.490±12.940 n.s. 30d 74.510±4.948 69.940±18.980 62.420±14.260 n.s. 60d 74.280±12.600 79.070±13.890 81.280±11.370 n.s. 90d 70.890±4.401 77.160±4.913 64.390±18.440 n.s. 120d 84.610±6.610 82.040±6.592 76.020±14.470 n.s. note: 1(con) – control group fed with commercial feed; t1 – experimental group fed 0.3% of natural mix supplement; t2 – experimental group fed 0.6% of natural mix supplement; 2p – value: n.s. = not signi�cant results and discussion �e dietary supplementation with the natural extracts mix did not cause any changes in the animal body weights, and it did not induce any evident clinical signs in rabbits over the 120-days of the experimental period. �e concentration of spermatozoa was not signi�cantly di�erent between experimental groups and the control group a�er 4 months of dietary treatment (tab. 1). mourvaki et al. (2010) also found no e�ect with use of �axseed dietary supplementation on the volume and spermatozoa concenfr an ce sc o v iz za rr i, m ar is a p al az zo m , d on at o c as am as si m a, c ar lo c or in o, s ar a c hi ap pa rin i, lu bo m ir o nd ru sk a, n ik ol a k ni za to va , m ar tin m as sa ny i, fi lip t irp ak , p et er m as sa ny i 94 tration in rabbit. however, okab et al. (2013), feeding rabbit bucks with dried seaweed (2%), showed a signi�cant decrease in spermatozoa concentration, the percentage of live spermatozoa, and ejaculate volume. �e spermatozoa motility parameters (motility and progressive motility; tab. 1) were not signi�cantly di�erent between the control group and experimental group with natural extracts mix supplementation. controversially, yousef et al. (2003) observed an improvement of spermatozoa motility parameters a�er the dietary supplementation of ascorbic acid and vitamin e, alone and in combination, in male rabbits. since a lack of e�ects has been observed in our experimental study, further research is needed in order to test di�erent doses of natural extracts mix supplements. tab. 3. antioxidant seminal plasma markers (x ± sd) of control (con, n = 5) and experimental (t1, n = 4; t2, n = 5) buck rabbits items dietary treatment1 con t1 t2 p–value2 sod [u × mg-1 tp] basal 0.408±0.085 0.502±0.117 0.477±0.091 n.s. 30d 0.422±0.097 0.664±0.575 0.353±0.059 n.s. 60d 0.296±0.194 0.443±0.268 0.346±0.150 n.s. 90d 0.381±0.123 0.296±0.102 0.421±0.088 n.s. 120d 0.264±0.114 0.429±0.240 0.311±0.094 n.s. gpx [u × g-1 tp] basal 42.517±10.225 40.120±0.311 39.370±2.221 n.s. 30d 44.630±15.370 36.330±0.476 30.370±9.601 n.s. 60d 23.280±12.330 41.760±28.010 35.790±16.640 n.s. 90d 32.960±12.300 32.750±14.000 44.420±15.620 n.s. 120d 26.600±11.2501 65.580±19.3102 30.530±4.5251 ** frap [µmol fe2+ × g-1 tp] basal 85.125±29.119 93.225±17.455 89.541±7.853 n.s. 30d 97.420±48.960 112.800±18.400 60.600±12.100 n.s. 60d 75.590±54.340 106.300±64.980 69.750±26.350 n.s. 90d 59.240±15.330 73.330±15.540 68.020±28.520 n.s. 120d 69.440±18.5101 103.000±36.1002 67.110±7.5271 * note: 1(con) – control group fed with commercial feed; t1 – experimental group fed 0.3% of natural mix supplement; t2 – experimental group fed 0.6% of natural mix supplement; 2 p–value: n.s. = not signi�cant; * (p < 0.05); ** (p < 0.01) tables 2–3 contain data regarding the tested oxidative markers in seminal plasma. at the end of the dietary treatment (a�er 120 days), all three parameters (sod, gpx and frap) were positively altered, although the statistical signi�cance was reached only for gpx (p < 0.01) and frap (p < 0.05) values. in fact, group t1 data showed the highest content of the two parameters when compared with group con. in literature, it has also been reported that sod activity survey in seminal plasma could be a useful tool d ietary supplem entation w ith algae and polyphenols in m ale rabbits: effects on sem en quality traits 95 for determining sperm fertilization potential and could improve the diagnosis of male infertility (shiva et al., 2011). in general, antioxidant dietary supplementation leads to an improvement of the antioxidant markers pro�le in the seminal plasma, in particular, when algae-based feed additive is supplemented in the diet (murphy et al., 2017). conclusion since oxidative stress is considered a biochemical process negatively a�ecting reproduction, the reduction of its e�ects is highly important for the promotion of animal welfare. �e use of polyphenols and tannins in rabbit diets is a source of natural antioxidants, and we can conclude that supplementation of 0.3% of natural mix did not signi�cantly negatively a�ect any of the studied reproductive parameters of male rabbits, but we have found some improvement in several antioxidant parameters. acknowledgment �is work was funded by the slovak research and development agency grants no. vega 1/0539/18, vega 1/0760/15, apvv-16-0289 and apvv-15-0544. references aitken, r.j., krausz, c. (2001). oxidative stress, dna damage and the y chromosome. reproduction, 122, 497–506. benzie, f.f., strain, j.j. (1996). �e ferric reducing ability of plasma (frap) as a measure of “antioxidant power”: the frap assay. analytical biochemestry, 239, 70–76. doi: 10.1006/abio.1996.0292 massanyi, p., chrenek, p., lukac, n., makarevich, a.v., ostro, a., zivcak j., bulla, j. (2008). comparison of di�erent evaluation chambers for analysis of rabbit spermatozoa motility parameters using casa system. slovak journal of animal science, 41(2), 60–66. mourvaki, e., cardinali, r., dal bosco, a., corazzi, l., castellini, c. (2010). e�ects of �axseed dietary supplementation on sperm qualità and on lipid composition of sperm subfractions and prostatic granules in rabbit. science direct, 73, 629–637. doi: 10.1016/j.theriogenology.2009.10.019 murphy, e.m., stanton, c., brien, c.o., murphy, c., holden, s., murphy, r.p., varley, p., boland, m.p., fair, s. (2017). �e e�ect of dietary supplementation of algae rich in docosahexaenoic acid on boar fertility. �eriogenology, 90, 78–87. doi: 10.1016/j.theriogenology.2016.11.008 okab, a.b, samara, e.m., abdoun, k.a., rafay, j., ondruska, l., parkanyi, v., pivko, j., ayoub, m.a., alhaidary, a.a., aljumaah, r.s., massanyi, p., lukac, n. (2013). e�ects of dietary seaweed (ulva lactuca) supplementation on the reproductive performance of buck and doe rabbits. journal of applied animal research, 41(3), 347–355. doi: 10.1080/09712119.2013.783479 rahal, a., kumar, a., singh, v., yadav, b., tiwari, r., chakraborty, s., dhama, k. (2014). oxidative stress, prooxidants, and antioxidants: �e interplay. biomed research international, id 761264. doi: 10.1155/2014/761264 russo, r., pucci, l., giorgetti, l., árvay, j., vizzarri, f., longo, v., pozzo, l. (2017). polyphenolic characterisation of plant mixture (lisosan® reduction) and its hypocholesterolaemic e�ect in high fat diet-fed mice. natural product research doi: 10.1080/14786419.2017.1402328 sas user’s guide: statistics. version 8 edition. cary (2003). nc: sas institute. shiva, m., gautam, a.k., verma, y., shivgotra, v., doshi h., kumar, s. (2011). association between fr an ce sc o v iz za rr i, m ar is a p al az zo m , d on at o c as am as si m a, c ar lo c or in o, s ar a c hi ap pa rin i, lu bo m ir o nd ru sk a, n ik ol a k ni za to va , m ar tin m as sa ny i, fi lip t irp ak , p et er m as sa ny i 96 sperm quality, oxidative stress, and seminal antioxidant activity. clinical biochemistry, 44, 319–324. doi: 10.1016/j.clinbiochem.2010.11.009 sikiru, a.b., alemede, i.c., egena, s.s.a., ijaiya, a.t. (2018). oxidative stress and reproductive ine�ciencies: �e science, evidences, and solutions. agricultural extension journal, 2(1), 17–26. yousef, m.i., abdallah, g.a., kamel, k.i. (2003). e�ect of ascorbic acid and vitamin e supplementation on semen quality and biochemical parameters of male rabbits. animal reproduction science, 76, 99–111. doi: 10.1016/s0378-4320(02)00226-9 abstract in recent years, many studies have been focused on natural substances that can a�ect the health of animals. a mix of di�erent extracts was used as a dietary supplement, and it consisted exclusively of natural products. its main components were polyphenols from terrestrial and marine origins and plant polysaccharides. �e e�ect of this supplement on reproduction has not been reviewed in the past, which is why its e�ect on the reproduction potential of male rabbits was tested. �e aim of the present study is to determine the e�ects of the natural mix during a 120-day in vivo experiment on selected reproductive traits of male rabbits. natural mix was supplemented in two di�erent concentrations (t1 – 0.3% and t2 – 0.6%) with the basal ingredients of the conventional rabbit feed in pellet form. in our experiments, emphasis was placed on both the spermatozoa concentration and its motility parameters as well as on the properties of seminal plasma and antioxidant activity. �e dietary supplementation with the natural extracts mix positively altered the quality traits of rabbit spermatozoa, but these changes were statistically not signi�cant. in experimental group t1, a signi�cant increase of gpx and frap content, both regarding the antioxidant markers pro�le in seminal plasma, was recorded. we can conclude that the supplementation of 0.3% of natural mix did not signi�cantly negatively a�ect any of the studied reproductive parameters of male rabbits, but some improvement in several antioxidant parameters was found. key words: extract, rabbit, spermatozoa, mobility, seminal plasma, antioxidants received: [2018.06.01] accepted: [2018.11.22] suplementacja diety algami i polifenolami u samca królika: wpływ na cechy jakościowe nasienia streszczenie w ostatnich latach wiele badań dotyczy substancji naturalnych, które mogą wpływać na zdrowie zwierząt. jako suplement diety wykorzystano mieszankę różnych ekstraktów, składającą się wyłącznie z naturalnych produktów. jej głównymi składnikami były polifenole pochodzenia lądowego i morskiego oraz polisacharydy roślinne. oddziaływanie tych suplementów na reprodukcję nie zostało w przeszłości poddane analizie, co było powodem dla którego zbadano ich wpływ na potencjał reprodukcyjny samców królików. celem niniejszego badania było określenie wpływu naturalnej mieszanki podawanej podczas 120-dniowego eksperymentu in vivo na wybrane cechy reprodukcyjne samców królika. naturalną mieszankę w dwóch różnych stężeniach (t1 – 0,3% i t2 – 0,6%) uzupełniono podstawowymi składnikami tradycyjnego pokarmu dla królików w postaci śrutu. w doświadczeniach nacisk położono, zarówno na koncentrację plemników, jak i na ich parametry ruchowe, a także na właściwości plazmy nasiennej oraz aktywność przeciwutleniającą. suplementacja diety mieszaniną naturalnych ekstraktów pozytywnie zmieniła cechy jakościowe plemników królika, ale zmiany te nie były statystycznie istotne. w grupie doświadczalnej t1 odnotowano istotny wzrost zawartości, zarówno gpx, jak i frap, pod względem pro�lu markerów antyoksydacyjnych w plazmie nasienia. można zatem stwierdzić, że suplementacja 0,3% naturalnej mieszanki nie wpłynęła znacząco negatywnie na żaden z badanych parametrów reprodukcyjnych samców królików, a nawet odnotowano tu pewną poprawę kilku parametrów antyoksydacyjnych. słowa kluczowe: ekstrakt, królik, plemniki, ruchliwość, plazma nasienna, przeciwutleniacze d ietary supplem entation w ith algae and polyphenols in m ale rabbits: effects on sem en quality traits 97 information on the authors francesco vizzarri http://orcid.org/0000-0003-1316-7937 fellow researcher at the university of bari aldo moro, researching animal welfare and the quality of animial production. marisa palazzo http://orcid.org/0000-0002-9116-9420 technician at the university of molise, researching animal welfare and the quality of animal production. donato casamassima http://orcid.org/0000-0002-3079-4249 full professor at the university of molise, researching animal welfare and animal production. carlo corino http://orcid.org/0000-0001-8146-0583 full professor at the university of milano, researching animal nutrition. sara chiapparini http://orcid.org/0000-0002-6484-3352 phd student at the university of milano, researching animal nutrition. lubomir ondruska http://orcid.org/0000-0002-8460-3137 director of institute of small farm animals, researching breeding, genetics, nutrition, controlled reproduction, technology, biotechnical methods, and their innovations in rabbit husbandry. nikola knizatova phd student at the university of agriculture in nitra, researching animal physiology. martin massanyi phd student at the university of agriculture in nitra, researching animal physiology. filip tirpak http://orcid.org/0000-0001-9293-9055 phd student at the university of agriculture in nitra, researching animal physiology. peter massanyi http://orcid.org/0000-0002-4216-0948 full professor at the slovak university of agriculture in nitra, researching animal physiology. 161 annales universitatis paedagogicae cracoviensis studia naturae, 4: 161–169, 2019, issn 2543-8832 doi: 10.24917/25438832.4.10 anna kocoń1*, sylwia janiczek1, natalia malejky-kłusek2 1department of zoology, institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland, *a_kocon@wp.pl 2department of ecology and environmental protection, institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland methods of protection against ticks (acari: ixodida) ticks are ruthless, dangerous ectoparasites that attack a large host circle. �ey are in the second place, a�er mosquitoes, in the rank of dangerous blood-drinking arthropods. �e increased pace of urbanization and weather variability in recent decades have created new, convenient conditions for ticks to be found among people and pets. it is particularly dangerous as ticks carry numerous pathogens and spread tick-borne diseases, such as: lyme borreliosis, tick-borne encephalitis (fig. 1a–b), babesiosis, granulocytic anaplasmosis. depending on the morphological structure, ticks can be divided into proper (hard) (ixodidae) and argasidae (so� ticks) (argasidae). of the 19 species of ticks permanently found in the polish fauna, ixodes ricinus l. is – the common tick (siuda, 1991) is the most common of the hard ticks (appendix 1a). it occurs in green areas – not only along forest paths, in glades, and forest roads, but also increasingly in home gardens, playgrounds, and recreational places. in addition to the fact that ticks can be found in nature, during trips, picnics, walks, we also cannot feel safe in farm buildings and our own homes, due to the possible occurrence of a so� tick, argas re�exus fabr., attacking mainly nocturnal rock pigeons in lo�s, attics of buildings, on church towers (appendix 1b). in terms of location of their occurrence, we can divide ticks into three ecological groups (siuda, 1991) (tab. 1): nest-burrow ticks – occurring in lo�s, attics, caves, basements, animal cavities and burrows, in the nests of mammals and birds, and in their hollows on trees or in rock shelters. �ese ticks are usually host-speci�c species, they spend their entire lives in hideouts, where they can feed on their host who lives, sleeps, breeds and hibernates there. in hidden habitats, there are small �uctuations in climatic conditions; non-nest ticks – found in litter, near bird nests built on trees, among vegetation. �ey occupy more open environments, feed on busy, migrating a nn a k oc oń , s yl w ia j an ic ze k, n at al ia m al ej ky -k łu se k 162 hosts, and are not attached to strictly located whereabouts and breeding of hosts. �e survival of a hungry tick in such conditions depends mainly on its tolerance to changing environmental conditions, such as temperature, light, humidity; non-nest-burrow ticks – larvae and nymphs occur at the entrance and inside the burrows of mammals, while adults are non-nest parasites. �e di�erences in protection against tick attacks are primarily related to our whereabouts, and thus the possibility of encountering this dangerous parasite on its way. �ere are several options for protection against tick attacks, but there is no 100% e�ective way to avoid these parasites. fig. 1. the number of cases of selected tick-borne diseases in poland; a – lyme borreliosis, b – tickborne viral encephalitis; *data for day 30.09.2019 (source: http://wwwold.pzh.gov.pl/oldpage/epimeld/index_p.html) m ethods of protection against ticks (a cari: ixodida) 163 tab. 1. ecological groups of ticks found in poland ecological group tick species nest-burrow argas polonicus siuda, hoogstraal, cli�ord et wassef a. re�exus fabricius carios vespertilionis latreille ixodes apronophorus latreille i. arboricola schulze & schlottke i. caledonicus nuttall i. crenulatus koch i. hexagonus leach i. lividus koch i. rugicollis schulze & schlottke i. simplex neumann i. vespertilionis koch non-nest haemaphysalis concinna koch h. punctata canestrini & fanzago ixodes frontalis panzer i. persulcatus schulze i. ricinus linneus i. trianguliceps birula dermacentor reticulatus fabricius – adults form non-nest-burrow d. reticulatus – larvae and nymphs ticks on the host’s body are looking for a suitable place, most o�en they are parts of the body with delicate skin, sweating: under the knees, groin, navel, armpits, at the base of the hair/neck, shoulders or behind the ears. in the case of pets, foraging of ticks is most o�en found around the neck, head, less o�en on the back and around the tail/ anus (siuda, 1991). to defend against tick attacks, the following simple guidelines should be observed: avoid places typical for ticks (forest edges, forest paths, mid-forest clearings, animal trails covered with tall grass and bushes), thus avoid resting on the grass or in places covered with tall grass, lush vegetation, bushes; put on appropriate clothing, suitable for going out into the �eld, which will prevent ticks from moving onto the naked body: long trousers, with socks rolled out, high shoes, a long-sleeved sweatshirt inserted into trousers, headgear, neck coverage; use tick repellents in the form of sprays, creams, impregnation of things that are exposed to tick attacks, e.g. clothing, tents, blankets, footwear; parasite repellents should also be used for pets/farm animals; check the body for any ticks on clothing or exposed areas of the body; the tick inserted into the body should be removed as soon as possible using hygienic tweezers or special pliers available at pharmacies, pet stores; a�er returning home, check the body and the removed clothing, because ticks can wander unnoticed on the body or clothing; if we take a dog or a cat out, we also check its whole body a�er returning home; in addition, a nn a k oc oń , s yl w ia j an ic ze k, n at al ia m al ej ky -k łu se k 164 taking a shower and thorough cleaning with a sponge can help get rid of ticks; people who o�en spend time actively in the nature, on trips or actively working professionally with the forest and green areas should receive protective vaccinations against tickborne encephalitis a�er consulting a doctor. �e most commonly used procedures in the human environment aimed at reducing the number of ticks are: draining of wetlands, cutting or moving vegetation; systematic mowing of grass, removal of bushy thickets; limiting access of rodents and other wild animals to human buildings: removal of hideouts, land�lls; in the case of so� ticks, systematic disinfection and protection of animal husbandry places: observation of farm animals, in lo�s, checking the technical condition of the rooms; not allowing farm animals to move to human housing; rotations of pastures for grazing animals. in recent years, attempts have been made to use natural organisms to �ght ticks. �ese experiments are currently carried out on selected tick species, they are not longterm studies, so there is no 100% certainty that the given examples of animals eliminating tick populations can be used on a larger scale. among the animals, we can mention several examples that can reduce the population of ticks in the environment – ants: �re ant (solenopsis invicta bur.) (castellanos et al., 2016); beetles: staphylinus caesareus ceder. – currently rare due to the use of a large number of spraying (samish, alekseev, 2001); bed bugs; butter�y larvae; �ies: ixodiphagus hookeri how., a small insect laying eggs in a tick larvae, which then becomes food for the hatched wasp larvae (collatz et al., 2010) (appendix 1 c-d); reptiles: sand lizard (lacerta agilis l.); birds: domestic hens (gallus gallus domesticus l.), ducks (anas sp.), turkeys (meleagris gallopavo l.), black grouse (lyrurus tetrix l.), pheasants (phasianus colchicus l.), partridges (perdix perdix l.), helmeted guineafowl (numida meleagris l.) – the fact that guineafowl eat adult ticks was con�rmed by scienti�c research in 1990 in the usa (du�y et al., 1992), red-billed oxpecker (buphagus erythrorhynchus stan.) (bezuidenhout, stutterheim, 1980), cattle egret (bubulcus ibis l.); rodents: sorex, mice – deer mice (peromyscus leucopus ra�n.) (shaw et al., 2003), rats (rattus sp. g. fischer). researchers (angelo et al., 2015) inform that also some viruses, bacteria and fungi, e.g. metarhizium anisopliae (metch.) sor. and beauveria bassiana (bals.-criv.) vuill. as well as nematodes (samish et al., 2000), have the ability to reduce and weaken tick populations. research conducted by scientists shows that some plant species have deterrent properties and limit the occurrence of ticks in the area of cultivation of these plants, e.g. due to the intense smell caused by the release of speci�c essential oils into the air. however, it should be kept in mind that these are only experimental ways of �ghting ticks so far. �e research is carried out on selected species of ticks, on small populations, so this is not a fully proven method of protection against ticks. �e plants listed m ethods of protection against ticks (a cari: ixodida) 165 by scientists in the literature include, among others: tansy (tanacetum vulgare l. – appendix 1e) which is distinguished by a sharp spicy smell due to the essential oil – it repels not only ticks, but also mosquitoes, �ies and aphids (pålsson et al., 2008); horseradish (armoracia rusticana p.gaertn., b.mey. et scherb.) – an extract from the roots of horseradish not only repels but also kills ticks; catnip (nepeta cataria l.) – contains nepetalactone – a compound that ticks and mosquitoes cannot tolerate (birkett et al., 2011); sweet �ag (acorus calamus l.) – this plant’s essential oil contains over 80% of azarone, which is a substance that repels ticks, mosquitoes, ants and �ies (ghosh et al., 2011); onion (allium cepa l.) – according to russian scientists, dried onion powder is one of the most e�ective ways to get rid of ticks (aboelhadid et al., 2013); garlic (a. sativum l.) (aboelhadid et al., 2013); wormwood (artemisia absinthium l.) (godara et al., 2015); wild garlic (a. ursinum l.); narrow-leaved lavender (lavandula angustifolia mill.) (pirali-kheirabadi, teixeira da silva, 2010); tanacetum cinerariifolium (trevir.) sch. bip. – the �owers of this plant contain cinerine and pyrethrin – toxic to insects (an extract from them is used in many insecticidal preparations); rosemary (rosmarinus o�cinalis l. – appendix 1f) (martinez-velazquez et al., 2011). repellents on sale are available in various forms: sprays, liquids in atomizers, lotions, gels, creams, wipes impregnated with a repellent substance, bracelets made of polyvinyl chloride (pvc) or silicone, plasters to stick on clothing or other surfaces, devices emitting ultrasonic sounds. in addition, pills, injections and spot-on are used for animals. repellents may contain active substances of synthetic or natural origin. �e most commonly used synthetic active ingredients are: n-n-diethyl-m-toluamide (deet); ethyl ester of 3-(n-acetyl-n-butyl) amino propionic acid (ir3535); 1-piperidine-carboxylic acid kbr3023 (icaridine) (przygodzka et al., 2019). in poland, in 2016, research was carried out to determine the repellent e�ect of dermacentor reticulatus fabricius (meadow tick) ticks, products containing deet, icaridine, ir3535 and a mixture of 3 substances: deet, ir3535 and geraniol. studies have con�rmed that a repellent containing 30% deet showed 90% repellence for ticks even a�er 7  hours from applying the repellent on human skin, while the activity of other substances decreased, the second best repelling compound was: 30% deet, 20% ir3535 and 0.1% geraniol (60% of ticks not entering the skin a�er 7 hours). products containing 20% of icaridine and 12% ir3535 repelled ticks e�ectively for 1.5 hour (gliniewicz et al., 2019). however, it should be remembered that the tick species included in the research rarely attack humans, so it is necessary to carry out such experiments on a larger scale on the tick species i. ricinus most threatening the human species. ticks are very dangerous parasites for humans and animals, which in addition to causing direct e�ects of foraging (skin damage) can also be the transmitters of tickborne diseases. currently, these parasites occupy many convenient habitats, have many hosts and the risk of an attack by ticks throughout poland is a common phea nn a k oc oń , s yl w ia j an ic ze k, n at al ia m al ej ky -k łu se k 166 nomenon. reducing the number of ticks will still remain the most sought a�er method for preventing these parasites from occurring in human and pet habitats, as well as from spreading dangerous tick-borne diseases. research on controlling ticks is based mainly on chemical insecticides, which are not completely safe for humans and animals, and long-term tests of the environmental impact of these preparations are needed. �ere are several species of animals that eliminate the presence of ticks to some extent, and several species of plants that repel these dangerous parasites. however, the most promising method of bio-control so far is the use of fungal pathogens, reducing the number of ticks as well as a�ecting the movement and e�ciency of parasites. all natural ways to prevent the occurrence of ticks are still sought a�er, improved and implemented in the environment, but it should be kept in mind that such actions do not cause changes in relationships in local trophic networks. acknowledgements �ank you prof. nowak-chmura magdalena for support and help in writing the article. con�ict of interest �e authors declare no con�ict of interest related to this article. references aboelhadid, s.m., kamel, a.a., arafa, w.m., shokier, k.a. 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(2015). acaricidal activity of ethanolic extract of artemisia absinthium against hyalomma anatolicum ticks. experimental and applied acarology, 65(1), 141–148. doi: 10.1007/s10493-014-9843-6 ixodes ricinus l. female: https://www.gmx.net/magazine/wissen/natur-umwelt/zecken-nutzen-33586218 ixodiphagus hookeri howard: http://www.zeckenhilfe.com/en/all-about-ticks/article/what-are-the-natural-enemies-of-ticks-; ixodiphagus hookeri attacking a fed female tick: https://alchetron.com/ixodiphagus-hookeri martinez-velazquez, m., rosario-cruz, r., castillo-herrera, g., flores-fernandez, j.m., alvarez, a.h., lugo-cervantes, e. (2011). acaricidal e�ect of essential oils from lippia graveolens (lamiales: verbenaceae), rosmarinus o�cinalis (lamiales: lamiaceae), and allium sativum (liliales: liliaceae) against rhipicephalus (boophilus) microplus (acari: ixodidae). journal of medical entomology, 48(4), 822–827. doi: 10.1603/me10140 pålsson, k., jaenson, t.g., baeckström, p., borg-karlson, a.k. (2008). tick repellent substances in the essential oil of tanacetum vulgare. journal of medical entomology, 45(1), 88–93. doi: 10.1603/0022-2585(2008)45[88:trsite]2.0.co;2 pirali-kheirabadi, k., teixeira da silva, j.a. (2010). lavandula angustifolia essential oil as a novel and promising natural candidate for tick (rhipicephalus (boophilus) annulatus) control. experimental parasitology, 126(2), 184–186. doi: 10.1016/j.exppara.2010.04.012 przygodzka, m., mikulak, e., chmielewski, t., gliniewicz, a. (2019). repellents as a major element in the context of prevention of tick-borne diseases. przegląd epidemiologiczny, 73(2), 269–280. doi: 10.32394/pe.73.25 samish, m., alekseev, e. (2001). arthropods as predators of ticks (ixodoidea). journal of medical entomology, 38(1), 1–11. samish, m., alekseev, e., glazer, i. (2000). biocontrol of ticks by entomopathogenic nematodes. research update. annals of the new york academy of sciences, 916, 589–594. doi: 10.1111/j.1749-6632.2000. tb05341.x shaw, m.t., keesing, f., mcgrail, r., ostfeld, r.s. (2003). factors in�uencing the distribution of larval blacklegged ticks on rodent hosts. american journal of tropical medicine and hygiene, 68, 447–452. doi: https://doi.org/10.4269/ajtmh.2003.68.447 siuda, k. (1991). kleszcze polski (acari: ixodida). część i. zagadnienia ogólne. warszawa, wrocław: wydawnictwo naukowe pwn. [in polish] tanacetum vulgare l.: http://www.nic.funet.�/pub/sci/bio/life/plants/magnoliophyta/magnoliophytina /magnoliopsida/asteraceae/tanacetum/vulgare-2.jpg �e number of cases of selected tick-borne diseases in poland. data for day 30.09.2019: http://wwwold.pzh. gov.pl/oldpage/epimeld/index_p.html a nn a k oc oń , s yl w ia j an ic ze k, n at al ia m al ej ky -k łu se k 168 appendix 1 a – ixodes ricinus l. female (source: https://www.gmx.net/magazine/wissen/natur-umwelt), b – argas re�exus fabricius (source: https://zeckenrollen.de/zecken/zeckenarten-in-deutschland/), c – ixodiphagus hookeri howard (source: http://www.zeckenhilfe.com/en/all-about-ticks/article/), d – ixodiphagus hookeri attacking a fed female tick (source: https://alchetron.com/ixodiphagus-hookeri), e – tanacetum vulgare l. (source: http://www.nic.funet.�/pub/sci/bio/life/plants), f – rosmarinus o�cinalis l. (photo. a. kocoń) m ethods of protection against ticks (a cari: ixodida) 169 metody ochrony przed kleszczami (acari: ixodida) streszczenie kleszcze (acari: ixodida) należą do roztoczy, pasożytujących najczęściej na gadach, ptakach i ssakach. ze względu na znaczenie epidemiologiczne, epizootiologiczne, jak również bezpośrednią szkodliwość wśród ludzi i zwierząt, zaliczają się do jednych z najgroźniejszych pasożytów zewnętrznych. na całym świecie stwierdzono występowanie około 850 gatunków kleszczy, w polsce, do tej pory stwierdzono 19 gatunków stale występujących w faunie naszego kraju. bytują one w różnych siedliskach od terenów nizinnych aż po tereny górskie, zajmując takie miejsca jak: lasy, tereny zieleni, strychy budynków w tym mieszkalnych i gospodarskich, nory, jamy zwierząt, jaskinie. coraz częstsze występowanie kleszczy w bliskim otoczeniu człowieka i zwierząt przydomowych oraz domowych stwarza idealne warunki do przenoszenia patogenów chorób odkleszczowych. nie ma wątpliwości, że stale poszukiwane są wszelkie sposoby ochrony osobistej i działania środowiskowe, chroniące przed atakami kleszczy oraz przed konsekwencjami jakie mogą wystąpić po żerowaniu pasożyta. key words: parasite, tick protection, ticks received: [2019.08.08] accepted: [2019.11.15] 106 annales universitatis paedagogicae cracoviensis studia naturae, 3 (supplement): 106–115, 2018, issn 2543-8832 14th international scientific conference “animal physiology” kraków, 13–15 july 2018, poland 14 międzynarodowa konferencja naukowa „animal physiology” kraków, 13–15 czerwiec 2018, polska on june 13–15, 2018, the 14th international scienti�c conference – “animal physiology” took place in kraków. �e main organisers of this year’s edition of the conference were representatives from the institute of biology of the pedagogical university of cracow (poland). as before, scientists from the department of animal physiology of the faculty of biotechnology and food sciences of the slovak university of agriculture in nitra (slovakia), the department of animal morphology, physiology and genetics of the faculty of agronomics of the mendel university in brno (czechia) and the institute of animal physiology of the slovak academy of sciences (slovakia) also took part in the organisation of the conference. �e chair of the organising committee was assoc. prof. krzysztof piksa and the secretaries were phd eng. renata muchacka and phd zo�a goc. �e scienti�c committee was composed of representatives of universities and scienti�c units, both from poland and abroad: phd laszlo bardos (szent istván university, gödöllő, hungary), phd łukasz j. binkowski (pedagogical uniw dniach 13–15 czerwca 2018 r. w krakowie odbyła się 14 międzynarodowa konferencja naukowa „animal physiology”. głównymi organizatorami tegorocznej edycji konferencji byli przedstawiciele instytutu biologii uniwersytetu pedagogicznego im. komisji edukacji narodowej w  krakowie (polska). w organizacji konferencji brali także udział pracownicy katedry fizjologii zwierząt wydziału biotechnologii i  nauk o  żywności uniwersytetu rolniczego w  nitrze (słowacja), zakładu morfologii, fizjologii i  genetyki zwierząt wydziału agronomicznego uniwersytetu im. mendla w  brnie (czechy) oraz instytutu fizjologii zwierząt słowackiej akademii nauk (słowacja). przewodniczącym tegorocznego komitetu organizacyjnego był dr hab. krzysztof piksa, a  sekretarzami dr inż. renata muchacka oraz dr zo�a goc. w składzie komitetu naukowego znaleźli się przedstawiciele uczelni i  jednostek naukowych, zarówno z  kraju, jak i  z  zagranicy: dr laszlo bardos (uniwersytet im. szent istván, gödöllő, węgry), dr łukasz j. binkowski (uniwersytet pedagogiczny w krakowie, polska), dr mar107 r eportversity of cracow, poland), phd martha valdivia cuya, (national university of san marcos, lima, peru), prof. štefan faix (slovak academy of sciences, košice, slovakia), phd zita faixová (university of veterinary medicine and pharmacy in košice, slovakia), assoc. prof. grzegorz formicki (pedagogical university of cracow, poland), assoc. prof. agnieszka greń (pedagogical university of cracow, poland), phd zdeněk havlíček, (mendel university in brno, czechia), assoc. prof. monika martiniaková (constantine the philosopher university, nitra, slovakia), assoc. prof. adriana kolesárová (slovak university of agriculture in nitra, slovakia), assoc. prof. jaroslav kováčik (slovak university of agriculture in nitra, slovakia), assoc. prof. norbert lukáč (slovak university of agriculture in nitra, slovakia), prof. peter massányi, (slovak university of agriculture in nitra, slovakia), assoc. prof. radoslav omelka (constantine the philosopher university, nitra, slovakia), phd aleš pavlík (mendel university in brno, czechia), assoc. prof. krzysztof piksa (pedagogical university of cracow, poland), phd barbara pinto (university of pisa, italy), phd shubhadeep roychoudhury (assam university, silchar, india), assoc. prof. robert stawarz (pedagogical university of cracow, poland), assoc. prof. waldemar szaroma (pedagogical university of cracow, poland), assoc. prof. krzysztof tokarski (polish academy of sciences, cracow, poland), phd francesco vizzarri (university of molise, campobasso, italy), prof. michal zeman (comenius university in bratislava, slovakia). on the �rst day of the conference (13.06.2018), the opening ceremony and tha valdivia cuya (narodowy uniwersytet w san marcos, lima, peru), prof. štefan faix (słowacka akademia nauk, koszyce, słowacja), dr zita faixová (uniwersytet medycyny weterynaryjnej i  farmacji w  koszycach, słowacja), dr hab. grzegorz formicki (uniwersytet pedagogiczny w  krakowie, polska), dr hab. agnieszka greń (uniwersytet pedagogiczny w  krakowie, polska), dr zdeněk havlíček (uniwersytet im. mendla w  brnie, czechy), dr hab. monika martiniaková (uniwersytet im. konštantína filozofa w  nitrze, słowacja), dr hab. adriana kolesárová (uniwersytet rolniczy w  nitrze, słowacja), dr hab. jaroslav kováčik (uniwersytet rolniczy w  nitrze, słowacja), dr hab. norbert lukáč (uniwersytet rolniczy w  nitrze, słowacja), prof. dr hab. peter massányi (uniwersytet rolniczy w nitrze, słowacja), dr hab. radoslav omelka (uniwersytet im. konštantína filozofa w  nitrze, słowacja), dr aleš pavlík (uniwersytet im. mendla w  brnie, czechy), dr hab. krzysztof piksa (uniwersytet pedagogiczny w krakowie, polska), dr barbara pinto (uniwersytet w pizie, włochy), dr shubhadeep roychoudhury (uniwersytet w assamie, silchar, indie), dr hab. robert stawarz (uniwersytet pedagogiczny w krakowie, polska), dr hab. waldemar szaroma (uniwersytet pedagogiczny w krakowie, polska), dr hab. krzysztof tokarski (polska akademia nauk, kraków, polska), dr francesco vizzarri (uniwersytet molise w campobasso, włochy) oraz prof. dr hab. michal zeman (uniwersytet comenius w  bratysławie, słowacja). ceremonia otwarcia i sesje w pierwszym dniu konferencji (13.06.2018) miały miejsce w  sali obrad rady miasta krakowa im. 108 r ep or t sessions took place in the stanisław wyspiański room, city council of kraków, at the wielopolski palace (kraków city hall at plac wszystkich świętych 3–4) (fig. 1). �e conference was o�cially opened by the vice-rector for the development of the pedagogical university in kraków assoc. prof. robert stawarz. on the second and third days of the conference (14–15.06.2018), sessions took place in the krzysztofory palace (rynek główny 35). �e inaugural lectures, on the �rst and second days, were presented by invited guests. on the �rst day, directly a�er the o�cial opening of the conference, the plenary lecture “�e e�ects of stress and corticosterone on synaptic transmission and neuronal plasticity in the rat motor cortex” by prof. grzegorz hess (department of neurophysiology and chronobiology, institute of zoology and biomedical research, stanisława wyspiańskiego w  pałacu wielopolskich (urząd miasta krakowa przy placu wszystkich świętych 3–4) (ryc. 1). uroczystego otwarcia konferencji dokonał prorektor ds. rozwoju uniwersytetu pedagogicznego w  krakowie dr hab. robert stawarz. sesje w  drugim i  trzecim dniu konferencji (14–15.06.2018) odbyły się w pałacu krzysztofory (rynek główny 35). wykłady inauguracyjne w  pierwszym i  drugim dniu konferencji zaprezentowali zaproszeni goście. w  pierwszym dniu, bezpośrednio po uroczystym otwarciu konferencji wykład plenarny pt. „wpływ stresu i kortykosteronu na transmisję synaptyczną i  plastyczność neuronalną w  korze ruchowej szczura” wygłosił prof. dr hab. grzegorz hess (zakład neuro�zjologii i  hronobiologii, instytut zoologii, uniwersytet jagielloński, kraków – polska), a  w  drugim dniu dr hab. jan rodriguez parkitna (zakład neufig. 1. participants of the poster session on the �rst day of the conference “animal physiology” in the stanisław wyspiański room at the wielopolski palace (photo. w. wojtaś) ryc. 1. uczestnicy sesji referatowej w pierwszym dniu konferencji „animal physiology” w sali im. stanisława wyspiańskiego w pałacu wielopolskich (fot. w. wojtaś) 109 r eport jagiellonian university, kraków – poland) was performed. on the second day, assoc. prof. jan rodriguez parkitna (department of molecular neuropharmacology, institute of pharmacology, polish academy of sciences, kraków – poland) presented relation between reward system of the brain: the pleasure of learning (fig. 2). �e �rst session of the conference was conducted by assoc. prof. robert stawarz and phd łukasz j. binkowski (pedagogical university of cracow, poland). as part of this session, lectures were presented by four speakers, and they concerned the following: �e impact of alcohol administration a�ects compact bone structure of mice a�er one remodelling cycle – assoc. prof. monika martiniaková (constantine the philosopher university, nitra, slovakia), dietary supplementation with algae and polypherofarmakologii molekularnej, instytut farmakologii, polska akademia nauk, kraków – polska) przedstawił zagadnienia związane z systemem nagradzania mózgu: przyjemnością uczenia się . pierwszą sesję konferencji prowadzili dr hab. robert stawarz oraz dr łukasz j. binkowski (uniwersytet pedagogiczny w  krakowie). w  ramach tej sesji referaty wygłosiło czterech prelegentów, a  dotyczyły one: wpływu podawania alkoholu na zwartą strukturę kostną myszy po jednym cyklu przebudowy – dr hab. monika martiniaková (uniwersytet im. konštantína filozofa w nitrze, słowacja), suplementacji diety algami i  polifenolami u  samca królika: wpływu na cechy jakościowe nasienia – dr francesco vizzarri (uniwersytet molise w campobasso, włochy) (ryc. 3), embriotoksycznego potencjału różnych typów insetycydów – dušan fig. 2. assoc. prof. jan rodriguez parkitna during the inaugural lecture on the second day of the conference “animal physiology” (photo. w. wojtaś) ryc. 2. dr hab. jan rodriguez parkitna w trakcie wykładu inauguracyjnego wygłoszonego w drugim dniu konferencji „animal physiology” (fot. w. wojtaś) 110 r ep or t nols in rabbit male: e�ects on semen quality traits – phd francesco vizzarri (university of molise, campobasso, italy) (fig. 3), �e embryotoxic potential of various types of insecticides – dušan fabian (slovak academy of sciences, košice, slovakiai, and answers on questions – does benzo[a]pyrene a�ect the heart embryonic development? – msc łukasz m. kołodziejczyk (pedagogical university of cracow, poland). on the same day, in the evening, the “underground market” in kraków museum was visited, and then in the hawełka restaurant as part of the banquet the integration meeting of all conference participants was organised. on the second day of the conference (14.06.2018), the morning poster session was conducted by assoc. prof. marko halo (university of agriculture in nitra, slovakia) and assoc. prof. jaroslav kováčik (university fabian (słowacka akademia nauk, koszyce, słowacja), oraz odpowiedzi na pytanie – czy benzo[a]piren wpływa na rozwój embrionalny serca? – mgr łukasz m. kołodziejczyk, uniwersytet pedagogiczny w  krakowie, polska). tego samego dnia w  godzinach wieczornych odbyło się zwiedzanie muzeum „podziemia rynku w krakowie”, a następnie spotkanie integracyjne wszystkich uczestników konferencji w ramach bankietu zorganizowanego w restauracji „hawełka”. w drugim dniu (14.06.2018), przedpołudniową sesję referatową prowadzili dr hab. marko halo (uniwersytet rolniczy w  nitrze, słowacja) oraz dr hab. jaroslav kováčik (uniwersytet rolniczy w  nitrze, słowacja). w  ramach tej sesji trzech prelegentów zaprezentowało referaty na temat: jak sposób wyboru kontrastujących poziomów testosteronu w  żółtku wpływa na oś rozrodczą samfig. 3. phd francesco vizzarri from university of molise, campobasso, italy during lecture in the �rst session (photo. w. wojtaś) ryc. 3. dr francesco vizzarri z uniwersytetu molise w campobasso podczas wykładu w pierwszej sesji referatowej (fot. w. wojtaś) 111 r eportof agriculture in nitra, slovakia). as part of this session, three lecturers presented papers on the following: how selection for contrasting yolk testosterone levels a�ects reproductive axis in male japanese quail? – assoc. prof. monika okuliarová (comenius university in bratislava, slovakia), deposition of immunoactive substances into egg and immune response of young japanese quail selected for shape of growth curve – phd zuzana kanková (comenius university in bratislava, slovakia) and organ toxicity of diethylnitrosamine and capsaicin in mice-in vivo study – msc vendula kuchařová (university of veterinary medicine and pharmacy, brno, czechia). a�er the co�ee break, the chairmanship of the session was taken over by assoc. prof. adriana kolesárová (agricultural university in nitra, slovakia) and prof. štefan faix (slovak academy of sciences, košice, slovakia). in this part of the session, �ve oral presentations were presented, including: e�ects of zinc supplementation in mineral status of farm animals – ľubomíra grešáková (slovak academy of sciences, košice, slovakia), e�ects of amygdaline on gene activity in cultivated human osteoblasts – msc veronika kováčová (uniwersytet im. konštantína filozofa w  nitrze, słowacja), and two sides of non-ionizing radiation in daily life use – reproductive approach – eng. filip tirpák (agriculture university in nitra, slovakia). a�er the lunch break, there was a  poster session in which 38 posters concerning the broadly understood animal physiology, the physiology of human nutrition and physiology at the cellular and molecular level were presented. �e poster session was chaired by assoc. prof. radoców przepiórek japońskich? – dr hab. monika okuliarová (uniwersytet komeńskiego w bratysławie, słowacja), osadzanie się substancji immunoaktywnych w jajach i odpowiedź immunologiczna młodych przepiórek japońskich – dr zuzana kanková (uniwersytet komeńskiego w  bratysławie, słowacja) i  toksyczność narządowa dietylotrojaminy i  kapsaicyny u  myszy w  badaniu in vivo – mgr vendula kuchařová (uniwersytet nauk weterynaryjnych i farmaceutycznych, brno, czechy). po przerwie kawowej przewodnictwo sesji przejęli dr hab. adriana kolesárová (uniwersytet rolniczy w  nitrze, słowacja) oraz prof. štefan faix (słowacka akademia nauk, koszyce, słowacja). w  tej części sesji referatowej przedstawiono pięć wystąpień ustnych, m.in.: wpływ suplementacji cynku na stan mineralny zwierząt gospodarskich (słowacka akademia nauk, koszyce, słowacja), wpływ amigdaliny na aktywność genową w  hodowanych ludzkich osteoblastach – mgr veronika kováčová (uniwersytet im. konštantína filozofa w  nitrze, słowacja), dwie strony promieniowania niejonizującego w  codziennym życiu – podejście reprodukcyjne – inż. filip tirpák (uniwersytet rolniczy w  nitrze, słowacja). po przerwie obiadowej odbyła się sesja posterowa, w  której zaprezentowano 38 plakatów dotyczących szeroko rozumianej �zjologii zwierząt, żywienia człowieka oraz �zjologii na poziomie komórkowym i  molekularnym. sesji posterowej przewodniczyli dr hab. radoslav omelka (uniwersytet im. konštantína filozofa w  nitrze, słowacja) oraz prof. dr hab. peter massányi (uniwersytet rolniczy w  nitrze, słowacja). późnym 112 r ep or t slav omelka (constantine the philosopher university, nitra, slovakia), and prof. peter massányi (agriculture university in nitra, slovakia). in the late a�ernoon, the participants of the conference took part in the “royal way” walk to the wawel hill. on the third and last days of the conference (june 15, 2018), the chairmanship of the morning lecture session was entrusted to assoc. prof. monika martiniaková (konštantín philosopher university in nitra, slovakia) and prof. michal zeman (comenius university in bratislava, slovakia). in this part, six lectures were presented, for example: �e identi�cation of glucocorticoid receptors transcripts in mouse oocytes and preimplantation embryos – phd alexandra špirková (slovak academy of science, košice, slovakia), di�erent macs sorting strategies for the enrichment of lin-(cd34 + cd45–) hematopoietic progenitor cell: preliminary study – phd jaromír vašiček (agriculture university in nitra, slovakia), and e�ects of taraxacum o�cinale root extract on murine �brosarcoma cells in vitro – phd jaroslava tomenendálová (university of veterinary medicine and pharmacy, brno, czechia). a�er the co�ee break, the chairmen of the last session were phd zita faixová (university of veterinary medicine and pharmacy in košice, slovakia) and assoc. prof. grzegorz formicki (pedagogical university of cracow, poland). �is time, six topics were also presented, including: �e effect of substances used in smart drugs on selected parameters of spermatozoa motility – msc katarzyna stachańczyk (pedagogical university of cracow, poland), �e relationship between bio�lm formation, genes of virulence, and iron metabolism in escherichia coli – phd lívia popołudniem uczestnicy konferencji wzięli udział w  spacerze „drogą królewską” na wzgórze wawelskie. w  trzecim i  ostatnim dniu konferencji (15.06.2018) przewodnictwo porannej sesji referatowej powierzono dr hab. monice martiniakovej (uniwersytet im. konštantína filozofa w  nitrze, słowacja) i  prof. dr hab. michalowi zeman (uniwersytet comenius w bratysławie, słowacja). w tej części wygłoszono sześć referatów dotyczących, np.: komunikacji embrionalno-matczynej w okresie rozwojowym przedimplantacyjnym: receptorów komórkowych przekazujących sygnały ze środowiska matczynego – dr alexandra špirková (słowacka akademia nauk, koszyce, słowacja), różnych strategii sortowania macs do wzbogacania hematopoetycznych komórek progenitorowych lin-(cd34+ cd45-) – dr jaromír vašiček (uniwersytet rolniczy w  nitrze, słowacja), czy też wpływu wyciągu z korzenia taraxacum o�cinale na komórki włókniako-mięśniaka myszy in vitro – dr jaroslava tomenendálová (uniwersytet nauk weterynaryjnych i  farmaceutycznych, brno, czechy). po przerwie kawowej, ostatnią sesję referatową prowadzili dr zita faixová (uniwersytet medycyny weterynaryjnej i farmacji w koszycach, słowacja) oraz dr hab. grzegorz formicki (uniwersytet pedagogiczny w  krakowie, polska). tym razem zaprezentowano również sześć tematów, m.in.: wpływ substancji stosowanych w  „lekkich narkotykach” na wybrane parametry ruchliwości plemników – mgr katarzyna stachańczyk (uniwersytet pedagogiczny w krakowie, polska), związek między powstawaniem bio�lmów, genami wirulencji i  metabolizmem żelaza w escherichia coli – dr lívia handrová 113 r eport handrová (center for biological sciences sas, košice, slovakia), and �e concentration of mercury in organs of whip�n silver biddy (gerres �lamentosus cuvier, 1829) and flathead grey mullet (mugil cephalus linnaeus, 1758) in coastal central vietnam – msc �iep vo van (pedagogical university of cracow, poland). around 12 o’clock, there was a �nal discussion, a  summary of the entire conference and the o�cial closing ceremony. �is year’s 14th international scienti�c conference “animal physiology” certainly ful�lled its task. as the organisers of the conference emphasized, its aim was to discuss and exchange experiences and scienti�c achievements in the broadly understood animal physiology (fig. 4). �e opportunity to share your achievements is undoubtedly an additional impulse for further fruitful studies, and getting to know the results of others can become an inspiration for new research. (centrum nauk biologicznych sas, koszyce, słowacja), czy stężenie rtęci w  narządach gerres �lamentosus cuvier, 1829 i  mugil cephalus linnaeus, 1758 w  przybrzeżnym centralnym wietnamie – mgr �iep vo van (uniwersytet pedagogiczny w  krakowie, polska). około godziny 12 odbyła się dyskusja końcowa i podsumowanie całej konferencji oraz o�cjalna ceremonia zamknięcia. tegoroczna, 14 międzynarodowa konferencja naukowa „animal physiology” z pewnością spełniła swoje zadanie. jak podkreślają organizatorzy konferencji, jej celem była dyskusja oraz wymiana doświadczeń i  osiągnięć naukowych z  szeroko rozumianej �zjologii zwierząt (ryc. 4). możliwość podzielenia się swoimi osiągnięciami jest niewątpliwie dodatkowym impulsem do dalszej owocnej pracy, a zapoznanie się z wynikami prac innych – może stać inspiracją do nowych poszukiwań badawczych. podczas fig. 4. participants of the “animal physiology” conference during informal discussions (photo. w. wojtaś) ryc. 4. uczestnicy konferencji „animal physiology” w podczas nieformalnych dyskusji w kuluarach (fot. w. wojtaś) 114 r ep or t during the intensive three days of the conference, 24 oral presentations were presented during the 5 scienti�c sessions. in total, 85 people from 15 scienti�c centres from 6 countries took part in the conference. abstracts of oral speeches and posters were prepared by renata muchacka and edyta kapusta in a  special edition book printed by the pedagogical university in kraków, “14th international scienti�c conference – animal physiology 2018 – book of abstracts, 13–15 june 2018, kraków, poland” – pp. 73, e-isbn 978-83-8084-152-9, doi 10.24917/9788380841529, while some of the full articles are included in this supplement annales universitatis paedagogicae cracoviensis studia naturae, as reviewed post-conference materials. intensywnych trzech dni obrad konferencji i trwania 5 sesji naukowych, zaprezentowano 24 wystąpienia ustne. ogółem w konferencji uczestniczyło 85 osób z 15 ośrodków naukowych z 6 państw. streszczenia wystąpień ustnych i  posterów zostały opublikowane pod redakcją renaty muchackiej i  edyty kapusty w  specjalnie wydanym przez uniwersytet pedagogiczny w  krakowie opracowaniu pt.: „14th international scienti�c conference – animal physiology 2018 – book of abstracts, 13–15 june 2018, kraków, poland” – ss. 73, e-isbn 978-83-8084-152-9, doi 10.24917/9788380841529, natomiast część pełnych artykułów znalazło się w niniejszym suplemencie annales universitatis paedagogicae cracoviensis studia naturae, jako recenzowane materiały pokonferencyjne. renata muchacka department of animal physiology and toxicology, institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland; renata.muchacka@up.krakow.pl; https://orcid.org/0000-0002-8255-006x 67 annales universitatis paedagogicae cracoviensis studia naturae, 4: 67–77, 2019, issn 2543-8832 doi: 10.24917/25438832.4.3 héctor m.j. lópez-castilla1, ángel j. ríos-oviedo1, william cetzal-ix1*, saikat kumar basu2 1tecnológico nacional de méxico, instituto tecnológico de chiná. calle 11 entre 22 y 28, colonia centro chiná 24050. campeche, méxico; *rolito22@hotmail.com 2ps, lethbridge ab canada t1j 4b3 construction of the nest of amazilia rutila de lattre (trochillidae) and its antipredatory defensive strategy in a medium deciduous forest in campeche, mexico introduction �e construction of nests by birds is a natural behavior, its size and composition varies depending on the species, and the selection of materials is opportunistic (deeming, mainwaring, 2015). in some cases, birds select fresh leaves of plants that have antimicrobial properties or that allow the abatement of ectoparasites in their nests (dubiec, mazgajski, 2013). as for the selection of habitats, birds that nest in the soil to achieve reproductive success tend to look for sites with heterogeneous vegetation and dense foliage, which also helps them counteract predation (martin et al., 1997). but nests with reproductive success depend on factors such as the availability of food and the presence of other individuals of the same species (danchin et al., 2004). in the case of hummingbirds, reproductive success can be a�ected by the isolation and fragmentation of vegetation, resulting in the reduction of their populations (feinsinger et al., 1987). however, predation in nests is common in ecosystems, being the main factors that cause reproductive failure, a�ecting population density and a�ecting abundance at the community level (angelstam, 1986; martin, 1988; buler, hamilton, 2000; jokimäki, huhta, 2000). in this sense, hummingbirds are used as indicator species of disturbance in tropical ecosystems; for example, in the yucatan peninsula (yp), mexico, the diversity of hummingbirds is related to a greater abundance of tree species (navarro et al., 2016). in the yp there are 12 species of hummingbirds, among these the cinnamon hummingbird amazilia rutila de lattre (trochillidae), which is distributed in the paci�c slope from the northeast of mexico to costa rica and the atlantic slope in the portion north and east of the yp and in belize; this species is found from tropical forests, urban areas, bushes to savannas (birdlife international, h éc to r m .j . l óp ez -c as til la , á ng el j . r ío so vi ed o, w ill ia m c et za l-i x, s ai ka t k um ar b as u 68 2016). in the o�cial mexican standard (dof-2010) and the birdlife international (2016) it is categorised as least concern (lc). in general, hummingbirds are considered as �ags because of their diversi�cation of plumage colours and �ight capabilities (sánchez-jasso, cebrián-abellán, 2015; medinavan berkum et al., 2016), these species function as symbols to attract governmental and non-governmental support, as well as a �ag for the implementation of conservation programs (noss, 1990; andelman, fagan, 2000; carignan, villard, 2002; caro et al., 2004; isasi-catalá, 2011). with the aim of contributing to the knowledge of the biology and reproductive strategies of a. rutila, the architecture, dimensions and vegetative materials of plants used for the construction of their nests and the hosts used as possible adaptations to counteract predation are described. material and methods study area �e �rst nest of amazilia rutila was located in the native flora conservation unit of the yucatan peninsula (nfcuyp) at the instituto tecnológico de chiná in campeche, mexico (19°46ʹ10.44ʹʹ n, 90°30ʹ16.24ʹʹ w) (cetzal-ix et al., 2017) (fig. 1–2). �e second nest was located at the residential farms cholul (rfc), campeche, mexico (19°42ʹ56.44ʹʹ n, 90°23ʹ6.59ʹʹ w) 14.3 km southeast of the nfcuyp in chiná, campeche, mexico (fig. 3). both sites have medium deciduous forest (pérez et al., 2017) with a sub humid warm climate (aw) and an annual precipitation of 1141 mm (garcía, 1988). however, rfc site has a vegetation disturbed by agriculture and livestock, restricted to agro-crops, citrus and pasture (fig. 2–3). observations and identi�cation of materials for the construction of the nest observations of behavioral activities in the hummingbird nests were made at time intervals of 12 to 23 minutes during each hour, between 8:00 a.m. to 1:00 p.m. (diurnal) and 8:00 p.m. to 12:00 p.m. (nocturnal) during 16 days, of 14 from march to april 1, 2019. �e characterisation of the nests was made according to hansell (2000) and with a millimeter precision vernier. in the laboratory of agroecosystems and conservation of biodiversity, the identi�cation of the host plants and the materials used for the construction of the nest was made, through specialised literature and illustrated guides of �ora of the yucatan peninsula (yucatán) (carnevali et al., 2010; cetzal-ix et al., 2017). 69 fig. 1. plant material for nest and study area: a – asclepias curassavica l.; b – seeds; c – �owers and seeds; d – pappus with seeds; e – native flora conservation unit of the yucatan peninsula (nfcuyp) at the instituto tecnológico de chiná in campeche, mexico (photo. w. cetzal-ix) c onstruction of the nest of am azilia rutila d e lattre (trochillidae) and its antipredatory defensive strategy in a m edium deciduous forest in c am peche, m exico h éc to r m .j . l óp ez -c as til la , á ng el j . r ío so vi ed o, w ill ia m c et za l-i x, s ai ka t k um ar b as u 70 results and discussion �e nest of hummingbird located in the nfcuyp presented a female of a. rutila with two eggs, one hatched (and the brood was not found) and the other did not hatch a�er 16 days of observation, due to the abandonment of the adult female (fig. 1). on the other hand, the hummingbird nest located in rfc presented an adult individual and a juvenile chick. regarding the sizes of nest, both presented a concave cup-shaped symmetric shape, but di�ered in their sizes (fig. 3); the nfcuyp nest has an external length of 61.27 mm, a cup diameter of 24.62 mm, an internal diameter of 23 × 23 mm and a depth of 19.76 mm. �e rfc nest had an external length of 52.49 mm, a cup diameter of 40.89 mm, an internal diameter of 23.37 × 26.07 mm and a depth of 27.94 mm. �e main vegetative material identi�ed for the construction of the nests in both locations, were made from pappus from asclepias curassavica l. (apocynaceae) (fig. 2a–d). �e pappus is a thin and cottony �lament that possesses the seeds for the dispersion (toro, briones, 1995). approximately between 10 and 14 m away from the host plants of the nests, respectively, a. curassavica was observed growing as a herbaceous from 0.5 to 1 m in height, in disturbed sites and without vegetation. in addition, we observed in both nests the presence of lichens in the lateral parts of the same and covered with cobweb, which gives them greater rigidity and support. �e web possibly belongs to peucetia viridans hentz, which was registered in the same host plants, in both sites (fig. 1e). in the nfcuyp, the identi�ed plant-host where the hummingbird nest was found was chaya bush cnidoscolus acotinifolius (mill.) i.m. johnst. (euphorbiaceae), known in the yp as “chaya de monte” which are shrubs with abundant white latex, trichomes in the form of sharp hairs and thorns (fig. 1). �is species is native to the yp and is widely distributed in the northern portion of the region, mainly in deciduous and medium-sized sub-deciduous forest (carnevali et al., 2010). �e nest in the host plant was found 1.45 m above the ground, at 37.76 mm in diameter of the main branch, 12.50 mm in diameter of the support branch, placed 10 branches of the host plant, 45 cm away from the stem of the host plant; the percentage of coverage of the leaves that provide the nest was 80% and with a percentage of 70% visibility, based on the criteria of observations indicated by ralph et al. (1996). stinging trichomes and the latex that possesses c. acotinifolius have been reported to act as defensive mechanisms for the plant (torres-gonzález, garcía-guzmán, 2014). in this sense, they can also represent a defense and protection mechanism for the nests of a. rutila, particularly during periods of drought (january to may) when the greatest loss of leaves (40–60%) occurs in the low and medium deciduous and sub caducifolious forests, respectively. the nest registered in rfc, was found in a mango tree mangifera indica l., a species cultivated and used for agriculture in tropical areas, the nest was observed at 71 fig. 2. cinnamon hummingbird and host plant: a – amazilia rutila de lattre in nest; b – nest, c – nest in cnidoscolus aconitifolius (mill.) i.m. johnst.; d – nest eggs; e – peucetia viridans hentz in c. aconitifolius (photo. h.m.j. lópez-castilla) c onstruction of the nest of am azilia rutila d e lattre (trochillidae) and its antipredatory defensive strategy in a m edium deciduous forest in c am peche, m exico h éc to r m .j . l óp ez -c as til la , á ng el j . r ío so vi ed o, w ill ia m c et za l-i x, s ai ka t k um ar b as u 72 fig. 3. nest and study area: a – nest on mangifera indica l.; b – nest with lichen and cobwebs; c – plants of asclepias curassavica l. near the nest; d – residential farms cholul (rfc) in chiná, campeche, mexico (photo. h.m.j. lópez-castilla) 73 1.70 meters from the ground, at 37.76 mm in diameter of the main branch, 12.50 mm in diameter of the support branch, placed 10 branches of the host plant, 45 cm away from the stem of the host plant; the percentage of coverage of the leaves that provide the nest was 80% and with a percentage of 70% visibility. �e mango tree is widely cultivated in the yp and they do not lose their foliage during the seasons of the year, therefore, they do not expose the hummingbird nests. �ese dimensions and data on host plants can be considered for further studies on nesting patterns in hummingbirds. di�erent species of birds use a wide variety of materials for the construction of their nests, from dry grasses (mainwaring et al., 2014) or dry leaves of pinus patula schltdl. & cham. (pinaceae) (morales-rozo et al., 2009) to cigarette butts in urban species as haemorhous mexicanus müll. and passer domesticus l. (suárezrodríguez et al., 2013). in some hummingbirds, recorded is the use of branches and dry leaves of calea urticifolia (mill.) dc. (asteraceae) for the construction of the nest (ortiz-pulido et al., 1998), which are decorated with lichens of the genus parmotrema sp. (garcía, botero, 2013) or mosses such as ancistrodes genu�exa (c. müll.) cros. and weymouthia cochlearifolia (schwägr.) dixon (meteoriaceae) (torres-dowdall et al., 2007). generally, the materials used for the construction of the nest are adhered with cobwebs (ortiz-pulido et al., 1998; garcía, botero, 2013). although plant materials for nest construction are described in some bird species, information on various groups of birds is still scarce (deeming, mainwaring, 2015; biddle et al., 2018; wesołowski, wierzcholska, 2018). as well as the identity of the plant species used and if they are selected for some function in the nest, as for example by their antimicrobial properties to counteract ectoparasites (dubiec, mazgajski, 2013). for example, in amazilia violiceps gould we reported the use of dry branches of plants (not determined species plants) for the construction of the outer part of the nest and whitish �bers of the fruit of ceiba aesculifolia (kunth) britt. & baker f. tree for the inner part of the nest; interwoven with cobwebs of possibly nephila sp. (desucre-medrano et al., 2016). on the other hand, amazilia cyanocephala lesson records the use of small grasses, small pieces of leaves and �owers of mimosa sp.; likewise, scales of the stems of the fern alsophila �rma (baker) d.s. conant, cyathea bicrenata liebm. and cyathea a�. fulva) (cyatheaceae); seeds of tillandsia deppeana steud. (bromeliaceae) and horse hairs. �e nests are decorated with pieces of liverworts calypogeia spp. (calypogeiaceae) and adhered with the cobweb of peucetia viridans (ornelas-rodríguez, 2010). �is same species of spider was identi�ed in amazilia rutila, with which it also adheres the materials to the nest. previous studies identi�ed as host plants of hummingbird nests species represent species like phyllostachys siebold & zucc. (poaceae) (ornelas-rodríguez, 2010), heliocarpus terebinthinaceus (dc.) hochr. (malyaceae) (desucre-medrano et al., 2016) and citrus limon (l.) osbeck (rutaceae) (garcía, botero, 2013). �e knowledge of nesting species of the genus amazilia is far from complete c onstruction of the nest of am azilia rutila d e lattre (trochillidae) and its antipredatory defensive strategy in a m edium deciduous forest in c am peche, m exico h éc to r m .j . l óp ez -c as til la , á ng el j . r ío so vi ed o, w ill ia m c et za l-i x, s ai ka t k um ar b as u 74 (ornelas-rodríguez, 1995). although a. rutila is permanently resident in the yp, its reproductive habits, composition and structure of its nests have not yet been fully studied. conclusion the cinnamon hummingbird individuals located at the two sites of campeche were selective-opportunistic with the vegetative materials used for the construction of their nests, because they collect plants that are close to where they establish their nests within their habitat. on the other hand, they select exclusively the pappus from the seeds of asclepias curassavica for the construction of the nest walls, and provide rigidity and support with lichen and cobwebs of peucetia viridans that are also living in the same host plant. the selection of the host plant occurs selectively and opportunistically for periods of drought when the eggs in the nests are mostly exposed to predation. the location of materials for the construction of the nest and host plants can help understand the composition and architecture of nests for the survival of tropical bird species. con�ict of interest �e authors declare no con�ict of interest related to this article. references andelman, s., fagan, w. 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(2013). incorporation of cigarette butts into nests reduces nest ectoparasite load in urban birds: new ingredients for an old recipe? biology letters, 9(1), 20120931. doi: 10.1098/rsbl.2012.0931 toro, g.j., briones, v.j. (1995). manejo de pulsiana (asclepias curassavica l.) en pastizales. portoviejo: iniap, estación experimental portoviejo, departamento de control de malezas. [in spanish] torres-dowdall, j., osorio, f., suárez, g.m. (2007). materiales utilizados por el pica�or rubí (sephanoides sephaniodes) para la construcción de nidos en la selva valdiviana, chile. ornitologia neotropical, 18(3), 433–437. [in spanish] torres-gonzález, d., garcía-guzmán, g. (2014). análisis del papel de los caracteres foliares de cnidoscolus (euphorbiaceae) en la defensa contra herbívoros y patógenos. tip. revista especializada en ciencias químico-biológicas, 17(2), 126–134. [in spanish] wesołowski, t., wierzcholska, s. (2018). tits as bryologists: patterns of bryophyte use in nests of three species cohabiting a primeval forest. journal of ornithology, 159(3), 733–745. doi: 10.1007/s10336 018–1535-2 abstract �e yucatan peninsula (yp) is part of a biogeographical area characterised by its diversity of �ora and fauna, among which are the birds, mainly hummingbirds, which are indicators of the state of conservation of the ecosystems. in birds, the site establishment and construction of nest plays a fundamental role for reproduction and survival rate, the selection of materials occurs opportunistically, but birds tend to use fresh leaves of plants with antimicrobial properties or that allow the depletion of ectoparasites in their nests. in this sense, for the �rst time we recorded for the cinnamon hummingbird (amazilia rutila de lattre), the materials used for the construction of its nest and the site of establishment of the nest in the host plants in two sites of a medium sub-deciduous forest in, mexico. we recorded the construction of nests of a. rutila in two locations in campeche; in the �rst site the nest was found in a chaya bush cnidoscolus aconitifolius (mill.) i.m. johnst. (euphorbiaceae); most possibly as an anti-predatory strategy for trichomes in the form of sharp hairs and spines that the plants possess in their stems and leaves. in the second site, the nest was found in a mango tree mangifera indica l. (anacardiaceae). �e main vegetative material identi�ed for the construction of the nests in both locations, were made from pappus (thin and cottony �lament that possess the seeds for the dispersion) from asclepias curassavica l. (apocynaceae). key words: asclepias curassavica, cnidoscolus aconitifolius, hummingbird, mangifera indica, yucatan peninsula received: [2019.08.05] accepted: [2019.11.25] 77 budowa gniazda amazilia rutila de lattre (trochillidae) i jego anty-drapieżna strategia obronna w lasach liściastych w campeche w meksyku streszczenie półwysep jukatan (yp) jest częścią obszaru biogeogra�cznego charakteryzującego się różnorodnością �ory i fauny. wśród fauny licznie występują ptaki, głównie kolibry, będące wskaźnikami stanu zachowania ekosystemów. dla ptaków zakładanie i budowa gniazda odgrywa podstawową rolę w rozmnażaniu i przeżywalności. wybór materiałów na gniazda odbywa się oportunistycznie, ale ptaki zwykle używają świeżych liści, o właściwościach przeciwdrobnoustrojowych lub pozwalających na odstraszenie pasożytów zewnętrznych z ich gniazd. w tym kontekście, dla kolibra cynamonowego (amazilia rutila de lattre) po raz pierwszy opisaliśmy materiały użyte do budowy gniazda oraz miejsce założenia gniazda w roślinach żywicielskich, w dwóch lokalizacjach lasu liściastego w meksyku. obserwowaliśmy budowę gniazd a. rutila w dwóch lokalizacjach w campeche. w pierwszym miejscu gniazdo znaleziono w zaroślach chaya cnidoscolus aconitifolius (mill.) i.m. johnst. (euphorbiaceae); ostre włoski i kolce, które mają rośliny z tego rodzaju na swoich łodygach i liściach, najprawdopodobniej występują tu jako anty-drapieżna strategia dla włosieni. w drugim miejscu gniazdo znaleziono na drzewie mango mangifera indica l. (anacardiaceae). główny materiał roślinny zidenty�kowany do budowy gniazd w obu lokalizacjach został wykonany z puchu – pappus (cienkiego i bawełnianego �lamentu służącego do dyspersji nasion) z asclepias curassavica l. (apocynaceae). słowa kluczowe: asclepias curassavica, cnidoscolus aconitifolius, koliber, mangifera indica, półwysep jukatan information about authors héctor mj lópez-castilla he is a bachelor student of biology at the tecnológico nacional de méxico, instituto tecnológico de chiná. he is interested in the diversity and conservation of avifauna in yucatan peninsula, mexico. ángel j ríos-oviedo he is a bachelor student of biology at the tecnológico nacional de méxico, instituto tecnológico de chiná. he is interested in the diversity and conservation of pollinators in yucatan peninsula, mexico. william cetzal-ix https://orcid.org/0000-0003-4276-6664 phd, focused in systematics, taxonomy and conservation of neotropical orchids and in �oristic studies of indicator species for conservation of forest of south-eastern mexico. alto interested in melliferous plants to increase honey in the apiculture industry in the yucatan peninsula, mexico. saikat kumar basu https://orcid.org/0000-0001-7305-4817 traditionally trained in botany (plant sciences) and specialising in microbiology, works actively in various areas of plant sciences and environmental conservation. �e author works extensively on forage crops like forage legumes and grasses, medicinal herb and spice crops like fenugreek. currently he is also working in areas of pollinator insect conservation, integrated habitat development and on establishing pollinator sanctuaries in various agroclimatic regions. c onstruction of the nest of am azilia rutila d e lattre (trochillidae) and its antipredatory defensive strategy in a m edium deciduous forest in c am peche, m exico 202 annales universitatis paedagogicae cracoviensis studia naturae, 4: 202–206, 2019, issn 2543-8832 28th meeting of the european vegetation survey “vegetation diversity and global change”, september 2–6, 2019, madrid, spain 28 spotkanie z europejskimi badaniami nad roślinnością „różnorodność roślinności i zmiany globalne”, 2–6 września 2019, madryt, hiszpania in madrid, from 2 to september 6, 2019, the 28th international conference was took place as part of the so-called 28th meeting of the european vegetation survey “vegetation diversity and global change”. �e main organiser of the conference was the institute of pharmacology, pharmacognisation and botany of the complutense university in madrid. �e members of the scienti�c committee were: phd emiliano agrillo and phd fabio attorre (department of plant biology, university of la sapienza in rome), phd andraž čarni and phd jovan hadži (institute of biology, research center of the slovenian academy of sciences and arts of ljubljana), prof. milan chytrý (department of botany and zoology, university of brno), phd monika janišová (institute of botany, slovak academy of sciences, banská bystrica), phd maría pilar rodríguez-rojo (institute of environmental sciences, university of castilla-la mancha, toledo), prof. john rodwell (environmental consultant, lancaster), prof. joop schaminée (group for the protection of nature and plant ecology, university of wageningen). w  dniach od 2 do 6 września 2019 roku, w  madrycie, odbyła się już po raz 28 międzynarodowa konferencja w  ramach tzw. spotkań dotyczących badań roślinności, pt. „różnorodność roślinności i zmiany globalne”. głównym organizatorem konferencji był instytut farmakologii, farmakognozji i  botaniki uniwersytetu complutense w madrycie. wśród członków komitetu naukowego znaleźli się: dr emiliano agrillo i dr fabio attorre (zakład biologii roślin, uniwersytet la sapienza w rzymie), dr andraž čarni i  dr jovan hadži (instytut biologii, centrum badawcze słoweńskiej akademii nauk i  sztuki z  ljubljana), prof. milan chytrý (zakład botaniki i zoologii, uniwersytet w brnie), dr monika janišová (instytut botaniki, słowacka akademia nauk, banská bystrica), dr maría pilar rodríguez-rojo (instytut nauk o  środowisku, uniwersytet castilla-la mancha, toledo), prof. john rodwell (konsultant ekologiczny, lancaster), prof. joop schaminée (grupa ochrony przyrody i ekologii roślin, uniwersytet wageningen). 203 r eportsduring the ceremonial opening of the conference (september 3, 2019) plenary lecture entitled “linking above and below ground plant community responses: a melting pot of interactions and soil heterogeneity” as presented by phd adrian escudo (rey juan carlos university of mostoles). a�er the of�cial greeting of the guests, the participants were invited to participate in the �rst lecture session on the subject of “sand-dune and halophilous vegetation”. �e speakers presented, among others, issues from research on coastal dunes ecosystems in central italy, the conservation status of dune habitats in two contrasting natura 2000 areas, overview of sandy vegetation of the pannonian and western pontic region and a syntaxonomic approach to halophytic communities from western europe. a�er a short co�ee break, in the second session entitled “high mountain vegetation”, the speakers discussed issues related to: the di�erentiation of observational errors and the e�ects of climate change in long-term monitoring of the composition and abundance of high-mountain plant species, biogeography of alpine plant communities in southern europe, the coenological and syntaxonomical features of relict populations of two salix species in the high apenin zone, with mountain tundra vegetation, as well as the e�ect of ozone on mediterranean alpine meadows in the guadarrama range (central spain). in the a�ernoon, the �rst poster session took place in which participants presented a wide thematic spectrum in the �eld of vegetation research. of particular interest were the issues concerning: development of datapodczas uroczystego otwarcia konferencji (3. 09. 2019) wykład plenarny pt. „połączenie nad i  podziemnych odpowiedzi zbiorowisk roślinnych: tygiel interakcji a hetreogeniczność gleby” wygłosił dr adrian escudo (uniwersytet rey juan carlos z mostoles). po o�cjalnym przywitaniu przybyłych gości, uczestnicy zostali zaproszeni do udziału w  pierwszej sesji wykładowej, obejmującej tematykę „roślinności wydm piaszczystych i halo�tów”. prelegenci zaprezentowali, m.in. zagadnienia z badań o ekosystemach wydm przybrzeżnych w  środkowych włoszech, stan ochrony siedlisk wydmowych na dwóch kontrastujących obszarach natura 2000, przegląd roślinności psamo�lnej w regionie panońskim oraz syntaksonomiczne podejście do halo�tycznych zbiorowisk z europy zachodniej. po krótkiej przerwie kawowej, w  drugiej sesji pt. „roślinność wysokogórska”, prelegenci omawiali zagadnienia związane: ze różnicowaniem błędów obserwatorskich i  skutków zmian klimatu w  długoterminowym monitorowaniu składu i liczebności gatunków roślin wysokogórskich, biogeogra�ą zbiorowisk roślin alpejskich w południowej europie, cechami cenotycznymi i  składniowymi reliktowych populacji dwóch gatunków salix w stre�e wysokogórskiej apeninów, z  roślinnością tundry górskiej, a także wpływem ozonu na śródziemnomorskie łąki wysokogórskie w  paśmie guadarrama (centralna hiszpania). w  godzinach popołudniowych, odbyła się pierwsza sesja plakatowa, w  której uczestnicy prezentowali szerokie spektrum tematyczne z  zakresu badań nad roślinnością. szczególnym zainteresowaniem cieszyły się 204 r ep or ts bases for assessing the conservation status of habitats on the example of the “vegfrance” database, how to improve remote sensing research using knowledge of vegetation, classi�cation and ecological diversity, on the example of dry grassland habitats of ukraine, as well as the development of a new system for describing diversity dry grasslands in poland. in the third plenary session entitled “assessment and conservation of european habitats” the lectures concerned: review of endangered and endemic species associated with di�erent types of habitats in europe and monitoring the status of natura 2000 habitats in flanders. a�er the co�ee break, in the second part of the session presented were: identi�cation of habitats using satellite images, implementation of the natura 2000 program in albania and the diversity of plant communities of the strazhata hill in central and northern bulgaria. �e next day of the conference (september 4, 2019) three thematic sessions took place: “vegetation patterns in the palaearctic”, “methods and databases for vegetation studies”, “mediterranean and thermophilous forests”. during the lectures presented were, among others, a new method and application for android as a probabilistic key to identify types of vegetation in the �eld, modeling the composition of vegetation and their richness in spatial scales and environmental gradients in the central apennines, native forest dominance patterns in the iberian peninsula, a review of types and habitats of thermophilic vegetation on the edge of ukrainian forests, as well as �oristic and coenotic diversity of vyatka-kama (tatarstan, russia). on the zagadnienia dotyczące: opracowania baz danych do oceny stanu zachowania siedlisk na przykładzie bazy “vegfrance”, jak ulepszyć badania teledetekcyjne wykorzystując wiedzę o  roślinności, klasy�kacji oraz zróżnicowaniu ekologicznym, na przykładzie suchych siedlisk trawiastych ukrainy, a  także opracowania nowego systemu opisu różnorodności suchych muraw w polsce. w  trzeciej sesji plenarnej zatytułowanej „ocena i  ochrona siedlisk europejskich” referaty dotyczyły: przeglądu zagrożonych i  endemicznych gatunków, związanych z  różnymi typami siedlisk w  europie oraz monitorowania stanu siedlisk natura 2000 we flandrii. po przerwie kawowej, w drugiej części sesji przedstawiono problematykę: identy�kacji siedlisk za pomocą zdjęć satelitarnych, realizacji programu natura 2000 w  albanii oraz różnorodności zbiorowisk roślinnych wzgórza strazhata w środkowej i północnej części bułgarii. następnego dnia konferencji (4. 09. 2019) odbyły się trzy sesje tematyczne: „wzory wegetacji w palearktyce”, „metody i  bazy danych do badań roślinności” oraz „lasy śródziemnomorskie i  ciepłolubne”. w trakcie wykładów zaprezentowano, m.in. nową metodę i aplikację na androida, jako probabilistyczny klucz do identy�kacji typów roślinności w terenie, modelowanie składu roślinności i  ich bogactwa w  skalach przestrzennych i  gradientach środowiskowych w  apeninach środkowych, rodzime wzorce dominacji lasów na półwyspie iberyjskim, przegląd rodzajów i  siedlisk roślinności termo�lnej na skraju ukraińskich lasów, a  także różnorodność 205 r eportssame day, in the second poster session, participants discussed topics such as: the importance of small-leaved forests in the vegetation cover in the central part of the russian plain, �oristic patterns in the altitude gradient in the mongolian part of altai, between steppe and high mountain belts, the state of conifers in the eastern alps and changes in water-peat vegetation in lower silesian forests (poland). on the penultimate day of the conference (september 5, 2019) in plenary sessions: “wetlands, riparian and aquatic vegetation” and “vegetation dynamics and succession in di�erent habitats” the speakers presented topics related to: the succession of peat bogs in the engure lake nature park in latvia, the mediswet project as an opportunity to improve the state of knowledge and methods of protecting wetlands in sicily and sardinia, research on vegetation in the güímar valley in tenerife (canary islands), phenological trends in plant communities dominated by grasses in mediterranean areas, the impact of drought on the changing composition of mediterranean forests and plant succession in post-industrial areas. in the evening, a�er the o�cial closing of the meeting, it was time for a celebratory dinner, during which scientists had the opportunity to make friends with colleagues from various centers, which will certainly result in joint research projects. �e dinner ended with a �amenco dance show and good fun. on friday morning (september 6, 2019), �eld sessions were organised for interested conference participants, which became both an opportunity to explore knowledge and to learn about the natural qualities of high mountain areas in the sierra de guadarrama �orystyczną i  cenotyczną vyatka-kama (tatarstan, federacja rosyjska). w  tym samym dniu, w  drugiej sesji plakatowej uczestnicy dyskutowali, m.in. na temat: znaczenia lasów drobnolistnych w  pokrywie roślinności w  środkowej części równiny rosyjskiej, wzorów �orystycznych w  gradiencie wysokościowym w  mongolskiej części ałtaju, między pasami stepowymi a wysokogórskimi, stanu drzew iglastych we wschodnich alpach oraz zmian roślinności wodno-torfowej w  borach dolnośląskich (polska). w przedostatnim dniu konferencji (5. 09. 2019) w sesjach plenarnych: „mokradła, roślinność nadbrzeżna i wodna” i „dynamika roślinności i  sukcesja w  różnych siedliskach”, prelegenci poruszali tematy związane z: sukcesją torfowisk w  parku przyrody engure lake na łotwie, projektem mediswet, jako szansą na poprawę stanu wiedzy i metod ochrony mokradeł na sycylii i  sardynii, badaniami roślinności w  dolinie güímar na tenery�e (wyspy kanaryjskie), trendami fenologicznymi w  zbiorowiskach roślinnych zdominowanych przez trawy na obszarach śródziemnomorskich, wpływem suszy na zmieniający się skład lasów śródziemnomorskich oraz sukcesją roślin na terenach poprzemysłowych. wieczorem, po o�cjalnym zakończeniu obrad, przyszedł czas na uroczystą kolację, podczas której naukowcy mieli możliwość nawiązywania nowych znajomości z  kolegami z  różnych ośrodków, co z  pewnością zaowocuje wspólnymi projektami badawczymi. kolacja zakończyła się pokazem tańców �amenco i wspólną zabawą. w  piątkowy poranek (6. 09. 2019), dla zainteresowanych uczestników konferencji 206 r ep or ts national park (central spain). during this session, various plant communities were admired – from open oak forests with quercus rotundifolia lam., through shelf formations of limestone or gypsum soils and cli�s in the miocene sediments near the el campillo lagoon, to di�erent groups of halophytes in the el salobral nature reserve. �e vegetation of west-iberian areas was also studied, including oak formations q. pyrenaica willd. and evergreen q. suber l. and q. rotundifolia. �is year’s 28th european meeting on vegetation research was attended by over 170 scientists from various european centres, among others from the czech republic, slovakia, poland, ukraine, lithuania, latvia, russia, italy, france, spain and germany. �is meeting was certainly an opportunity to present the results of research on the current state of vegetation in eurasia. it gave the opportunity to exchange views on topics of modern research methodology, collection and processing of phytosociological data in relation to their habitats. for the purposes of this conference, information on the conference has been published in an electronic version at: http://evs2019madrid.es/. zorganizowano sesje terenowe, które stały się, zarówno okazją do zgłębiania wiedzy, jak i  do poznania walorów przyrodniczych terenów wysokogórskich w  parku narodowym sierra de guadarrama (centralna hiszpania). w  trakcie tej sesji, podziwiano różne zbiorowiska roślinne – od otwartych lasów dębowych z  quercus rotundifolia lam., poprzez szelfowe formacje wapiennych lub gipsowych gleb i klifów w osadach mioceńskich w  pobliżu laguny el campillo, do różnych grup halo�tów w rezerwacie przyrody el salobral. zapoznano się również z roślinnością terenów zachodnio-iberyjskich, w tym m.in. formacjami dębu q. pyrenaica willd. oraz zimozielonych q. suber l., i q. rotundifolia. w  tegorocznym 28 europejskim spotkaniu, dotyczącym badań roślinności, wzięło udział ponad 170 naukowców z  różnych ośrodków europy, m.in. z  czech, słowacji, polski, ukrainy, litwy, łotwy, rosji, włoch, francji, hiszpanii i  niemiec. spotkanie to z  pewnością było okazją do prezentacji rezultatów badań nad obecnym stanem roślinności w  eurazji. dało możliwość wymiany poglądów na tematy współczesnej metodyki badań, gromadzenia i  przetwarzania danych �tosocjologicznych, w powiązaniu z ich siedliskami. na potrzeby niniejszej konferencji, informacje dotyczące zjazdu zostały opublikowane w  wersji elektronicznej pod adresem: http://evs2019madrid.es/. ingrid turisová department of biology and ecology, faculty of natural sciences, matej bel university, tajovského 40, banská bystrica 974 01, slovakia, *ingrid.turisova@umb.sk 25 annales universitatis paedagogicae cracoviensis studia naturae, 5: 25–33, 2020, issn 2543-8832 doi: 10.24917/25438832.5.2 anna sołtys-lelek1*, wojciech gruszka2 1ojców national park, 32-045 sułoszowa, ojców 9, poland; *ana_soltys@wp.pl 2department of biological sciences, faculty of physical culture in gorzów wielkopolski., poznań university school of physical education, estkowskiego 13, 66-400 gorzów wielkopolski, poland occurrence of rosa blanda ait. (rosaceae) in poland introduction rosa blanda ait. (sect. cinnamomeae) is a north american species of rose, growing in canada and the united states. its natural range extends from quebec to ontario, south to kansas, and east to missouri and ohio (stephens, 1973; gleason, cronquist, 1991; lewis et al., 2014). however, it has been cultivated and grown wild in poland, finland, austria, germany, and france (tutin et al., 1968; zieliński, 1987; czarna, 2016; rosa blanda ait.…). mirek et al. (2002) granted r. blanda the status of an anthropophyte permanently established in polish �ora. fig. 2. flower of rosa blanda ait. (photo. 2013, w. gruszka, dendrological garden in przelewice) a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 26 r. blanda is morphologically similar to the native species r. majalis herrm., for which it is a vicarious species (zieliński, 1987). characteristic features of this rose are needle-like thorns at the base of long shoots; �owering stems usually without thorns; lea�ets blade elliptic or ovate, sometimes obovate; margins 1-serrate; teeth 10–26 per side; acute; and in�orescences corymbs, 1–5(–10)-�owered (stephens, 1973; zieliński, 1987; lewis et al., 2014; minnesota wild�owers a �eld…; fig. 1 – appendix 1, fig. 2). �ere is almost no information on the distribution of this species in the polish botanical literature, probably due to the fact that, outside of cultivation in botanical gardens or arboreta, few specimens are seen. in 2017, during �eld research carried out in the vicinity of gorzów wielkopolski, the authors came across a site where r. blanda was growing in 17 di�erent-sized clusters. �is �nding provided the impetus to undertake this research study aimed at presenting all the r. blanda sites in poland so far identi�ed and determining its ability to establish and spread. materials and methods in order to establish the distribution of spontaneous sites of this species in the country, the authors, in addition to their own observations, consulted collections in the largest national herbariums via e-mail. information was obtained from the following herbariums: the herbarium of the białowieża geobotanical station at the university of warsaw (bsg), the herbarium of the institute of dendrology of the polish academy of sciences in kórnik (kor), the scienti�c herbarium of the university of silesia (ktu), the herbarium of the institute of botany at the jagiellonian university in cracow (kra), the herbarium of vascular plants at the institute of botany of the polish academy of sciences (kram), the herbarium of the maria curie-skłodowska university (lbl), the herbarium of the faculty of biology at the university of warsaw (wa), and the herbarium of the department of taxonomy and plant geography at the nicolaus copernicus university in toruń (trn). data published in papers by zieliński (1977, 1987) and czarna (2009, 2011, 2016) and unpublished information made available by researchers, prof. błażej gierczyk from the adam mickiewicz university in poznań and lucjan rutkowski, phd from the nicolaus copernicus university in toruń, were also included. all the collected data are shown on the map (fig. 3). �e list of sites and the distribution map includes only sites that have been created spontaneously or those that are remnants of an old local cultivation (feral). sites of specimens in private collections, botanical gardens, arboreta, etc. have not been recorded. �e sites are located in relation to atpol (atlas of poland) squares, 10 × 10 km, (zając, zając, 2001) and the nearest village. abbreviations used: obs. – observation, com. – compartment. 27 o ccurrence of r osa blanda a it. (r osaceae) in p oland glossary of geographical names in article �e list includes geographical coordinates for the villages in article: białowieża 52°42ʹ04,9ʺn; 23°52ʹ10,2ʺe (gc65) budachów 52°9ʹ14,88ʺn; 15°5ʹ3,56ʺe (ad26) dobrosułów 52°11ʹ37ʺn; 15°07ʹ28,99ʺe (ad26) konin 52°13ʹ39ʹʹn; 18°15ʹ41ʹʹe (cd27) poznań 52°24ʹ52ʹʹn; 16°55ʹ16ʹʹe (bc98, bc99, bd08, bd18, bd19) puck 54°42‘59,93“n; 18°24‘27,21“e (ca49) pyzdry 52°10ʹ13ʹʹn; 17°41ʹ24ʹʹe (cd24) skwierzyna 52°35ʹ46,58ʺn; 15°30ʹ10,38ʺe (ac89) strzelce krajeńskie 52°52ʹ43,22ʺn; 15°31ʹ58,16ʺe (ac49) fig. 3. current distribution of rosa blanda ait. in poland based on the atpol grid square system; ● – squares in which single sites were recorded for the species, ▲ – squares in which two sites were recorded for the species, ■ – squares in which four sites were recorded for the species a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 28 szczecin 53°26‘n; 14°34‘e (ab74) śrem 52°05‘n; 17°01‘e (bd39) toruń 53°02ʹn; 18°37ʹe (dc30) wielisławice 52°53ʹ51,28ʺn; 15°27ʹ58,91ʺe (ac49) zemsko 52°16‘20,0“n; 16°34‘45,4“e (ac89) results unpublished sites in atpol grid squares ac4944, ac4955: between strzelce krajeńskie and wielisławice, roadside trench, 2017, w. gruszka, ad2615: dobrosułów near torzym, 2010, a. czarna, (kor 52896), bc9897: poznań, 100 m from the intersection of solidarności avenue and dojazd street, roadside, obs. 2019, b. gierczyk (e-mail information), bc9961: poznań, bożywoja street, hedges, 2010, j. zieliński, (kor 48896), bd0814: poznań, near żwirki and wigury ii family allotment gardens, 800 m from bukowska street, obs. 2019, b. gierczyk (e-mail information), bd0831: poznań, at owczej street, near marceliński forest, com. 85lx, obs. 2019, b. gierczyk (e-mail information), bd1883: poznań, greater poland national park, com. 76c, woodland, 2004, a. purcel, 2004 (kor 47092), cd27: konin, roadside slope, 1975, k. balcerzak, 1975 (kor 8975), dc3021: toruń, pawia street, remnant of former local cultivation, obs. 2019, l. rutkowski (e-mail information), and dc3061: toruń, sienkiewicza street, remnant of former local cultivation, obs. 2019, l. rutkowski (e-mail information). published sites in atpol grid squares ab74: szczecin area, (zieliński, 1987), ac89: between skwierzyna and zemsko, remnant of former local cultivation (zieliński, 1977), ad2662: budachów, on the road from osiecznica (zieliński, 1977), ad3624: lubuskie lakeland, before dobrosołów from budachowo, remnant of an old hedge, 1974, j. zieliński (kor 6633, zieliński, 1977), bd0845: poznań, old górczyński cemetery, remnant of former local cultivation (czarna et al., 2011; czarna 2016), bd0853: poznań, near junikowska street, side of fence of plot no. 54, b. gierczyk 11.06.2006 (rosa blanda aiton…), bd19: poznań area, (zieliński, 1987), 29 bd39: śrem, hedges at the “śrem” iron foundry (czarna, 2009), ca49: puck area, (zieliński, 1987), cd2400: pyzdry, old cemetery, remnant of former local cultivation (czarna, 2016), and gc6508: białowieża, białowieski national park, polana białowieska, 1991, ł. łuczaj, herbarium bsg vascular plants (catalogue number a20192) published in: global biodiversity information facility database (rosa blanda ait.…). species were previously generally reported in the białowieska forest (zieliński, 1987). discussion rosa blanda ait. has the status of an established, non-invasive kenophyte in poland. its introduction into poland is estimated to be 1817 (tokarska-guzik et al., 2012). since the second half of the 20th century, information about its sporadic spread beyond cultivation in gardens, arboreta, or botanical gardens has appeared in the literature (zieliński, 1977, 1987). r. blanda, as a foreign escapee species, is a rare species in poland. a total of 22 sites, created spontaneously or as a remnant of old local cultivations, were identi�ed. its current range is mainly restricted to central and north-western poland, with isolated sites in the north and east of the country (fig. 3). however, it can be assumed that the distribution of the species in poland is probably in�uenced by insu�cient �eld research, particularly in the central and eastern part of the country. �erefore, it may be expected that the list of published sites will be supplemented with additional records in the future. �e spread of this species within the country may be facilitated by its wide tolerance, both in terms of climate and habitat conditions. r. blanda ait. is a frost-resistant species; it tolerates temperatures to −42.8°c (usda zones 2 to 6; united states department of agriculture). it is also shade resistant. in polish climatic conditions it blooms abundantly and bears fruit; it also propagates very e�ectively by stolons (authors’ observation). usually occurring in sunny, dry to moist sites (minnesota wild�owers a �eld…), it tolerates dry, sandy habitats that are poor in nutrients, such as roadsides (stephen, 1973) and heavy clay soils. in terms of ph, it grows on acidic, alkaline, and neutral soils (plants for a future…). most o�en, it occurs in anthropogenic, partially transformed habitats (tokarska et al., 2012). although rose has been included in the alien species list for poland (gatunki obce w polsce…) its negative impact on native species of �ora or other elements of the habitats in which it grows has not yet been described. nevertheless, the possibility that such impacts exist cannot be excluded. for many alien species a so-called delay phase, o ccurrence of r osa blanda a it. (r osaceae) in p oland a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 30 also known in the literature as a lag phase, has been observed between the appearance of the species and the discovery of its invasive behaviour (hobbs, humphries, 1995; richardson, pyšek, 2006). �erefore, it is important to know the current distribution of the plant’s spontaneous sites and, in the long term, to undertake research aimed at determining its impact on the native �ora. furthermore, it has been proven that the species has a tendency to spontaneously cross with other species, for example r. acicularis lindl., r. carolina l., and r. virginiana mill. (lewis, 2016; lewis, elvin-lewis, 2017). a dangerous phenomenon may have been described by mercure and bruneau (2008) when they con�rmed spontaneous crossing between r. blanda and r. rugosa �unb. �ese are two species with great potential for range expansion that could pose a serious threat to the native �ora. although r. rugosa is widespread throughout poland (and its abundance is increasing), as mentioned earlier, r. blanda occurs in few sites outside of cultivation. however, it should be noted that the hybrid form between r. rugosa and most probably r. blanda was observed by zieliński (2014) in poland in 1988; not as an f1 hybrid but as a next-generation segregant. in this context, it should be mentioned that r. blanda occurs in two places together with r. rugosa in the site between strzelce krajeńskie and wielisławice, so the emergence of a hybrid between these species seems possible. acknowledgements �e authors would like to thank prof. jerzy zieliński for the data sent from the herbarium of the institute of dendrology pas in kórnik. we would also like to thank lucjan rutkowski, phd from the nicolaus copernicus university in toruń and prof. błażej gierczyk, from the adam mickiewicz university in poznań for making available their unpublished data. con�ict of interest �e authors declare no con�ict of interest related to this article. references czarna, a. (2009). rośliny naczyniowe środkowej wielkopolski (vascular plants of central wielkopolska). poznań: wydawnictwo uniwersytetu przyrodniczego. 184 pp. [in polish] czarna, a. (2016). roses (rosa spp.) in old cemeteries in the wielkopolska region (poland). annales universitatis mariae curie-skłodowska, sectio c, 71(2), 7–31. https://doi.org/10.17951/c.2016.71.2.7 czarna, a., woźnicka, a., maj, m., morozowska, m. (2011). flora of vascular plants of selected poznań cemeteries. acta agrobotanica, 64, 123–140. https://doi.org/0.5586/aa.2011.054 gatunki obce w polsce (alien species in poland). http://www.iop.krakow.pl/ias [in polish] gleason, h.a., cronquist, a. (1991). manual of vascular plants of north-eastern united states and adjacent canada (2nd ed.). new york: �e new york botanical garden. 910 pp. hobbs, r.j., humphries, s.e. (1995). an integrated approach to the ecology and management of plant invasions. conservational biology, 9, 761–770. https://doi.org/10.1046/j.1523-1739.1995.09040761.x lewis, w.h. (2016). nomenclatural novelties in rosa (rosaceae) subgenus rosa recognized in north america. novon: a journal for botanical nomenclature, 25(1), 22–46. https://doi.org/10.3417/2016018 lewis, w.h., elvin-lewis, m. (2017). new records and range extensions for rosa (rosaceae) in north america. journal of the botanical research institute of texas, 11(1), 185–191. 31 lewis, w.h., ertter, b., bruneau, a. (2014). rosa. in: flora of north america editorial committee (eds.), 1993+. flora of north america north of mexico. 19+ vols. new york and oxford. vol. 9. mercure, m., bruneau, a. (2008). hybridization between the escaped rosa rugosa (rosaceae) and native r. blanda in eastern north america. american journal of botany, 95(5), 597–607. https://doi. org/10.3732/ajb.2007385 minnesota wild�owers a �eld guide to the �ora of minnesota. https://www.minnesotawild�owers.info mirek, z., piękoś-mirkowa, h., zając, a., zając, m. (2002). flowering plants and pteriodophytes of poland. a checklist.. kraków: władysław szafer institute of botany. polish academy of sciences. 442 pp. plants for a future, earth, plants, people https://pfaf.org richardson, d.m., pyšek, p. (2006). plant invasions: merging the concepts of species invasiveness and community invasibility. progress in physical geography: earth and environment, 30, 409–43. https:// doi.org/10.1191%2f0309133306pp490pr rosa blanda ait. in: global biodiversity information facility (gbif) database. https://www.gbif.org/species/3002372 rosa blanda aiton, róża labradorska. w: flora polski atlas-roslin.pl. rośliny naczyniowe dziko rosnące, uprawne, ozdobne gruntowe, mszaki (rosa blanda aiton, a smooth rose. in: polish �ora atlas-roslin. pl. vascular plants growing wild, cultivated, ornamental, ground, bryophytes). https://atlas-roslin.pl/ gatunki/rosa_blanda.html [in polish] stephens, h.a. (1973). woody plants of the north central plains. lawrence: university of kansas press. 530 pp. tokarska-guzik, b., dajdok, z., zając, m., zając, a., urbisz, a., danielewicz, w., hołdyński, c. (2012). rośliny obcego pochodzenia w polsce ze szczególnym uwzględnieniem gatunków inwazyjnych (plants of foreign origin in poland with particular emphasis on invasive species). warszawa: generalna dyrekcja ochrony środowiska. tutin, t.g., heywood, v.h., burges, n.a., moore, d.m., valentine, d.h., walters, s.m., webb, d.a. (1968). flora europaea. rosaceae to umbelliferae. volume 2. cambridge: cambridge university press. 454 pp. united states department of agriculture. https://planthardiness.ars.usda.gov/phzmweb/ (access: 2020). zając, a., zając, m. (2001). distribution atlas of vascular plants in poland. kraków: laboratory of computer chorology, institute of botany, jagiellonian university. zieliński, j. (1977). roses of the lubusz upland. fragmenta floristica et geobotanica, 23(2), 125–140. zieliński, j. (1987). rosa l. in: a. jasiewicz (ed.), flora of poland, 5, 48 pp. kraków: institute of botany, polish academy of sciences. zieliński, j. (2014). dwa różne kultywary o nazwie rosa ‘poznań’ (two di�erent cultivars called rosa ‘poznań’). rocznik polskiego towarzystwa dendrologicznego, 62, 63–66. [in polish] abstract �e article presents the smooth rose, rosa blanda ait. distribution in poland based on literature data, available herbarium materials, and shared unpublished data. r. blanda is a north american species that is cultivated and has become wild throughout poland, where it has the status of a domestic, non-invasive kenophyte. �is species is rare in poland. only 22 sites have been identi�ed (mainly in the central and northwestern part of the country), as created spontaneously or as remaining from old, local cultivations. however, it cannot be excluded that this species will become invasive in the future and will negatively a�ect native �ora species, especially through its ability to cross with other rose species (including invasive r. rugosa �unb.). key words: rosaceae, distribution of stands, invasive potential, kenophytes received: [2020.07.02] accepted: [2020.09.06] o ccurrence of r osa blanda a it. (r osaceae) in p oland a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 32 występowanie róży labradorskiej, rosa blanda ait. (rosaceae) w polsce streszczenie w artykule przedstawiono rozmieszczenie w polsce róży labradorskiej rosa blanda ait. na podstawie danych z literatury, dostępnych materiałów zielnikowych, a także udostępnionych danych niepublikowanych. r. blanda jest gatunkiem północnoamerykańskim, który jest uprawiany i zdziczał na terytorium całej polski, gdzie posiada status zadomowionego, nieinwazyjnego keno�ta. gatunek ten jest rzadki w polsce. zidenty�kowano tylko 22 stanowiska (głównie w środkowej i północno-zachodniej części kraju), powstałe prawdopodobnie spontanicznie lub będące pozostałością starych, lokalnych upraw. nie można jednak wykluczyć w przyszłości zachowania inwazyjnego i negatywnego wpływu na rodzime gatunki �ory, zwłaszcza że gatunek ten ma tendencję do krzyżowania się z innymi gatunkami (w tym inwazyjną r. rugosa �unb.). słowa kluczowe: rosaceae, rozmieszczenie stanowisk, potencjał inwazyjny, keno�ty information on the authors anna sołtys-lelek https://orcid.org/0000-0002-9595-3167 author of numerous scienti�c and popular science studies in the �eld of botany and environmental protection. her research interests relate particularly to the critical type of rose (rosa) and hawthorn (crataegus). member of the polish and slovak botanical society. wojciech gruszka http://orcid.org/0000-0002-6229-8397 author and co-author of scienti�c and popular science studies in lichenology and botany. his main research interests relate to the ecology and protection of lichen. in addition, he participates in the research on the distribution of representatives of the rose (rosa) and hawthorn (crataegus) species in poland. 33 fig. 1. smooth rose, rosa blanda ait. (photo. 2018, a. sołtys-lelek, specimen from poland, between strzelce krajeńskie and wielisławice, 2017, w. gruszka); a – part of long shoot, b – part of fruiting short shoot, c – stipule, d, e – fruit, f – fruit with glandular sepal, g – leaf. solid bar = 1 cm appendix 1 o ccurrence of r osa blanda a it. (r osaceae) in p oland 91 annales universitatis paedagogicae cracoviensis studia naturae, 4: 91–102, 2019, issn 2543-8832 doi: 10.24917/25438832.4.5 roland kopaliani1, temur gvinianidze1*, rezo jabnidze2 1akaki tsereteli state university, kutaisi, georgia; *temurigvinianidze@gmail.com 2shota rustaveli state university, batumi, georgia the bio-flavanoid concentrate of vitis vinifera l. ‘red aladasturi’ introduction flavanoids are the largest group of phenolic compounds, and owing to their high biological activity, they are o�en referred to as bio�avanoids. de�ciency of �avonoids in the human body manifests with the following symptoms: the general weakness and chronic fatigue, nasal hemorrhage, reduced immunity recurrent colds and infections, the formation of hematomas and vesication, the reduction in vascular conductance and elasticity, pains in the upper and lower extremities during movement, and so on (kurkin et al., 2013; yilmaz, тоledo, 2004; gvinianidze et al., 2019). �ere is extensive literature on high antioxidant activity of bio�avonoid-rich coloured grape seed and skin hydrophilic extract and red and white wine produced from it, as well as on inactivation of free radcals (demrow et al., 1995; gvinianidze, gvinianidze, 2018). in 2011, the vital (fred hutchinson cancer research center, seattle, washington) published a study on prostate cancer, and 35.239 men aged 50–76 volunteered for this study. it was found that patients regularly consuming grape-seed hydrophilic extracts were 41% less likely to su�er from prostate cancer than patients taking other drugs such as chondroitin, coenzyme q10, �sh oil, ginseng, ginkgo biloba, garlic, and glucosamine and palmetto (zharskaya et al., 2014). vitis vinifera l. ‘red aladasturi’ is a georgian, aboriginal, late-ripening, industrial cultivar, mostly common in the viticulture and winemaking zones of imereti and guria. grapes ripen in late october and early november, and in full maturity, sugar content reaches 19.5–24.5%, and titrable acidity varies in the range of 8.0–9.3 g/dm3 (ketskhoveli et al., 1960). it has been established that grape raw materials grown in di�erent micro-zones di�er in their sensory characteristics, uvological and chemical composition, as well as in antioxidant, antiradical and antimicrobial properties (darra et al., 2012; kvesitadze et al., 2019). secondary resources accrued from the processing of coloured grapes (in r ol an d k op al ia ni , t em ur g vi ni an id ze , r ez o ja bn id ze 92 the form of skin and stone), by the contents of biologically active compounds have barely analogs in the autotrophic organisms, and they are not of less value products than wine itself. only 9–12% of the total amount of phenolic compounds is contained in grape juice and pulp, accounting for 75–81% of the total mass of raceme, while the remaining 88–91% of phenolic compounds is mostly localised in the skin and stone, the mass of which is only 18–25% of raceme. �is clearly shows how rich the biologically active compounds are in the solid parts of colored grapes, as well as how big is their role in the production of powerful antioxidant polyphenolic concentrates. accordingly, research in this �eld is of high relevance. �e aim of the study was to investigate a polyphenolic complex and antioxidant activity of secondary resources remained a�er the initial processing of v. vinifera ‘red aladasturi’ grapes growing in imereti and di�erent micro-zones, as well as to explore the possibilities of using them for the production of drastic, antioxidant polyphenolic concentrates. �e solid parts of colored grapes, with the content of biologically active compounds are the best raw materials for the production of therapeutic extracts and concentrates to treat various pathologies (gvinianidze et al., 2017, 2018; morandi vuolo et al., 2019). materials and methods object of study research covered the raw materials of vitis vinifera ‘red aladasturi’ grape from different vineyards of the imereti viticulture and winemaking zone, particularly: sample n1 – lifnari vineyards (rokhi village, baghdati district, 120–160 m above sea level), sample n2 – sviri vineyards (sviri village, zestafoni district, 230–250 m above sea level) and sample n3 – bagineti vineyards (bagineti village, vani district, 580–600 m above sea level). research also covered hydrophilic extracts of grape skin and stone thickened by the vacuum of ‘aladasturi’ coloured grapes raw materials, as well as the concentrates produced from their composition. research methods for research, there were used gravimetric, extractive, spectral and chromatographic methods (singleton et al., 1999; palomino et al., 2000; giusti, ronald, 2001; mensor et al., 2001; kammerer et al., 2004; prior et al., 2005; gómez-alonsoet al., 2007; rajha et al., 2013; benmeziane et al., 2016; gvinianidze et al., 2018). in test samples, we determined: the moisture and solid matter contents by heat-gravitational (gost 28561 90) and refractometric methods. 93 the bio-flavanoid concentrate of vitis vinifera l. ‘r ed a ladasturi’ quantitative analysis of total phenols quantitative analysis of total phenols was performed spectrophotometrically, by folin-ciocalteu reagent. in particular, we extracted the crushed test samples with 75– 81% ethyl alcohol at the temperature of 72–75°c and under conditions of periodic stirring for 6–7 hours. 1 ml of extract obtained, we placed into a 25 ml �ask and added 0.5 ml of h2o, 1 ml of folin-ciocalteu reagent, and settled for 8 minutes at room temperature, then we added 10 ml of 7% na2co3, �lled the �ask with h2o, and settled it for 2 hours at room temperature. the determination was carried out at 750 nm. as a control, we took 1 ml of the appropriate extracting agent and went through the same process. calculation of the data obtained from the determination was carried out on the calibration curve of gallic acid. the total phenol content shall be calculated in accordance with the formula: x = (d × k × v × f) × 1000 /m, where x – the total phenol content, mg/kgგ; d – optical density; k – gallic acid conversion factor; f – solubility; v – the total volume of extract, ml; m – raw materials mass taken for extraction, g. antioxidant activity antioxidant activity in test samples was determined by one of the most common methods – dpph method. dpph is a rapid, simple and accurate test method for determining antioxidant activity. dpph – (c18h12n5o6 m = 394.33) is a stable free radical with maximum absorption at 515–517 nm, and purple-violet coloration of its methanol extracts changes to bright yellow as a result of the recovery. �e reaction occurs in accordance with the following pattern: dpph. + ah → dpph-h + a. dpph. + r. → dpph-r, where ah is an antioxidant and r is a free radical. quanti�cation of total �avonoids was carried out with alcl3 reagent by spectral method – test sample was extracted with 80% ethyl alcohol at the temperature of 70–75°c. 1 ml of extract obtained from the total volume was placed into a 10 ml �ask, then we added 5 ml of h2o, 0.3 ml of 5% nano2 was settled for 5 minutes, and then we added 0.3 ml of 10% alcl3 and settled for 6 minutes, then we added 2 ml of 1n naohr and the determination was performed at 510 nm. as a control, we took 1 ml of the appropriate extracting agent and then went through the same process. r ol an d k op al ia ni , t em ur g vi ni an id ze , r ez o ja bn id ze 94 calculation of the data obtained from the determination was carried out on the rutin calibration curve. �e total �avonoid content shall be calculated in accordance with the formula: x = (d × k × v × f) × 1000 /m; where x – the total �avonoid content, mg/kg; d – optical density; k – rutin conversion factor; f – solubility; v – the total volume of extract, ml; m – raw materials mass taken for extraction, g. monomeric anthocyanins �e course of the ph-di�erential method for quanti�cation of monomeric anthocyanins was as follows: we take test sample from 1 to 5 grams and carry out extraction with 45% ethyl alcohol. �e volume of extract was reduced to 50 or 100 ml according to the extraction quality. from the total volume of extract, we take in two test-tubes 1 ml of extract in each, and add 4 ml of bu�er solution in each. in one test-tube, we add 0.025 m of potassium chloride, and in the other test-tube, we add 0.4 m of sodium acetate, and 20 minutes later, we determine the optical density of the test solutions at 520 nm and 700 nm. quanti�cation of leucoanthocyanins and catechins by spectral method quanti�cation of leucoanthocyanins and catechins by spectral method – extraction of test sample was carried out with 80% ethyl alcohol at the temperatures of 70–75°c. 1 ml taken from the total volume of extract was added with 3 ml of vanillin reagent and, 3 minutes later, we determine the optical density of red test sample at 500 nm. as a control, we shall take 1 ml or 3 ml of vanillin reagent. calculation of the data obtained from the determination was carried out on the (+)catechin calibration curve. �e catechin content shall be calculated in accordance with the formula: x = (d × k × v × f) × 1000 /m; where x – the catechin content, mg/kg; d – optical density; k – 35.0 (+) catechin conversion factor; f – solubility; v – the total volume of extract, ml; m – raw materials mass taken for extraction, g. results and discussion vitis vinifera ‘red aladasturi’ is a late-ripening colored grape cultivar with a very special aroma that reaches full maturity in the second half of november, and the range of aromatic compounds in it increases in proportion with the increase in the sugar content (ketskhoveli et al., 1960). �e area of our concern was represented by polyphenolic compounds, and we were less interested in the sugar and aroma compound contents. accordingly, the grape raw materials 95 were taken during the period of their technical maturity, while phenolic compounds were present in grapes to the extent possible. grape samples were taken on 16 october 2018. �e analysis of the uvological characteristics of individual samples of grape raw materials is given in table 1. tab. 1. uvological characteristics of individual samples of grape raw materials characteristics samples n1 n2 n3 parts of the cluster of grapes [%] juice and �esh 78.60 79.67 79.83 grape stalk 4.71 4.74 4.69 skin 11.87 10.85 10.82 stone 4.48 4.44 4.39 number of seeds in the grain 1–4 solid remains (grape stalk + grape skin + grape stone) 21.06 20.03 19.90 structural indicator 3.74 3.98 4.02 �e study of the uvological characteristics of selected samples showed that structural indicators of all three samples of grapes (the ratio of �esh and juice to solid waste), at both stages of the grape harvest, were almost similar (relatively smaller for sample n1, and relatively larger for sample n3), indicating small di�erences in the quantitative phenolic complex contents in these samples (gvinianidze et al., 2018). we processed samples of grapes raw materials according to the following pattern: (1) identifying qualitative indicators of grapes raw materials; (2) passing grapes raw materials through the dmcsi-type grape clustercomb divider; (3) pressing-out the comb-less must in a basket press and separation of juice; (4) vacuum sublimation drying of juice-less sweet pomace with an initial moisture content of 45–65% to a �nal moisture content of 9–10%; (5) separation of the ‘aladasturi’ cultivar’s skin and stone dried to the moisture content of 9–10%, using tea sorting machine; (6) crushing separately the skin and stone in a micro-mill (tp2 hammer mill) until the fraction of 50–100 µm. �e crushed grape-stone was extracted by two di�erent methods. �e �rst method (grape-stone i – extract): as an extracting agent for extraction of the grape-stone micropowder, we have selected a complex hydrophilic solvent – ethanol containing 40% volumetric alcohol, which was diluted with mineral drinking water “borjomi” whose ph = 3.6–6.3 and mineralisation is in the range of 7–14 g/ dm3. �is mineral water contains sodium (potassium) hydrogen carbonate and boric acid. preliminary experiments have demonstrated that the extracting agent of ethanol diluted with mineral water can successfully replace the extracting agent diluted with water of ethanol containing 40% volumetric alcohol, which is oxidised by hydrochloric acid. the bio-flavanoid concentrate of vitis vinifera l. ‘r ed a ladasturi’ r ol an d k op al ia ni , t em ur g vi ni an id ze , r ez o ja bn id ze 96 tab. 2. biologically active compounds of the grape-stone �uid extract b.a.c. [mg / 100 g/ dry weight basis] stages of super�uid extraction total 1 2 3 4 5 6 7 8 sample n1 phenolic compounds 131.6 977.66 782.9 395.9 344.6 114.1 137.5 95.1 2979.3 flavonoids 290.8 505.6 421.9 310.3 243.6 144.6 219.4 89.0 2225.2 flavan-3-ols 120.6 293.7 414.4 284.9 192.5 104.2 100.4 84.5 1594.2 leukoanthocyanins – 123.4 253.0 148.37 – – – – 524.7 sample n2 phenolic compounds 123.8 943.0 762.1 382.7 332.4 184.9 129.5 87.9 2946.3 flavonoids 289.6 500.2 418.1 308.7 243.4 146.4 219.6 91.8 2217.8 flavan-3-ols 118.0 287.6 406.0 279.0 188.4 101.8 97.2 82.6 1560.6 leukoanthocyanins – 130.6 257.7 153.2 – – – – 541.5 sample n3 phenolic compounds 132.4 953.3 764.2 388.7 343.9 201.9 142.0 99.8 3026.1 flavonoids 292.9 501.1 421.8 310.5 247.7 149.9 219.3 98.5 2242.7 flavan-3-ols 119.7 288.6 403.8 271.1 190.4 105.6 101.1 86.6 1566.9 leukoanthocyanins – 114.3 249.1 147.9 – – – – 511.3 we have determined experimentally the mass ratio of the extracting agent and the grape-stone microdispersed powder, which is 5 l/kg. we have also determined experimentally the extraction parameters: temperature 54–57°c, duration 180–210 minutes, pulsation 4 sec–1 and the pulsation amplitude 2–3 mm. grape-stone ethanol extract at the initial stage, at the temperature of 4–5°c, is subject to sedimentation for 7–9 hours, removal from sediment and �ltration with a wine �lter with plates. tab. 3. biologically active compounds and antioxidant activity of grape-stone extracts with 61–63% of solid matter content composition of hydrophilic extracts biologically active compounds [mg / 100 g on dry weight basis] aoa [%] (f = 100)phenolic compounds flavonoids flavan-3-ols leukoanthocyanins sample n1 n2 n3 3043.76 3014.78 3181.23 2293.94 2276.10 2308.65 1643.90 1597.70 1603.80 567.20 585.90 549.80 51.50 50.60 52.30 �e second method (grape-stone ii – extract): extraction of a bio�avanoid complex from the grape-stone micro-powder was carried out using a supercritical super-�uid extractor (sfe – 100-2-c10) produced by water corporation, where the extracting agent was present together with co2 ethyl alcohol. for maximal extraction of the bio�avanoid complex, we have determined experimentally the optimal �uid 97 extraction parameters: pressure –95 bar, co2 delivery rate – 6.5 kg/h. in addition, the extraction quality was also a�ected by 72% ethanol as co-solvent, whose ratio to co2 was 21–22%. grape-stone �uid extract at the initial stage, at the temperature of 4–5°c, is subject to sedimentation for 7–9 hours, removal from sediment and �ltration with a wine �lter with plates. �e data of the studies of biologically active compounds of the grape-stone super�uid extract are shown in table 2. we have blended the grapestone extracts obtained by both methods at a ratio of 1:1. �e �ltered extract contained 5.2–6.3% of solid matters, and it was concentrated using a vacuum-rotary evaporator at the temperature of 54–57°c to the solid matter content of 63%. �e composition of the concentrated grape-stone hydrophilic exracts was pumped over into the enameled collecting tank, from which test samples have been taken for the analysis on the biologically active compound content and antioxidant activity (tab. 3). from the crushed grape skin, we obtained a hydrophilic liquid extract rich in bio�avonoids in accordance with the following technological scheme (grape skin extract): to e�ectively carry out extraction of anthocyanins from the grape skin, we processed the grape skin micropowder in advance to 0.4% with potassium metabisulphate. as an extracting agent, we selected 36–45% volumetric ethanol processed by 2% citric acid. �e optimal ratio of microdispersed raw materials and the extracting agent we determined experimentally at 3 l/kg. we determined experimentally the extraction optimal parameters: temperature 54–57°c; duration 180–210 minutes; the extraction mass pulsation 4 minutes; the amplitude 5 mm. prior to sedimentation and �ltration, the obtained grape skin exfig. 1. chromatogram of anthocyanins (sample n1) the bio-flavanoid concentrate of vitis vinifera l. ‘r ed a ladasturi’ r ol an d k op al ia ni , t em ur g vi ni an id ze , r ez o ja bn id ze 98 tract was processed by potassium bicarbonate (khco3 – potassium bicarbonate) for correcting 0.7–0.9 g/dm3 excessive acidity. tab. 4. biologically active compounds and antioxidant activity of grape-skin hydrophilic extracts grape skin hydrophilic extract biologically active compounds [mg / 100 g on dry weight basis] aoa [%] (f = 100)phenolic compounds flavonoids flavan-3-ols leukoanthocyanins sample n1 3178.5 646.9 1295.9 2106.1 46.6 n2 3098.8 396.0 1484.5 1302.4 45.3 n3 3265.3 520.6 1667.8 1954.8 47.1 �e obtained extract, at the temperature of 4–5°c, is subject to sedimentation for 7–9 hours, removal from sediment and �ltration with a wine �lter with plates. �e composition of the �ltered grape skin exracts contained 4.5–5.2% of solid matters, and it was concentrated using a vacuum-rotary evaporator at the temperature of 54–57°c to the solid matter content of 61–63%, and then we assessed biologically active compounds and antioxidant activity (tab. 4). figure 1 illustrates the chromatogram of anthocyanins of extract containing 61–63% of solid matters of the micro-dispersed skin of lifnari’s ‘red aladasturi’ cultivar, and �gure 2 illustrates the chromatogram of �avonoids. we have blended the obtained grape-stone ethanol and �uid extracts containing 61–63% of solid maters at an equal ratio (1:1:1) and assessed biologically active compounds and antioxidant activity in this composition (tab. 5). fig. 2. chromatogram of �avonoids (sample n1) 99 tab. 5. biologically active compounds of grape-stone and skin ethanol and �uid extracts with 61–63% of solid matter content sample number biologically active compounds [mg / 100 g on dry weight basis] aoa [%] (f = 100)phenolic compounds flavonoids flavan-3-ols anthocyanins leukoanthocyanins sa m pl e n1 3089.8 1746.3 1529.1 2131.9 572.5 51.4 n2 3044.7 1651.8 1562.6 1332.7 591.0 50.3 n3 3210.6 1714.2 1625.9 2011.8 554.9 52.2 �e second stage of concentration was implemented by method of vacuum-sublimation or lyophilization to 74–75% of the solid matter content and pumped over into the enameled collecting tank, from which test samples have been taken for the analysis. �e results of the assessment of biologically active compounds and antioxidant activity of bio-�avonoid liquid concentrate ‘red aladasturi’ are shown in table 6. tab. 6. biologically active compounds and antioxidant activity of vitis vinifera l. ‘red aladasturi’ biologically active compounds [mg / 100 g on dry weight] sample n1 n2 n3 phenolic compounds flavonoids flavan-3-ols anthocyanins leukoanthocyanins dry matter [%] aoa, (f = 100) [%] 3401.8 1921.2 1682.2 2348.3 578.9 74–75 56.6 3351.1 1808.4 1719.0 1467.9 597.1 74–75 55.31 3533.3 1886.7 1788.9 2213.2 560.6 74–75 57.45 �e studies have shown that the bio-�avonoid concentrates containing 74–75% solid matters of ‘red aladasturi’ obtained from di�erent samples of colored grapes are slightly di�erent from each other in the biologically active compound contents, but all three samples produce the bio-�avonoid concentrates with high antioxidant activity. conclusion it has been studied that the grape-stone and skin hydrophilic extracts of ‘aladastur’ colored grape cultivar’s raw materials taken in the separate viticulture and winemaking micro-zones of imereti and the liquid bio-�avonoid concentrates are characterised by high antioxidant activity (n1 – 56.60%; n2 – 55.31% and n3 – 57.45%). �e bio-�avanoid liquid concentrates obtained from sample n1 are characterised by a high anthocyanin content, while the conentrates obtained from sample n2, are characterised by a high leucoanthocyanin content, and the bio-�avanoid liquid concentrates obtained from sample n3 are characterised by the content and antioxidant the bio-flavanoid concentrate of vitis vinifera l. ‘r ed a ladasturi’ r ol an d k op al ia ni , t em ur g vi ni an id ze , r ez o ja bn id ze 100 activity of phenolic compounds and �avan-3-ols. anthocyanins in samples of ‘red aladasturi’ cultivar are localised in the grape skin. acknowledgement �is study was supported by shota rustaveli georgia national science foundation (srnsf) [n216752, developing innovative technologies of drastic antioxidant polyphenol concentrates. con�ict of interest �e authors declare no con�ict of interest related to this article. references benmeziane, f., cadot, y., djamai, r., djermoun, l. 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[in russian] abstract �is paper dwells on the uvological characteristics of cultivar vitis vinifera l. ‘red aladasturi’ grape raw materials growing in the viticulture and winemaking zone of imereti (georgia), as well as biologically active compounds and antioxidant activity of hydrophilic extracts and liquid concentrates of its solid matters (stone and skin). research also covered hydrophilic extracts of grape skin and stone thickened by the vacuum of ‘red aladasturi’ grapes raw materials, as well as the concentrates produced from their composition. for research, there were used gravimetric, extractive, spectral and chromatographic methods. we processed samples of grapes raw materials according to the following pattern: identifying qualitative indicators of grapes raw materials; passing grapes raw materials through the dmcsi-type grape clustercomb divider; pressing-out the combless must in a basket press and separation of juice; vacuum sublimation drying of juiceless sweet pomace with an initial moisture content of 45–65% to a �nal moisture content of 9–10%; separation of the ‘red aladasturi’ cultivar’s skin and stone dried to the moisture content of 9–10%, using tea sorting machine designed by g. lominadze; crushing separately the skin and stone in a micro-mill (tp2 hammer mill) until the fraction of 50–100 µm. we have blended the obtained grape-stone ethanol and �uid extracts containing 74–75% of solid maters at an equal ratio (1:1:1) and assessed biologically active compounds and antioxidant activity in this composition. it has been established that the bio-�avanoid liquid concentrate ‘red aladasturi’ is strong antioxidant (55.31–57.45%), and one tablespoon or 8–9 ml of it contains 110–127 mg of �avanoids, which is 105–110% of a full day of rations per person per day. key words: anthocyanins, antioxidant activity, grapes, phenolic compounds, vitis vinifera ‘red aladasturi’ received: [2019.09.17] accepted: [2019.11.20] the bio-flavanoid concentrate of vitis vinifera l. ‘r ed a ladasturi’ r ol an d k op al ia ni , t em ur g vi ni an id ze , r ez o ja bn id ze 102 koncentrat bio-flawonoidów z vitis vinifera l. ‘red aladasturi’ streszczenie w artykule omówiono właściwości odmian winogron vitis vinifera l. ‘red aladasturi’, rosnących w stre�e uprawy winorośli i  winiarstwa w  imereti (gruzja), a  także związki aktywne biologicznie i  aktywność przeciwutleniającą ekstraktów hydro�lowych oraz płynnych koncentratów z ich ciał stałych (pestka i skórka). badania obejmowały również hydro�lowe ekstrakty ze skórki winogron i  pestek, zagęszczone przez sublimaty surowca z  gron „aladasturi”, a  także wytwarzane z  nich koncentraty. do badań wykorzystano metody grawimetryczne, ekstrakcyjne, spektralne i chromatogra�czne. próbki surowców winogronowych przetwarzano według następującego szablonu: identy�kacja wskaźników jakościowych surowców winogronowych; przepuszczanie surowców do produkcji winogron przez dzielnik kombajnu do zbioru winogron typu dmcsi; wyciskanie moszczu w prasie koszowej i oddzielanie soku; sublimacja próżniowa – suszenie słodkich wytłoków bez soku, o  początkowej zawartości wilgoci 45–65% do końcowej zawartości wilgoci 9–10%; oddzielenie skórek i pestek odmiany ‘red aladasturi’, wysuszonych do wilgotności 9–10%, za pomocą maszyny do sortowania herbaty, zaprojektowanej przez g. lominadze; oddzielne mielenie skórek i pestek w mikro-młynie (tp2 hammer mill), do frakcji 50–100 µm. zblendowano otrzymany etanol z pestek winogron i płynne ekstrakty owoców, zawierające 74–75% substancji stałych w równym stosunku (1: 1: 1) i oceniono w tym składzie związki aktywne biologicznie oraz aktywność przeciwutleniającą. ustalono, że płynny koncentrat bio-�awanoidów ‘red aladasturi’ jest silnym przeciwutleniaczem (55,31–57,45%), a jedna łyżka stołowa lub jego 8–9 ml zawiera 110–127 mg �awanoidów, co stanowi 105–110% pełnej racji żywnościowej dziennie na osobę. słowa kluczowe: antocyjany, aktywność przeciwutleniająca, winogrona, związki fenolowe, vitis vinifera ‘red aladasturi’ information on the authors roland kopaliani he is a specialist of the agricultural sciences, especially of subtropical crops. he works at the akaki tsereteli state university in georgia, as a pro�esor. temur gvinianidze he is interested in di�erent issues conected with food technology. he works in georgia at the akaki tsereteli state university, as a pro�esor. rezo jabnidze his research subject is connected with broadly understood issue of subtropical crops. he works at the shota rustaveli state university (batumi, georgia), as a pro�esor. 61 annales universitatis paedagogicae cracoviensis studia naturae, 5: 61–65, 2020, issn 2543-8832 doi: 10.24917/25438832.5.4 valerián franc1*, vladimír hemala2 1department of biology and ecology, faculty of natural sciences, matej bel university, tajovského 40, 97401 banská bystrica, slovakia; *valerian.franc@umb.sk 2department of botany and zoology, faculty of science, masaryk university, kotlářská 2, cz-611 37 brno, czechia new remarkable records of myrmecophilous spiders (araneae) in slovakia introduction myrmecophilous spiders rank among little known and ‘hidden’ arthropods, deserving of increased attention (nelson, jackson, 2009). myrmecophiles are arthropods that live in or near ant nests and are considered true symbionts. myrmecomorphy in spiders is generally considered a type of batesian mimicry in which spiders gain protection from predators through their resemblance to aggressive or unpalatable ants. selection pressure from spider predators and egg-sac parasites may trigger greater integration of myrmecophilous spiders into ant colonies. �e adaptations by which this integration is possible are not well understood, although it is hypothesised that most spider myrmecophiles are chemical mimics and some are even phoretic on their hosts (cushing, 1997, 2012). �e aim of our research was to present the distribution of three rare myrmecophilous spiders in slovakia. methods over several years, we carried out occasional research on myrmecophilous arthropods in slovakia. we applied current methods of sampling, particularly individual exploration under stones and si�ing the detritus of formica genus ant colonies. old and undated records were not included in the record summary. �e geographic coordinates of the sites were included only for thoroughly localised records. �e four-digit numeric code, dfs (databank of fauna of slovakia), is only cited for several inaccurately localised records, where the coordinates were not clearly determinable. other records with a three-digit numeric code were cited according to the code of the geomorphological units of slovakia. v al er iá n fr an c, v la di m ír h em al a 62 results and discussion linyphiidae 1) acartauchenius scurrilis (o. p.-cambridge, 1873) – occurs sporadically and scarcely in xerothermic grasslands and also live near ants (nentwig et al., 2020). it was documented also warm dry oak forests, sand habitats, stone habitats, etc. and from anthropogenic habitats (e.g. vineyards). host ants: lasius niger l., l. �avus fabricius, formica rufa l. and tetramorium caespitum l. (cushing, 1997; buchar, růžička, 2002). obenberger (1949) and miller (1971) mentioned only t. caespitum. apparently, it highly prefers the last ant species (franc, 2007). recent record: domaníky (48°15ʹ35.30” n, 18°58ʹ44.08” e, 208 m a. s. l.), andesite rocky grassland, in the colony of t. caespitum, may 4, 2020, 3 ♂ and 2 ♀. a relatively large number of records from slovakia are available (gajdoš et al., 1999; franc, 2007). �e further recent record: jurský šúr surroundings (7769), april 2009 – 2011, 2 specimens (dankaninová, gajdoš, 2012); malé kršteňany (7376), april 2015 (franc, fašanga, 2017); dolné vestenice (7276), april – may 2010 (melicherčíková, 2010). 2) �yreosthenius biovatus (o. p.-cambridge, 1875) – occurs sporadically and rarely together with ants (nentwig et al., 2020) especially in forests and grasslands of warmer areas, though it may be locally abundant. host ants: formica polyctena foerster, f. pratensis retzius, f. fusca l. and f. sanguinea latreille (cushing, 1997; buchar, růžička, 2002), also f. lugubris zetterstedt (robinson, 1998). miller (1971) mentioned the same ant species except as f. polyctena, while obenberger (1949) noted only f. pratensis and f. rufa. it has also occasionally been recorded away from ant nests (roberts, 1987). recent records: hajnáčka (48°12ʹ46.74” n, 19°56ʹ17.93” e, 290 m a. s. l.) in the colony of formica rufa in the older deciduous forest (beech, oak and hornbeam) february 17, 2019, more than 10 ♀ v. franc and v. hemala lgt.; banská štiavnica – the ‘little water damʹ (48°28ʹ05.48ʺ n, 18°53ʹ03.94ʺ e, 778 m a. s. l.), in the colony of f. rufa in a mixed forest february 14, 2020, 8 ♀, v. franc and v. hemala lgt. et coll. only a few records from slovakia are available, for example: the town of banská bystrica (7280) (franc, 2007) and surroundings (franc, 2005); the hronská pahorkatina hills (804), dudich et al. 1940 (in gajdoš et al., 1999); malanta near the town of nitra (7674d) (gajdoš, 1993) and in the town žilina (6778) (kratochvíl, miller, 1937). it is a little-known species, due to its hidden way of life, but it actually seems to be rather abundant. n ew rem arkable records of m yrm ecophilous spiders (a raneae) in s lovakia 63 hahniidae 3) mastigusa arietina (� orell, 1871) – this scattered species occurs rarely in warm habitats, especially open forests (fig. 1) and on xerothermic slopes. � ey live under bark or in formicaries (nentwig et al., 2020). host ants: in the nests of the ants lasius brunneus and l. fuliginosus (brian, 1977). � ey are not speci� ed in the literature, it obviously may live together with a large range of ant species. new recent records: jalovec (49°09ʹ28.00ʺ n, 19°37ʹ54.70ʺ e, 768 m a. s. l.), in the colony of formica rufa near the chapel (fig. 1), march 10, 2019, 1 ♀ and 2 juveniles v. franc and v. hemala lgt. � is mountain record at the foot of the západné tatry mts is highly notable!; banská štiavnica – the ‘little water dam’ (48°28ʹ05.48ʺ n, 18°53ʹ03.94ʺ e, 778 m a. s. l.), in the colony of f. rufa in mixed forest february 14, 2020, 1 ♀ and 1 juvenile, v. franc and v. hemala lgt. et coll. � e following records from slovakia are available, for example: the rohy national nature reserve near the town of detva (7482a/c), in a under-stone colony of lasius niger, march 3, 1991, 1 ♂; the village of plášťovce (7879b), in the colony of messor muticus nylander, on a xerothermic slope, april 1, 1994, 2 ♀ [together with the very rare leiodid-beetle attaephilus arenarius hampe!]; the rykynčice village (7779d), in the same circumstances, april 9, 1994, 2 ♂, 1 ♀, all records v. franc and a. hanzelová lgt.; the village of nedelište (7682a/b), in a colony of lasius alienus foerster in a xerothermic oak forest, april 22, 2000, 1 ♂ v. franc lgt. (franc, 2007); the zobor national nature reserve near the town of nitra (7674), april 29, 1978 (gajdoš, krumpál, 1987); fig 1. � e colony of red wood ant formica rufa l. near the village of jalovec in the západné tatry mts (photo. v. hemala) v al er iá n fr an c, v la di m ír h em al a 64 jurský šúr surroundings: malé nové hory (7769), april 2009 – 2011 (dankaninová, gajdoš, 2012); ruské (49°7.970ʹ n, 22°20.443ʹ e, 568 m), 1 ex (žila, gajdoš, 2015). identi�ed also in undated records from the slovenský kras mts. (060) j. svatoň lgt. (gajdoš et al., 1999) and the urpín hill near the town of banská bystrica (7280d) (svatoň, 1985). con�ict of interest �e authors declare no con�ict of interest related to this article. references brian, m.v. (1977). ants. london: collins new naturalist. 223 pp. buchar, j., růžička, j. (2002). catalogue of spiders of the czech republic. praha: peres publisher. 351 pp. cushing, p.e. (1997). myrmecomorphy and myrmecophily in spiders: a review. florida entomologist, 80(2), 165–193. cushing, p.e. (2012). spider-ant associations: an updated review of myrmecomorphy, myrmecophily, and myrmecophagy in spiders. psyche: a journal of entomology, ants and �eir parasites – special issue, id 151989, 23 p. https://doi.org/10.1155/2012/151989 dankaninová, l., gajdoš, p. (2012). epigeic spider communities in historical structures of agricultural landscape (vineyard landscape in the svätý jur area). folia faunistica slovaca, 17(3), 275–290. franc, v. (2005). contribution to the knowledge on spiders (araneae) in the surroundings of banská bystrica (slovakia). entomofauna carpathica, 17, 48–54. franc, v. (2007). prevailingly trophic relations between spiders (araneae) and ants (formicoidea). in: k. tajovský, j. schlaghamerský, v. pižl (eds.), contributions to soil zoology in central europe, české budějovice: institute of soil biology, biology centre, academy of sciences of the czech republic. p. 41–45. franc, v., fašanga, m. (2017). spiders (araneae) of the abandoned pasture near the village of malé kršteňany (western slovakia). annales universitatis paedagogicae cracoviensis studia naturae, 2, 39–56. gajdoš, p. (1993). research on epigeic communities in agricultural landscape of malanta (south-west slovakia). bollag accademia gioenia di scienze naturali in catania, 26, 135–144. gajdoš, p., krumpál, m. (1987). pavúky (araneae) zobora 1. ochrana prírody, úšop liptovský mikuláš (spiders (araneae) zobora 1. nature protection, úšop liptovský mikuláš). príroda, 8, 309–328. [in slovak] gajdoš, p., svatoň, j., sloboda, k. (1999). katalóg pavúkov slovenska (catalog of spiders of slovakia). bratislava: ústav krajinnej ekológie sav. 337 pp. [in slovak] kratochvíl, j., miller, f. (1937). k poznání myrmeko�lních pavouků československa (to get to know the myrmecophilous spiders of czechoslovakia). entomologické listy, 1(1), 5–13. [in czech] melicherčíková, m. (2010). prírodné pomery širšieho okolia npr rokoš a využitie v terénnom vyučovaní (natural conditions of the wider surroundings of npr rokoš and use in �eld teaching). msc., diploma �esis depone in matej bel university library in banská bystrica. 79 pp. [in slovak] miller, f. (1971). pavouci (araneida) (spiders (araneida)). in: m. daniel, v. černý (eds.), klíč zvířeny čsr 4. praha: academia. p. 51–306. [in czech] nelson, x.j., jackson, r.r. (2009). �e in�uence of ants on the mating strategy of a myrmecophilic jumping spider (araneae, salticidae). journal of natural history, 43(11–12), 713–735. n ew rem arkable records of m yrm ecophilous spiders (a raneae) in s lovakia 65 nentwig, w., blick, t., bosmans, r., gloor, d., hänggi, a., kropf, c. (2020). araneae – spiders of europae. version 16.2.2020. https://www.araneae.nmbe.ch obenberger, j. (1949). ze života mravenců (from the life of ants). praha: vyšehrad, 221 pp. [in czech] roberts, m.j. (1987). �e spiders of great britain and ireland. linyphiidae and check list. vol. 2. colcheste: harley books. 204 pp. robinson, n.a. (1998). two new records of the myrmecophile spider �yreosthenius biovatus cambridge in nests of formica rufa l. british journal of entomology and natural history, 11, 72. svatoň, j. (1985). náčrt fauny pavúkov (araneida) navrhovaného chráneného náleziska urpín pri banskej bystrici. stredné slovensko (sketch of the spider fauna (araneida) of the proposed protected site urpín near banská bystrica. central slovakia). zborník stredoslovenského múzea v banskej bystrici, 4, 237–259. [in slovak] žila, p., gajdoš, p. (2015). spider fauna (aranea) in non-forest habitats of the evicted area of the horná cirocha. folia faunistica slovaca, 19(3), 269–285. abstract �e authors of this article provide data on the occurrence and ecology of three little-known myrmecophilous spiders (acartauchenius scurrilis o.p.-cambridge, �yreosthenius biovatus o.p.-cambridge and mastigusa arietina �orell) in slovakia. �e mountain record of m. arietina at the foot of the západné tatry mts is especially remarkable. key words: araneae, faunistic, myrmecophiles, spiders received: [2020.07.07] accepted: [2020.09.08] nowe i niezwykłe notowania myrmecofilnych pająków (araneae) na słowacji streszczenie autorzy tego artykułu podają dane o występowaniu i ekologii trzech mało znanych pająków myrmeko�lnych (acartauchenius scurrilis o. p.-cambridge, �yreosthenius biovatus o. p.-cambridge i mastigusa arietina �orell) na słowacji. zwłaszcza górskie notowanie m. arietina u podnóża zachodnich tatr jest bardzo niezwykłe. słowa kluczowe: araneae, faunistyczny, myrmeko�le, pająki information on the authors valerián franc he is a lecturer of general zoology and systematic invertebrate zoology. he has deal with the research of beetles and spiders for more than 30 years, with special regard to the nature conservation and the problems of biological indication and the factors of endangerment of separate animal taxa. vladimír hemala he is interested in phenology and distribution of bugs (hemiptera sp.) in europe. 239 annales universitatis paedagogicae cracoviensis studia naturae, 5: 239–241, 2020, issn 2543-8832 popularyzacja nauki poprzez środowisko naukowe odbywa się na wiele różnych sposobów. zaliczyć do nich można różne publikacje popularnonaukowe w czasopismach ogólnodostępnych, takich jak: „kosmos”, „świat wiedzy”, czy national geographic, ale też różnego rodzaju wydarzenia, np. festiwale nauki i sztuki, małopolskie noce naukowców, czy noce biologów. co roku kolejne edycje tych wydarzeń przyciągają coraz większą liczbę uczestników, co świadczy o zwiększającym się zainteresowaniu społeczeństwa nauką oraz jej osiągnięciami. w zorganizowanej 10 stycznia 2020 roku kolejnej edycji nocy biologów pt. „globalne zmiany środowiska” już po raz 9 uczestniczył wydział biologii uniwersytetu jagiellońskiego w krakowie. różnorodne zajęcia przygotowali pracownicy naukowi, doktoranci, studenci oraz koła naukowe z trzech instytutów: instytutu nauk o środowisku, instytutu zoologii i badań biomedycznych oraz instytutu botaniki. na uczestników tegorocznej edycji czekały liczne pokazy, warsztaty, wykłady, wystawy oraz gry trwające cały dzień w budynkach wydziału na iii kampusie unithe biologists’ night 2020 in the faculty of biology of the jagiellonian university in krakow (poland), january 10th, 2020 noc biologów 2020 na wydziale biologii uniwersytetu jagiellońskiego w krakowie (polska), 10 stycznia 2020 popularisation of science through the scienti�c community takes place in many different ways. �ese include various popular science publications in magazines, such as: “cosmos”, “world of knowledge” or national geographic as well as various types of events, e.g. festivals of science and arts, researchers night in małopolska or biologists’ night. every year, subsequent editions of these events attract an increasing number of participants, which proves the growing public interest in science and its achievements. �is year edition of the biologists’ night organized on january 10th, 2020 titled “global environmental changes” was attended by the faculty of biology of the jagiellonian university in cracow for the 9th time. various activities have been prepared by academics, phd candidates, students, and science clubs from three institutes: �e institute of environmental sciences, the institute of zoology and biomedical research and the institute of botany. �e participants of this year’s edition had the opportunity to participate in numerous demonstrations, workshops, lectures, exhibitions, and games that lasted all day in the 240 r ep or ts wersytetu jagiellońskiego przy ulicy gronostajowej w krakowie. przygotowane zajęcia dotyczyły różnorodnej tematyki, m.in. zmian środowiska, zarówno globalnych, jak i  lokalnych. wśród zróżnicowanych warsztatów znalazły się, np. „adaptacje do nowych warunków środowiska” zaprezentowane przez weronikę antoł, joannę palka, paulinę kosztyła z  zespołu genomiki i ewolucji eksperymentalnej; „rośliny oczyszczające powietrze” poprowadzone przez koło przyrodników studentów uj, czy „myśl globalnie, działaj lokalnie” autorstwa dr hab. pauliny kramarz prof. uj oraz dr hab. zo�i prokop z  zespołu ekologii fizjologicznej wraz z  aktywistami i  aktywistkami miejskimi, a  także społecznymi ruchami oddolnymi, działającymi na terenie polski. wśród licznych wykładów zaprezentowano, np. „konsekwencje globalnych zmian środowiskowych u owadów” wygłoszone przez dr hab. stanisława knutelskiego z  zakładu entomologii; „brudne myśli – czyli jak zanieczyszczenie środowiska wpływa na nasze samopoczucie” zaprezentowane przez izabelę ciurej z  koła naukowego neuronus, czy „wirusy i  bakterie w  obliczy globalnych zmian klimatu” wygłoszone przez karola wadasa z  koła naukowego biologii komórki. nie zabrakło również innych interesujących tematów, m.in. o dinozaurach mezozoiku (eryk sroka); o krakowskiej awifaunie (koło przyrodników studentów uj), o tym co kryje się w miodzie (artur górecki), czy o  zmienności owadów (witold morek, piotr gąsiorek). w  sumie w  tegorocznej edycji zaprezentowano: jedną wystawę, dwie gry, jedenaście pokazów, dwadzieścia sześć wykładów oraz osiemdziesiąt siedem warsztatów. w  ciągu całego faculty’s buildings on the iii campus of the jagiellonian university at gronostajowa st. in cracow (kraków). �e prepared classes concerned various topics related to environmental changes, both global and local. �e prepared diverse workshops included, among others “adaptations to new environmental conditions” presented by weronika antoł, joanna palka, paulina kosztyła from the genomics and experimental evolution group; “air purifying by plants” led by the student naturalist society at the jagiellonian university or “�ink globally, act locally” by assoc. prof. paulina kramarz and assoc. prof. zo�a prokop from the physiological ecology team together with urban activists and grassroots activists in poland. among the numerous lectures were, e.g. “consequences of global environmental changes in insects” delivered by assoc. prof. stanisław knutelski from the department of entomology; “dirty thoughts – how environmental pollution a�ects our well-being” presented by izabela ciurej from the student neuroscience society neuronus or “viruses and bacteria in the calculation of global climate change” delivered by karol wadas from the student association of cell biology of the jagiellonian university. �ere were also other interesting topics, including e.g.: mesozoic dinosaurs (eryk sroka); cracow’s avifauna (student naturalist society at the jagiellonian university); what is hidden in honey (artur górecki) or insect variability (witold morek, piotr gąsiorek). in total, this year’s edition included: one exhibition, two games, 11 demonstrations, 26 lectures and 89 workshops. �roughout the day, two competitions with attractive prizes were also held: a 241 r eportsdnia przeprowadzono również dwa konkursy z  atrakcyjnymi nagrodami: konkurs na plakat, dotyczący tegorocznego tematu nocy biologów dla różnych klas wiekowych oraz konkurs „biologiczne koło fortuny”. wszystkie przygotowane zajęcia spotkały się z dużym zainteresowaniem uczestników co umożliwiło rozpropagowanie wiedzy przyrodniczej wśród wszystkich klas wiekowych, w szczególności wśród dzieci młodzieży ze szkół podstawowych i średnich. o dużym zaciekawieniu świadczy nie tylko duża liczba odbiorców na przygotowanych zajęciach, ale również liczne pytania do prowadzących. z niecierpliwością będziemy wyczekiwać kolejnej edycji, która miejmy nadzieję również przyciągnie rzeszę uczestników. poster competition for this year’s biologists’ night theme for di�erent age classes and the “biological wheel of fortune” competition. all prepared activities met with great interest of the participants, which enabled the dissemination of knowledge of science among all age classes, in particular among participants from primary and high schools. great curiosity of society is evidenced not only by the large number of recipients in the prepared classes, but also by numerous questions to the lecturers. we are looking forward to the next edition, which will hopefully also attract many participants. additional information about this year’s biologists’ night can be found at/ dodatkowe informacje o przeprowadzonych zajęciach tegorocznej nocy biologów można znaleźć na stronie: http://www.nocbiologow.pl/index.php?id=jednostka&nazwa=krakow&idp=opis. rita rakowska institute of botany, faculty of biology, jagiellonian university, gronostajowa 3 st. 30-387 kraków, poland, rita.rakowska@doctoral.uj.edu.pl 135 annales universitatis paedagogicae cracoviensis studia naturae, 4: 135–145, 2019, issn 2543-8832 doi: 10.24917/25438832.4.8 jiří kupka1*, hana švehláková1, rostislav poláček2 1vsb-technical university of ostrava, faculty of mining and geology 17. listopadu 15 st., 70833 ostrava-poruba, czechia; *jiri.kupka@vsb.cz 2ministry of the environment of the czech republic vršovická 1442/65 st., 100 10 praha 10, czechia selected environmental issues of the landscape of shale (nízký jeseník mt., czechia) – preliminary results introduction for our purposes, we understand the environment as a  set of natural, arti�cial and social components of the world that are (or may be) in direct contact with man. natural components include, for example, climate, water and soil conditions. �e arti�cial components can include buildings, production and transport facilities, communications and also terrain shapes such as mine heaps. finally, social components include interpersonal relationships, culture, laws, economic conditions. �e interaction between man and (his) environment then occurs in the landscape. �e very content of the term landscape then has a  very wide range, which can moreover be understood from various points of view. �e landscape structure is variable over time and determined by its composition elements, which perform their own functions and are more or less dynamic. in the landscape we can distinguish the primary structure formed by the physical-geographic complex, i.e. natural elements such as geology, climate, water, natural terrain, etc. �e secondary structure is the land use and material elements of this use (settlements, communications, mining remains, real vegetation cover, etc.). �e tertiary structure of the landscape then includes intangible phenomena that can be re�ected in the primary and secondary landscape structures. �ese include, for example, administrative units, ownership relationships, protection zones and regimes. �e tertiary structure can also include the so-called magical places, which are connected with various legends and folklore traditions and the natural and arti�cial point of view. �e primary, secondary and tertiary structure of the landscape is shaped by its typical character. an important landscaping agent are human activities. �e ‘shale landscape’ is a  distinctive environmental and landscape phenomenon that can be seen from many points of view, as already mentioned above. it is located 136 ji ří k up ka , h an a š ve hl ák ov á, r os tis la v p ol áč ek in the geomorphological unit nízký jeseník (2.894 km²) (vencl, strohalm, 2005). �e mining of roo�ng shale tiles was concentrated in a part of the lower carboniferous of the moravian-silesian region in terms of the geological division of the bohemian massif (hrušecký, 1946; zapletal et al., 1989) – fig. 1. here, for more than two centuries, shale mining has also le� a distinct mark on local architecture and urbanism. with the resettlement of the original german-speaking population in 1945–1946, not only the roo�ng shale mining, but also the typical architectural elements that signi�cantly contributed to the landscape character of the area were lost were reduced. among these typical architectural features of the ‘shale landscape’ were, in particular, the original roo�ng – shale tiles attached with copper nails, which is now being replaced by another material or shale from import. �e social composition of the population has also changed. new residents of abandoned settlements a�er the expulsion of germans can be considered basically the �rst human colonisers of the territory. �ey began to transform the landscape and settlement structure, o�en without a  deeper understanding of its speci�c dynamics and of course without following the ancestors traditions and respect to the genius loci. we can characterise the ‘shale landscape’ as a post-industrial landscape; remnants of shale mining as landscape elements of post-industrial landscape. �ese include quarries and quarry lakes, adits and sha�s, mine heaps and remnants of various buildfig. 1. localisation of analysed region on map of czechia (source of map: czech environmental information agency – changed) 137 ings. in total, these elements can be found in the cadastre of more than 50 villages and towns of nízký jeseník mts. an interesting �nding of recent decades is the fact that some habitats, which are traditionally perceived as a  symbol of degradation of the natural environment (for example abandoned quarries or mine heaps), are inhabited by quite unique communities of plants and animals (tropek, řehounek, 2012). in the case of post-mining landscape, it is possible to speak of a biological colonisation, formed by plant and animal communities in various stages of succession and a cultural colonisation, formed by man and his activities. �e subject of our research is a  comprehensive understanding of the landscape a�er shale mining, which includes the colonisation of landscape mining elements by plants, vegetation and animals, including humans. material and methods �e following text presents only some (most important) representatives of animals whose presence in the landscape a�er shale mining is related to post-mining landscape elements. �e fauna survey started here in 2017 and can be divided into two phases. in the �rst phase, it is an inventory survey, which is focused especially on species protected, endangered or otherwise important from di�erent taxonomic groups. in the second phase selected taxa (e.g. amphibians in quarry ponds, hymenoptera insects on heaps or bats in underground spaces), the so-called speci�c zoological survey, are systematically studied. various types of methods were used in the inventory survey (observation, shearing using a hard skid net on islets of vegetation, individual collection in heaps of tailings material, etc.). �ey were in the underground in vertebrates and vertebrates studied by using conventional �ashlights while browsing underground space each mine. in this way animals were examined in particular on the walls and ceilings (spiders, butter�ies and bats). on the �oor animals were surveyed under various objects (stones, remains of timbering, etc.). it was necessary in some taxa to take specimens for accurate determination. animals were collected using entomological tweezers or exhauster, and �xed in 70% alcohol or killed by vapours of ethyl acetate. in the case of speci�c zoological research suitable methods were chosen for the study of individual taxa (e.g. aquatic animals net for amphibian studies, mörick dish for hymenoptera, bat-detector for bats). �e �ora and vegetation survey was launched in 2017 with orientation terrain walks to �nd out the basic physiognomy of vegetation and the nature of the relief. in the �rst phase, an inventory survey was carried out focusing on rare and protected species according to act no. 114/1992 coll. and the corresponding red lists. currently a phytosociological survey according to the rules of the zurich – montpellier school 138 (braun-blanquet, 1964). in the �atter parts of the heaps, reléves had the shape of 100 m2 squares. on slopes, terraces and trailers, the shape of a rectangle was chosen as the more suitable one to maintain the recommended area. phytosociological survey was performed at available slopes and exposures. species were recorded in phytosociological tables during the season. due to the abundance of individual localities and sub-areas spread over a  large area (including jakartovice – 49°54′54″ n, 17°41′3″ e, surroundings of hrubá voda – 49°40′13″ n, 17°26′11″ e, břidličná – 49°54′42″ n, 17°22′16″ e, moravice – 49°51′28″ n, 17°43′13″ e, zálužné – 49°49′23″ n, 17°42′59″ e and others), the phenomenon of ‘colonisation’ of shale landscape cannot be a�ected by tramps and campers but at least on the basis of some selected localities to describe the characteristics of this post-industrial colonisation and to outline issues that will be developed in the future. �e existing survey is based primarily on published literature, including tramp texts and �eld research, which included both observation and documentation of informants’ statements contacted on the spot and through links to well-known camp organisations in the ostrava and opava regions. in the future, it would be possible to use, for example, archival sources for individual localities. preliminary results and discussion when anthropogenic activity is terminated or signi�cantly reduced, natural processes will prevail and species with speci�c demands can be encountered at these post-industrial sites (sometimes at industrial sites), including a signi�cant presence of rare and endangered species or the occurrence of organisms, which we consider to be unusual in our nature. in the czech republic, some species occur only at post-industrial sites. �is is particularly so because post-industrial habitats create speci�c conditions that are typical to the present central european landscape. man has traditionally stopped farming in the landscape. of course, the most common types of species are less demanding, common. however, in both cases, there are plants and animals that can �nd optimal conditions at post-industrial sites (konvička et al., 2005). landscape elements a�er shale mining represent a varied mosaic of micro-sites in terms of both botanical and zoological aspects (appendix 1a-b). for example, acidophilous grasses and sub-xerothermic plant species grow on the tops of mine heaps: hieracium bauhini schult. asteraceae, sedum acre l. rosaceae and potentilla argentea agg. l. rosaceae. some species, such as chamaenerion palustre scop. (= epilobium dodonaei vill.) oenotheraceae, filago arvensis l. asteraceae or lepidium campestre (l.) r. br. brassicaceae, are found directly on the mine heaps created by shale fragments. some of these taxa belong to species from the red list of vascular plants in the czech republic. ji ří k up ka , h an a š ve hl ák ov á, r os tis la v p ol áč ek 139 for the remains of buildings from indigenous peoples, we can see massive linden trees, or ornamental plants such as dwarf periwinkle (vinca minor l.) apocynaceae, poet’s narcissus (narcissus poeticus l.) amaryllidaceae, and yellow �gwort (scrophularia vernalis l.) scrophulariaceae. �ere are endangered grass annual fescue (vulpia myuros (l.) c. c. gmel.) poaceae on dry warm places and the most endangered species round-leaved wintergreen (pyrola rotundifolia l.) pyrolaceae on unstable, shaded areas of mine heaps. from the animals, there is a  xerothermal snake in the same places – the endangered smooth snake (coronella austriaca laur.) colubridae, the endangered green tiger beetle (cicindela campestris l.) carabidae or rare spider ozyptila claveata walck �omisidae (appendix 1b). adits represent a  signi�cant wintering ground for bats, for example western barbastelle (barbastella barbastellus schreb.) vespertilionidae or very abundant greater mouse-eared bat (myotis myotis borkh.) vespertilionidae and lesser horseshoe bat (rhinolophus hipposideros bech.) rhinolophidae. in �ooded quarry lakes there is a rich population of critically endangered species european cray�sh (astacus astacus l.) astacidae. amphibians are represented by common toad (bufo bufo l.) bufonidae, smooth newt (lissotriton vulgaris l. =triturus vulgaris l.) salamandridae, alpine newt (ichthyosaura alpestris laur. =triturus alpestris laur.) salamandridae and common frog (rana temporaria l.) ranidae. interesting �ndings include the �nding of niphargus tatrensis wrześ. niphargidae in the leachate of mining galleries. it follows from published and unpublished materials that a�er the expulsion of the original population, the colonisers of the above-mentioned elements of the mining landscape were campers followed by tramps who, unlike the new permanent residents, approached the landscape with respect and interest, based on the philosophy of the tramp movement. for example, the 49th section of junák in ostrava had summer camp in mokřinky near melč – 49°51′0″ n, 17°45′28″ e, in 1947 (bvú, 2005). new, seemingly temporary and occasional ‘residents’ came to the country. �e landscape features of the mining landscape have played and still play an important role in the life of campers and tramps as “colonisers”. one of the possibilities to demonstrate this is the use of tramp toponyms and their spread. some of the names refer directly to a particular landscape element (for example, a sha� with a tunnel called “rodriguez’s tomb” on the cadastre of the defunct village of nové oldřůvky – 49°45′4″ n, 17°40′38″ e, or a quarry with a quarry pond ‘na špici’ in jakartovice), others include a wider area where several di�erent mining elements are found (for example, a place called ‘el fuego’ in the cadastre of the extinct lesy). sometimes a name given initially for only one particular landscape element was used for a much wider territory, as is the case with an abandoned quarry with a quarry pond in jakartovice, which is named ‘horse mine’ and it functions under this name even in the mining register. 140 appropriately situated quarry, not �ooded or �ooded only in part, with various terraces and heaps, provided ideal conditions for establishing a permanent campsite. o�en, such a ‘romantic’ place has become the goal of traditional clubs, and in some cases, the centre of group games. quarry lakes themselves were attractive for the possibility of their exploration, bathing, or to build ra�s and other simple vessels. �e walls of operational and other buildings provided a suitable refuge a�er a minor modi�cation (in some cases, the roof was made of shale tiles), �replaces and sleeping bunk beds were built. shale tiles served as ‘guestbook sheets’ for example to record visitor names or nicknames. from the shale itself were built camp circles and speci�c �replaces (stacked to a height of 50 cm or more), various inscriptions visible from above, as well as towers and mounds. an important role was played by the fact that stay in these localities was not signi�cantly regulated (kupka, pohunek, 2017). conclusion �e issues presented here are only a brief introduction to the wider elaborate of fauna, �ora and vegetation of this region. analysed colonisation processes, taking place in such a speci�c post-mining environment, should be considered in spontaneous and induced aspects. in the latter case, an important role is played by anthropopression of a di�erent nature than it used to be, which is currently consequence of the increase in tourism in this area. acknowledgements �e research was co-funded by �nancial subsidy from the student grant competition sp2018/15 “selected environmental aspects of landscape a�er shale mining”. con�ict of interest �e authors declare no con�ict of interest related to this article. references biolib https://www.biolib.cz/cz/taxon/ [in czech] bvú (2005). kompas. kompas tom bvú 1945-2005 vydaný k 60. výročí založení oddílu, 65p. braun-blanquet, j. 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[in czech] 142 appendix 1 preliminary list of plant (a) and animal (b) species recorded in the habitats of the shale post-mining landscape (jakartovice village 49°54′54″ n, 17°41′3″ e): nomenclature of plants: according polish flora (www.atlas-roslin.pl) and other internet sources. (a) bryophytes: abietinella abietina (hedw.) m.fleisch., amblystegium serpens (hedv.) schimp., atrichum undulatum (hedw.) p.beauv., brachythecium rutabulum (hedw.) schimp., b. salebrosum (ho�m. ex f.weber et d.mohr) schimp., brachytheciastrum velutinum (hedw.) ignatov et huttunen (= brachythecium velutinum w. p. schimp.), bryum argenteum hedw., ceratodon purpureus (hedw.) brid., cynodontium polycarpon (hedw.) schimp., dicranella heteromalla (hedw.) schimp., d. varia (hedw.) schimp., dicranum polysetum sw. ex anon., d. scoparium hedw., hylocomium splendens (hedw.) schimp., hypnum cupressiforme hedw., h. revolutum (mitt.) lindb., lophocolea bidentata (l.) dumort., lophozia ventricosa (dicks.) dumort., orthotrichum a�ne schrad. ex brid., o. anomalum hedw., oxyrrhynchium hians (hedw.) loeske (= eurhynchium hians (hedw.) sande lac.), plagiomnium a�ne (blandow ex funck) t.j.kop., plagiothecium denticulatum (hedw.) schimp., p. succulentum (wilson) lindb., pleurozium schreberi (willd. ex brid.) mitt., pohlia nutans (hedw.) lindb., polytrichastrum formosum (hedw.) g. l. sm. (= polytrichum formosum hedw.), polytrichum juniperinum hedw., p. piliferum hedw., pseudoscleropodium purum (limpr) m. fleisch. ex broth., ptilidium ciliare (l.) hampe, p. pulcherrimum (weber) vain., racomitrium lanuginosum (hedw.) brid., rhizomnium punctatum (hedw.) t.j. kop, rhytidiadelphus squarrosus (hedw.) warnst., rosulabryum laevi�lum (syed) ochyra (= bryum moravicum podp.), sanionia uncinata (hedw.) loeske, scapania nemorea (l.) grolle, schistostega penata (hedw.) f.weber et d.mohr, syntrichia ruralis (hedw.) f. weber et d. mohr (= tortula ruralis (hedw.) gaertn., meyer, & scherb.), tortula muralis hedw. vascular plants: acer campestre l., a. platanoides l., a. pseudoplatanus l., achillea millefolium l., agrostis stolonifera l., alnus glutinosa (l.) gaertn., anthriscus sylvestris (l.) hoffm., arenaria serpyllifolia l., arrhenatherum elatius (l.) p. beauv. ex j. presl et c., artemisia vulgaris l., athyrium �lix femina (l.) rot, betula pendula roth, bidens frondosa l., bromus sterilis l., calamagrostis epigejos (l.) rot, calystegia sepium (l.) r.br., carex praecox schreb., carpinus betulus l., centaurea cyanus l., cerasus avium (l.) moench (= prunus avium l.), chamaenerion palustre scop. (= epilobium dodonaei vill.), chelidonium majus l., chenopodium album agg., cirsium arvense (l.) scop., conyza canadensis (l.) cronquist, cornus alba l. (= swida alba (l.) opiz), c. sanguinea l. (= swida sanguinea opiz), corylus avellana l., crataegus monogyna jacq., crepis biennis l., daucus carota l., digitaria sanguinalis (l.) scop., dryopteris ji ří k up ka , h an a š ve hl ák ov á, r os tis la v p ol áč ek s elected environm ental issues of the landscape of shale (n ízký jeseník m t., c zechia) – prelim inary results 143 �lix-mas (l.) schott, echium vulgare l., epilobium adenocaulon hausskn. (= e. ciliatum raf.), erigeron annuus (l.) pers., eupatorium cannabinum l., fagus sylvatica l., festuca rubra l., festuca sp., filago arvensis l., fragaria vesca l., fraxinus excelsior l., galeopsis pubescens besser, g. speciosa mill., galium aparine l., geranium robertianum l., geum urbanum l., glechoma hederacea l., hedera helix l., hieracium bauhini schult., h. murorum l., h. sabaudum l., hypericum maculatum crantz, h. perforatum l., impatiens parvi�ora dc., inula conyza dc., juglans regia l., lepidium campestre (l.) r. br., ligustrum vulgare l., malus domestica borkh., matricaria perforata mérat (= tripleurospermum inodorum (l.) sch. bip.), melandrium album (mill.) garcke (= silene latifolia poir.), mycelis muralis (l.) dumort., myosotis sylvatica ehrh. ex ho�m., oenothera biennis l., oxalis acetosella l., padus avium mill. (= prunus padus l.), pastinaca sativa l., picea abies (l.) h.karst., picris hieracioides l., pinus nigra j.f. arnold, p. sylvestris l., poa compessa l., p. pratensis l., populus tremula l., potentilla argentea agg. l., p. erecta (l.) raeusch., prunus spinosa l., pyrola rotundifolia l., quercus robur l., ribes uva-crispa l. (= grossularia uva-crispa (l.) mill.), robinia pseudacacia l., rosa sect. caninae dc., rubus caesius l., r. idaeus l., rumex acetosa l., r. acetosella l., r. obtusifolius l., salix caprea l., s. purpurea l., sambucus nigra l., scrophularia vernalis l., senecio jacobaea l., s. ovatus (p. gaertn., b. mey. et scherb.) willd., s. vulgaris l., solidago canadensis l., s. gigantea aiton, sorbus aucuparia l. em. hedl., stellaria graminea l., s. media (l.) vill., symphytum o�cinale l., tanacetum parthenium (l.) sch. bip., taraxacum sect. ruderalia kirsch., h.øllg. & štěpánek, tilia cordata mill., t. platyphyllos scop., urtica dioica l., verbascum thapsus l., veronica chamaedrys l., viola odorata l., vulpia myuros (l.) c. c. gmel. nomenclature of animals: according biolib (https://www.biolib.cz/cz/taxon/), fauna europaea (https://fauna-eu.org/) and other internet sources. (b) snails: aegopinella nitens mich. (= hyalinia nitens mich.), alinda biplicata mont. (= balea biplicata mont.), arion distinctus j. mabil., arion vulgaris moquin-tand., boettgerilla pallens simr., cochlicopa lubrica o. f. müll., deroceras agreste l., d. reticulatum o. f. müll., discus rotundatus o. f. müll., helix pomatia l., limax cinereoniger wolf, monachoides incarnatus o. f. müll., oxychillus glaber rossm., radix labiata rossm. crustaceans: armadillidium vulgare latre., astacus astacus l., ligidium hypnorum cuv., niphargus tatrensis wrześ., oniscus asellus l., porcellio scaber latre. spiders: amaurobius fenestralis ström, cicurina cicur fabr., coelotes pabulator sim. (= c. terretris wild.), drassyllus prae�cus l. koch, meta menardi latre., ozyptila claveata walck., pirata hygrophilus �or., pisaura mirabilis cler. grasshoppers and crickets: pseudochorthippus parallelus zetter. (= chorthippus parallelus zetter.), gomphocerippus rufus l., oedipoda caerulescens l., phaneroptera falcata poda, pholidoptera griseoaptera de geer, tetrix subulata l., tettigonia cantans fuess. 144 butter�ies and moths: aglais urticae l., anthocharis cardamines l., apatura ilia denis & schi�., aphantopus hyperantus l., araschnia levana l., argynnis paphia l., autographa gamma l., coenonympha pamphilus l., colias hyale l., gonepteryx rhamni l., inachis io l. (= aglais io l.), lasiommata megera l., leptidea reali reiss., lycaena dispar haw., maniola jurtina l., melanargia galathea l., nymphalis antiopa l., pararge aegeria l., pieris brassicae l., p. napi l., p. rapae l., polygonia c-album l., polyommatus icarus rottem., scoliopteryx libatrix l., �ymelicus lineola ochsen., triphosa dubitata l., vanessa atalanta l., v. cardui l. beetles: cantharis rustica fall., carabus violaceus l., chrysomela fastuosa scop., cicindela campestris l., anoplotrupes stercorosus scri. (=geotrupes stercorosus l.), leptura quadrifasciata l., nicrophorus vespillo l., oiceoptoma thoracicum l., phosphuga atrata l., psyllobora vigintiduopunctata l., rhagonycha fulva scop., silpha obscura l., staphylinus caesareus cederh., tachyura parvula deje., �anatophilus rugosus l. hymenoptera: agenioideus cinctellus spin., ammophila sabulosa l., andrena fulva müll., andrena haemorrhoa fabr., a. minutula kir., a. nigroaenea kir., a. nitida müll., a. strohmella stöck., a. varians kir., anoplius infuscatus van der lind., apis mellifera l., arachnospila anceps wesm., bombus campestris panz., b. pascuorum scop., b. rupestris fabr., b. terrestris l., halictus maculatus smith, h. tumulorum l., hylaeus signatus panz., lasioglossum calceatum scop., l. morio fabr., l. pauxillum schen., melecta albifrons foer., nomada conjungens herrich-schäf., n. �avoguttata kir., n. fulvicornis fabr., n. succincta panz., osmia bicornis l., polistes nimpha christ, priocnemis perturbator harr., rhopalum clavipes l., sphecodes gibbus l., trypoxylon minus beaum., vespula germanica fabr., v. vulgaris l. amphibians: bufo bufo l., ichthyosaura alpestris laur. (=triturus alpestris laur.), lissotriton vulgaris l. (= triturus vulgaris l.), pelophylax esculentus l., rana temporaria l. reptiles: anguis fragilis l., coronella austriaca laur., lacerta agilis l., natrix natrix l. bats: barbastella barbastellus schreb., myotis emarginatus é. geo�., m. myotis borkh., m. mystacinus kuhl, m. nattereri kuhl, plecotus auritus l., rhinolophus hipposideros bech. ji ří k up ka , h an a š ve hl ák ov á, r os tis la v p ol áč ek s elected environm ental issues of the landscape of shale (n ízký jeseník m t., c zechia) – prelim inary results 145 abstract �e area of the nízký jeseník mts. is, among other things, well known for its shale roo�ng tiles since the 18th century. in places where shale was intensively or extensively exploited until 1945, abandoned areas a�er mining works remained. in general, every mining is perceived as a activity of landscape degradation by the public. however, these indelible traces of shale mining in the form of various mining-related objects (e.g. abandoned quarries, quarry ponds, sha�s, drains etc.) are also gradually becoming places that are colonized by unique plant and animal communities. �ere are very interesting species bond to speci�c environmental conditions of post-mining landscape, with frequent rare and endangered species. people have also become ‘new’ colonisers in the case of the shale landscape. key words: post-industrial landscape, abandoned quarries, mine heaps, fauna colonisation, vegetation colonisation, cultural colonisation received: [2019.09.05] accepted: [2019.11.19] wybrane zagadnienia środowiskowe krajobrazu łupków ilastych (nízký jeseník, czechy) – wstępne rezultaty streszczenie obszar gór niskiego jesionika znany jest od xviii wieku, m.in. z  łupków dachówkowych. w  miejscach, w których łupek był intensywnie lub ekstensywnie eksploatowany do 1945 r., pozostały wyrobiska górnicze. generalnie, każda działalność wydobywcza jest postrzegana przez społeczeństwo jako działanie degradujące krajobraz. jednak te nieusuwalne ślady wydobycia łupków ilastych, w postaci różnych obiektów związanych z  górnictwem, np. opuszczone kamieniołomy, stawy kamieniołomowe, szyby, dreny itp., również stopniowo stają się miejscami kolonizowanymi przez unikalne ugrupowania roślin i zwierząt. interesujące gatunki wiążą się ze specy�cznymi warunkami środowiskowymi krajobrazu pogórniczego, a  często występują tam gatunki rzadkie i  zagrożone. w  przypadku krajobrazu łupkowego, także sami ludzie stali się „nowymi” kolonizatorami. słowa kluczowe: krajobraz poprzemysłowy, opuszczone kamieniołomy, hałdy min, kolonizacja fauny, kolonizacja roślinności, kolonizacja kulturowa information about authors jiří kupka �e author is focusing on ecological and environmental applications in post-industrial landscape (especially fauna study at post mining sites, including underground spaces), biological researches related to biogeochemistry and environmental geochemistry, brown�elds reuse and didactics of natural sciences. hana švehláková �e author is a geobotanist with a focus on plant communities a�ected by industry and coal mining. she also deals with the relationship between soil seed bank and above-ground vegetation and the spread of invasive plant species in the post-mining landscape. rostislav poláček �e author is engaged in research of hymenoptera insects with a focus on wasps, bumble bees and solitary bees in post-industrial landscape. �e research is carried out mainly at localities (heaps) a�er mining of mineral resources. 170 annales universitatis paedagogicae cracoviensis studia naturae, 4: 170–179, 2019, issn 2543-8832 doi: 10.24917/25438832.4.11 mohamad hesam shahrajabian*, wenli sun, qi cheng biotechnology research institute, chinese academy of agricultural sciences, beijing 100081, china nitrogen fixation laboratory, qi institute, building c4, no.555 chuangye, jiaxing 314000, zhejiang, china; *hesamshahrajabian@gmail.com, chengqi@caas.cn measures to achieve a stable farming system in sustainable agriculture – a short review introduction �e rapid increases in human population and exploitative use of non-renewable resources have worsened food shortages (amini et al., 2012; esfandiary et al., 2012; soleymani et al., 2012a–b; shahrajabian et al., 2017, 2018; ogbaji et al., 2018; soleymani, shahrajabian, 2018; yong et al., 2018). in the context of improving the global food situation, chemical fertilisers play a dominant role (yazdpour et al., 2012; shahrajabian, soleymani, 2017a–b). most scientists believe that increasing yield per ha is a major way for increasing crops yield (soleymani et al., 2016; soleymani, shahrajabian, 2017; yong et al., 2017). due to high costs and poor accessibility of inorganic fertilisers to resource-poor farmers, other inputs are o�entimes proposed as alternatives (abedi et al., 2010; shahri et al., 2011; soleymani, shahrajabian, 2012a; shahrajabian et al., 2013). it is believed that much of de�cient plant nutrients could be supplied to soils through organic matters while small shortage are made up with mineral fertilisers (oluleye, akinrinde, 2010; ogbaji et al., 2013). in forage production, considering chemical position of forage crop is important (rezaeifard et al., 2010). farmyard manure (fym) contains very small amount of major nutrients and involves transportation. but, it maintains the soil physical and chemical condition and improves the overall ecological balance of the crop production system. fym reduces the external inputs and can on self-regulating ecosystem process. �e aim of this research is to review intercropping, its importance and comparison of fertilisers, organic manures, and green manures. intercropping intercropping is known as a  practice, which can improve the utilisation of available resources and cause yield advantages and increases yield stability compared to sole m easures to achieve a stable farm ing system in sustainable agriculture – a short review 171 cropping (soleymani et al., 2011e; soleymani, shahrajabian, 2012a). it is a  sustainable practice used in many developed and developing countries and an essential element of agricultural sustainability (singh et al., 2010). in intercropping system, there is normally one main crop and one or more added crops, with the main crop being of primary importance for food and forage production. �e most important aim and advantage of intercropping is to produce a higher yield on a given piece of land by appropriate use of the available growth resources that may not be utilised by each single crop grown alone. �ere are di�erent types of intercropping but the most important types are, row intercropping, strip intercropping, mixed intercropping and relay intercropping. intercropping system may lead to soil conservation, improvement of soil fertility, and improvement of forage quality, reduction of pest and diseases. �e intercropping systems are old and widespread applications in low-input agricultural systems, and they were common for many countries before the modernisation of agriculture. �ere are both direct and indirect facilitative interactions of intercropping systems. intercropping systems can cause more e�ective use of resources by providing symbiotic nitrogen from legumes or making available inorganic phosphorus �xed in soil because of lowering of ph via nitrogen �xing legumes. also, more e�cient water usage in intercropping systems was suggested by numerous researchers. intercropping practices are the most productive when intercrops of di�erent growth period are used so that their maximum requirements for growth resources occur at di�erent times. �ey are the best way of introducing more biodiversity into agro-ecosystems and results have shown that increased crop diversity may increase the number of ecosystem service provided. �ese practices are the best way to ecological balance, more utilisation of resources, increase in the quantity and quality of agricultural products and signi�cant reduction of damage and loss by pests, diseases and weeds. on the basis of multiple advantages of intercropping especially in the terms of sustainable agriculture and organic farming, it is clear that intercropping is more reasonable than sole cropping systems. �e agricultural use of a living cover crop during a crop growth cycle (relay intercropping) may help to preserve biodiversity, increase soil organic matter content and carbon sequestration and provide nutrient recycling (shili-touzi et al., 2010). leguminous as a cover crops are extensively used in the tropics for soil conservation in plantation crops, maintaining it fertility. �ese plants have good potential for replacing many unwanted weeds (olorynmaiye, 2010). for example, in potato and corn intercropping, land equivalent ration (ler) reached 1.58 (ebwongu et al., 2001). bekele and sommartya (2006) noticed that in intercropping of potato with garlic ler was more than one. according to dua et al. (2005) the intercropping treatments increased yield as compared to sole-cropping. ler was higher than one in the intercropping of potato and pinto bean (nasrollahzadeh asl et al., 2009). ghanbari et al. (2010) reported that m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 172 land equivalent ration values were higher in all intercropping systems with di�erent planting ration of maize-cowpea which indicated the yield advantage of intercropping over sole cropping maize. bilalis et al. (2005) reported that in the maize-bean intercrop system, ler values were statistically higher than in maize-cowpea. �e intercropping shows the bene�cial e�ect on the quality and quantity of growth of crop plants. for example, soleymani et al. (2012c) reported that in iran there has been a  rapid increase of fertiliser application in recent years to achieve high yields. mix cropping legumes with cereal and grasses species were used for enhance nutrition value, supply energy and protein on both crops. �is mixture o�ers a sustainable alternative to maintain e�cient farming systems with reduced environmental impacts. �e studies showed that intercropping causes yield advantage and better nutrition uptake. for suitable ways to animal’s grazing were intercropping of berseem clover and forage corn in low input farming system and nitrate accumulation in clover. �e intensive cropping system, heavy input technology, environmental degradation and other related problems again encouraged to use green manuring in plant nutrient supply system. residue burning accompanied with usage of triticale as a green manure was the best choice to achieve high quality. for obtaining the most fresh forage yield and biological yield of forage corn, triticale plantation can be replaced by barley cultivation. four weeks of residue retention accompanied by using of barley as green manure led to the highest yield and yield components of forage corn. �at is why, the green manuring is an age-old practice used for supplying nitrogen to crop plants. fertiliser low-input farming systems such as arable organic farming, o�en have limited access to nitrogen and decreased the productivity of these systems (marsalis et al., 2010; soleymani et al., 2010, 2011e, 2012a; soleymani, shahrajabian, 2012b; abdollahi et al., 2018). �e minimal or no fertiliser input causes serious nutrient depletion, which coupled with the low fertility status of soil is the major limiting factor to crop production. increasing nitrogen supply enhances both growth of shoot and root of plants (shahrajabian et al., 2011; soleymani, shahrajabian, 2011). to optimise plant production and minimise production cost needed is supplemental nitrogen application (ahmadi moghaddam et al., 2010; kayan, 2010; soleymani et al., 2013). inadequate amount of nutrient availability can show de�ciency symptom and in�uence on the quality and quantity of yield of crops. in most commercially available fertilisers, the concentrations of active ingredients rapidly decreased due to chemical, photochemical and biological degradation, volatilisation, leaching, adsorption or immobilisation in soil (broumand et al., 2010; xiong et al., 2010). farmers o�en intercrop on soils without adequate knowledge of the right quality of fertilisers to be applied. by human activity nitrate found naturally at moderate m easures to achieve a stable farm ing system in sustainable agriculture – a short review 173 concentrations in many environments o�en rises to dangerous level. for example, nitrogen fertilisers a�ect yield and nutritive value of corn (marsalis et al., 2010). combined organic and inorganic fertilisation enhances organic matter in soil and increases yield of sweet maize (e�himiadou et al., 2010). however, the application of excessive amounts of nitrogen can cause the accumulation of toxic levels of nitrate (no3 -) in plants (gulmezoglu et al., 2010; khoshkharam et al., 2010; soleymani, shahrajabian, 2013). nitrate toxicity in forage plants can cause chronic or acute stress in livestock. manure organic farming, which evolved in the 1980s, is one way to solve the current farming problems. in this method, manure and green manure is used instead of chemical fertilisers (soleymani et al., 2011b–d; 2012b). because of this substitution, the food and environment become safer. manures are very variable products, o�en di�cult to apply accurately and release nutrients in the soil at a desirable rate. some studies have shown that farmyard manure applied alone or in combination with inorganic fertilisers was e�ective in maintaining soil fertility under continuous cultivation. applying farm manure increased cation exchange capacity (cec), organic carbon and water holding capacity of the soil and nutrient availability. for example, a dairy manure is an excellent source of nitrogen for crops and can easily ful�l the nitrogen requirement. to get satisfactory results well-composted manure must be used, because it is usually free of weed seeds and has a better nutrient balance. barmaki et al. (2008) reported that the yields of potato plots in which manure was used were 0.4 kg m-2 higher than in plots that receive only chemical fertilisers. it has been noted that application of organic manure has a more lasting bene�cial residual e�ect that can remain signi�cant up to four seasons when compared with inorganic fertilisers whose residual bene�t do not last beyond season (babaji et al., 2010). liu et al. (2010) reported that sheep manure had no signi�cant e�ect on rice’s characteristics. long term application of npk and pig manure together with straw return to �eld produced highest rice grain yield. future of agriculture lies in the development of organic based fertilisers. fym improves plants production better than mineral fertilisers as the crop is not capable of optimising single application of inorganic fertilisers but prefers slow continuous release of nutrient that is possible with the use of organic manure. green manure symbiotic n2 �xation (snf) in legumes is a fundamental process for maintaining soil fertility continued productivity of organic cropping systems (singh et al., 2010). it is very o�en used in inter-row crops. one of the bene�ts of this kind of crop is having high potential of extrapolation. cultivation of legumes together with non-legume m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 174 plants is a  common practice in the world. mixture of annual legumes and cereals is intensively cultivated in the world as a  forage. recently increasing interest of intercropping as an attempt to substantiate functional biodiversity for agriculture and reduce chemical inputs use was observed. for example, sulc et al. (1993) concluded that ryegrass-alfalfa mixtures cultivate in north-central usa can provide a  good forage. �e typical organic production is characterised by extended rotations involving leguminous crop green manure and organic amendments utilisation (soleymani et al., 2011a–b; soleymani, shahrajabian, 2012c). canali et al. (2010) noted that supply of nitrogen from the soil, which consist of nitrogen mineralised from organic soil matter and crop residue is an important and variable contributor of nitrogen to potato crop production. without organic farming, food security will be hampered (sarker, itohara, 2010). legumes are o�en grown for incorporation into soil as a green manure providing bene�ts such as o�-season soil cover, stimulated soil biological activity and improved plant nutrition (soleymani et al., 2011c). most interest has been attached to the legume’s ability to furnish subsequent crops with readily available nitrogen (n). some plants used for the production of green manure can signi�cantly increase in yield of crops (singh et al., 2010). conclusion sustainable agriculture means a  shi� from monoculture to intercropping. in other words, intercropping means the agricultural cultivation of two or more crops in the same space and at the same time. sustainable farming also means self-sustaining, low-input and energy-e�cient agricultural systems. biodiversity is the main key and strategy for sustainable agriculture. application of organic and synthetic fertilisers to soil would provide multiple bene�ts for improvement of soil chemical, physical and biological properties leading to improved crop yield. integrated use of synthetic and organic fertilisers leads to development of sustainable crop production. also, this may improve the e�ciency of synthetic fertilisers and reduce their usage. integrated use of organic and synthetic fertilisers is a good method to improve crop productivity and sustain soil quality and fertility. in sustainable agricultural system, fertilisers, livestock manure and cover crops are important parameters in productive agricultural systems to have stable food. con�ict of interest �e authors declare no con�ict of interest related to this article. m easures to achieve a stable farm ing system in sustainable agriculture – a short review 175 references abdollahi, m., soleymani, a., shahrajabian, m.h. 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(2018). changes in dry matter, protein percentage and organic matter of soybean-oat and groundnut-oat intercropping in di�erent growth stages in jilin province, china. acta agriculturae slovenica, 111(1), 1–7. doi: 10.14720/ aas.2018.111.1.04 środki służące do uzyskania stabilnego systemu rolnego w zrównoważonym rolnictwie – krótki przegląd streszczenie zrównoważony system rolny jest najlepszym sposobem na zaspokojenie potrzeb dzisiejszych i przyszłych pokoleń. w systemie tym wielkość plonu wzrasta wraz z zastosowaniem upraw międzyrzędowych, poprzez wyższy współczynnik wzrostu roślin, redukcję nasion chwastów, ograniczenie ilości szkodników i chorób oraz bardziej efektywne wykorzystanie zasobów. uprawa międzyrzędowa jest jednym z  najważniejszych sposobów zwiększenia różnorodności w  ekosystemie rolniczym. systemy międzyplonowe mogą być barm easures to achieve a stable farm ing system in sustainable agriculture – a short review 179 dziej stabilnymi systemami praktyk rolniczych niż uprawy monokulturowe. najważniejszymi zaletami uprawy międzyrzędowej są: zwiększenie produkcji rolnej i  większe wykorzystanie zasobów środowiska. zintegrowane stosowanie nawozów syntetycznych i  organicznych może również prowadzić do rozwoju zrównoważonej produkcji roślinnej. metoda ta poprawia wydajność działania nawozów chemicznych i jednocześnie ogranicza ich stosowanie. zielony nawóz z roślin strączkowych, takich jak: koniczyna, lucerna i inne, które są bogatym źródłem azotu uwalnianego w glebie, jest w stanie zmniejszyć znacząco zapotrzebowanie na azot syntetyczny. tego rodzaju biologiczne wiązanie azotu odgrywa bardzo ważną rolę w zrównoważonych systemach rolniczych. w zrównoważonym systemie rolnym nawozy, odchody zwierzęce i rośliny uprawne oraz chwasty są ważnymi parametrami dla zapewnienia stabilnej produkcji żywności. key words: intercropping, chemical fertiliser, manure, sustainable agriculture, stable system received: [2019.06.12] accepted: [2019.11.14] 103 annales universitatis paedagogicae cracoviensis studia naturae, 4: 103–118, 2019, issn 2543-8832 doi: 10.24917/25438832.4.6 aleksandra mazur cracow institute of development and education, wielicka 42/105 st., 30-552 kraków, poland; aleksandra.mazur.krk@gmail.com the role of seed coat in the germination and early stages of growth of bean (phaseolus vulgaris l.) in the presence of chickweed (stellaria media (l.) vill.) introduction human activity is one of the main factors a�ecting soil, water, air and living organisms pollution. increasing amounts of chemical compounds used in agriculture, construction and industry contribute to the increase of pollution and degradation of the natural environment. �erefore, more and more o�en production attempts are madein ecological systems, which are an opportunity for sustainable development and protectionof environmental biodiversity. in ecologic farming, synthetic chemical compounds are replaced by natural substances produced by plants, which is why research into the practical use of allelopathy is desirable. �e term ‘allelopathy’ comes from the greek language and is a combination of two words allelon (mutual) and pathos (su�er, harm). in present times, allelopathy was described by hans molish (1937), who de�ned the phenomenon as the interaction of adjacent plants (or microorganisms), of both harmful and bene�cial biochemical nature (gniazdowska et al., 2004). during the �rst world allelopathy congress “allelopathy – a science for the future” in 1996, deliberations were made to create a de�nition of the phenomenon described, treating allelopathy as any process involving secondary metabolites produced by plants, microorganisms and fungi that a�ect the growth and development of biological systems and farming, excluding from these transformations animals (oleszek, 1996; wójcik-wojtkowiak et al., 1998). however, the phenomenon of allelopathy cannot be regarded as a form of direct in�uence of one plant on another, because metabolites secreted into the environment undergo various transformations. �e substance in its original form secreted by the donor plant does not always have to reach the acceptor plant. �e level of toxicity of allelopathic compounds is determined by retention, transport and transformation processes (rice, 1984; oleszek, 1992). pos104 a le ks an dr a m az ur itive allelopathic interactions can have a  practical aspect for plant growth. as part of biological competition, they can also perform a protective function against pests, weeds and diseases, increasing plant resistance (nowiński, 1961). a bothersome weed of many crops is the chickweed (stellaria media (l.) vill.). �is species from the family of caryophyllaceae juss., is characteristic of weed communities of arable �elds and ruderal areas (matuszkiewicz, 2006). it is an annual or biennial plant, cosmopolitan and nitrophilous. it blooms o�en all year round and shows germination at low temperatures (van der vegte, 1978). it grows in segetal areas, land�lls, roadside and wastelands. s. media propagates both by seeds and vegetatively. it forms low, dense clusters covered with pale yellow leaves and white �owers (parus, 2015). beans (phaseolus vulgaris l.) belong to the beans family (fabaceae lindl., =papilionaceae giseke). it comes from central and south america. currently, it is widely grown in more than 200 cultivars on the old continent, as well as in africa and asia. its popularity in crops is due to seeds that are rich in protein, contain folic acid, vitamin b6, iron. bean seed coat is a rich source of biologically active ingredients, among others: amino acids, �avonoids, triterpenes, sugars, steroidal saponins, trace elements, guanidine derivatives, organic acids, vitamins c and e (kuchanowicz et al., 2017). �e interest in seed germination biotests increases every year, which are easy to observation, easy to perform and does not require large �nancial outlays. in germination biotests, it is important to determine what concentrations of chemicals adversely a�ect seed germination, plant growth and development. �e aim of the study was to investigate the role of seed coat in the germination process and in the early stages of growth of bean seeds (p. vulgaris) in the presence of aqueous extracts from dry shoots of chickweed (s. media). �e in�uence of chickweed extracts on the values of germination index of bean seed with seed coat and seed without it was determined (1), the growth inhibition rate of seedlings was determined (2), the values of fresh and dry matter and the percentage of water content in 7 days seedlings of p. vulgaris, grown from seeds with seed coats, as well as without them (3). material and methods plant material bean seeds (phaseolus vulgaris) from the horticultural company polan (kraków, poland) were used for the experiments. herbaceous parts of chickweed (stellaria media) in form of fresh shoots were harvested in southern poland near kraków and dried in laboratory conditions. �en they were stored in the dark, so as to avoid microbiological destruction of allelopathic compounds contained in them. 105 the role of seed coat in the germ ination and early stages of grow th of bean (phaseolus vulgaris l.) in the presence of chickw eed (stellaria m edia (l.) v ill.) extracts preparation aqueous extracts from s. media dry shoots were prepared in three percentage concentrations: 5, 10 and 15%. grinded in a  grinder (braun 4045, germany) shoots of chickweed depending on the concentration were �ooded with the appropriate amount of distilled water (5% extract = 5 g dry material + 95 ml cold boiled water, 10% extract = 10 g dry material + 90 ml cold boiled water, 15% extract = 15 g dry material + 85 ml cold boiled water). �e extracts prepared in this way were le� for 24 hours in the dark at a temperature of about 25°c to extract the chemical compounds contained in them. a�er one day, extracts from chickweed dry shoots were strained through a  double layer of gauze and stored in the refrigerator for the duration of the experiment. seeds germination bean seeds, counted 10 for each petri dishes, were rinsed under running water for 30 minutes, and then 3 times with distilled water and divided into two groups. �e �rst experimental group were beans with seed coats, and the second were seeds without them. in order to easily remove the coat from the seeds, some of them were le� for 2–3 hours in distilled water until the coat clearly wrinkled, and then it was removed with a knife so as not to damage the inside of the seed. �e seeds of p. vulgaris prepared in this way, both with seed coats and without them, were lined with tweezers on petri dishes, 9 cm in diameter, with a triple layer of �lter paper moistened with 5 ml of appropriate aqueous extract from s. media shoots, at concentrations of 5, 10 and 15%. �e control group consisted of bean seeds, both those with seed coats and seeds without them, put on petri dishes with �lter paper moistened with 5 ml of distilled water. for the duration of the experiment, all seed on petri dishes were placed in the dark at room temperature. every 24 h for 7 days the number of germinated seeds was checked. germinated seeds were those which germinal root was half the size of the seed. germination indexes a�er 7 days of the experiment, the e�ect of aqueous extracts from s. media dry shoots on the germination indexes of p. vulgaris seeds with and without coats were assessed. �ese formulas of germination indexes are in table 1. biometric analysis �e length of whole p. vulgaris seedlings was measured with a ruler with an accuracy of 1 mm. �e impact of s. media dry shoots extracts on bean seedling growth on length was determined according to the formula proposed by islam and kato-noguchi (2012): ip = (1 – (le/lc)) × 100; where: ip – growth inhibition index [%]; le – seedlings length (mm) treated with emitter data; lc – seedlings length (mm) from the control. 106 tab. 1. germination index formulas germination index formula author gp (germination percentage) gp = [number of germinated seeds at everyday/ total number of seeds sets for bioassay] × 100 global method se (speed of emergence) se = (number of germinated seeds at the starting day of germination/number of germinated seeds at the �nal days of measurement) × 100 islam et al. (2009) svi (seedling vigour index) svi = (seedling length(mm) × germination percent)/100 fresh and dry mass, water content �e fresh mass of 7-day-old bean seedlings with and without seed coat was determined on the laboratory scale (ohaus adventurer pro, usa). to obtain a dry mass, the plant material was dried for 48 hours at 105°c in an dryer (wamed sup 100, poland) and then weighed. on the basis of the mass values obtained, the percentage water content was determined according to the formula: 100 – [(dry mass × 100) / fresh mass]. statistical analysis �e results were developed in microso� excel based on data collected from observations carried out during the experiment. additionally, statistical analysis was performed using the one-way anova / manova analysis of variance test. duncan’s test at p ≤ 0.05 was used to assess the signi�cance of di�erences between the means ± sd tested. �e data was analysed in statso�, inc. (2018). statistica (data analysis so�ware system), version 13.1. www.statso�.com. results germination indexes germination capacity of seeds phaseolus vulgaris, with seed coat and without it, in the presence of aqueous extracts of dry shoots of stellaria media with di�erent concentrations of 5, 10, 15% and control conditions was varied (fig. 1–2). among seeds with seed coat, no seeds germinated on the �rst day of the experiment. on the third day, the highest percentage of germinated seeds was observed on the 10% extract, where the germination value was 40%. seeds watered with a 5% extract and those from the control sample reached value of 30%, while no seeds germinated in the presence of 15% extract. on the seventh day, the percentage of germinated seeds increased in three cases, reaching the highest value of 80% for seeds a le ks an dr a m az ur 107 fig. 1. percent of germinated seeds phaseolus vulgaris l. grown from seed with coat in the presence of aqueous extracts from the dry shoots of stellaria media (l.) vill. at various concentrations of 5, 10 and 15% and control conditions (distilled water) fig. 2. percent of germinated seeds phaseolus vulgaris l. grown from seed without coat in the presence of aqueous extracts from the dry shoots of stellaria media (l.) vill. at various concentrations of 5, 10 and 15% and control conditions (distilled water) germinating in the presence of 10% extract, 50% for seeds watered with 5% extracts, and 40% for the control sample. only on the 15% extract of chickweed, no germinated seeds were observed (fig. 1). the role of seed coat in the germ ination and early stages of grow th of bean (phaseolus vulgaris l.) in the presence of chickw eed (stellaria m edia (l.) v ill.) 108 for p. vulgaris seeds without coat, germination began only on the third day of observation. �e largest number of newly germinated bean seeds was on the seventh day of experiment for seeds from the control sample. a  slightly smaller number of germinated seed, compared to the control, was observed on dishes with 5% chickweed extract. in the other two cases, in the presence of 10 and 15% extracts, no bean germination was noted until the end of the experiment (fig. 2). in the case of seed emergence (se), the highest, though uneven, results were obtained for bean seeds watered with 5% aqueous extract from s. media shoots (at 60% for seeds with coat and 80% for seeds without it). similar values of se were achieved by bean seeds with coat watered with distilled water (control test). lower values of the discussed coe�cient apply to seeds of p. vulgaris with coat, germinating in the presence of 10% extracts and seeds without seed coat from the control sample. compared to the control conditions, the extract of 15% from chickweed shoots, regardless of the presence or absence of seed coat, inhibited the germination of bean seeds in 100% (fig. 3–4). seed vigour index (svi) has a similar values, both for seeds germinating with and without seed coat (fig. 5–6). in both cases, both on the 3rd and the last 7th day, it was observed that the higher the concentration of the aqueous extract from s. media, the higher the seedling viability (up to 10% for seeds with coat and to 5% without coat). in the studied sample of seed with coat, the highest value of the svi index was recorded for seedlings p. vulgaris watered with 10% extracts, in relation to the control. for seeds without coat, the highest value of this parameter was observed in the case of the control sample. at 15% of the extract concentration, the bean svi index, for both seed with and without coat, had 0 value. fig. 3. seed emergence (se) of seeds phaseolus vulgaris l. grown from seed with coat in the presence of aqueous extracts from dry shoots of stellaria media (l.) vill. at various concentrations of 5, 10, 15% and control conditions (distilled water); mean values ± sd marked with letters a, b, c di�er signi�cantly according to duncan’s test p ≤ 0.05 a le ks an dr a m az ur 109 fig. 4. seed emergence (se) of seeds phaseolus vulgaris l. grown from seed without coat in the presence of aqueous extracts from dry shoots of stellaria media (l.) vill. at various concentrations of 5, 10, 15% and control conditions (distilled water); mean values ± sd marked with letters a, b, c di�er signi�cantly according to duncan’s test p ≤ 0.05 fig. 5. seed vigour index (svi) of phaseolus vulgaris l. grown from seeds with and without coat, on the 3th germination day, in the presence of aqueous extracts from dry shoots of stellaria media (l.) vill. at various concentrations of 5, 10 and 15% and control conditions (distilled water); mean values ± sd marked with letters a, b, c di�er signi�cantly according to duncan’s test p ≤ 0.05 biometric analysis biometric analysis of p. vulgaris seedlings germinated from seed with coat showed that both extracts at 5 and 10% concentrations stimulated the growth of them, compared to control. in the case of bean seeds germinating without coat, the growth inhibition the role of seed coat in the germ ination and early stages of grow th of bean (phaseolus vulgaris l.) in the presence of chickw eed (stellaria m edia (l.) v ill.) 110 fig. 6. seed vigour index (svi) of phaseolus vulgaris l. grown from seeds with and without coat, on the 7th germination day, in the presence of aqueous extracts from dry shoots of stellaria media (l.) vill. at various concentrations of 5, 10 and 15% and control conditions (distilled water); mean values ± sd marked with letters a, b, c di�er signi�cantly according to duncan’s test p ≤ 0.05 had already occurred in the presence of 10% extracts. regardless of the presence or absence of seed coat, clear inhibition of the growth in length of p. vulgaris seedlings was caused by extracts of 15% concentration from s. media, compared to seedlings grown in control (fig. 7–8). fig. 7. inhibition percentage index of growth (ip) (expressed as a percentage of control value) of phaseolus vulgaris l. germinated seed with coat in the presence of aqueous extracts from stellaria media (l.) vill. at various concentrations of 5, 10 and 15% and control (distilled water); a negative (-) value on the y axis indicates growth, and a positive (+) value indicates growth inhibition a le ks an dr a m az ur 111 fig. 8. inhibition percentage index of growth (ip) (expressed as a percentage of control value) of phaseolus vulgaris l. germinated seed without coat in the presence of aqueous extracts from stellaria media (l.) vill. at various concentrations of 5, 10 and 15% and control (distilled water); a negative (-) value on the y axis indicates growth, and a positive (+) value indicates growth inhibition fresh and dry mass, water content analysing the values of fresh mass of bean seedlings watered with aqueous extracts from s. media shoots, signi�cant di�erences were found compared to the control (tab.  3). �e highest increase in fresh mass was observed in p. vulgaris watered with 5% extracts (for seeds with coat) and for seedlings watered with 5 and 10% extracts (for seeds without coat). in the case of dry mass, the only signi�cant statistical di�erences in the values of this parameter noticed for bean seedlings grown with seed coat on 5% extracts, compared to the dry mass of seedlings watered with distilled water (tab. 3). in other cases, regardless of the type of seeds analysed, dry mass values changed slightly. tab. 2. changes in the length of phaseolus vulgaris l. seedlings [cm] grown from seeds with seed coat – (a) and without seed coat – (b) in the presence of aqueous extracts from dry shoots of stellaria media (l.) vill. at di�erent concentrations of 5, 10 and 15% and control (distilled water), marked on di�erent days of the experiment (days 1, 3 and 7) day stellaria media extract [%] control 5 10 15 a b a b a b a b 1 1.25 b 1.30 b 1.45 b 1.35 b 1.35 b 1.25 b 1.15 b 1.25 b 3 1.40 b 1.55 b 1.45 b 1.60 b 1.55 b 1.25 b 1.15 b 1.25 b 7 2.45 a 2.55 a 2.65 a 2.25 a 2.00 ab 1.25 b 1.15 b 1.25 b mean values ± sd marked with letters a, b, c di�er signi�cantly according to duncan’s test p ≤ 0.05 the role of seed coat in the germ ination and early stages of grow th of bean (phaseolus vulgaris l.) in the presence of chickw eed (stellaria m edia (l.) v ill.) 112 �e percentage of water content in the control samples was 35.37% for bean seed with coat and 44.56% for seed without it. both in the �rst and second analysed samples, the value of the parameter increased signi�cantly in seedlings watered with 5% extracts of chickweed extracts, compared to the control sample (tab. 3). however, as the concentration of allelopathic compounds increased in aqueous extracts from s. media, a decrease in the water content of bean seedlings was observed in each of the studied samples. tab. 3. fresh and dry mass values and percentage of water content in phaseolus vulgaris l. seedlings grown from seed with seed coat – (a) and without seed coat – (b) in the presence of aqueous extracts from shoots of stellaria media (l.) vill., with di�erent concentrations of 5, 10 and 15% and control (distilled water) parametr stellaria media extract [%] control 5 10 15 a b a b a b a b fresh mass [g] 0.2944 b±0.06 0.2534 b ±0.06 0.4880 a ±0.11 0.4142 a ±0.11 0.2187 b ±0.06 0.5435 a ±0.15 0.2007 b ±0.04 0.2229 b ±0.03 dry mass [g] 0.1877 bc±0.04 0.1393 a ±0.04 0.2041 a ±0.08 0.1479 a ±0.04 0.1316 c ±0.03 0.1600 a ±0.05 0.1641 bc ±0.03 0.1494 a ±0.04 water content [%] 35.37 b ±0.04 44.56 c ±0.04 59.09 a ±0.08 63.68 b ±0.04 38.75 b ±0.03 70.49 a ±0.05 18.25 c ±0.03 33.30 cd ±0.04 mean values ± sd marked with letters a, b, c di�er signi�cantly according to duncan’s test p ≤ 0.05 discussion modern agriculture pays great attention to the skilful and environmentally safe increase in the quantity and quality of crops (górecki et al., 1994). �e �rst important stage in obtaining high-quality crop plants is to provide them with the right conditions for seed germination. changes occurring during germination are regulated at the molecular, cellular and whole seeds levels (grzesiuk, kulka, 1981; higashiyama et al., 2003). in the �rst stage of germination an important role is played by the seed coat (herse, 1982). seed coat accounts for 12 to 20% of the mass of seeds, and the remaining seed components of the embryo are: cotyledon and germinal root, which constitute respectively 75 to 83% and 5% of the mass (byszewski, 1972). �e chemical composition of the seed coat and embryo is di�erent, depending on the physiological functions performed, and the stage of seed development (grzesiuk, kulka, 1981; niewiadomski, 1990). �e di�erences in germination process and early stages of phaseolus vulgaris growth observed during the experiment showed the allelopathic properties of stellaria media, as well as the protective e�ect of the seed coat in the interaction of seeds with allelopathic substances. previous studies on allelopathic properties of a le ks an dr a m az ur 113 chickweed show that the presence of this weed causes a decrease of brassica napus l. var. napus crops (lutman et al., 2000), reduced production of triticum aestivum l. grains (lutman, 2002), and also depending on the concentration of the extract favourably or adversely e�ects on the germination and growth of di�erent cultivars of zea mays l. (zandi et al., 2019). however, s. media does not a�ect every plant in the same negative way. �is thesis was con�rmed by this experiment (fig. 1–8; tab. 2–3). seed germination is a complex process that includes both catabolic and anabolic reactions and biochemical transformations (grzesiuk, kulka, 1981). �e seed viability index accepted on an international scale is germination capacity (dąbrowska, 1998). in this study it was observed that the percentage values of germinated p. vulgaris seeds with and without seed coat were the largest for seeds watered with 5% aqueous extracts from s. media dry shoots, compared to seeds germinated on distilled water (fig. 1–2). in addition, a high number of germinated seeds was observed on 10% extracts in the group of bean seeds with seed coat. �e opposite e�ect was for seeds without seed coat, where 10% extracts completely inhibited the germination of bean seeds (fig. 2). seed germination studies that take place in laboratory conditions do not exactly re�ect crop conditions (faligowska et al., 2012). �erefore, the vigour of seeds to additionally assess is proposed (conreras, barros, 2005). in this experiment, the seed emergence (se) index values for both tested samples (seeds with and without coat) were characterised by a various seed reaction to aqueous extracts from s. media shoots (fig. 3–4). in the case of seed with seed coat, the relationship between se index and the value of the aqueous concentration of s. media can be noticed. compared to the se from the control, it was observed that the higher concentration of the extract, the faster the se index values decreased. compared to the control, in the case of seeds without coat, only 5% extract of chickweed caused an increase in the se index. signi�cantly higher values of seed vigour index (svi) for bean seeds with seed coat were noted. in contrast, seeds without seed coat were more sensitive to aqueous extracts from s. media (fig. 5–6). probably, these results were connected with the presence or absence of seed coat. �e seed coat has a protective function of the seed against harmful external factors, including: bacterial and fungal infections, as well as drying out, the in�uence of toxic chemical compounds and mechanical damage (grzesiuk, kulka, 1981; możdżeń, rzepka, 2016). biometric analysis showed the stimulating e�ect of 5 and 10% aqueous extracts on the growth of 7-day-old p. vulgaris seedlings, germinated with seed coat. for seeds without seed coat, quite the opposite results were obtained (fig. 7–8). both, in one and the second experimantal group of seeds in the presence of 15% of extracts from s. media, a total inhibition of seedlings growth was found (tab. 2). �e results obtained are consistent with the literature data of many authors. for example: możdżeń et al. the role of seed coat in the germ ination and early stages of grow th of bean (phaseolus vulgaris l.) in the presence of chickw eed (stellaria m edia (l.) v ill.) 114 (2016) showed the inhibitory e�ect of aqueous extracts from capsella bursa-pastoris (l.) medik. on germination and growth of lactuca sativa l. cv ‘maryna’, puła et al. (2016) studied the e�ect of aqueous extracts of galium aparine l. on various cultivars of zea mays l., możdżeń et al. (2018) observed the allelopathic activity of galinsoga parvi�ora cav. and oxalis fontana bunge on early growth stages of di�erent cultivars of raphanus sativus l. var. radicula pers, zandi et al. (2018) con�rmed the allelopathic properties of s. media on r. sativus var. radicula and others. �e increase in fresh and dry mass of bean seedlings, grown from seed with seed coat, was signi�cantly the largest on substrates with 5% extracts, compared to the control (tab. 3). at 10 and 15% extract concentrations, the mass values were near to the control. �e values of fresh mass in seedlings grown from seeds without seed coat were clearly higher in the case of extracts 5 and 10%, in relation to the control. in the case of dry mass, no statistically signi�cant changes were found in the values of this parameter. it can be supposed that the di�erences in mass increase are due to the disruption by the s. media extracts of the capture, transport and use of ions of calcium (ca2+), magnesium (mg2+), potassium (k+) and water (das et al., 1997). according to bieżanowska-kopeć and pisulewski (2006), bean seeds have antioxidant properties due to the polyphenols content, which are part of the plant’s natural protective ingredients against harmful compounds. another protective barrier for seeds with a seed coat may be cuticle, which, depending on the thickness and saturation with waxes, is an impermeable barrier to water and gases (russi et al., 1992; zeng et al., 2005). �e chemical composition of seeds, their size and variety play an important role in mass growth and seed swelling (rice, 1984). �e percentage of water content increased signi�cantly in seeds without a  seed coat – from 44.56 to 63.68% for seeds watered with a  5% extract, up to 70.49% for seeds grown on a 10% extract (tab. 3). for seeds with seed coat grown in the presence of 5% extract, the percentage increase in water was smaller and amounted to 59.09%. seed coat protected the seeds against the negative allelopathic e�ects of the aqueous extract from s. media – inhibition of germination was observed only at a  concentration of 15%. it also did not allow for high absorption of the aqueous extract, as evidenced by the lower percentage of water in seeds with seed coat. �e di�erences obtained most likely result from the presence in the extracts of chemical compounds that modify the properties of cell membranes and contribute to a low degree of water absorption, thereby reducing the germination rate of seeds (siwek, 2008). rut et al. (2012) report that allelopathic compounds inhibit roots hair formation, reducing their active surface, and thus reduce water uptake. �e essence of the allelopathy process is the secretion by the plants (donors) of speci�c chemical compounds that favourably or adversely a�ect other plants (acceptors) (wójcik-wojtkowiak, politycka, 1998). allelopathic reactions of one plant to anotha le ks an dr a m az ur 115 er are the result of interactions of mixtures of di�erent compounds, not just a single substance (blum, 1996; veronneau et al., 1997). in crops, the antagonistic e�ects of various groups of chemical compounds are more pronounced at low concentrations, compared to the action of individual substances. herb of s. media contains oligosaccharides, saponins, �avonoids, proteins, provitamins, vitamins b1, b2, c, e, pp, triterpene glycosides, sugar alcohols and mineral salts (vanhaecke et al., 2006; hu et al., 2009), which can cause this kind of complex action. �e analysed indexes show clearly that water extracts from dry s. media shoots show allelopathic properties for germination and early stages of bean growth. while, seed coat plays an important role in protecting seeds from the e�ects of aqueous extracts of chickweed. experiment carried out here con�rmed, that seeds throughout the entire life cycle of plants, despite the best protection against various stress factors, during germination become sensitive to any type of stress (weiqiang et al., 2005). conclusion (1) germination indexes for seeds of phaseolus vulgaris showed that seeds without seed coats were more sensitive to aqueous extracts from stellaria media at low allelopathic compounds concentrations the seeds germinated similarly to distilled water, however at higher concentrations of this substances the process of germination was inhibited. (2) �e increase in the length of seedlings grown from seeds with coats and without them decreased with increasing allelopathic substances concentrations; in the presence of 15% of s. media extracts, inhibition of seedling growth was observed. (3) �e values of fresh and dry mass and the percentage of water content di�ered for each of the examined group of seeds, depending on the concentration of the extract used. con�ict of interest �e author declares no con�ict of interest related to this article. references bieżanowska-kopeć, r., pisulewski, p.m. (2006). wpływ procesów termicznych i  biologicznych na pojemność przeciwutleniającą nasion fasoli (phaseolus vulgaris l.). żywność, 3(48), 51–64. [in polish] blum, u. (1996). allelopathic interactions involving phenolic acids. journal of nematology, 28, 259–267. byszewski, w. (1972). surowce roślinne. warszawa: wydawnictwo pwn. [in polish] contreras, s., barros, m. 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(1998). allelopatia. poznań: wydawnictwo akademii rolniczej im. augusta cieszkowskiego w poznaniu. [in polish] zandi, p., barabasz-krasny, b., stachurska-swakoń, a., puła, j., możdżeń, k. (2018). allelopathic e�ects of stellaria media (l.) vill. on germination and early stages of growth of raphanus sativus var. radicula. annales universitatis paedagogicae cracoviensis studia naturae, 3, 90–99. doi: 10.24917/25438832.3.7 zandi, p., możdżeń, k., barabasz-krasny, b., puła, j., stachurska-swakoń, a., wang, y. (2019). �e in�uence of aqueous extracts from stellaria media l. on the growth of zea mays l. cultivars. notulae botanicae horti agrobotanici cluj-napoca, 47(3), 921–928. doi: 10.15835/nbha47311597 zeng, l.w., cocks, p.s., kailis, s.g., kuo, j. (2005). structure of the seed coat and its relationship to seed so�ening in mediterranean annual legumes. seed science and technology, 33, 351–362. abstract �e aim of the study was to determine the role of the seed coat in the presence of aqueous extracts from stellaria media (l.) vill. on germination and early growth stages of bean seeds phaseolus vulgaris l. dry shoots of the chickweed aqueous extracts were prepared, with which the bean seeds with coat and without coat were treated. �e control group was seeds watered only with distilled water. a�er 7 days of the experiment, seed germination parameters, seed germination rate (se), seed vitality index (svi), seedling growth inhibition index (ip), fresh and dry mass values and percentage water content were determined. �e experiment showed the germination capacity of bean seeds was varied, in relation to seeds from the control. with increasing concentrations of extracts, a signi�cant reduction in the seed germination rate was observed, both for those with seed coat and without seed coat. �e seed vitality index increased only in seeds with coat, and decreased in each of the applied concentrations of extracts in seeds without seed coat. �e seedling growth inhibition index reached negative values in both groups of seeds tested only at a concentration of 5%. in comparison to the control, ip was positive for seedlings watered with 15% extracts. for p. vulgaris seedlings grown on 5% of extracts the highest increase in the fresh mass was observed, in relation to the value of control mass. for seedlings grown from seeds with seed coat the di�erences in the dry mass values primarily were noted. �e percentage of water content in bean seedlings varied depending on the group of seeds studied and the concentration of allelopathic substances in the chickweed aqueous extracts. �e examined indexes of seed germination and seedling growth showed that in the case of p. vulgaris seeds without seed coat the role of seed coat in the germ ination and early stages of grow th of bean (phaseolus vulgaris l.) in the presence of chickw eed (stellaria m edia (l.) v ill.) 118 were more sensitive to aqueous extracts from dry shoots of s. media. compared to the control group, in low concentrations of allelopathic substances the seeds germinated similarly to the distilled water, and at higher concentrations, the seeds germination, the seedlings growth and fresh and dry mass values were inhibited. key words: allelopathy, seed coat, fresh and dry mass, germination indexes received: [2019.08.05] accepted: [2019.11.20] rola łupiny nasiennej w kiełkowaniu i wczesnych stadiach wzrostu fasoli (phaseolus vulgaris l.), w obecności gwiazdnicy pospolitej (stellaria media (l.) vill.) streszczenie celem badań było określenie roli łupiny nasiennej w obecności wodnych ekstraktów z stellaria media (l.) vill. na kiełkowanie i wczesne etapy wzrostu nasion fasoli żółtostrąkowej (phaseolus vulgaris l.). przygotowano wodne ekstrakty z suchych pędów gwiazdnicy pospolitej, którymi podlewano nasiona fasoli z łupiną nasienną i bez łupiny nasiennej. grupę kontrolną stanowiły nasiona podlewane wyłącznie wodą destylowaną. po upływie 7 dni eksperymentu wyznaczono parametry zdolności kiełkowania nasion, szybkość kiełkowania nasion (se), wskaźnik żywotności nasion (svi), wskaźnik hamowania wzrostu siewek (ip), a także określono przyrost świeżej i suchej masy oraz procentową zawartość wody. eksperyment wykazał zróżnicowaną zdolność kiełkowania nasion fasoli, w stosunku do nasion z próby kontrolnej. wraz ze wzrostem stężeń ekstraktów obserwowano wyraźne zmniejszenie wartości wskaźnika szybkości kiełkowania nasion, zarówno tych z łupiną nasienną, jak i bez łupiny nasiennej. wskaźnik żywotności nasion wzrastał jedynie u nasion z łupiną nasienną, a malał w każdym z zastosowanych stężeń wyciągów u nasion pozbawionych łupiny nasiennej. wskaźnik hamowania wzrostu siewek osiągał wartości ujemne w  obu grupach nasion jedynie przy stężeniu 5%. w porównaniu z kontrolą, ip był dodatni w przypadku siewek podlewanych 15% ekstraktami. największy przyrost świeżej masy, w stosunku do wartości mas z kontroli, zauważono u siewek p. vulgaris wyrosłych na podłożach z 5% wyciągami. różnice w przyroście suchej masy odnotowano dla siewek wyrosłych z nasion z łupiną nasienną. procentowa zawartość wody w siewkach fasoli zmieniała się w  zależności od grupy nasion oraz koncentracji związków allelopatycznych w  wodnych wyciągach z  gwiazdnicy. analizowane wskaźniki kiełkowania nasion i  wzrostu siewek wykazały, iż w  przypadku p. vulgaris bardziej wrażliwe na wodne wyciągi z  suchych pędów s. media były nasiona pozbawione łupiny nasiennej. w niskich stężeniach allelopatin nasiona kiełkowały podobnie jak na wodzie destylowanej, a w wyższych, kiełkowanie, przyrost na długość oraz wartości mas były wyraźnie hamowane. słowa kluczowe: allelopatia, łupina nasienna, świeża i sucha masa, wskaźniki kiełkowania information on the author aleksandra mazur she is interested in an allelopathic interaction between aqueous extracts from weeds and di�erent seeds during germination and growth. a le ks an dr a m az ur 248 annales universitatis paedagogicae cracoviensis studia naturae, 5: 248–252, 2020, issn 2543-8832 koło naukowe przyrodników „biosfera”, działające aktualnie przy uniwersytecie pedagogicznym (up), zostało powołane w październiku 2017 roku. powstało z  połączenia kół naukowych dotychczas działających w ramach instytutu biologii up. nazwa koła – biosfera, strefa obejmująca całą przestrzeń zajętą przez organizmy żywe, została przyjęta ze względu na jego interdyscyplinarność. członkami koła są bowiem studenci pasjonujący się, zarówno dydaktyką, jak i biologią, chemią oraz bioinformatyką. działalność tego koła wzorowana jest na działalności dawnego koła naukowego przyrodników, założonego w  1955 roku. wśród członków dawnego koła byli doktorzy i  profesorowie, którzy aktualnie są nauczycielami akademickimi w instytucie biologii up. obecne koło „biosfera” liczy 20 członków – studentów z  kierunków: biologia, ochrona środowiska, bioinformatyka, a także edukacja techniczno-informatyczna. głównym celem działalności koła jest zdobywanie i rozwijanie wiedzy z zakresu nauk biologicznych oraz nauk im pokrewnych, nawiązywanie współpracy z innymi jednostkami naukowymi, organizowanie spotkań naukowych, scientific group of naturalists “biosphere” at the pedagogical university of krakow (poland) koło naukowe przyrodników „biosfera”, przy uniwersytecie pedagogicznym w krakowie (polska) �e “biosphere” scienti�c group of nature, currently operating at the pedagogical university (pu), was established in october 2017. it was created from the merger of scienti�c groups until now operating within the framework of the institute of biology of the up. �e name of the scienti�c group – the biosphere, the zone covering all the space occupied by living organisms, was adopted because of its interdisciplinary nature. members of the “biosphere” group are students passionate about both didactics and biology, chemistry and bioinformatics. �e activity of this scienti�c group is modelled on the activity of the former naturalists’ science group, founded in 1955. among the members of the former group were doctors and professors, who are currently academic teachers at the institute of biology pu. �e current “biosphere” scienti�c group has 20 members – students from the biology, environmental protection, bioinformatics, as well as technical and it education. �e main aim of this scienti�c group is to acquire and develop knowledge in the �eld of biological sciences and related sciences, establish cooperation with other scienti�c units, organ249 r eports warsztatów i  konferencji, rozszerzanie i  pogłębianie ogólnej wiedzy oraz umiejętności, promowanie instytutu biologii up w  polsce i  na świecie, a  także popularyzacja nauki. opiekunem koła jest dr hab. małgorzata nodzyńska, a  przewodniczącą studentka ii roku biologii justyna mikołajczyk. w  ubiegłym roku akademickim działalność koła naukowego przyrodników „biosfera” skupiła się na wielu zadaniach organizacyjnych, badawczych i  edukacyjnych, z udziałem studentów z instytutu biologii up oraz z  innych jednostek naszego uniwersytetu. nie zabrakło również gości z zewnątrz. w murach instytutu biologii goize scienti�c meetings, workshops and conferences, expanding and deepening general knowledge and skills, promoting the institute of biology pu in poland and on as well as popularising science. �e mentor is assoc. prof. małgorzata nodzyńska, and the chairwoman is justyna mikołajczyk 2nd year biology student. last academic year, the activity of the “biosphere” scienti�c group of naturalists focused on many organisational, research and educational tasks, with the participation of students from the institute of biology of up and from other units of our university. �ere were also external guests. within the fig. 1. students from the “biosphere” scienti�c group and children taking part in activities promoting biological knowledge, as part of the project “towards science – develop passions and interests” (photo. p. cieśla) ryc. 1. studenci z koła „biosfera” oraz dzieci biorące udział w zajęciach popularyzujących wiedzę biologiczną, w ramach projektu „w kierunku nauki – rozwijać pasje i zainteresowania” (fot. p. cieśla) 250 r ep or ts ściliśmy uczniów szkół podstawowych, ponadpodstawowych, a  nawet zaszczyciła nas swoją obecnością wspaniała grupa „misiów”, z  przedszkola samorządowego 36 w  krakowie. jednym z wydarzeń były zajęcia z dziećmi i  młodzieżą, zrealizowane przy wsparciu władz województwa małopolskiego i stowarzyszenia kulturalno-oświatowego – „razem, bliżej, nowocześniej”, w ramach projektu „w kierunku nauki – rozwijać pasje i zainteresowania”. członkowie koła przygotowali z  tej okazji najróżniejsze zajęcia praktyczne, o tematyce biologicznej i chemicznej. razem z  dziećmi i  młodzieżą badali tajniki układu nerwowego, pokazywali w  jaki sposób organizm człowieka chroni się przed bólem fig. 2. students from the “biosphere” naturalists’ scienti�c group, involved in last year’s sos action – colleges for animal shelters (photo. k. romanowska) ryc. 2. studentki z koła przyrodników „biosfera”, zaangażowane w ubiegłoroczną akcję: akcja sos – uczelnie schroniskom zwierząt (fot. k. romanowska) walls of the institute of biology, we hosted primary and secondary school students, and even honoured us with their presence a wonderful group of “bears” from the local government kindergarten 36 in kraków. one of the events was classes with children and youth, carried out with the support of the authorities of the lesser poland voivodship and the cultural and educational association – “together, closer, more modern” as part of the project “towards science – develop passions and interests”. on this occasion, members of the “biosphere” group prepared various practical classes on biological and chemical topics. together with children and young people they studied the secrets of the nervous 251 r eportsoraz obserwowali pod mikroskopem różne interesujące preparaty (ryc. 1). oprócz zajęć praktycznych, studenci z  koła „biosfera” zaprosili dr mariusza gogól, ze stowarzyszenia rzecznicy nauki instytutu zootechniki (państwowy instytut badawczy), do dyskusji na temat „popularyzacja nauki – jak to robić?”. na spotkaniu, które cieszyło się dużym zainteresowaniem, został poruszony m.in. temat wykorzystania nowoczesnych technik przekazu informacji. warto także wspomnieć, że koło naukowe przyrodników, przy wsparciu władz uniwersytetu oraz samorządu studentów, zorganizowało konkurs fotogra�czny pod hasłem „natura to za mało?”. celem tego konkursu było wsparcie pasji studentów i pracowników uniwersytetu w  odkrywaniu piękna i  magii otaczającego świata oraz uwiecznianie przyrody na fotogra�ach. z  ponad 60 nadesłanych prac fotogra�cznych, jury konkursu wyróżniło cztery najciekawsze ich zdaniem, które wraz 30 innymi fotogra�ami można było podziwiać na wernisażu, zorganizowanym w budynku głównym up. ponadto, koło naukowe starało się wspierać pasję teatralną dzieci i  młodzieży, poprzez organizację tzw. spektakli chemicznych. jednym z  nich był spektakl pt. „o doktorze, który na piękno pomoże” – wystawiony w maju 2019 roku, w ramach xix małopolskiego festiwalu nauki i  sztuki w  krakowie. członkowie koła naukowego „biosfera” wsparli także bezdomne zwierzęta, biorąc czynny udział w  zbiórce karmy, datków �nansowych i  innych potrzebnych akcesoriów, podczas listopadowej międzyuczelnianej akcji: akcja sos – uczelnie schroniskom zwierząt (ryc. 2). w  tym czasie studenci prowadzili zbiórkę karmy dla system, showed how the human body protects against pain and observed various interesting preparations under a microscope (fig. 1). in addition to practical classes, students from the “biosphere” scienti�c group invited phd mariusz gogól from the association of science advocates of the institute of animal production (national research institute) to discuss the topic “popularisation of science – how to do it?”. at the meeting, which enjoyed great interest, it was discussed by topic of using modern information transfer techniques. it is also worth mentioning that the naturalists’ scienti�c group, with the support of the university authorities and the student government, organised a photo competition under the slogan “is nature not enough?”. �e aim of this competition was to support the passion of students and employees of the university in discovering the beauty and magic of the surrounding world and capturing nature in photographs. from over 60 submitted photographic works, the jury of the competition distinguished four most interesting in their opinion, which together with 30 other photographs could be admired at the vernissage, organised in the pu main building. in addition, the scienti�c group tried to support the theatrical passion of children and youth by organising the so-called chemical performances. one of them was the spectacle “about a doctor who will help for beauty” – performed in may 2019, as part of the xix festival of science and arts in kraków. members of the “biosphere” scienti�c group also supported homeless animals by actively participating in the collection of food, �nancial donations and other accessories needed during the november inter-university action: 252 r ep or ts izabela wiśniowska*, justyna mikołajczyk institute of biology, pedagogical university of krakow, podchorążych 2 st. 30-084 kraków, poland; *izabela.wisniowska.1999@gmail.com sos action – colleges for animal shelters (fig. 2). at that time, the students were collecting dog and cat food, and collected money for the kraków homeless animal shelter. �anks to all these actions, members of the “biosphere” scienti�c group tried to include in active activities that support the popularisation of natural knowledge in its various aspects and to stimulate students to act. what counts above all in this group is the community and the opportunity to establish relationships with other people. psów i  kotów oraz kwestowali na potrzeby krakowskiego schroniska dla bezdomnych zwierząt. dzięki tym wszystkim akcjom członkowie koła „biosfera” starali się włączyć do aktywnych działań, wspierających popularyzację wiedzy przyrodniczej w  różnych jej aspektach oraz pobudzić studentów do działania. w  tym kole naukowym tym liczy się przede wszystkim społeczność oraz możliwość nawiązywania relacji z  innymi ludźmi. 180 annales universitatis paedagogicae cracoviensis studia naturae, 4: 180–190, 2019, issn 2543-8832 doi: 10.24917/25438832.4.12 sylwia śliwińska-wilczewska institute of oceanography, university of gdansk, av. piłsudskiego 46, 81-378 gdynia, poland; ocessl@ug.edu.pl cyanobacteria and cyanometabolites used in the pharmaceutical and medical industry introduction cyanobacteria from marine and freshwater habitats are known to produce a diverse array of active compounds (cyanometabolites). �ese include low molecular weight peptides, polysaccharides, fatty acids, phenols and alkaloids (burja et al., 2001; mazur-marzec et al., 2015). some of them are a threat to human and environmental health. but many of these natural products possess considerable interest due to their potential applications (berry et al., 2008; leão et al., 2012; almeida et al., 2015). it was estimated that out of the 660 new compounds identi�ed in marine bacteria in the years 1997–2008, till 33% were derived from cyanobacteria (imho� et al., 2011). �ese compounds could be used to obtain commercial algaecides, herbicides, and insecticides. furthermore, some of these chemical substances demonstrated antifungal, antibacterial, antiviral and even antitumor activity, which could lead to the development of new drugs from them (berry et al., 2008). �us, the issue of commercial application of cyanobacteria and their cyanometabolites requires more attention and investigation. �e �rst discovery of the healing properties of cyanobacteria took place in 1500 b.c. where the nostoc sp. species was used to treat several forms of cancer (singh et al., 2011). in recent years, many studies have been conducted on the use of cyanobacteria as a potential source of new biologically active substances e.g., microginine, cyanopeptin, aeruginosine and spumigine (głowacka et al., 2007). �e last two can be used to treat hypertension, cardiovascular diseases, and viral infections. in turn, microginine is used to cure hypertension, and cyanopeptin for asthma and viral infections (singh et al., 2011). bouillomides a and b from lyngbya bouillonii l.ho�. & v.dem. are strong inhibitors of serine protease, elastase, and chymotrypsin (rubio et al., 2010). particularly valuable are such compounds that have of antiviral (cyanovirin, scytovirin), antifungal (�scherelin, cryptophycin, calophycin), antibacterial c yanobacteria and cyanom etabolites used in the pharm aceutical and m edical industry 181 (microviridine, muscoride, nostocin a), antitumor (apratoxins a and d, dolastine) and antimalar (ambigol c) nature (wright et al., 2005; głowacka et al., 2007). in the years 1981–2002, over 60% of anti-cancer and anti-infectious drugs were of natural origin. currently, due to the high costs of introducing new products to the market by the pharmaceutical industry (500–2000 million dollars), the number of new drugs is decreasing. currently, drugs are becoming less e�ective because the resistance of pathogens to antibiotics is increasing. �erefore, it is important to explore new biologically active compounds to produce new drugs (lam, 2007). �e main aim of this study was to present the knowledge on active compounds of cyanobacteria, which may have potential applications in the pharmaceutical and medical industries. �is topic is very important but is still not su�ciently understood. for centuries, human diseases have been treated with natural products because these plant-based, natural drugs are much healthier than their chemical counterparts. in this paper, we showed the positive aspects of cyanobacteria cultivation and possibilities of its commercial use. algae nomenclature was used here according to algaebase (https://www.algaebase.org/) and other microbes from di�erent sources. medical and pharmaceutical use of cyanobacteria initial knowledge of the properties of cyanobacteria enabled their application on an industrial, pharmaceutical, and medical scale. �ey produce many biologically active cyanometabolites which have, among others, anticancer, antifungal, antiviral, anti-in�ammatory, and antimalarial properties (gupta et al., 2013; fig. 1). cyanobacteria also contain pigments that can strengthen the immune system, and even reduce the risk of heart disease multiple sclerosis, cataracts, and age-related diseases, as well as prevent cancer. pigments can be used both as medicines and cosmetics, as well as natural pigments for products including ice cream, sweets, so� drinks, and milk products. extract from blue pigment, which is phycocyanin, obtained from the species of e.g., arthrospira sp., is used in eye shadow, lipsticks and eyeliner. moreover, this pigment inhibits pancreatic lipase and also, depending on the dose, the growth of ehrlich cancer cells (el-baky, 2003). carotenoids, which are antioxidants, are also anti-cancer drugs substances (sheih et al., 2009). chlorophyll together with pigments such as phycocyanin and phycoerythrin have a protective e�ect against uv radiation on the skin, thus delaying the aging process. in addition, cyanobacteria produce polysaccharides that have the ability to stabilise emulsions and suspensions, form gels, etc., and these can be used in the cosmetics industry as ingredients in nutrients, creams and many other similar products, as well as in the pharmaceutical industry as ingredients in medicines (głowacka et al., 2007; tab.1). s yl w ia ś liw iń sk aw ilc ze w sk a 182 microand macroelements from cyanobacteria perhaps, cyanobacteria arthrospira platensis gomont can be the richest source of vitamins as well as macroand microelements. �ey contain nutrients, i.e. β-carotene (with antioxidant properties which protect the organism against free radicals), iodine, selenium, zinc, iron, magnesium, manganese, copper and γ-linolenic acid, derived from the group of omega-6 fatty acids, glycolipid h-b2, which is an inhibitor of pancreatic lipase and vitamin b12, which is necessary for the proper functioning of nerve tissue, b1, b2, b3 and e (klasik et al., 2010; gupta et al., 2013). isolated compounds of the arthrospira sp. species have nourishing, strengthening and detoxifying properties, and the extract from its cells has antiallergic, antiviral properties, inhibits carcinogenesis processes, and also reduces blood cholesterol (głowacka et al., 2007). arthrospira sp. is also rich in proteins – contain their about 60% (ishimi et al., 2006). it also helps alleviate the occurrence of anemia during pregnancy (nuhu, 2013). cyanobacteria compounds with antiviral activity viral diseases, including hiv, a�ects many people around the world. according to who and unaids, 36.7 million people lived with hiv at the end of 2016, of which 1 million died due to it. �ousands of marine organisms, including cyanobacteria, have been tested for antiviral properties (yasuhara-bell, lu, 2010). such an antiviral compound is cyanovirin produced by nostoc ellipsosporum rabenh. ex born. & flah. fig. 1. examples of cyanobacteria as a potential source for commercial applications: arthrospira sp. (a), lyngbya sp. (b), leptolyngbya sp. (c), nostoc cf. commune vauch. ex born. & flah. (d), nostoc muscorum c.ag. ex born. & flah. (e), and pseudanabaena sp. (f). scale bar = 10 µm (photo. s. śliwińskawilczewska) c yanobacteria and cyanom etabolites used in the pharm aceutical and m edical industry 183 �is protein inhibits proliferation of hiv-1, hiv-2, acquired immunode�ciency virus fiv cats and siv monkeys (głowacka et al., 2007). nosto�an polysaccharide from n. �agelliforme (born. & flah.) elen. also has activity against the herpes-1 virus (singh et al., 2011). scytovirin isolated from the aqueous extract of scytonema varium kütz. ex born. & flah. is also an antiviral protein. it acts similarly to the already mentioned cyanovirin, because both inhibit the process of virus absorption on the surface of host cells. it has been proven that if scitovirin is given up to 8 hours a�er infection with the virus, it successively prevents the development of infection (bokesch et al., 2003). in 2002, water extracts from arthrospira maxima setchell & n.l.gardner cells were tested and they were shown to inhibit the development of infections caused by the hsv-2 and hsv-1 herpes virus, cmv cytomegalovirus and the virus causing aujeszky’s prv disease. polysaccharides produced by cyanobacteria including a. platensis, demonstrated the ability to inhibit the replication of hsv, hiv-1, hiv-3, in�uenza a, and mumps virus, which were cultured in suspensions of human cell cultures (głowacka et al., 2007). zainuddin et al. (2002) researched the aqueous and methanolic extract from cyanobacterial cultures of the genus calothrix, microcystis, nodularia, oscillatoria, lyngbya and scytonema to check their activity against in�uenza a virus in a dog’s kidney cells. �e most e�ective extract turned out to be obtained from cyanobacteria of the genus microcystis. a reduction of approximately 90% in viral replication was observed due to protease inhibiting activity. none of the methanol extracts was cytotoxic. besides, lau et al. (1993) tested the ability of aqueous cyanobacterial extracts to inhibit reverse transcriptase rt of the avian myeloblastoma virus amv and hiv and it was proved that 18 (2.0%) extracts showed this possibility in the range of over 50%. on this basis, it was concluded that cyanobacteria can be a promising source of compounds used for viral therapies. cyanobacteria compounds with antibacterial activity kreitlow et al. (1999) studied hydrophilic and lipophilic extracts of cyanobacteria for their antibacterial activity. in the case of gram (-) bacteria, no inhibitory activity was found. while for gram (+) seven species of cyanobacteria from twelve (anabaena lemmermannii p.g.richt., a. solitaria kleb., limnothrix sp., microcystis ichthyoblabe (g.kun.) kütz., nodularia sp., oscillatoria rubescens dc ex gomont, o. tenuis c.ag. ex gomont) showed high activity against at least one of the bacteria such as: bacillus subtilis ehren., micrococcus �avus cohn and staphylococcus aureus f.j. rosenbach. in terms of antibacterial activity, oufdou et al. (2001) tested the benthic species of cyanobacteria pseudanabaena sp. and they showed that the extracts secreted by these organisms inhibited the growth of bacteria escherichia coli t. esch., salmonella sp. and staphylococcus aureus. in turn, antibacterial activity against bacillus cereus frank. & s yl w ia ś liw iń sk aw ilc ze w sk a 184 frank., escherichia coli and staphylococcus epidermidis evans have a compound called noscomin obtained from nostoc commune vauch. ex born. et flah. (singh et al., 2011). gutiérrez et al. (2008) isolated two abietane diterpenes from microcoleous lacustris desikachary and proved that they showed activity against bacteria salmonella typhi (=s. enterica (ex kau�. & edw.) le minor & popo� serovar typhi), staphylococcus aureus, s. epidermidis and vibrio cholerae pacini. another species of cyanobacteria that produces antimicrobial peptides, including microviridine and cavaguchipeptin is microcystis aeuroginosa kütz. in addition, antibacterial activity was discovered from muscoride isolated from nostoc muscorum c.ag. ex born. & flah., bastadine from anabaena basta, microsporins from nostoc commune, nostocycline a from nostoc sp. and nostocin a from n. spongiaeforme c.ag. ex born. & flah. all these metabolites have antibacterial properties because they can destabilise bacterial cell walls (głowacka et al., 2007). cyanobacteria compounds with antifungal activity many cyanobacteria also exhibit properties to inhibit fungal life processes. �e methanol extract from anabaena solitaria kleb. has antifungal activity against alternaria alternate (fr.) keissl., botrytis cinereal pers., and colletotrichum gloeosporioides (penz.) penz. & sacc. similarly is with cells from cyanobacteria nostoc commune, which, in addition to the fungi mentioned above, also act against fusarium oxysporium schltdl., phytophthora capsica leon., pythium ultimum trow and rhizopus stolonifer (ehrenb.) vuill. (kim, 2006). �e anti-fungal activity was also shown by ambigol a and b from fischerella ambigua (kütz. ex born. & flah.) gomont (falch et al., 1995), �scherellin from f. muscicola gomont (srivastava et al., 1998) and tanicolide cryptophycins, and a-d majuskulamides from lyngbya majuscula harv. ex gomont (głowacka et al., 2007). cyanobacteria compounds with anti-cancer activity it was found that some cyanometabolites have apoptotic properties. such type of action have got, among others, dolastine, which was initially isolated from sea hares (family aplysiidae), but later it was discovered that it is also produced by cyanobacteria (costa et al., 2012). �us, this compound can be used to suppress unnecessary and potentially harmful cells. dolastine 10, a pentapeptide produced by symploca sp., induces apoptosis of human lymphoblastic leukemia cells (wall et al., 1999) and also has an inhibitory e�ect on lung cancer cells (kalemkerian et al., 1999). apatoxin d isolated from lyngbya sp. has similar cytotoxic properties concerning lung cancer (gutiérrez et al., 2008). apratoxin a, extracted from l. majuscula harv. ex gomont cells, inhibits bone c yanobacteria and cyanom etabolites used in the pharm aceutical and m edical industry 185 sarcoma cancer cells (liu et al., 2009). microcyclamide from microcystis aeuroginosa has cytotoxic properties on murine p388 leukemia cells (ishida et al., 2000). calothrixin from cyanobacteria calothrix sp. inhibits the growth of hela cervical cancer cells (rickards et al., 1999). majngulamide c from lyngbya majuscula has inhibitory activity against lung cancer, large intestine cancer and glioblastoma cells (vijayakumar, menakha, 2015). coibamide from leptolyngbya sp. is a cyclic depsipeptide that causes cell cycle inhibition in the g1 phase for mda-mb-435 breast cancer (costa et al., 2014). cryptophycin-1 from nostoc linckia born. ex born. & flah. has a cytotoxic e�ect on cancer cells of the large intestine, prostate, solid tumors, colon cancer ht-29, caco-2 and gc3, breast cancer mcf-7 and mda-mb-231, hela cervical cancer, and also leukemia u937, ccrf-cem and hl-60 (shih, teicher, 2001; singh et al., 2011; vijayakumar, menakha, 2015). nodularins and microcystin from cyanobacteria are toxic inhibitors of protein phosphatases pp1 and pp2a therefore, a�er appropriate chemical modi�cation they can be used to produce analogues of potential drugs used to inhibit the processes leading to the formation of cancer (łukomska et al., 2002). costa et al. (2014) tested the anti-cancer activity of marine cyanobacterial strains. five of them were the most interesting in terms of bioactive compounds: leptolyngbya fragilis (gomont) anag. & kom., l. halophila (hansgirg) anag. & kom., l. mycoidea (frémy) anag., nodosilinea nodulosa (z.li & j.brand) perkerson & casamatta and synechocystis salina wislouch. it was also shown that two of them – l. fragilis and s. salina – are the most bioactive against cancer cells. cyanobacteria compounds with antiprotozoal activity tropical diseases such as malaria, cholera, leishmaniasis, african coma, schistosomiasis caused by protozoa are equally dangerous. according to who, over a billion people struggle with one or more of these diseases (simmons et al., 2008). cyanobacteria have been found to contain compounds that inhibit protozoa that cause these diseases. viridamide a is a compound isolated from oscillatoria nigrovirdis �waites ex gomont that acts against trypanosoma cruzi chagas, leishmania mexiana garnham, and plasmodium falciparum schaudinn (singh et al., 2011). ambigol c from fischerella ambigua (kütz. ex born. & flah.) gomont acts against trypanosoma rhodesiense and p. falciparum (wright et al., 2005). in addition, a compound called nostocarboline extracted from nostoc sp. also exhibits activity against protozoa such as leishmania donovani ross, p. falciparum, trypanosoma brucei plimmer & bradford and t. cruzi chagas (singh et al., 2011). s yl w ia ś liw iń sk aw ilc ze w sk a 186 tab. 1. examples of cyanobacterial with potential use in medicine and pharmaceutical industry cyanobacteria e�ect references arthrospira platensis gomont antiviral activity (hiv type 1, herpes simplex, polio) głowacka et al. (2007) dichothrix baueriana born. & flah. antiviral activity (herpes simplex type 2) larsen et al. (1994) fischerella muscicola gomont fungicides hagmann and jüttner (1996) lyngbya lagerheimii (gomont ex gomont) anag. & kom. antiviral activity (hiv type 1) gustafson et al. (1989) lyngbya sp. antitumor activity simmons et al. (2005) nostoc sp. cholinesterase inhibitor (alzheimer’s disease) blom et al. (2006) oscillatoria agardhii gomont larvicidal activity harada et al. (2000) phormidium tenue gomont antiviral activity (hiv type 1) gustafson et al. (1989) phormidium sp. fungicides (oral candidiasis) garima et al. (2013) pseudoanabaena sp. antibacterial activity (e. coli, salmonella sp.) głowacka et al. (2007) scytonema ocellatum lyngb. ex born. & flah. fungicides patterson and bolis (1995) symploca hydnoides kütz. ex gomont antiparasitic activity (malaria, chagas disease) linington et al. (2008) westiellopsis sp. larvicidal activity (malaria, meningitis) rao et al. (1999) conclusions cyanobacteria have a wide range of occurrences and occupy many habitats, including oceanic areas, freshwater lakes, and even extreme habitats such as deserts, coastal rocks, glacial lakes or hot springs. in eutrophic and hypertrophic waters, cyanobacteria o�en dominate the phytoplankton in the summer period, creating a massive and harmful bloom. some cyanobacteria produce toxins that can signi�cantly a�ect human health. however, even though cyanobacteria contain toxic substances, there are species whose secreted organic compounds can be used as a potential source for commercial applications. �e properties of cyanobacteria discussed in this work emphasize how these organisms can be potentially used by humans in many areas of their life. cyanometabolites can serve as drugs for incurable diseases such as cancer or other diseases caused by bacteria, viruses, fungi or protozoa. however, the composition and functional role of many cyanometabolites remain unknown, therefore, the issue of commercial application of cyanobacteria and their cyanometabolites require more attention and investigation. acknowledgements �is study was supported by bmn grants, poland, no. 538-g245-b116-18. con�ict of interest �e author declares no con�ict of interest related to this article. c yanobacteria and cyanom etabolites used in the pharm aceutical and m edical industry 187 references algaebase https://www.algaebase.org/. 1996–2019 m.d. guiry. almeida, j., freitas, m., cruz, s., leão, p., vasconcelos, v., cunha, i. 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(2002). cyanobacteria a potential source of antiviral s yl w ia ś liw iń sk aw ilc ze w sk a 190 substances against in�uenza virus. medical microbiology and immunology, 191(3–4), 181–182. doi: 10.1007/s00430-002-0142-1 cyjanobakterie i cyjanometabolity stosowane w przemyśle farmaceutycznym oraz medycznym streszczenie związki bioaktywne sinic wykazują różnorodne właściwości, które potencjalnie mogą być wykorzystane w wielu sektorach przemysłu. w artykule tym szczególny nacisk położono na wykorzystanie sinic i ich cyjanometabolitów, zarówno w przemyśle farmaceutycznym, jak i medycznym. scharakteryzowano związki wyizolowane ze szczepów sinic, które można stosować do wytwarzania leków o działaniu przeciwwirusowym, przeciwgrzybiczym, przeciwnowotworowym, przeciwdrobnoustrojowym oraz przeciwbakteryjnym. pokazano również pozytywne aspekty hodowli sinic i możliwości ich komercyjnego wykorzystania. key words: applications, blue-green algae, cyanobacteria, cyanometabolites, industry received: [2019.11.05] accepted: [2019.12.05] 149 annales universitatis paedagogicae cracoviensis studia naturae, 4: 149–160, 2019, issn 2543-8832 doi: 10.24917/25438832.4.9 andrzej danel1*, joanna puła2 1faculty of food technology, department of chemistry, �e h. kołłataj university of agriculture, balicka st. 122, kraków, poland; *rrdanela@cyf-kr.edu.pl 2 faculty of agriculture and economics, department of agroecology and plant production, �e h.kołłataj university of agriculture, mickiewicza 2 ave, kraków, poland plants as a treasury of fragrant substances for food industry and perfumery introduction fragrant substances play an important role in our life. every day we accidentally or deliberately smell hundreds or even thousands of natural and arti�cial compounds. some of them exhibit pleasant aroma like vanilla in ice creams or constituents of top perfumes. before we start to drink a good wine or whisky our noses are attacked with dozens of volatile chemical molecules preparing our tongues for subsequent �avour experiences. on the other hand sweat components or volatiles released from some mould cheeses or fermented food like ‘surströmming’ arouse disgust in some persons but a small amount of very bad smelling organic sulphur compounds like tetrahydrothiophene in natural gas we use in our kitchen can save somebody life in case of gas leakage. it would be di�cult to imagine our existence without various aromas. it seems that eating is one of the biggest pleasures we can experience in our life. even in the holy bible there is a  statement “�ere is nothing better for a  man than taking meat and drink, and having delight in his work”. eating is a multisensory experience. �e very �rst contact is with eyes, next the senses of taste and smell are engaged. in the paper published in science the authors claim that people are able to recognise more than one trillion of olfactory stimuli (bushdid et al., 2014). �ese results are breathtaking and in the strong contrast with many previous statements sometimes not proven by experiments. in popular literature we can still �nd relatively old information that people can distinguish just about 3.000–10.000 odours (crocker, henderson, 1927). food technologists very o�en encounter the problem of destroying food aroma during thermal food processing. �e volatile compounds are lost and the �avour of a nd rz ej d an el , j oa nn a p uł a 150 the resulting product may be di�erent from the starting components. at present due to the fragrant food additives we are able to a certain extent restore these aromas. addition of some synthetic or natural aromas to food products increases their tastiness and sensory attractiveness. it should be remembered that there are two ways in which we smell – orthonasal and retronasal route (spence, 2017). �is short review is devoted to plant-derived aromatic substances applied as food additives and in some cases as materials for perfume industry as well. essential oils – preparation essential oils belong to the natural fragrant materials widely applied in food industry and in pharmaceutical and perfumery ones too. �ese materials can be obtained in many ways. �e most important ones are extractions with various solvents like n-hexane c6h14, ethyl acetate ch3cooet, methylene chloride ch2cl2, �orasol (1,1,1,2-tetra�uoroethane) or supercritical carbon dioxide. �e last solvent is de�nitely environmental friendly and safe as far as residues in the �nal product. in food industry it is used for removal of ca�eine from co�ee beans and green tea (kim et al., 2010). another example is the extraction of essential from lavandula ×hybrida rever. (lavandin) �owers with supercritical carbon dioxide. �e two most important components of this oil are linalool and linalyl acetate applied in food industry and as biocides as well (kamali et al., 2015). supercritical carbon dioxide was also applied in isolation of essential oils from flixweed, eucalyptus globulus labill., mentha ×piperita l. (zekovic et al., 2009; mahdavi et al., 2015; singh et al., 2016). other examples can be found in some review papers on this subject (xu et al., 2011; capuzzo et al., 2013; manjare et al., 2019). �e second important method of essential oil isolation is steam distillation. �e steam �ow is passed through the plant material (bark, �owers, roots, leaves, peel, berries, rhizome) placed in a glass �ask or steel container and removes volatile compounds with subsequent condensation. �e distillate is collected in receiver and an essential oil is separated from the water phase. �e resulted oil can be subjected to fractional distillation and recti�cation. in some cases the cooling of the resulted oil results in crystallisation of some constituents. steam distillation process can be supported with microwave heating of the plant material/water mixture for more e�ective isolation of fragrant material (chemat et al., 2006; sahraoui et al., 2008; moradi et al., 2018). another group of researchers applied solvent free microwave extraction sfme of essential oils from plant material (lucchesi et al., 2018). �e authors signi�cantly reduced the time of oil extraction in comparison with traditional hydro-distillation method. at present a microwave reactor is a standard tool for chemists. beside it ultrasound assisted reactions are more and more popular in organic and analytical chemisp lants as a treasury of fragrant substances for food industry and perfum ery 151 try (capelo-martinez, 2009; cravotto et al., 2018). �is technique is a valuable tool for extraction of essential oils and other plant metabolites. �e details of this procedure and its application in food industry, cosmetics and pharmacy can be found in a recent review paper (chemat et al., 2017). it would be good to mention probably the oldest method of essential oils preparation based on the cold pressing of plant material like olives or citrus peels. �e most important products obtained in this way are orange, lemon, grapefruit and bergamot essential oils which are widely employed in food and cosmetic industry. in case of danger that some constituents of essential oils can be decomposed at the temperature of steam distillation the procedure of maceration can be applied. one of the oldest and very e�ective though very tedious is en�eurage. �is method is based on extracting of fragrant compounds from plant material (usually �ower petals or whole �owers) with animal fat like lard or tallow and leaving the whole for a few days. a�er this time the �owers are removed and new ones are mixed with lipids. �is procedure is repeated up to the saturation of fat with essential oils. a�er removing the �owers the fat is mixed with ethyl alcohol. �e components of essential oils are dissolved in it and the insoluble fat is separated o�. �e residue resulted a�er the evaporation of the alcohol is called an absolute. �e technique of en�eurage is almost abandoned and replaced with solvent extraction (surburg, penten, 2006). such an example is jasmine absolute, a very valuable ingredient for perfumery industry. at present it is prepared by double or triple extraction of jasmine blossoms with n-hexane. one needs to collect manually 8.000 000 blossoms to obtain 1 kg of jasmine absolute (konopski, koberda, 2003). �e chemical constituents of essential oils essential oils applied as food additives or products for fragrant composition in perfume industry are complicated mixtures of many compounds. for example the recent investigations on volatile and semi-volatile compounds in various citrus oils (ex. citrus limon (l.) burm., c. sinensis (l.) osbeck, c. medica l.) based on gas chromatography coupled to mass spectrometry (gc-ms) revealed the presence of 200–400 compounds (bozkurt et al., 2017; gonzales-mas et al., 2019). �e essential oils extracted from rosa ×centifolia l. or r. ×damescena mill. are one of the most expensive ingredients applied in cosmetics industry. �e amount of volatile compounds varies depend on the literature source. some authors reported 32 compounds based on gc and gc-ms chromatography. among them the most abundant were citronellol, geraniol and nerol (ahmad et al., 2009). �e other group reported 50 volatiles in damascene rose oil (naquvi et al., 2014). a recent review on rose essential oil or ‘liquid gold’ constituents suggests that investigation on this subject is far to be �nished (nunes, graca, 2017). a nd rz ej d an el , j oa nn a p uł a 152 fig. 1. chemical structures of some isolates from essential oils: 1) anethole, 2) benzaldehyde, 3) camphor, 4) cinnamaldehyde, 5) citral a, 6) citral b, 7) citronellal, 8) (s)-carvone, 9) (r)-carvone, 10) eucalyptol, 11) eugenol, 12) (r)-(+)-limonene, 13) linalyl acetate, 14) menthol, 15) pinene (source: rutkowski et al., 2003; kołodziejczyk, 2013) in some cases the content of some fragrant materials is quite high so some essential oils are subjected to fractional distillation or crystallisation to obtain valuable materials for food industry, perfumery or organic synthesis. �e signi�cant majority of them are terpenes hydrocarbons or their monofunctional derivatives. beside them we can �nd esters, aromatic aldehydes, phenols, ketones, ethers and nitrogen, oxygen and sulphur containing heterocycles. p lants as a treasury of fragrant substances for food industry and perfum ery 153 some important isolates 1–15 from essential oils are listed in �gure 1 (rutkowski et al., 2003; kołodziejczyk, 2013). �e worldly production of anise essential oil is estimated over 400 tonnes and the content of trans-anethol 1 is ca. 70–95% depending on the plant source. anethol can be prepared in a synthetic way too. it is used as a food additive to some alcoholic beverages like absinth, anisette and raki. �e last one is an alcoholic drink popular in turkey (ashurst, 1999). benzaldehyde 2 is obtained either synthetically or from natural cinnamaldehyde 4 in the reaction with sodium hydroxide (wiener, pittel, 1985; surburg, panten, 2006). it can be also prepared from amygdalin extracted from some kernels like apricot or bitter almonds (passos, ribeiro, 2010). cinnamaldehyde 4 is obtained from cinnamon bark cinnamomum verum j.presl or c. cassia (l.) j.presl through steam distillation. �ere are some alternative synthetic pathways to prepare this compound like aldol condensation of benzaldehyde and acetic aldehyde but due to the high worldly production of cinnamic bark the isolation is more economic. benzaldehyde 2 and cinnamaldehyde 4 are applied in food industry as �avours in chewing gums, cakes, bakery aromas or ice creams. camphor 3 can be isolated from c. camphora ness et eberm tree grown in china, vietnam, japan and taiwan. as a  food additive natural camphor is popular in india (aguilar et al., 2008). it can be also synthesised from natural pinene 15. beside it this terpene is used for synthesis of other fragrant substances like terpineol or verbenone. �ese substances can be prepared either via chemical or microbial oxidation of pinene (rozenbaum et al., 2006; praskoso et al., 2018). citronella grass cymbopogon nardus (l.) rendle is a rich source of citral a (geranial) 5, citral b (neral) 6 and citronellal 7. �ese compounds are used in food industry as lemon, lime and orange �avorings for ice creams, candies and baked goods (winter, 2009). geranial and neral were also found in hop essential oil. �ey are reduced by yeast into geraniol and nerol and in part these compounds are responsible for beer �avour (tressl et al., 1987). two carvone enantiomers 8 and 9 are used as food additives, insects repellents and building blocks in asymmetric organic synthesis (cravalho, fonesca, 2006). �ese compounds can be isolated from carum carvi l. and mentha spicata l., respectively. �e major application of eucalyptol 10 involves candies, aromatic balms or mouthwash (cameron, easton, 2000). just recently eucalyptol was applied as solvent in synthesis of heterocyclic compounds (campos et al., 2019). eugenol 11 is known to possess antiseptic properties and is used in dentistry. its application as food additive is relatively limited due to the very strong odour. �is compound is obtained from oil of clove derived from steam distillation of �ower buds, leaves and steam of syzygium aromaticum (l.) merr. & perry. chatterjee and paramita described investigations on application of eugenol as natural antioxidant in mayonnaise (chatterje, bhattacharje, 2015). moreover it can be a  starting material for synthesis of vanillin – a valuable natural fragrant substance (lampman et al., 1977). (r)-(+)-lia nd rz ej d an el , j oa nn a p uł a 154 monene 12 is produced on the biggest scale in the world. �e annual production of this fragrant compound in brazil and florida is estimated at 75.000 t (taylor, 2002). due to this abundant amount this compound with the smell of orange or lemon is applied not only in food industry and perfumery but it is used as a solvent for grease removal and a paint stripper too. �e bergamot essential oil derived from bergamot oranges contains limonene 12 (30.7%) and linalyl acetate 13 (30.1%), respectively (sawamura et al., 2006). �e last compound found a vast application in perfume industry (fahlbush et al., 2003). essential oils from mentha arvensis l. and m. ×piperita l. contain menthol 14 which found multiple applications in many consumer products like cosmetics, drugs and tobacco �avour additive (aldadyan, samet, 2018). diamonds among fragrant materials in some essential oils the content of a single fragrant substance can be very high. such an example is (r)-(+)-limonene. in citrus, orange or lemon essential oils its amount varies within 65–90%. on the other hand there are valuable aromatic substances which exist in very small amounts in plant material. some of them are depicted in �gure 2. one of the examples is nootkatone 16 which can be found in grapefruit, orange and mandarin oranges essential oils. it can be isolated from these sources but due to tiny amount of 0.01–0.5% the price of the natural compound is very high and varies between 4.000–6.500 euro/kg. pure compound found an application as insect repellent and a  food additive approved by fda (food and drug administration) as well (jordan et al., 2012). to avoid high costs of natural compound, alternative synthetic methods were developed including biotechnological based ones (fratz et al., 2009). even a more expensive food �avour additive and a diet supplement is raspberry ketone 17. it is a natural compound which can be found in cranberries, blackberries and raspberries. in the last example it occurs in the amount of 1–4 mg/kg so the extraction from natural resources is useless from economical point of view because the estimated price is ca. 20.000$/kg (beekwilder et al., 2007). �ere is a big demand for this �avour ingredient as a  food additive and a  diet supplement as well. �us raspberry ketone can be synthesised from simple starting materials such as p-hydroxbenzaldehyde 22 and acetone 23 resulting chalcone 24 which can be reduced with cheap reagents like nickel boride and hydrogen ni2b/h2 under atmospheric pressure yielding the �nal compound 17 (fig. 3) (bandarenko, kovalenko, 2014). unfortunately this product cannot be recognised as natural in spite of the identical properties with the raspberry ketone obtained from natural resources. to ful�l these demands, methods based on natural substrates and preparation processes as natural as possible are in great demand. joulain and fuganti published a procedure employing a baking yeast for reduction of a double bond in 4-(p-hydroxyphenyl)but-3-en-2-one 24 yielding raspberry ketone 17 in 56% yield (joulian, fuganti, 1999). acetone 23 can p lants as a treasury of fragrant substances for food industry and perfum ery 155 fig. 2. chemical structures of some expensive fragrant materials: 16) nootkatone, 17) raspberry ketone, 18) rose oxide, 19) cis-jasmone, 20) trans-jasmone, 21) vanillin (source: ruzicka, pfei�er, 1933; berger, 2007; fratz et al., 2009; alsters et al., 2010; bandarenko, kovalenko, 2014) fig. 3. synthesis of raspberry ketone: a) naoh, r.t. 24–48 hrs; b) ni2b or baker yeast (source: joulain, fugati, 1999; bandarenko, kovalenko, 2014) be obtained from microbial synthesis (sauer, 2016). natural p-hydroxybenzaldehyde 22 can be extracted from sorghum shoots (sorghum vulgare pers.) (bove, conn, 1961). rose oxide 18 is one of the constituent of rose essential oil obtained from rosa ×damascena and r. ×centifolia as well. due to the very high price of the previously mentioned essential oils, some synthetic procedures were developed to prepare this compound because it is used in contemporary perfume compositions (alsters et al., 2010). rose oxide was found also in some muscat wines and is one of the components responsible for �oral-green �avour of those wines (boelens et al., 1993). �e similar situation is with one of the constituent of the jasmine essential oil namely cis-jasmone 19 used in the creation of high quality perfumes. �e content of a nd rz ej d an el , j oa nn a p uł a 156 this compound in jasmine essential oil varies at 2.6–3.4% so the isolation is de�nitively unpro�table (clarke, 2008). it was discovered over 100 years ago and since that time many synthetic pathways have been developed to ful�l demand of perfume producers (hesse, 1899). �e synthetic jasmone is frequently a  mixture of cis/trans 19/20 isomers (ruzicka, pfei�er, 1933). �e last, but not the least example in this small gallery of natural fragrant materials is vanillin 21. �e natural compound can be extracted from vanilla planifolia andrews grown in madagascar. unfortunately the natural resources are not su�cient to satisfy the demands of the market so many synthetic procedures were developed to supply this valuable product for food and perfumery industry. �us synthetic vanillin can be prepared on industrial scale either from eugenol, guaiacol or from lignin – a  side product from cellulose industry (berger, 2007). �ere is also a  technological procedure of vanillin preparation from cow dung. �e author of the procedure – japanese scientist mayu yamamoto was awarded with ig-nobel prize for this outstanding achievement in 2008 (yamamoto et al., 2008). natural vanillin extracts are expensive so there is a danger that synthetic vanillin can be used to adulteration of natural extract. to avoid these problems numerous analytical techniques are applied including authentication with plant dna (philippe et al., 2019). conclusions �is very short review gives a slight glimpse on the application of fragrant substances of plant origin in contemporary food and perfume industry. at present as a  whole 2.000–3.000 �avouring chemicals – both natural and synthetic are used commercially. �e search and analysis of natural fragrant substances is especially challenging due to the fact that their content in plant or animal material is sometimes on the verge of a  homeopathic one. on the other hand the rapid progress in modern analytical techniques gives birth to hope that many new aromatic molecules will be isolated and characterised in the future. moreover we have to remember that a lot of organic chemists from academic and industrial laboratories constantly modify known structures and synthesise new molecules with interesting fragrant properties which do not exist in nature, so we can expect new olfactory experiences. con�ict of interest �e authors declare no con�ict of interest related to this article. references aguilar, f., autrup, h., barlow, s., castle, l., crebelli, r., dekant, w., engel, k.-h., gontard, n., gott, d., grilli, s., gürtler, r., larsen, j.ch., leclercq, c., leblanc, j.-c., malcata, x., mennes, w., milana, m.r., pratt, i., rietjens, i., tobback, p., toldrá, f. 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(2008). novel production method for plant polyphenol from livestock excrement using subcritical water reaction. international journal of chemical engineering, 603957. doi: 10.1155/2008/603957 zeković, z., lepojević, z., milić, s., aadamivić, d., mujić, i. (2009). supercritical co2 extraction of mentha (mentha piperita l.) at di�erent solvent densities. journal of the serbian chemical society, 74(4), 417–425. a nd rz ej d an el , j oa nn a p uł a 160 rośliny jako skarbnica substancji zapachowych dla przemysłu spożywczego i perfumeryjnego streszczenie artykuł poświęcony jest niektórym aspektom substancji zapachowych pochodzenia roślinnego, stosowanych jednocześnie w  przemyśle spożywczym i  perfumeryjnym. od starożytności opracowano wiele technik ekstrakcji w  celu uzyskania olejków eterycznych. niektóre z  nich są nadal stosowane. nowe ekstrakcje, takie jak: mikrofalowe lub ultradźwiękowe, są coraz bardziej popularne i pozwalają zaoszczędzić czas oraz koszty. niezależnie od procedury powstałe olejki eteryczne są źródłem wielu związków chemicznych, tzw. izolatów. mogą one być stosowane jako dodatki do żywności lub jako materiały wyjściowe do syntezy organicznej. niektóre substancje zapachowe występują w bardzo małych ilościach w materiale roślinnym, dlatego ekstrakcja nie jest opłacalna ekonomicznie, ale po ustaleniu ich struktur chemicznych i opracowaniu procedur syntetycznych, w niektórych przypadkach są one pozyskiwane na skalę przemysłową. substancje opisane poniżej to tylko niewielka część z 2000–3000 pachnących cząsteczek, które sprawiają, że nasze życie jest przyjemniejsze, zarówno w  jedzeniu, jak i perfumach. key words: essential oils, extraction techniques, food additives, fragrant substances. received: [2019.05.06] accepted: [2019.11.22] 242 annales universitatis paedagogicae cracoviensis studia naturae, 5: 242–247, 2020, issn 2543-8832 virtual open day at the institute of biology pedagogical university of krakow (april 21 2020) wirtualny dzień otwarty instytutu biologii uniwersytetu pedagogicznego w krakowie (21 kwiecień 2020) in the event of closure of schools and universities due to the sars-cov-2 coronavirus pandemic, for the sake of health and a sense of security, employees, doctoral students and students of the institute of biology at the pedagogical university of krakow invited high school graduates and other groups of young people to actively participate in the virtual open day. �e ‘on-line’ meeting was took place on april 21, 2020 in the a�ernoon on the o�cial facebook pro�le of the pedagogical university and the institute of biology. as in the stationary mode of such meetings, virtually, representatives of the university and the institute of biology wanted to familiarise future students with the didactic and scientific o�er of our unit. �e event was o�cially opened by the vice-rector for development, assoc. prof. robert stawarz. �en the live meeting was joined by a representative of the recruitment o�ce – msc rafał birgiel, who answered on questions about the limits of places for individual �elds of study, admission criteria, point thresholds, etc. a�er an hour from the beginning, the meeting was joined by the president of the student w  sytuacji zamknięcia szkół i  uczelni z  powodu pandemii koronawirusa sars-cov-2, w  trosce o  zdrowie i  poczucie bezpieczeństwa pracownicy, doktoranci i  studenci instytutu biologii uniwersytetu pedagogicznego w  krakowie zaprosili maturzystów oraz inne grupy młodzieży do czynnego udziału w  wirtualnym dniu otwartym. spotkanie ‘on-line’ odbyło się 21 kwietnia 2020 w  godzinach popołudniowych na o�cjalnym pro�lu facebooka uniwersytetu pedagogicznego oraz instytutu biologii. podobnie jak w trybie stacjonarnym tego rodzaju spotkań, tak i  wirtualnie, przedstawiciele uniwersytetu i  instytutu biologii chcieli zapoznać przyszłych studentów z  ofertą dydaktyczno-naukową naszej jednostki. uroczystego otwarcia tego wydarzenia dokonał prorektor ds. rozwoju dr hab. robert stawarz, prof. up. następnie do spotkania na żywo dołączył przedstawiciel biura rekrutacji – mgr rafał birgiel, który odpowiadał na pytania, dotyczące limitów miejsc na poszczególne kierunki studiów, kryteriów przyjęć na studia, progów punktowych itp. po godzinie od rozpoczęcia do spotkania dołączyła przewodnicząca samorządu studentów 243 r eportsgovernment and a computer science student – kamila kowalczyk. during the meeting, she answered all questions raised by participants of this event. �e questions concerned both subjects related to studying and student life outside the university walls. she also talked about full-time and �eld classes, methods of passing subjects, cooperation with lecturers, student activities in scienti�c groups, national and international exchange of students, co-creation of student organisations by the academic community. during this event, participants in correspondence with representatives of individual university scienti�c units. employees, doctoral students and students of the institute of biology also took an active part in this meeting. �ey presented the full o�er of �elds of study implemented at the institute of biology. to this end, short information (posts) with descriptions of the o�ered courses have been posted on the facebook portal of the institute of biology. for example, in the �eld of biology one could read that undergraduate or graduate studies (full-time and part-time) special attention is paid to the need to acquire interdisciplinary knowledge in the �eld of research issues and methodology in order to explain complex phenomena and biological processes as well as to learn and apply modern research methods and techniques used in modern biological sciences. during their studies, biology students, in addition to theoretical knowledge, acquire skills that will allow them to �nd jobs in science laboratories, i  studentka kierunku informatyka – kamila kowalczyk. podczas spotkania odpowiadała ona na wszystkie pytania zgłaszane przez uczestników tego wydarzenia. zapytania dotyczyły, zarówno tematów związanych ze studiowaniem, jak i życiem studenckim poza murami uczelni. opowiadała również o zajęciach stacjonarnych oraz terenowych, sposobach zaliczeń, współpracy z  wykładowcami, działalności studentów w kołach naukowych, wymianie krajowej i  międzynarodowej studentów, współtworzeniu organizacji studenckich przez społeczność akademicką. w  trakcie tego wydarzenia, uczestniczy korespondowali z  przedstawicielami poszczególnych jednostek naukowych uniwersytetu. pracownicy, doktoranci i studenci instytutu biologii również wzięli czynny udział w  tym spotkaniu. zaprezentowali oni pełną ofertę kierunków studiów realizowanych w instytucie biologii. w tym celu na portalu społecznościowym facebook instytutu biologii zostały zamieszczone krótkie informacje (posty), z  opisami oferowanych kierunków. na przykład, o kierunku biologia można było przeczytać, że na studiach licencjackich, czy magisterskich (stacjonarnych i  niestacjonarnych) zwraca się szczególną uwagę na konieczność zdobycia wiedzy interdyscyplinarnej, w zakresie problematyki i metodologii badawczej, celem wyjaśniania złożonych zjawisk oraz procesów biologicznych oraz poznanie i  stosowanie nowoczesnych metod i  technik badawczych, wykorzystywanych we współczesnych naukach biologicznych. studenci biologii w  trakcie studiów, oprócz wiedzy teoretycznej, nabywają umiejętności, które w  przyszłości pozwolą im znaleźć pracę w  laboratoriach naukowych, parkach 244 r ep or ts national parks, and for those interested in the teaching specialty, also in primary and secondary school. for those interested not only in biology, but also in computer science, bioinformatics was the second direction proposed by the institute of biology. it is a progressive �eld of study created for the needs of the modern labour market. it is a combination of computer science and biology, which enables the education of future experts, mainly dealing with the use of tools in the �eld of information science in biological research. bioinformatics is certainly an attractive direction for all those who are interested in new mathematical and natural technologies. engineering studies assume the education of future graduates in biology, chemistry, computer science, mathematics and physics. �ey are useful in solving tasks regarding the processing and interpretation of data obtained, both in the laboratory and in the �eld. students develop here their analytical competences and the ability to think logically, thanks to which they can easily �nd themselves in professional work. �e interdisciplinary nature of bioinformatics provides broad education and allows �nding employment in interesting areas determined by the labour market. environmental protection is the third course proposed by the institute of biology. �ese are �rst degree engineering studies, enabling the acquisition of knowledge and skills in the �eld of natural and technical sciences. �e study program includes issues related to human impact on the environment as well as legal and economic foundations for environmental protection, promoted in the aspect of sustainable development. �e stunarodowych, a dla zainteresowanych specjalnością nauczycielską, także w szkole podstawowej i ponadpodstawowej. dla zainteresowanych nie tylko biologią, ale i  informatyką, drugim kierunkiem proponowanym przez instytut biologii była bioinformatyka. jest to progresywny kierunek studiów stworzony na potrzeby nowoczesnego rynku pracy. stanowi połączenie informatyki z  biologią, które umożliwia kształcenie przyszłych ekspertów, zajmujących się głównie zastosowaniem narzędzi z  obszaru nauk informatycznych w  badaniach biologicznych. bioinformatyka to na pewno kierunek atrakcyjny dla wszystkich tych osób, które są zainteresowane nowymi technologiami matematyczno-przyrodniczymi. studia inżynierskie zakładają wykształcenie przyszłych absolwentów w  zakresie biologii, chemii, informatyki, matematyki oraz �zyki. są użyteczne w rozwiązywaniu zadań, dotyczących przetwarzania i  interpretacji danych pozyskanych, zarówno w laboratorium, jak i w terenie. studenci rozwijają tu swoje kompetencje analityczne i  umiejętność logicznego myślenia, dzięki czemu bez trudu odnajdują się w pracy zawodowej. interdyscyplinarność kierunku bioinformatyka zapewnia szerokie wykształcenie oraz pozwala na znalezienie zatrudnienia w  interesujących obszarach, wyznaczanych przez rynek pracy. trzecim proponowanym przez instytut biologii kierunkiem jest ochrona środowiska. są to studia inżynierskie i-go stopnia, umożliwiające zdobycie wiedzy i  umiejętności w  zakresie nauk przyrodniczych oraz technicznych. program studiów obejmuje zagadnienia dotyczące oddziaływania człowieka na środowisko oraz podstawy prawne 245 r eportsdent acquires practical skills in monitoring the environment, using physical, chemical and biological methods that can be used in future work. to understand the processes occurring in nature and the impact of man on the environment, it is worth applying to one of two specialties: (1) geographical environment management and (2) renewable energy sources, which specialties include subjects closely related to their speci�city. �e institute of biology of the pedagogical university in the o�er of study �elds also invited to study of biological wellness. �ese types of second-cycle studies in extramural mode give an opportunity to learn what is the physiology of e�ort, what are the physiological parameters of health and disease, how does the wellness system work and what is the organisation and management of spa & wellness centres. �ey help to understand what customer service is in the �eld of wellness and psychotherapy services and relaxation techniques. classes in unconventional medicine, dietetics as well as management and marketing provide the basis for development in the �eld of personal trainer. a novelty in the o�er of study at the pu institute of biology are �rst-cycle studies – chemistry-chemistry teacher, implemented as part of the european union project “a competent teacher – master and educator”. studying within this project is an opportunity to obtain the quali�cations of a chemistry teacher and guarantees the support of experts as part of individual classes. studies are based on the principles of so-called ‘green chemistry’ – a concept that assumes designing and conducting chemical shows in a way that limits the use and formation of harmful subi  ekonomiczne ochrony środowiska, promowane w  aspekcie zrównoważonego rozwoju. student zdobywa praktyczne umiejętności monitorowania środowiska, przy użyciu metod �zycznych, chemicznych i biologicznych, które może wykorzystać w  przyszłej pracy. aby zrozumieć procesy zachodzące w  przyrodzie i  wpływ człowieka na środowisko, warto aplikować na jedną z  dwóch specjalności: (1) zarządzanie środowiskiem geogra�cznym i (2) odnawialne źródła energii, które to specjalności obejmują przedmioty ściśle, związane z ich specy�ką. instytut biologii up w  ofercie kierunków zapraszał także do studiowania na kierunku odnowa biologiczna. tego rodzaju studia ii stopnia w trybie niestacjonarnym, dają okazję do poznania, m.in. czym jest �zjologia wysiłku, jakie są �zjologiczne parametry zdrowia i  choroby, jak funkcjonuje system odnowy biologicznej oraz na czym polega organizacja i  zarządzanie ośrodkami spa & wellness. pomagają one zrozumieć, czym jest obsługa klienta w  zakresie odnowy biologicznej oraz usług psychoterapii i  technik relaksacyjnych. zajęcia z  zakresu medycyny niekonwencjonalnej, dietetyki oraz zarządzania i marketingu, dają podstawę do rozwoju w zakresie trenera personalnego. nowością w  ofercie kierunków studiów w instytucie biologii up, są studia i stopnia – chemia-nauczyciel chemii, realizowane w ramach projektu unii europejskiej „kompetentny nauczyciel – mistrz i  wychowawca”. studiowanie w ramach tego projektu jest szansą na uzyskanie kwali�kacji nauczyciela chemii i  gwarantuje wsparcie ekspertów w  ramach zajęć indywidualnych. studia są oparte o zasady tzw. ‘zielonej chemii’ – kon246 r ep or ts stances. as part of the studies, an innovative program and modern teaching methods are proposed, e.g. tutoring, practical laboratory and �eld classes, research projects, auto-evaluations, creation of thematic lapbooks and others. as part of the studies, two specialties were prepared: a chemistry and nature teacher, a chemistry teacher and a nature culture animator. graduates of the teaching specialisation will be prepared to act as a teacher of chemistry and nature in primary school. in addition, they will have appropriate quali�cations in the �eld of psychology, pedagogy and didactics of chemistry. �e nature culture animator specialisation will enable you to take up work, e.g. in museums and science centres, as well as in units dealing with non-formal education. in addition to the o�er of �elds of study, students from the “biosphere” naturalist scienti�c group presented their activities. members of the scienti�c group talked about how they broaden their passions in biology and what their activities are. �ey talked about cooperation with schools, organising classes for children and youth on biological and chemical topics, about active participation in the collection of food, funds and other accessories for homeless animals as part of the action: sos action – colleges of shelters, about supporting passion for immortalising nature in photos as part of the competition and the opening of “nature is not enough?”, as well as exploring theatrical passions in the framework of chemical performances being prepared. students of all faculties of the pu institute of biology actively participate not only in scienti�c and didactic life, but also in orcepcji, zakładającej projektowanie i  przeprowadzanie pokazów chemicznych w  sposób, ograniczający użycie i  powstawanie szkodliwych substancji. w  ramach studiów proponowany jest innowacyjny program oraz nowoczesne metody kształcenia, np. tutoring, praktyczne zajęcia laboratoryjne i  terenowe, projekty badawcze, autoewaluacje, tworzenie tematycznych lapbooków i  inne. w  ramach studiów przygotowano dwie specjalności: nauczyciel chemii i przyrody, nauczyciel chemii i  animator kultury przyrodniczej. absolwenci specjalności nauczycielskiej będą przygotowani do pełnienia roli nauczyciela chemii i  przyrody w szkole podstawowej. dodatkowo będą posiadać odpowiednie kwali�kacje z  zakresu psychologii, pedagogiki i dydaktyki chemii. specjalizacja animator kultury przyrodniczej umożliwi podjęcie pracy, np. w  muzeach i centrach nauki oraz w jednostkach zajmujących się edukacją pozaformalną. oprócz oferty kierunków studiów, czynnie zaprezentowali swoją działalność studenci z  koła naukowego przyrodników „biosfera”. członkowie koła opowiadali, w  jaki sposób poszerzają swoje zamiłowania i pasje do biologii i na czym polega ich działalność. mówili o  współpracy ze szkołami, o  organizowaniu zajęć dla dzieci i  młodzieży o  tematyce biologicznej i  chemicznej, o  czynnym udziale w zbiórce karmy, środków �nansowych i innych akcesoriów dla bezdomnych zwierząt w ramach akcji: akcja sos – uczelnie schroniskom, o  wspieraniu pasji do uwieczniania przyrody na zdjęciach w  ramach konkursu i  wernisażu „natura to za mało?”, a  także o zgłębianiu pasji teatralnych w ramach przygotowywanych spektakli chemicznych. 247 r eportsganisational activities. �ey actively participate in the implementation of various types of initiatives for the social environment, in the preparation and implementation of the science and art festival, the małopolska night of scientists, the open day of the pedagogical university, the małopolska festival of innovation. �ey are invited to participate in the erasmus+ and “most” mobility programs, in national and international conferences, lectures of foreign guests, scienti�c and didactic meetings, during which they establish contacts and cooperation, which o�en results in their subsequent employment. �e organisers of this year’s virtual open day at the pu institute of biology expressed the hope that this initiative brought everyone interested in the o�er of studies and other opportunities to deepen their knowledge. at the same time, they cordially invited you to join our academic community. all details regarding recruitment for biological sciences can be found on the websites of the institute of biology and the main website of the pedagogical university and on the facebook social portal of the institute of biology. studenci wszystkich kierunków instytutu biologii up aktywnie uczestniczą nie tylko w życiu naukowym i dydaktycznym, ale także w  działalności organizacyjnej. czynnie biorą udział w  realizacji różnego rodzaju inicjatyw na rzecz otoczenia społecznego. aktywnie uczestniczą w przygotowaniach i realizacji festiwalu nauki i sztuki, małopolskiej nocy naukowców, dniu otwartym uniwersytetu pedagogicznego, małopolskim festiwalu innowacji. zapraszani są do uczestniczenia w programach mobilności erasmus+ i  „most”, w  konferencjach krajowych i  międzynarodowych, wykładach gości zagranicznych, spotkaniach naukowych, i  dydaktycznych, podczas których nawiązują kontakty i  współpracę, co niejednokrotnie owocuje ich późniejszym zatrudnieniem. organizatorzy tegorocznego wirtualnego dnia otwartego instytutu biologii up wyrazili nadzieję, że inicjatywa ta przybliżyła wszystkim zainteresowanym ofertę studiów i innych możliwości pogłębiania wiedzy. jednocześnie, serdecznie zapraszali do przyłączenia się do naszej społeczności akademickiej. wszelkie szczegóły, dotyczące rekrutacji na kierunki biologiczne, można znaleźć na stronach instytutu biologii oraz głównej stronie uniwersytetu pedagogicznego i na portalu społecznościowym facebook instytutu biologii. grzegorz rut https://orcid.org/0000-0001-6719-3060 institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland; grzegorz.rut@up.krakow.pl 83 annales universitatis paedagogicae cracoviensis studia naturae, 5: 83–95, 2020, issn 2543-8832 doi: 10.24917/25438832.5.6 bent al-hoda asghari1, mohsen yousefi2, katarzyna możdżeń3, joanna puła4, peiman zandi5,6*, wang yaosheng6 1department of agronomy, science and research branch, islamic azad university, tehran, iran 2department of agronomy, takestan branch, islamic azad university, takestan, iran 3institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland 4faculty of agriculture and economics, university of agriculture, kraków, poland 5international faculty of applied technology, yibin university, yibin, sichuan, 644600, p. r. china 6institute of environment and sustainable development in agriculture, chinese academy of agricultural sciences, beijing 100081, p. r. china; *peiman.zandi@yibin.edu.cn agronomic performance of autumnand winter-cropped indian mustard (brassica juncea l.) in response to varying levels of nitrogen fertiliser introduction currently, a huge gap in supply and demand for oilseed calls for a considerable increase in oilseed crop production around the globe (radha kumari et al., 2004). based on reports by the food and agriculture organization of the united nations (2019), the world average yield of oilseed crops is 350 mt, which is 875 times more than iran’s oilseed production (400,000 t). generally, oilseed crops are grown in many countries, and in iran they are especially cultivated for oil production (shirani rad, zandi, 2012). indian mustard (brassica juncea l. czern) is an opportune crop for regions with short seasons and low rainfall (burton et al., 1999), and its use and cultivation are increasing in the world as a medicinal plant with oily seeds and high nutritional value. �e noted mustard-growing countries are india, canada, china, pakistan, poland, bangladesh and sweden (iraddi, 2008). currently, iran has insu�cient agronomic knowledge for mustard (locally known as khardal) production, and this problem has restricted its conventional production; yet, it can be found as a wild plant throughout a wide range of the country. proper sowing time is a climate-dependent factor that provides su�cient growth and development and plays a central role in the satisfactory production and productivity of any crop (pandey et al., 1981; verma et al., 2012), as it provides optimum growth conditions such as temperature, light, humidity and rainfall (gul, ahmad, 2007; iraddi, 2008). �e role of sowing time in mustard growth has been investigated by many researchers, as shown in table 1. most of them concur that sowb en t a l-h od a a sg ha ri, m oh se n y ou se fi, k at ar zy na m oż dż eń , j oa nn a p uł a, p ei m an z an di , w an g y ao sh en g 84 ing time is crucial for mustard production (rahman et al., 1988; ra�ei et al., 2011). early sowing of crops in semi-dryland conditions, like the takestan region, may be confronted by drought stress; however, the risk of frost loss increases if sowing is delayed in autumn cultivation. a later sowing date for mustard can reduce the adequate growth period due to high temperatures during the reproductive phase, which can subsequently decrease yield (radha kumari et al., 2004), especially in winter-sown crops. various sowing dates and varieties provide variable environmental conditions within the same region in terms of crop growth and development and yield stability (daneshian et al., 2008). bhuiyan et al. (2008) pointed out that environmental conditions a�ected the seed yield and maturity of mustard to a large extent. tab. 1. �e recommended proper sowing dates for mustard production growth qualitative and quantitative parameters sowing dates for optimum crop performance plant height august 30 (ra�ei et al., 2011), 10th and 30th october (angrej et al., 2002), october 14 (singh, singh, 2002), october 16 (panda et al., 2004), 2nd and 3rd week of october (singh et al., 2001), november 15 (aziz et al., 2011), and november 17 (kurmi, 2002) number of siliques per plant august 30 (ra�ei et al., 2011), september 25 and october 5 (shivani, sanjeev, 2002),10th and 30th october (angrej et al., 2002), october 14 (singh, singh, 2002), october 16 (panda et al., 2004), 2nd and 3rd week of october (singh et al., 2001), november 15 (aziz et al., 2011), and november 17 (kurmi, 2002) seed yield august 30 (ra�ei et al., 2011), during ii fortnight of september (iraddi, 2008), october 1 (radha kumari et al., 2004), october 14 (singh, singh, 2002), october 15 (awasthi et al., 2008), october 16 (panda et al., 2004), 2nd and 3rd week of october (singh et al., 2001), 3nd week of october (gajendra, 2001; raj singh et al., 2001), october 17 (yadav et al., 1999), october 20 (khichar et al., 2000; khan, tak, 2002; yadav yogesh et al., 2011), october 24 (khushu, singh, 2005), 30th october (bhuiyan et al., 2008) and november 15 (aziz et al., 2011) biomass yield october 5 (singh et al., 2002), october 14 (singh, singh, 2002), october 15 (sihag et al., 2003), october 16 (panda et al., 2004), october 18 (jadhav, singh, 1992) and october 24 (khushu, singh, 2005) oil content october 10 (bishnoi, singh,1979), 10th and 30th october (angrej et al., 2002), and october 27 (das, 1998), november 17 (kurmi, kalita ,1992) �e di�erences between the reported dates may come from the various environmental regions under which these experiments have been undertaken nitrogen management is one of the critical focal points in the cropping system (maresma et al., 2019). it is o�en a demanding task to strike a balance between levels su�cient for normal plant growth and those that are approved for human consumption (maereka et al., 2007). usually, plants take up nitrogen through fer85 a gronom ic perform ance of autum nand w inter-cropped indian m ustard (brassica juncea l.) in response to varying levels of nitrogen fertiliser tiliser application. allen and morgan (1972) stated that rapid growth, increased seed and fruit production and enhanced leaf quality in oilseed crops are, together, highly dependent on nitrogen supply. long-term �eld experiments on mustard revealed that su�cient levels of nitrogen progressively led to an increase in crop performance (yadav et al., 1994; bhalerao, 2001; garg et al., 2001; premi, manoj, 2004; mckenzie et al., 2006; verma et al., 2012). nitrogen application during the sowing and �owering stages resulted in rapid leaf area development; prolonged life of leaves; improved duration of leaf area a�er �owering; enhanced number and weight of siliques, seeds and �owers per stand and increased overall crop assimilation, which as a result led to increased seed yield and quality in most stand crops (wright et al., 1988; geetha et al., 2011). it was also shown that increasing available nitrogen prolonged the vegetative growth and increased dry matter accumulation (šidlauskas, tarakanovas, 2004). however, a considerable decrease in physiological yield potential and slowing of the �nal growth stage through bending stems (or lodging) was noticed by wright et al. (1988) in response to crop supplementation with a large amount of nitrogen fertiliser. �is paper aims to evaluate the e�ects of �ve levels of nitrogen fertiliser, supplied in the form of urea, during two planting seasons, on the growth parameters, seed yield and agronomic properties of indian mustard in the agro-climatic conditions of takestan (iran). materials and methods study site �e �eld experiments were conducted at the research station of takestan azad university in south-west qazvin plain (36°03ʹn; 49°42ʹe) from 2009 to 2010. �is site is 1,283.4 m above sea level, accentuated by a thermo-mediterranean (semi-arid) climate, according to de martonneʹs classi�cation, which is summarised as hot and dry summers and cool and wet winters. �e average from a 30-year climatic data record shows a mean annual rainfall of 312 mm with uneven distribution during the year, which is mostly concentrated in late autumn and early spring. in the region, the winter temperature can fall below 8.2°c and in the summer it can rise above 38.7°c. meteorological data records were acquired from the agrometeorological station in the university. �e total precipitation that occurred during the study (october 2009–july 2010) was 326.3 mm. moreover, during the 2009 mustard-growing period, the total rainfall was lower than 2010 (56.2 mm and 270.1 mm, respectively), especially during the vegetative phase for the �rst sown treatments (fig. 1). b en t a l-h od a a sg ha ri, m oh se n y ou se fi, k at ar zy na m oż dż eń , j oa nn a p uł a, p ei m an z an di , w an g y ao sh en g 86 soil properties tab. 2. physical and chemical analysis of soil of the experimental �eld (2009–2010) physical properties soil texture 0–60 depth [cm] clay loam clay [%] 29 silt [%] 45 fine sand [%] 26 chemical analysis [mg/1000 g soil] exchangeable k 330 available mn++ 2.90 cu++ 0.90 fe++ 7.80 zn+ 1.20 p 26.60 total nitrogen n [%] 0.089 organic carbon c [%] 1.03 ec [ds/m/25°c] 1.43 ph [h2o] 8.10 �e presented data are the mean values for two soil depths of ‘0–30’ and ‘30–60’ cm; ec – electrical conductivity fig. 1. monthly minimum temperature (tmin), maximum temperature (tmax) and rainfall during the �rst (october 2, 2009–2010 to june 15, 2010) and the second (march 1, 2009–2011 to july 16, 2010) crop cycle of mustard 87 soil samples to a depth of 60 cm were collected from 15 cores in each block prior to cultivation, and the composite samples were prepared for soil fertility analysis. �e soil had a low content of both nitrogen (n) (0.08–0.09%) and organic carbon (oc) (1.03%), was slightly alkaline (ph (h2o) = 8.10) and had a soil textural class of clay loam (tab. 2). experimental design a split-plot trial arrangement based on a randomised complete block design with four replicates was established, with two planting seasons (ps1 = october 2, 2009 and ps2 = march 1, 2010) as the main plots and nitrogen levels (0, 50, 100, 150, and 200 kg n ha–1), supplied as urea, as subplots. each subplot consisted of 8 rows, 30 cm apart and 4 m long × 3 m width (plot size: 27.5 m long × 4 m width). �e trial �eld was initially irrigated and, at the stage of soil puddling, it was ploughed once deeply via a mouldboard plough. next, to fragment clods and to create uniform soil conditions, a perpendicular disk was used. �is was followed by use of a harrowing thrice and smoothing with a wooden board to supply a �ne seedbed – the furrows and ridges were constructed with a furrower and the distance between the ridges was 30 cm. for treatment, a standard dose of phosphorus, i.e. 70 kg ha–1 single super phosphate along with one-third of the nitrogen treatment, was incorporated and added to the soil at the time of �nal land preparation prior to drilling by disking. pre-sowing watering was done to ensure normal germination and isomorph plant stands. irrigations was provided when required to preserve the soil’s moisture at an optimum condition. �e mustard cultivar ‘landrace’ was used in the trial. �e pre-soaked seeds were sown using a single row hand drill with a seeding rate of 5 kg ha–1 at 2 cm depth in rows with 30 cm spacing. a thinning operation was performed about two weeks a�er sowing to leave more vigorous stands and to �nally maintain plant-to-plant distance at 5 cm. �e weeds were eradicated by hand-hoeing as necessary. a basal half dose of non-applied nitrogen was given at the 4–6 leaves phase and the remaining dose was top-dressed at the start of �owering. all other recommended crop management practices were carried out uniformly and regularly for all of the treatment groups during the probationary period to ensure proper crop growth. �e plots were harvested in late spring (10 to 15 june, 2010) and early summer (11 to 16 july, 2010) for the �rst and second seeding dates, respectively. �e shoots of �ve randomly selected and tagged plants belonging to subplots were hand-harvested at the end of the growing period and the plant height and the number of siliques per plant were recorded. �e mean height of �ve randomly selected plants from the base to the tip of the main stem was measured and expressed in centimetre (cm). when the majority of stands were at full maturity in all plots, an area of 3.6 m2 (the inner �ve rows) was evaluated to determine the biomass and seed yield. simultaneously, the quadrate samples were cut by hand a gronom ic perform ance of autum nand w inter-cropped indian m ustard (brassica juncea l.) in response to varying levels of nitrogen fertiliser b en t a l-h od a a sg ha ri, m oh se n y ou se fi, k at ar zy na m oż dż eń , j oa nn a p uł a, p ei m an z an di , w an g y ao sh en g 88 at approximately 5 cm above the ground and le� in the �eld for sun drying until they reached a constant weight. a�er one week, when the plant water content was less than 12%, the samples were weighed and recorded as biomass yield [t ha–1] per subplot. next, they were threshed using a kurt pelz stationary thresher and hand-cleaned. �e separated seeds were air-dried to 10–12% humidity and weighed with a digital balance (kaifeng group co., ltd., china). �e seed yield was then determined and expressed in kg seed ha–1. a sample of 100 seeds was taken from every seed lot related to subplots, oven-dried at 105°c for 1.30 h and ground into �ne particles. �e oil content in powdered samples [%] was determined using a soxhlet instrument with carbon tetrachloride as an organic solvent (250 cc; 1:1 ratio; at 70°c). statistical analysis data were analysed statistically using proc glm (sas inc., 2001) to detect signi�cant di�erences (p ≤ 0.05) among treatments and the comparisons of means were carried out applying the method of duncan’s multiple range test (dmrt) at a 5% level of signi�cance. �e ombrothermic diagram was drawn with ms excel ver. 2007. results and discussion results revealed that the simple e�ect of cultivation season and nitrogen rate was significant for plant height, biomass yield, the number of siliques per plant, seed oil content and seed yield (p ≤ 0.01). �e interaction of cultivation season and nitrogen rates also had a signi�cant e�ect on plant height and the number of siliques per plant (tab. 3). �e means comparison of planting season and nitrogen rate interaction showed that 150 and 200 kg n ha–1 treatments were signi�cantly more e�ective than other treatments in terms of plant height, for autumn planting on october 2. �e lowest plant heights occurred with the zero dose of nitrogen in both the winter (81.7 cm) and autumn (99.5 cm) cultivation season (tab. 3). increasing nitrogen consumption increases the amount of protein in the cells, a subsequent increase in cell size and larger leaf area, resulting in greater photosynthetic activity and ultimately leading to an increase in plant height in rapeseed (wysocki et al., 2007). greater plant heights are due to having a longer in�orescence axis, or in other words, having the largest number of �owers and pods on the in�orescence of the stem. additionally, leaf fall during the �lling of siliques with seeds requires plant photosynthesis to be maintained by the siliques and the stems. �erefore, having a longer stem means having a higher photosynthetic surface and producing more metabolic material to �ll siliques and seeds (norton et al., 1991). �e means comparison of planting season e�ect on the amount of biomass showed that the autumn planting season of october 2, with an average of 12.64 t ha–1, result89 ed in statistically higher levels of biomass than the winter planting season of march 1. �e results also showed that treatments of 150 and 200 kg n ha–1 resulted in the highest biomass yield, while the lowest amount of biomass was found for the control treatment (tab. 3). tab. 3. plant parameters of indian mustard (brassica juncea l. czern) in response to di�erent cultivation seasons and rates of nitrogen fertiliser application (mean 2009–2010) treatments plant height [cm] biomass yield [t ha–1] number of siliques per plant [no.] seed oil content [%] seed yield [kg ha–1] autumn cultivation 144.5a 12.64a 118.60a 42.57a 3394a winter cultivation 116.2b 9.58b 89.00b 41.13b 2459b nitrogen fertilisation (n) n1: 0 kg ha–1 (ck) 90.60d 8.26d 61.20e 41.25c 1506e n2: 50 kg ha–1 121.60c 10.46c 93.10d 42.59b 2549d n3: 100 kg ha–1 139.10b 11.81b 112.50c 43.51a 3162c n4: 150 kg ha–1 147.70a 12.40a 123.40b 41.86bc 3624b n5: 200 kg ha–1 152.70a 12.62a 128.80a 40.03d 3794a planting season (ps) autumn cultivation (ps1: october 2, 2009) n1 99.5e 9.93 72.6g 41.87 1985 n2 130.6c 12.00 107.2d 43.23 2976 n3 152.8b 13.23 127.4c 44.16 3562 n4 165.7a 13.94 139.2b 42.76 4123 n5 173.8a 14.12 146.6a 40.85 4325 winter cultivation (ps2: march 1, 2010) n1 81.7f 6.58 49.8h 40.63 1026 n2 112.6d 8.93 79.0f 41.95 2122 n3 125.3c 10.39 97.6e 42.86 2761 n4 129.7c 10.86 107.6d 40.97 3125 n5 131.6c 11.13 111d 39.22 3262 source of variations analysis of variance ps ** ** ** ** ** 1n ** ** ** ** ** ps×n ** ns * ns ns c.v. [%] 4.72 2.35 2.77 1.46 3.88 means with the same letters are not signi�cantly di�erent at p < 0.05; 1n: nitrogen fertiliser as urea was applied in three equal portions: 1) at the pre-sowing stage, 2) on 4–6 leaves growing phase and as top-dress fertiliser 3) in the reproductive stage; ps×n: represents interaction terms between treatment factors; c.v. – coe�cient of variation; ns, f test not signi�cant, *f test signi�cant at p ≤ 0.05, **f test signi�cant at p < 0.01 comparison of the mean e�ects of planting season and nitrogen levels showed that the application of 200 kg n ha–1 during autumn cultivation on october 2 resulted in the highest number of siliques (146.6) in the plant. however, a lack of nitrogen fertilisation decreased the average number of siliques (49.8) for winter cultivation (tab. 3). a gronom ic perform ance of autum nand w inter-cropped indian m ustard (brassica juncea l.) in response to varying levels of nitrogen fertiliser b en t a l-h od a a sg ha ri, m oh se n y ou se fi, k at ar zy na m oż dż eń , j oa nn a p uł a, p ei m an z an di , w an g y ao sh en g 90 it follows from the results that nitrogen application caused a signi�cant increase in the number of branches per plant (data not shown) (raghuvanshi et al., 2018), and subsequently led to the production of more siliques in mustard (bilsborrow et al., 1993). it has been reported previously that the main reason for a decrease in the number of siliques per plant is associated with a decrease in the number of sub-branches (tayo, morgan, 1979). clark and simpson (1978) argue that the numbers of branches in a plant, as well as the number of seeds in a small silique, are in�uenced by environmental conditions. �e results showed that the autumn planting season of october 2 provided the highest content of seed oil, with an average of 42.57%, which was signi�cantly higher than the winter planting season. �e reason for the decrease in the percentage of seed oil in winter cultivation is likely due to temperature changes during the stage of seed �lling and a reduction in net photosynthesis (daneshian et al., 2008). in this case, less overall material is produced, and those carbohydrates are converted to oil. �e longer the �owering to ripening period is, the greater the time for oil synthesis and, ultimately, the percentage of oil increases (gecgel et al., 2007). �e highest seed oil content was 100 kg n ha–1, with an average of 43.51% oil, followed by an increase in nitrogen application, which reduced seed oil content (tab. 3). seed yield results showed that the autumn planting season, with an average of 3,394 kg ha–1, had a signi�cant advantage over the winter planting season, with an average of 2,459 kg ha–1. increasing the length of the growing season, along with the proper conditions for germination and optimal plant establishment, can increase seed yield. planting date is an important factor that a�ects seed yield and seed oil content (koutroubas, papasoska, 2005). johnson et al. (1995) compared di�erent planting dates in canola and concluded that the delay in planting date signi�cantly reduced seed yield, which was due to a decrease in the number of siliques in the plant and a decrease in harvest index. increasing nitrogen consumption to 200 kg n ha–1 increased seed yield. �e treatment of 200 kg n ha–1 resulted in an average of 3,794 kg ha–1, the highest seed yield (tab. 3). kazemeini et al. (2010) showed that increasing nitrogen levels led to an increase in yield components and, ultimately, seed yield of rapeseed genotypes. elewa et al. (2014) also examined new rapeseed cultivars at di�erent nitrogen levels and demonstrated that increasing the amounts of nitrogen increased the seed yield for the studied cultivars. conclusions cultivation season and nitrogen level signi�cantly a�ected plant height, biomass yield, number of siliques per plant, seed oil content and seed yield of indian mustard (brassi91 ca juncea l. czern). �e mean comparison of the interaction between planting season and nitrogen level showed that 150 and 200 kg n ha–1 treatments resulted in higher and superior values for the parameters of interest, with autumn planting, than the other treatments. �e lowest values were observed in control plants. �e use of nitrogen had a positive e�ect on the growth and development of mustard. appropriate fertilisation, climatic conditions and plant care allow for high quality and high quantity of crops. acknowledgements we are very thankful to those who funded us during the study, especially the research board of science and research branch, islamic azad university, tehran, iran. con�ict of interest �e authors declare no con�ict of interest related to this article. references allen, e.j., morgan, d.g. 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(1979). factors in�uencing �ower and pod development in oil seed rape. journal of agricultural science, 92, 363–373. https://doi.org/10.1017/s0021859600062894 verma, n.k., pandey, b.k., singh, u.p., lodhi, md 2012. e�ect of sowing dates in relation to integrated nitrogen management on growth, yield and quality of rabi maize (zea mays l.). �e journal of animal & plant sciences, 22(2), 324–329. wright, g.c., smith, c.j., woodro�e, m.r. (1988). the effect of irrigation and nitrogen fertilizer on rapeseed (brassica napus) production in south-eastern australia. irrigation science, 9(1), 1–13. https://doi.org/10.1007/bf00292139 wysocki, d.j., corp, m., horneck, d.a., lutcher, lk (2007). nutrient management guide: irrigated and dryland canola. oregon state university em-8943-e. https://catalog.extension.oregonstate.edu/sites/ catalog/�les/project/pdf/em8943.pdf yadav yogesh, c., khichar, m.l., bishnoi, o.p., niwas, r. (2011). e�ect of sowing dates on energy balance and yield in raya (brassica juncea). agricultural science digest, 21(2), 118–120. yadav, k.s., rajput, rl, agarkar, m.s. (1999). e�ect of sowing dates and irrigation schedules on yield and water use e�ciency of indian mustard (brassica juncea). indian journal of agronomy, 44(1), 145–150. yadav, s.k., chandar, k., singh, d.p. (1994). response of late-sown mustard (brassica juncea) to irrigation and nitrogen. �e journal of agricultural science, 123(2), 219–224. https://doi.org/10.1017/ s0021859600068489 abstract indian mustard (brassica juncea l. czern) cultivation is suggested for regions with short seasons and low rainfall. although there have been many studies conducted on agronomic production of mustard in iran, the information regarding the interactive impact of cropping seasons and nitrogen fertiliser on growth characteristics and yield quality of mustard plant is still insu�cient and requires further investigation. �is study focused on the possible implications of di�erent cropping seasons and di�erent nitrogen levels on selected agronomic traits in mustard. in this experiment, �ve di�erent doses of nitrogen and two sowing periods were used to assess for their combined e�ects on the growth parameters, seed yield and agronomic characteristics of mustard in the semi-arid climatic conditions of takestan. �e results revealed that cultivation seasons and nitrogen rates had a signi�cant e�ect on plant height, biomass yield, number of siliques per plant, seed oil content and seed yield. key words: indian mustard, nitrogen fertilisation, planting season, seed oil content, seed yield received: [2020.06.11] accepted: [2020.09.07] agronomiczna wydajność jesienno-zimowej uprawy gorczycy sarepskiej (brassica juncea l.), w odpowiedzi na różne dawki nawozu azotowego streszczenie gorczyca sarepska = kapusta sitowata (brassica juncea l. czern), jest charakterystyczna dla upraw w regionach o krótkich porach roku oraz mniejszych opadach. obecnie w iranie brakuje wystarczającej wiedzy rolniczej do wydajnej produkcji gorczycy. w artykule tym, będącym formą pracy przeglądowej i badawczej jednocześnie, skoncentrowano się na produkcji gorczycy w półsuchym regionie oraz ocenie jakości jej oleistych nasion. celem było również porównanie nawożenia pięcioma różnymi dawkami azotu (w  postaci mocznika), w  trakcie dwukrotnego wysiewu oraz zbadanie wpływu tych dawek na wzrost, plon nasion i niektóre właściwości agronomiczne gorczycy, w warunkach agroklimatycznych takestanu (iran). wyniki pokazały, że sezon uprawy oraz dawki azotu, miały znaczący wpływ na wzrost roślin, plon biomasy, liczbę owoców – łuszczyn przypadających na roślinę, zawartość oleju siewnego i  ogólny plon ziarna. data sadzenia, warunki środowiskowe, nawożenie są ważnymi 95 czynnikami wpływającymi na jakość i wielkość plonów gorczycy. słowa kluczowe: gorczyca indyjska, nawożenie azotem, sezon sadzenia, zawartość oleju w nasionach, plon nasion information on the authors bent al-hoda asghari she studied at the islamic azad university, science and research branch, tehran, iran. during her master studies, she worked under direct supervision of associate prof. dr amir hussain shirani rad. particularly; she was investigating the interaction of cropping seasons and nitrogen fertiliser on some of the agronomic characteristics of indian mustard. mohsen youse� https://orcid.org/0000-0003-4602-713x he has a phd in agro-ecology from the islamic azad university, takestan branch. currently he is an agricultural expert at the jahad agricultural organisation of qazvin province. he is focusing on agronomic aspect of agricultural studies with giving special attention to fertlisers, planting dates and spatial density of crop plants. katarzyna możdżeń https://orcid.org/0000-0002-5695-4474 her scienti�c interests concentrate on the e�ects of di�erent environmental factors (light, ozone, heavy metals, allelopathic extracts) on the morphology and physiology cultivated plants, protected and invasive species. joanna puła https://orcid.org/0000-0002-3672-5690 her research is connected with agrotechnology in plant cultivation and plant ecology. presently, she is interested in use of the biomass of plants and other organic fertiliser like biochar in agriculture. peiman zandi https://orcid.org/0000-0003-3520-3994 he was deeply trained in agronomy (crop science) and specialising in stress physiology, biotic/abiotic stresses and agroecology. he is also interested in working in di�erent areas of agroecology, plant nutrition, bioremediation, botany, plant breeding and genetics. wang yaosheng https://orcid.org/0000-0002-2657-7057 he is interested in plant ecophysiology at di�erent levels molecular, tissue, organ, whole plants) under biotic and abiotic stress. �e main research aim is to utilise resources e�ciently and develop strategies and technologies. a gronom ic perform ance of autum nand w inter-cropped indian m ustard (brassica juncea l.) in response to varying levels of nitrogen fertiliser 159 annales universitatis paedagogicae cracoviensis studia naturae, 5: 159–176, 2020, issn 2543-8832 doi: 10.24917/25438832.5.11 grażyna sroka university of józef dietl in kraków, legnicka 5 st., 31-216 kraków, poland; grazyna.sroka73@gmail.com prevention and care of diabetic foot syndrome – an assessment of respondents’ competence for many diseases, complications arise that could have o�en been prevented by earlier intervention. �is is particularly relevant to complications in chronic diseases. many disease complications start with skin damage, especially in metabolic diseases, of which diabetes is one of the most common (king et al., 1998; karvonen et al., 2000; zieliński et al., 2014; benoit et al., 2020). such damage can be countered by taking various types of preventive measures. �is refers to a preventive orientation of diagnostics and treatment of complications in large disease populations. early protection of the skin of the feet relates signi�cantly to the prevention of disease syndromes in the course of diabetes complications (ho�stad et al., 2015). for this reason, there is a need for planned, multi-directional skin protection. preventive foot skin protection requires, �rst and foremost, quality diabetes treatment and specialised, planned leg skin hygiene (tatoń et al., 2013). pathological conditions of the lower limbs, that develop within the arterial, venous and lymphatic vessels, can lead to disorders in the functioning of cells, then tissues and, consequently, all of the limb structures (digiovani, greisberg, 2010; holman et al., 2012; czeleko et al., 2014). diabetic foot syndrome (dfs) is an infection of the lower extremities with ulceration and/or destruction of deep tissues, occurring in conjunction with neurological disorders and peripheral vascular diseases of varying degrees. �e �rst symptoms of dfs are numbness and tingling in the foot, followed by swelling, discoloration of the skin and changes in the appearance of nails. �e feet become dry; small, di�cult to heal wounds and numerous corns and calluses form on them (wujczyk, 2009). �e main risk factors for ulceration include improper levels of glucose in the body, nephropathy, improper nutrition, impaired vision, older age and, above all, improper foot hygiene (kasperczyk, 2004). �e etiopathogenesis of dfs, one of the most common complications of diabetes, is complex. �e main pathogenetic factors of this disease syndrome are peripheral isg ra ży na s ro ka 160 chemic changes, peripheral neuropathy, mechanical factors and various types of infections. depending on the main etiological factor, diabetic foot may be ischemic, neuropathic or mixed (bernas, 2003; głuszek et al., 2007). ischemic dfs is the result of progressive diabetic angiopathy (disease of a group of arterial vessels) (kozek, 2002). �e resulting disorders are the result of atherosclerosis, sclerosis in small arterioles and degenerative changes in the capillaries of the foot tissues (koblik, 2005; tuttolomondo et al., 2015). �e diabetic neuropathic foot syndrome develops as a result of lesions of both the somatic and autonomic peripheral nervous system. a�er injuries, it is the second most common cause of nerve damage (boucek, 2006; boulton et al., 2004; boulton, 2006; levy, valabhji, 2008). whereas, mixed dfs combines ischemic and neuropathic diabetic foot features. it results from the formation of both neuropathy and angiopathy, dependent on similar clinical factors, such as duration of diabetes, degree of metabolic control and genetic background (koselak, 2014). �e risk of developing dfs according to koblik (2005, 2008) is a concern in all patients with diabetes. tatoń et al. (2008, 2013) are convinced that the prevention of dfs begins with the diagnosis of diabetes. �ey claim that early prevention of diabetic foot will protect against the risk of amputation of the lower limbs. �ey pay attention primarily to preventive measures, e.g. foot exercises, careful skin care and controlling blood supply and innervation. koselak (2014) also wrote about the basis of prevention, i.e. the patient’s education in the proper treatment of the dfs case. a huge role in this prevention is assigned to the podologist, who ensures proper control of the progressive destruction of the skin of the limbs (świderska, 2012). his knowledge of various �elds of medicine allows for the introduction of appropriate treatments and care products. �erefore, he should be the �rst to inform the patient about possible complications and to indicate methods of proper foot care. �e aim of this study was to (1) assess the practical knowledge of respondents from three age groups on the subject of prevention and care in a dfs and to (2) identify the most important areas of the podologist’s work in promoting the prevention of this disease. research methodology evaluations, regarding knowledge about the care of diabetic foot, were made among 86 clients with diabetes randomly selected from a cosmetic salon in krakow (southern poland) within six months of visiting the salon. �e research group was divided into three age categories (tab. 1). unequal numbers in di�erent age categories con�rmed the fact that diabetic complications are more common in older people (tatoń et al., 2013), which more o�en forces them to visit a podologist. p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 161 tab. 1. characteristics of the statistical population taking part in the survey on feet care in diabetic foot syndrome age range gender ∑ [%] male female i – 30–50 years old 3 6 9 10 ii – 51–70 years old 14 19 33 38 iii – over 70 years old 9 35 45 52 ∑ 26 60 86 100 �e study was conducted using a survey technique. in the chosen method, in order to obtain the necessary information, a research tool was used, which was appropriately designed the questionnaire of the survey. �e questionnaire consisted of 35 questions investigating knowledge about feet care – how to wash, moisturise, massage, cut nails, remove layers of epidermis and use footwear, and awareness of how to control glucose, normal body weight, blood pressure and other items. �e survey contains suggested questions (appendix 1 – survey template). it was modelled on the survey of july 31, 2014, national health fund as annex no. 10 to ordinance no. 88/2013/dsoz. all respondents were informed about the aim of the research. �e content of the survey and how to complete it was explained in detail to the respondents. �e survey was anonymous. in the statistical study, individual questions were assigned the following score: “+1” point – if the answer was correct, “−1” point – if the answer was incorrect and “0” points – if the respondent selected the answer “i do not know”. by providing all correct answers, a respondent could receive a maximum of 35 points, while answering all questions incorrectly would result in the lowest score of −35 points. based on the surveys, a comprehensive summary of all results was created in tabular form. for the statistical analyses, the following were compared: correct, incorrect and “i don’t know” answers given by respondents to survey questions in terms of numbers and percentages, average survey scores for men and women in the analysed statistical population, average survey score for the three age categories of respondents, the total score achieved by all respondents to survey questions, the total score of the respondents separately in the gender groups and the total score obtained by the respondents for each of the three age groups. to test the statistical signi�cance of the average results for the female and male groups, the student’s t-test for independent groups was used. statistical di�erences between the average results in three distinguished age groups of respondents were assessed using the parametric one-way anova test, using tukey’s post hoc test. statistical signi�cance for all tests was assumed at p ≤ 0.05. statistical tests were performed using the statistica v10.0 for windows package. g ra ży na s ro ka 162 results of study �e numerical and percentage comparison of correct, incorrect and “i don’t know” answers given by respondents to individual survey questions revealed that most people correctly believe that an important element preventing diabetic foot is to control glucose, blood pressure and maintain a healthy weight (94% of respondents). respondents know about the need for thorough drying of the space between the toes in the prevention of diabetic foot (91% of them) and about checking the shoe before wearing it in order to avoid mechanical injuries (93% of them). �ey also know that smoking can lead to earlier development of dfs (92% of respondents) (appendix 2 – survey summary table). however, almost all respondents mistakenly believed that, for cutting the nail plate, it is best to use a metal clipper (91% of respondents); it is actually easy to damage the skin surfaces with this type of tool, which is very dangerous with a diabetic foot. respondents were also not aware of how many times a day they can change dressings fig. 1. comparison of the total score obtained by all respondents on subsequent survey questions (1–35 question numbers) regarding of diabetic foot care; n (number of respondents) = 86 p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 163 in the presence of fresh ulceration (84% of respondents); they did not know that it is best to walk around the house in closed-toe slippers (88% of respondents), that they should not wear so�-soled shoes (80% of respondents) or that it is not advisable to moisten the space between the toes of feet with dfs (84% of respondents) (appendix 2 – survey summary table; fig. 1). on questions such as: hydro massagers should be used for foot massage; for removal of nail cuticles a special preparations should be used to dissolve them; �e most important thing when checking your feet is checking the dorsal parts – almost half of the respondents answered “i don’t know” (appendix 2 – summary survey table). a comparison of the average scores of surveys for males and females in the analysed statistical population showed that males averaged 6.27 points (sd ± 5.57) and females 4.87 (sd ± 4.45) (fig. 2; tab. 2). �e results of the student’s t-test showed no statistically signi�cant di�erences in the average value of points between female and male groups (p = 0.22). fig. 2. comparison of the average scoring of surveys on diabetic foot care for the groups: fe – females and ma – males, in the analysed statistical population; statistical signi�cance between the examined groups was tested by the student’s t test; di�erences were considered signi�cant at p ≤ 0.05 g ra ży na s ro ka 164 tab. 2. student t-test results for independent groups: fe – females, ma – males, t – test value; df – degrees of freedom; p – probability value; n – sample count, sd – standard deviation; the di�erences were considered signi�cant at p ≤ 0.05 variable test-t average fe average ma t df p n fe n ma ±sd fe ±sd ma quotient f(w) p(w) number of points 4.87 6.27 –1.24 84 0.22 60 26 4.45 5.57 1.57 0.16 fig. 3. comparison of the average scoring of surveys on the care of diabetic foot in three age categories of respondents; i – from 30 to 50 years, ii – from 51 to 70, iii – above 70 however, the average scores for the three age groups were 6.00 (sd ± 4.90) for age group i, 7.53 for group ii (sd ± 4.71) and 3.72 (sd±4.37) for group iii (fig. 3; tab. 3). tab. 3. �e results of the parametric anova test made to examine the signi�cance of statistical di�erences between the average scores of respondents in the three age categories analysed; the di�erences were considered signi�cant at p ≤ 0.05 e�ect one-dimensional signi�cance tests for “number of points” ss degrees of freedom ms f p intercept 1796.97 1 1796.97 87.079 0.000 age category 270.86 2 135.43 6.562 0.002 error 1712.87 83 20.64 p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 165 �e anova test, used to determine statistical signi�cance in the average survey score, showed that there were di�erences between the compared age groups (p = 0.002). tukey’s post hoc test showed that these di�erences only applied to age groups iii and ii (tab. 4). tab. 4. tukey’s post hoc test results regarding statistical signi�cance between average scoring obtained in three age categories of respondents; i – from 30 to 50 years, ii – from 51 to 70, iii – above 70; the di�erences were considered signi�cant at p ≤ 0.05 no. variable – ‘number of points’; approximate probabilities for post hoc tests; error: ms intergroup = 20.637, df = 83; uneven n age category {1}3.723 {2} 7.533 {3} 6.000 1. i 0.539 0.754 2. ii 0.005 0.755 3. iii 0.005 0.539 a comparison of the total score for individual questions by group, females and males, showed that the surveyed females remembered to dry the space between the �ngers (56 points). �ey knew that important elements in avoiding the formation of diabetic foot are to control glucose, blood pressure and strive to maintain a healthy weight (55 points) and to check shoes (55 points) before wearing. �ey were aware that smoking can lead to earlier development of diabetic foot (55 points). however, the female respondents did not know that they should not cut nails with diabetic foot with metal clippers (−55 points) or how many times a day they should change the dressings in the case of foot ulceration (−55 points), etc. (fig. 4a). �e same comparison in the male group revealed similar answers. males also knew that controlling glucose, blood pressure and striving to maintain a healthy weight (24 points) are part of avoiding diabetic foot formation. in addition, they knew that a�er washing, it is best to dry your feet with a so� towel (24 points) and that they should purchase socks made only from natural �bres (22 points). males, like females, did not know that they should not cut nails with diabetic foot with metal clippers (−23 points) or that it is best to walk around the home in closed-toe slippers (−22 points) (fig. 4b). a comparison of the total score of the respondents by age group showed that, in age group i – i.e. 30 to 50 years old, respondents knew that a�er buying new shoes they should allow their feet to gradually become accustomed to them, by daily walking for 10–15 minutes, and that a�er washing it is best to dry feet with a so� towel. �ey were aware that controlling glucose and blood pressure and maintaining a healthy weight are important preventive elements in dfs. �is group of respondents knew to dry the space between the toes and that they should not wear shoes on bare feet (9 points for all answers). however, they were not aware that people with dfs should not walk barefoot on the beach. �ey made the mistake of thinking that a�er buying new shoes, g ra ży na s ro ka 166 if there is skin abrasion, they should wait for healing and then the same shoes can worn again (−9 points for all answers) (fig. 5a). in age group ii – i.e. 51 to 70 years old, the respondents knew that only socks made from natural �bers should be purchased (28 points). other results in this group were similar – i.e. respondents were aware of needing to control of glucose and blood pressure and maintain normal body weight in the prevention of diabetes (28 points), they knew that smoking can lead to earlier development of diabetic foot (28 points) and they knew that it is best to dry feet with a so� towel (26 points). �ey also remembered the need to dry the space between the toes (26 points). �e weakest results for this group were for the questions: is it good to use metal clippers to cut the nail plate? (−26 points), should you walk in shoes with a so� sole at home? (−24 points), is it good to walk around the home in open slippers? (−24 points) and how many times can daily dressings be changed in the presence of fresh ulceration? (−24 points) (fig. 5b). in age group iii – i.e. over 70 years of age, respondents knew that smoking can lead to an earlier development of dfs (45 points) and to check before putting on shoes that there was nothing in them (44 points). �ey were aware of needing to control glucose and blood pressure and maintaining normal body weight in diabetic prophylaxis (42 points). �ey performed most poorly on the questions: what is the best way to cut the nail plate? (−43 points), how many times a day can the dressing be changed if there is a fresh uncleration (−43 points), and is it advisable to moisturise the space between the toes in a diabetic foot? (−37 points) (fig. 5c). discussion �e study results were used to illustrate the most common mistakes in dfs care among respondents of di�erent genders and from di�erent age ranges. �ere are research studies that point to the essentiality of prevention in the treatment of diabetic foot but they are from the point of view of the role of a diabetologist (e.g. mrozikiewicz-rakowska et al., 2013; tatoń, 2014). to date, there are few studies that demonstrate the essential role of a podologist against the background of the real knowledge of respondents about feet care in dfs. many diabetologists believe that early prevention of diabetic foot will prevent amputation of the lower limbs (bernas, 2003; valabhji et al., 2010; holmanet al., 2012; ho�stad et al., 2015). �ey agree that from the initial stages of diabetes the elimination of smoking, for example, is necessary, and that regular monitoring of basic diagnostic parameters important for diabetes is needed. an important role in this respect is served by, for example, foot exercises, careful skin care and controlling blood supply and innervation (bowering, 2001; boulton, 2006). p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 167 fig. 4. comparison of the total score obtained by respondents in the survey (1–35 question numbers) regarding diabetic foot care, for individual questions separately in groups of gender: a) females (n = 60), b) males (n = 26) g ra ży na s ro ka 168 fig. 5. comparison of the total score obtained by respondents in the survey (1–35 question numbers) regarding diabetic foot care, for individual questions separately in groups of age: a) i – from 30 to 50 years (n = 9), b) ii – from 51 to 70 (n = 30), c) iii – above 70 (n = 47) p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 169 �e survey showed that respondents were aware of the risk of smoking in diabetic disease (92% of respondents). �ey (77% of respondents) also knew that gymnastics has a signi�cant impact on foot health and that you need to regularly check your blood sugar and body weight (tab. 2). �e research also showed that respondents had some knowledge about foot care but in other respects it was insu�cient, hence the need for more frequent visits to the podologist. when caring for nails, most respondents did not know that metal clippers cannot be used to cut nails (tab. 2; fig. 1, 4–5). in diabetic foot patients, even the smallest wounds can develop in�ammation and even into ulcerations, leading, in extreme cases, to tissue necrosis (margolis et al., 2002; potaczek, 2005; ho�stad et al., 2015). respondents were not aware that, for ulcerations, the number of dressing changes depends on a doctor’s recommendation; ulcerative conditions di�er and should be treated strictly according to a doctor’s instructions, especially for diabetic foot patients (rosiński, jasik, 2001; catanese, 2002; koblik, 2005; głuszek et al., 2007; wujczyk, 2007). another risk is the ignorance of respondents regarding the wearing of open-toe slippers in patients with diabetic foot. open-toe slippers do not protect against various types of mechanical injuries, which are not always easy to avoid, and slippers should not be made of plastic materials because it makes ventilation di�cult and promotes moisture between the toes. similarly, a so� sole for home footwear is also not recommended (koselak, 2014). keeping foot skin healthy is very important for people with diabetes because their skin is more prone to cracking (tatoń et al., 2008, 2013). respondents knew about this issue and care for proper moisturisation of foot skin (tab. 2; fig. 1, 4–5). �ey knew that they need to dry the spaces between the toes because any humidity remaining there can be conducive to fungal infections, which are very common in dfs (rosiński, jasik, 2001; catanese, 2002; zieliński et al., 2014). �e survey showed that there were no statistically signi�cant di�erences in the level of knowledge about the care of diabetic foot between females and males (tab. 3; fig. 2). both males and females agreed that smoking can lead to earlier development of dfs (fig. 4). �ey believed that controlling glucose and blood pressure and maintaining a healthy body weight are an important element in avoiding diabetic foot. �ey made similar mistakes for responses on the care of feet: they were not aware of what tool is the best for cutting nails, they moisturized the spaces between the toes with cream, they did not know how many times a day they should change dressings in the event of fresh ulcerations, etc. in this case, gender did not matter because dfs equally a�ects both females and males, although, as the study showed, females more o�en decided to visit a cosmetic salon and undergo podological consultations (tab. 1). �is is probably due to the fact that statistically more females than males have diabetes (king et al., 1998; tatoń et al., 2013). g ra ży na s ro ka 170 �e results of surveys among respondents divided into three age categories revealed many similarities but statistically signi�cant di�erences were found between the age group ii and age group iii (tab. 3–4; fig. 3). generally, the highest average score in the surveys was achieved by people from the age group ii – from 51 to 70 years of age. perhaps a greater interest in the care of diabetic foot at this age is due to the fact that this is the group in which diabetes most o�en begins to appear (koselak, 2014; benoit et al., 2020). �ey are middle-aged people, willing to independently oppose the e�ects of this disease. it is probable that people over 70 are less willing to perform foot care procedures on their own, hence their signi�cantly lower level of knowledge on this topic (fig. 5). respondents from the 51–70 years age group proved that they are aware of basic recommendations in the prevention of diabetes, and the irregularities they commit are similar to those in other analysed age categories (fig. 5a–c). people in the 30–50 years age range did not know that with dfs there is an absolute contraindication for walking barefoot on the beach (fig. 5a), due to the possibility of foot damage from shells. for similar reasons, it is impossible to wear the same shoes again a�er foot skin abrasions are caused by new footwear. �is can quickly lead to a renewal of wounds, and with longer duration, and can be the cause of infection and di�cult-to-heal ulcers (rosiński, jasik, 2001; bernas, 2003). however, respondents from this age group knew that feet should become accustomed to new shoes and fee should be dried with a so� towel. it can, therefore, be concluded that in the analysed statistical population many negligences in the care of diabetic foot could be avoided because they are primarily the result of lack of contact with a podologist. of course, they can also be the result of a lack of interest in self-care for diabetic foot by respondents themselves. �e basic task of a podologist is to implement care recommendations that protect patients from complications of dfs, but perhaps even more important is their function to make patients aware of the need for individual foot care (świderska, 2012; koselak, 2014). such care is very important in the prevention of this age-old disease (catanese, 2002; tatoń, 2002; tatoń et al., 2008, 2013; zieliński et al., 2014). conclusion (1) most respondents correctly believed that an important element in avoiding the formation of dfs is to control glucose and blood pressure and maintain a healthy weight; relatively poor knowledge was observed by all respondents concerning nail care and, in the case of ulcerations, how many times a day dressing should be made; there were no statistically signi�cant di�erences in the level of knowledge about the care of diabetic foot between females and males; survey results in the age groups distinguished that respondents made similar mistakes in knowledge of the methods of p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 171 care for the diabetic foot, and statistically signi�cant di�erences were found for age categories ii and iii; the highest scores for knowledge of dfs care were achieved by people 51–70 years of age who want to perform foot skin care themselves. (2) �e survey also showed that respondents made basic mistakes during foot care that could have been avoided if they had contacted a podologist; the role of the podologist is therefore not only to perform foot care procedures but to make patients aware and implement simple recommendations that will protect patients from complications. references benoit, s.r., hora, i., pasquel, f.j., gregg, e.w., albright, a.l., imperatore, g. (2020). trends in emergency department visits and inpatient admissions for hyperglycemic crises in adults with diabetes in the u.s., 2006–2015. diabetes care, 43(5), 1057–1064. https://doi.org/10.2337/dc19-2449 bernas, m. (2003). patogeneza i klinika zespołu stopy cukrzycowej – współpraca z chirurgiem (pathogenesis and the clinic of diabetic foot syndrome – cooperation with a surgeon). przewodnik lekarski, 6, 169–175. [in polish] boucek, p. 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[in polish] p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 173 appendix 1 educational survey – diabetic foot syndrome (template) below are 35 terms for foot care. please assess their correctness by putting a cross in the appropriate box – yes, no or i don’t know no. ascertainment yes no i don’t know 1. it is best to use metal clippers to cut the nail plate 2. feet should be washed at temperatures above 37 °c 3. a�er washing, moisturising cream should be to applied to your feet between your toes 4. it is best to walk barefoot on the beach 5. at home, one should walk in shoes with a so� sole 6. gymnastics (pre-planned exercises performed 2-3 times a week) has a signi�cant impact on foot health 7. toes nails should be cut as short as possible 8. feet should be washed very thoroughly and soaked in a bowl for at least 15 minutes 9. in case of excessive sweating of the feet, talcum powder should be used 10. it is best to walk around the home in open-toe slippers 11. a�er buying new shoes, a foot should gradually get used to them, by daily walking for 10–15 minutes 12. plaster is the best dressing to feet ulceration 13. socks, tights made only of natural �bres (cotton) should be purchased; wool socks should be avoided 14. �e nails should be cut without rounding on the sides 15. hydro massagers should be used for foot massage 16. intensive moisturising creams should not be used 17. �e best shoes are natural leather, no seams inside 18. if there is a skin abrasion a�er buying new shoes, you should wait for healing and wear them again 19. you should inform your doctor about any feet ulceration 20. you should limit physical activity associated with direct im-pact on feet 21. to remove nail cuticles, you should use special preparations to dissolve them 22. you should dry the feet with a so� towel a�er washing 23. you should remove layered hard skin by yourself 24. shoes should be purchased in the morning when the foot is rested 25. always measure your foot before buying shoes 26. �e feet should be examined daily g ra ży na s ro ka 174 27. an important element to avoid the formation of diabetic foot is to control glucose and blood pressure and strive to maintain a healthy weight 28. using a �le, your nails should be �led in one direction 29. you should remember to thoroughly dry the space between your toes 30. to warm feet, hot water bottles should be used 31. it is advisable to wear shoes with bare feet 32. before wearing on shoes, you should make sure that there is nothing in them (sand, stones, small items) 33. �e most important thing when checking your feet is to check the dorsal parts 34. if a fresh ulceration occurs, dressings should be changed twice a day 35. smoking can lead to earlier development of diabetic foot syn-drome gender: female ............. male………. please circle the age range: 30–50 years old, 51–70 years old, over 70 years old. p revention and care of diabetic foot syndrom e – an assessm ent of respondents’ com petence 175 appendix 2 summary survey table comparison of the number and percentage of “correct”, “incorrect” and “don’t know” answers given by 86 respondents to the questionnaire regarding diabetic foot care; bold has the highest percentage of responses no. of question in survey correct answers incorrect answers no answer – ”don’t know” numerically [%] numerically [%] numerically [%] 1 0 0 78 91 8 9 2 35 41 39 45 12 14 3 11 13 72 84 3 3 4 24 28 62 72 0 0 5 3 4 69 80 14 16 6 66 77 11 13 9 10 7 39 45 38 44 9 11 8 17 20 54 63 15 17 9 63 73 15 18 8 9 10 8 9 76 89 2 2 11 62 72 6 7 18 21 12 41 48 20 23 25 29 13 77 89 4 5 5 6 14 27 31 43 50 16 19 15 19 22 19 22 48 56 16 12 14 43 50 31 36 17 70 82 8 9 8 9 18 20 23 65 76 1 1 19 61 71 9 10 16 19 20 48 56 8 9 30 35 21 25 29 19 22 42 49 22 78 91 8 9 0 0 23 27 31 55 64 4 5 24 32 37 40 47 14 16 25 60 70 11 13 15 17 26 70 81 13 15 3 4 27 81 94 2 2 3 4 28 42 49 8 9 36 42 29 78 91 2 2 6 7 30 54 63 12 14 20 23 31 60 70 24 28 2 2 32 80 93 4 5 2 2 33 14 16 26 30 46 54 34 1 1 72 84 13 15 35 79 92 3 3 4 5 g ra ży na s ro ka 176 profilaktyka i pielęgnacja stopy cukrzycowej – ocena kompetencji respondentów streszczenie w  roku 2015 przeprowadzono badania wśród 86 klientów losowo wybranego gabinetu kosmetycznego w  krakowie (południowa polska). celem pracy była ocena wiedzy respondentów na temat pielęgnacji i  pro�laktyki stopy cukrzycowej. dodatkowym celem było wskazanie roli podologa w  pro�laktyce chorób stóp. grupę badawczą stanowiło 36 kobiet oraz 26 mężczyzn powyżej siedemdziesięciu lat, 19 kobiet i  14 mężczyzn w  przedziale wiekowym od 51–70 lat i  6 kobiet 3 mężczyzn w  przedziale wiekowym od 30–50 lat. przeprowadzona ankieta była anonimowa i składała się z 35 pytań. dotyczyła wiedzy na temat pielęgnacji stóp: ich sposobu mycia, nawilżania, masowania, obcinania paznokci, usuwania nawarstwionego naskórka, używanego obuwia, a  także świadomości kontroli podstawowych parametrów, ważnych w pro�laktyce cukrzycy. badania pokazały, iż respondenci podczas pielęgnacji stóp popełniają podstawowe błędy, których można byłoby uniknąć w  przypadku kontaktu z  podologiem. najbardziej zainteresowaną grupą w samodzielnej pielęgnacji stóp okazali się chorzy z przedziału 51–70 lat, którzy mieli stosunkowo największą wiedzę na ten temat. ankieta pokazała, że rola podologa polega na uświadomieniu i wdrożeniu u pacjenta prostych zaleceń, które uchronią go przed powikłaniami oraz nauczą właściwej pielęgnacji kończyn dolnych w zespole stopy cukrzycowej. key words: diabetic foot syndrome (dfs), podology, questionnaire method received: [2020.03.28] accepted: [2020.05.18] annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832 doi: 10.24917/25438832.7.x wojciech w. a. kowalski department of botany and nature conservation, west pomeranian university of technology in szczecin, j. słowackiego 17 st., 71-434 szczecin, poland; e-mails: wojciech.wakowalski@wp.pl, botanika@zut.edu.pl, desmatractum bipyramidatum (chodat) pascher 1930 (chlorophyceae, chlorococcales) – a new locality in poland introduction genus desmatractum west et g.s. west 1902 (= bernardinella chodat, calyptobactron geitler) includes species found in various habitats on almost all continents of the eart. according to guiry, guiry (2022) currently eight species are recognised (desmotractum bipyramidatum (chodat) pascher; d. delicatissimum korshicov; d. elongatum pascher; d. indutum (geitler) pascher; d. nyanzae, (wołoszyńska) g.s. west ex printz; d. obtusum pascher; d. plicatum weste & g.s. west, and d. spryii nicholls) of which only four has been found in poland: d. bipyramidatum, d. delicatissimum, d. indutum, and d. nyanzae (classified earlier by wołoszyńska (1914) as a peniococcus nyenzae). a characteristic feature of the cells of this genus are thick protrusions of the cell wall running along the long axis of the cell, in the form of ribs. this feature is more pronounced in most species. the cells are bipolar, with a spherical or oval protoplast with one pyrene. towards the pole, the cells are more or less narrowing, having a spindle-shaped form, with a spike of various lengths. habitat and plant material registered in the goleniów forest in western pomerania (fig. 1), in the habitat of the mid-forest peat bog desmatractum bipyramidatum, it is the second place of occurrence of the species. earlier, its occurrence in poland was noted only by mrozińska (1984). on the analysed site, a floristic list of vascular plants was made, for the general characteristics of the habitat, as well as a list of other algae species accompanying d. bipyramidatum. fig. 1. the location of the “wrzosiec” nature reserve (53°37′03″n 14°57′41″e) and the location of the habitat of desmatractum bipiramidatum; a – roads, b – forest roads, c – borders of forest divisions, d – borders of forest areas, e – border of nature reserve, f – location of species locality d. bipyramidatum cells occur on the flooded organic substrate, covering the synusions in caricetum elatae koch 1926. small pits in area, 0.5–1.5 m2, trough-shaped pits between the carex elata all. clumps are covered with an organic substance consisting mainly of dead shoots with varying degrees of decomposition of shoots c. elata and bryophytes. mineral substances deposited by the peatland flowing from the periphery also have a small share in the length. the habitat is characterised by poor vegetation. locally, the substrate is inhabited by small grasses of bryophytes: sphagnum auriculatum schimp., s. rufescens (nees & hornsch.) warnst., fossombronia foveolata lindb., drepanocladus exannulatus (schimp.) warnst. and a few vascular plants: juncus bulbosus l., hydrocotyle vulgaris l., drosera intermedia hayne, less often d. rotundifolia l. coverage of the vascular layer is 5–20%, and the moss layer 5–50%. the low position of depressions between the carex elata clumps, in relation to the neighbouring plant communities of the remaining parts of the fen, causes constant runoff and stagnation of waters in the habitat. hence, in most of the year they are flooded with a 1.0–1.5 cm layer of water, or it stagnates evenly with their surface, giving them the character of a bush. from spring to late autumn, this habitat is a place of mass development of various systematic groups of algae covering the surface of the substrate with a gelatinous layer. the development of algae in synusias is favoured by favourable light conditions. the leaves of the tall clumps of sedges diffuse the sun's rays, but without limiting the good illumination of the synapse substrate. d. bipyramidatum cells are accompanied by various cyanobacteria and algae taxa represented mainly by chlorophyceae, cyanoprokaryota and bacillariophyceae species. the absolutely dominant taxonomic group are chlorophyceae species of the desmidiales order, among which the largest numbers are: netrium digitus (ehrenb.) itzigs. et rothe, tetmemorus granulatus (bréb.) ralfs ex ralfs, euastrum ansatum ralfs var. ansatum, closterium baileyanum (bréb.) bréb., closterium dianae (ehrenb.) ralfs, pleurataenium ehrenbergii (bréb.) de bary, euastrum pectinatum (bréb.) ex bréb. the cells are singly present: cylindrocystis brebissonii (ralfs) de bary, penium exiguum w. west var. glaberrimum grönbl., p. spirostriolatum bark var. spirostriolatum, closterium costatum corda ex ralfs var. costatum, cl. gracile bréb. ex ralfs, cosmarium debary arch. in pritchard, c. margaritiferum menegh. ex ralfs var. margaritiferum, c. portianum arch. var. portianum, c. quadratum ralfs ex ralfs var. quadratum, euastrum gayanum de toni var. gayanum, e. verrucosum ehrenb. ex ralfs, micrasterias thomasiana arch. var. notate (nordst.) grönbl., m. truncate (corda) ex bréb. var. truncata, staurastrum teliferum ralfs. diatoms represent: frustulia rhomboides var. saxonica (rabenhorst) de toni = frustulia saxonica rabenhorst, eunotia exigua (brébisson ex kützing) rabenhorst, e. lunaris = eunotia bilunaris (ehrenberg) schaarschmidt, tabellaria flocculosa (roth) kützing var. flocculosa, pinularia sp. div. less common along with d. bipyramidatum cells are the species: merismopedia glauca (ehrenb.) kützing, chroococcus turgidus (kützing) nägeli, oscillatoria sp. (cyanobacteria), pediastrum tetras (ehrenb.) ralfs, p. angulosum (ehrenb.) (chlorophyceae) and pseudostaurastrum enorme (xanthophyceae). cell description desmatractum bipyramidatum (chodat) pascher 1930 basionym: bernardinella bipyramidata chodat 1921. bull, soc. bot. geneva, ser. 2, 12; fusiform cells (fig. 2), widened in the equatorial plane into a ring surrounding the cell, tapering conically towards the apexes and stretched on both sides into short appendages, sharply rounded at the ends. the total length of cells 23.6–28.6 µm, width in the equatorial plane 10.2–12.9 µm, length of the apical processes 4.2–4.5 µm. cell wall smooth, colourless or slightly reddish with 5–8 ribbed projections along the long axis of the cell. in plan, the cells are round with slightly concave walls between the ribbed projections. the inside of the cell is a spherical protoplast 7.7– 9.7 µm in diameter, with a chloroplast containing one clearly visible pyrenoid and several (usually 4–6) distinct grains with a diameter of 0.6–0.7 µm. the shape of the cells is identical to d. bipyramidatum sensu skuja (1964). taper more or less evenly conically towards the ends, drawn into a short spike, rounded at the top. in the collected material, d. bipyramidatum cells were observed only in two samples collected during the summer period, when the water level is stabilised and not too high. w. their occurrence, however, was sporadic. these were 6–8 cell clusters. in spring and autumn, the depressions are flooded with water and are characterised by completely different ecological conditions. reporting the occurrence of the species on different continents indicates that it is a taxa with a cosmopolitan range, but very rarely observed. fig. 2. desmatractum bipyramidatum sensu skuja (1964) from a high atlantic peat bog in the goleniów forest (original) distribution distribution in poland according to the information contained in the study (siemińska, wołowski, 2003) and in the icon library and species records of the department of algology, ib pan in krakow, the occurrence of desmatractum bipyrimidatum has been recorded so far only in the tatra mountains from the peat bog in the valley dolina pięciu stawów (mrozińska, 1984). in unpublished materials from the peat bogs of the śnieżka region in karkonsze, it is mentioned by matuła. the site from western pomerania is therefore the second documented place of its occurrence in poland. general distribution d. bipyramidatum cells have so far been found in marshy habitats of peat bogs in central europe; this species should be considered cosmopolitan in the area (tsarenko et al., 2011; guiry, guiry, 2022). there are known localities from this region in bulgaria (vodeničarov, 1960), sweden (suja, 1964), great britain (lund, 1942), and the tyrolean alps (ettl, 1968). according to komarek and fott (1983), who give the positions of d. bipyramidatum also from denmark, germany, france, czechoslovakia and the territory of the former ussr (koršikov, 1953). outside europe, single d. bipyramidatum sites are also known from the state of michigan in the usa (taft, 1939, prescott, 1962), new zealand (skuja, 1976); asia (china, japan, russia); south west asia (bangladesh, india); africa (ivory coast); australia, papua new guinea (guiry, guiry, 2022). prescott (1962) believes that the taxon is definitely related to the acidic habitats of bog bogs. summary and conclusion registered in the goleniów forest in western pomerania (fig. 1), in the habitat of the mid-forest peat bog desmatractum bipyramidatum mesar fen, it is the second place of occurrence of the species. earlier, its occurrence in poland was noted only by mrozińska (1984). d. bipyramidatum cells occur on the flooded organic substrate, covering the synusions in caricetum elatae koch 1926. small pits in area, 0.5–1.5 m2, trough-shaped pits between the carex elata clumps are covered with an organic substance consisting mainly of dead shoots with varying degrees of decomposition of c. elata and bryophytes. the habitat is characterised by poor vegetation. coverage of the vascular layer is 5–20%, and the moss layer 5–50%. the location of the depressions between the c. elata clumps, in relation to the neighbouring plant communities of the remaining parts of the fen, causes a constant runoff and stagnation of water in the habitat, which is the nature of the marshes. in terms of shape and other diagnostic features, the cells are identical to the described d. bipyramidatum sensu skuja (1964). they taper more or less evenly conical towards the ends stretched out in a short squeeze, rounded at the top. their smooth, colourless or slightly reddish cell wall is covered with 5–8 ribbed bulges along the long axis of the cells. the interior of the cell is occupied by a spherical protoplast with a chloroplast containing 1 pyrene and a few (usually 4– 6) distinct granules. conflict of interest the author declare no conflict of interest related to this article. references ettl, h. (1968). ein beitrag zur kenntnis der algenflora tirols. berichte des naturwissenschaftlichen-medizinischen verein innsbruck, 56, 177–354. [in german] guiry, m.d., guiry, g.m. (2022). algaebase. listing the world’s algae. national university of ireland, galway. https://www.algaebase.org komarek, j., fott, b. (1983). chlorophyceae (grünalgen), ordnung chlorococcales. in: g. huber-pestalozzi (ed.), das phytoplankton des süsswassers, die binnengewässer, 16(7/1), 1–1044. [in german] koršikov, a.a. (1953). pidklas protokokovi (protococcinae). viznačnik prisnovodnich vodorostej ukrainskoj rsr. akad. nauk ursr, 5, 1–439. [in russian] lund, j.w.g. (1942). contribution to our knowledge of british algae – viii. journal of botany (london), 80, 57– 73. mrozińska, t. (1984). botryosphaerella sudetica (lemmermann) silva (botryococcaceae, chlorophyta), a green alga new to tatra mts. archiv fur hydrobiologie, supplement 67, 3(algological studies 36), 245–249. https://doi.org/0.1127/algol_stud/67/1984/245 prescott, g.w. (1962). algae of the western great lakes area. iowa: wm. c brown comp. publ. dubuque. pp. 977. siemińska, j., wołowski, k. (2003). catalogue of polish prokaryotic and eukaryotic algae. katalog glonów prokariotycznych i eukariotycznych polski. ser. biodiversity of poland. in: z. mirek (ed.), różnorodność biologiczna polski, 5, kraków: institute of botany polish academy of science. skuja, h. (1964). grundzüge der algenflora und algenvegetation der fjeldgegenden um abisko in schwedischlappland. nova acta regiae societatis scientiarum upsaliensis. ser. 4, 18(3), 1–465. [in german] skuja, h. (1976). zur kenntnis der algen neuseeländischer torfmoore. nova acta regiae societatis scientiarum upsaliensis, ser. 5(2), 1–158. [in german] taft, c.e. (1939). additions to the algae of michigan. bulletin of the torrey botanical club, 66, 77–85. tsarenko, p.m., wasser, s.p., nevo, e. 2011. algae of ukraine. diversity, nomenclature, taxonomy, ecology and geography. 3. chlorophyta. ruggell a.r.a. gantner verklag k.-g. vodeničarov, s. (1960). beitrag zur algenflora bulgariens – iii. izvestiya na botaniceskiya institut, 1, 137–155. [in bulgarian] wołowszyńska, j.(1914). zapiski algologiczne – algologische notizen. sprawozdania z posiedzeń towarzystwa naukowego warszawskiego, wydział nauk matematycznych i przyrodniczych, 7(1), 1-4. [in german] abstract in the materials collected during phycological studies of bog bogs in the goleniów forest in western pomerania, cells of desmatractum bipyramidatum (chodat) pascher 1930, rarely administered from poland, were found. and bryophytes. in terms of shape, the cells are identical and refer to the described d. bipyramidatum sensu skuja (1964). d. bipyramidatum cells are accompanied by various algae taxa represented mainly by species of the chlorophyceae, cyanoprokaryota and bacillariophyceae classes. the absolutely dominant taxonomic group in the habitat are chlorophyceae species of the order desmidiales key words: chlorococcales, chloropohyceae, new locality, western pomerania received: [2022.01.19] accepted: [2022.02.05] desmatractum bipyramidatum (chodat) pascher 1930 (chlorophyceae, chlorococcales) – nowe stanowisko w polsce streszczenie w materiałach zebranych w trakcie badań fykologicznych torfowisk mszarnych w puszczy goleniowskiej na terenie pomorza zachodniego, znaleziono komórki rzadko podawanej z terenu polski zielenicy desmatractum bipyramidatum (chodat) pascher 1930. komórki d. bipyramidatum występowały na przepojonym wodą substracie organicznym, utworzonym z obumarłych szczątków roślin naczyniowych i mszaków. pod względem kształtu komórki są identyczne i nawiązują formą do opisanego d. bipyramidatum sensu skuja (1964). komórkom d. bipyramidatum towarzyszą różne taksony glonów reprezentowane głównie przez gatunki klasy chlorophyceae, cyanoprokaryota oraz klasy bacillariophyceae. bezwzględnie dominującą grupą taksonomiczną, występującą w siedlisku są gatunki chlorophyceae, rzędu desmidiales. słowa kluczowe: chlorococcales, chloropohyceae, nowe stanowisko, pomorze zachodnie. information about the author wojciech w. a. kowalski the author is a specialist in the field of algology. his research interests concern both single species of algae and whole groups of marine and freshwater algae, with particular emphasis on rare and endangered taxa. a special taxonomic group of interest are the taxa associated with the ecosystems of bog bogs, as well as freshwater red algae. 48 annales universitatis paedagogicae cracoviensis studia naturae, 6: 48–59, 2021, issn 2543-8832 doi: 10.24917/25438832.6.3 lidia tasenkevich*, marya seniv, krystyna skrypec ivan franko national university of lviv, 4 hrushevsky st., lviv 79005, ukraine; *tasenkevich@gmail.com rare and endangered vascular plant species of male opillya (lviv region, ukraine) introduction one of the e�ective measures for the conservation of biota in general and phytodiversity in particular is the red books and red lists, in which information on the state and conservation status of endangered plant species globally, regionally and locally is published. although this method is passive, its use is one of the most e�ective means to raise the level of environmental education and awareness of the general public of the need to protect and preserve phytodiversity. studies of the �ora of galicia and volhyn-podillya was started by willibald besser more than 200 years ago (koczwara, 1925). in the �rst works published by besser, 1215 species were listed, a number of which were mentioned for the territory of opillya (besser, 1809, 1822). in general, beginning in the second half of the 19th century, the �oristic study of galicia, and roztocze and opillya in particular, intensi�ed. �us, in 1868 the list of plants from galicia was published by wojciech grzegorzek (1868). in 1880 bronisław gustawicz (1880) gave a list of species of �ora of bibrka county. bronisław błocki (1881) published �oristic lists of plant species in galicia, including species from the territory of opillya, specimens of which are now stored in the herbarium of ivan franko national university of lviv (acronym: lw) (tasenkevich et al., 2018). many �oristic �nds from opillya were noticed by hugo zapałowicz (1906–1911) in his critical overview of galicia’s �ora. �e studies of jan grochmalicki and władуsław szafer (1910) are devoted to the �ora of opillya and roztocze. most of the research in the western part of opillya was conducted by the polish naturalist szymon wierdak (1916, 1923, 1926, 1932), who, in addition to collecting �oristic data, studied chorological characteristics, including rare species. rare and steppe components of the �ora of opillya were explored by aniela kozłowska (1931) and szafer (1935), and wacław gajewski (1937) performed a geographical analysis of the �ora of podillya, covering the �ora of opillya part. r are and endangered vascular plant species of m ale o pillya (lviv region, u kraine) 49 in the 1960s intensive study of the �ora of di�erent parts of opillya was resumed during �oristic and phytocenological research in volhyn-podillya (bradis, rubtsov, 1966; zaverukha, 1965, 1976, 1978, 1981, 1982, 1985; kukovytsia, 1970, 1972, 1976; shelyag-sosonko, 1970; shelyag-sosonko, kukovytsia, 1970, 1974; shelyag-sosonko et al., 1981; shelyag-sosonko, zhyzhyn, 1983; zhyzhyn et al., 1990; kukovytsia et al., 1994, 1998). it is paradoxical that male opillya, with its complex mosaic of landscapes, which led to the diversity of habitats and, consequently, to the diversity of plant species, located in the neighbourhood of lviv – one of the botanical centres, in which �oristic studies of galicia and opillya were initiated, was overlooked by botanists in the 19th and 20th century. herbarium collections from male opillya are practically absent, and in the literature – except the paper of gustawicz (1880), there are only a few indications of the localities of a few species from this area (zelenchuk, bednarska, 1998; danylyk, 2001; borsukevych, 2008, 2009; borsukevych et al., 2016). �e aim of this study was the inventory of male opillya �ora, allowed to establish its species composition and to identify the presence and evaluate the state of rare and endangered species. general characteristics of the study area within the western part of the podillya upland – 49°1′39″ n, 27°51′22.32″ e (that is a part of the eastern european plain) is one of its area with most complex relief – opillya. �is area is located southeast of lviv within three administrative regions: lviv, ivano-frankivsk and ternopil. among the numerous schemes of opillya regionalisation, we used the julian czyżewski’s (1925) scheme of geomorphological zoning, practically forgotten in ukraine for 80 years (palienko et al., 2004). czyżewski considered the extreme south-western part of opillya within the lviv region as a separate district of male opillya (fig. 1). �e territory of male opillya, with an area of about 58.6 ha or 586 km2 (between bibrka – 49°37′0″ n, 24°14′0″ e, mуkolaiv – 46°58 0″ n, 32°0′0″ e, and novyi rozdil – 49°28′11″ n, 24°8′12″ e), is characterised by a complex relief: in the extreme west of male opillya there is a ridge of steep hills, which rise on average 120–150 m above the bottom of river valleys, and some reach maximum heights up to 400 m. part of the district is a plain, that descends to the dnister and its tributaries (tsys’, 1962; pavlyuk, haskevych, 2011). in the territory of the long-inhabited and well-developed opillya, few forests have survived to this day: they cover only 10.6% of the territory. oak, hornbeam-oak and hornbeam forests predominate. oak-pine forests make up only 1.2% of the land covered with forest vegetation. li di a ta se nk ev ic h, m ar ya s en iv , k ry st yn a s kr yp ec 50 in the recent past, the valley of the upper plain of the dnister and its tributaries was signi�cantly swampy. decreased groundwater levels due to land reclamation in the 1960s–1970s, changed the vegetation on the riverbed terraces. much of the territory (up to 80%) has been turned into arable lands, hay�elds and pastures. material and methods field research to inventory male opillya’s �ora was carried out during 2013–2020. in order to possibly identify collections from male opillya, research was conducted in herbaria of lviv and kyiv (acronyms: lw, lws, lwks and kw). a few literature sources fig. 1. �e schematic map of male opillya region (adjusted according to czyżewski, 1925) r are and endangered vascular plant species of m ale o pillya (lviv region, u kraine) 51 (zelenchuk, bednarska, 1998; danylyk, 2001; borsukevych, 2008, 2009; borsukevych et al., 2016), mentioning the plant species of male opillya were analysed. �e list of rare and endangered species of �ora includes: • species listed in the third edition of the red book of ukraine (2009) (while maintaining the categorisation adopted in this document), • species subject to protection at the regional level (decision of the regional council ..., 2015; tasenkevich et al., 2015), • species subject to protection at the international level – bern convention (1979), european red list of vascular plants (bilz et al., 2011), �e iucn red list ... (2021) and cites appendix №2 list (1963). nomenclature of species listed is given in accordance with the databases wfo and �e plant list, version 1.1 (http://www.world�oraonline.org; http://www.theplantlist. org). statistical data processing was performed using microso� excel 2010. results and disscusion of the 1192 species of vascular plants that make up the �ora of male opillya, as a result of �eld research, processing of herbarium materials and analysis of literature sources, 143 species and one subspecies of vascular plants (which is about 12% of the total �ora of this area) are endangered or rare and have di�erent conservation status (fig. 2a). among them, 55 species are listed in the red book of ukraine (2009). �ey are divided between threat categories as follows: endangered category (en) – 6 species: botrychium virginianum, carex strigosa, epipogium aphyllum, juncus subnodulosus, hemipilia cucullata (syn. neottianthe cucullata (l.) schltr.), spiranthes spiralis; vulnerable category (vu) – 25 species: anacamptis coriophora, a. morio, atropa belladonna, carex buxbaumii, c. chordorrhiza, c. davalliana, c. dioica, cypripedium calceolus, cytisus albus, dactylorhiza incarnata, d. maculata, epipactis atrorubens, e. palustris, festuca heterophylla, fritillaria meleagrіs, gladiolus imbricatus, iris sibirica, liparis loeselii, malaxis monophyllos, orchis mascula, o. militaris, pedicularis sylvatica, pinguicula vulgaris, utricularia intermedia, u. minor; rare category (r) – 9 species: cephalanthera damasonium, c. longifolia, c. rubra, corallorhiza tri�da, cytisus blockianus, dactylorhiza majalis, d. viridis, epipactis purpurata, lathyrus laevigatus; not evaluated category (ne) – 15 species: allium ursinum, colchicum autumnale, epipactis helleborine, dactylorhiza fuchsii, galanthus nivalis, huperzia selago, leucojum vernum, lilium martagon, listera ovata, neottia nidus-avis, platanthera bifolia, p. chlorantha, salvinia natans, scopolia carniolica, trapa natans. li di a ta se nk ev ic h, m ar ya s en iv , k ry st yn a s kr yp ec 52 fig. 2. percentage categorisation plant species of male opillya region, based in the red book of ukraine (a) and the list of regionally rare plant species of lviv region (b); vu – vulnerable, ne – not evaluated, r – rare, en – endangered, nt – near-threatened, lc – least-concern, dd – data de�cient, re – regionally endangered �e last version of the list of rare plant species of lviv region was approved by the lviv regional council by decision of june 15, 2015 no. 1370 (decision of the regional council ..., 2015; tasenkevich et al., 2015). 127 species and one subspecies of this list were found in male opillya, of which 22 species belong to the near-threatened category (nt), 13 species – to the least-concern category (lc), 13 species – to the r category, 10 species – to the ne category, and eight – to the data de�cient category (dd). among threatened there are 8 species (en category), 52 species are categorised as vu, and 2 species are regionally endangered (re) (fig. 2b). r are and endangered vascular plant species of m ale o pillya (lviv region, u kraine) 53 according to the iucn list (2021) – as of the beginning of 2021, 100 species and one subspecies are part of rare and endangered species of �ora of male opillya. in particular: lc category – 74 species and one subspecies (not endangered) dd category – 15 species: anthyllis vulneraria, bromus japonicus, campanula patula, glechoma hederacea, g. hirsuta, jacobaea vulgaris, klasea lycopifolia, malus sylvestris, ostericum palustre, populus nigra, rosa gallica, r. villosa, scirpus radicans, ulmus glabra, u. laevis; nt category – 9 species: anacamptis morio, fraxinus excelsior, galanthus nivalis, iris sibirica, liparis loeselii, malaxis monophyllos, marrubium vulgare, sonchus arvensis; vu category – 2 species: botrychium virginianum, luzula campestris; en category – 1 species: hemipilia cucullata. �e european red list of vascular plants (bilz et al., 2011; korotchenko, 2016) includes 55 species and one subspecies of a rare component of the �ora of male opillya (according to the iucn categorisation). in particular: lc category – 40 species and one subspecies (not endangered); dd category – 9 species: camelina sativa, carex atherodes, klasea lycopifolia, malus sylvestris, medicago falcata, ostericum palustre, scirpus radicans, utricularia intermedia; nt category – 6 species: anacamptis morio, cypripedium calceolus, galanthus nivalis, liparis loeselii, malaxis monophyllos, sparganium natans; en category – 1 species: hemipilia cucullata. 27 species of male opillya �ora (26 species of the family orchidaceae juss. and galanthus nivalis) are protected in accordance with cites appendix №2 list (1963). five species are protected by the bern convention (1979): liparis loeselii, luronium natans, ostericum palustris, salvinia natans, trapa natans. �e comparison of the number of species with the protected status from the male opillya area according to all the lists of protected and endangered taxa included in the analyses is illustrated in �g. (3). rare and endangered species of vascular plants of male opillya belong to 100 genera, 46 families and four divisions (tab. 1). li di a ta se nk ev ic h, m ar ya s en iv , k ry st yn a s kr yp ec 54 fig. 3. species that have conservation status in the territory of male opillya; rbu – red book of ukraine, rr – rare species for lviv region, bc – bern convention, erl – european red list, iucn red list, cites appendix №2 list tab. 1. quantitative characteristics of the main systematic units, which include rare and endangered species of male opillya �ora no. taxon number of families [%] number of genera [%] number of species [%] 1. lycopodiophyta 1 2.2 1 1.0 1 0.7 2. equisetophyta 1 2.2 1 1.0 1 0.7 3. polypodiophyta 5 10.8 6 6.0 6 4.2 4. magnoliophyta 39 84.8 92 92.0 136 94.4 magnoliоpsida 27 58.7 55 55.0 69 47.9 liliopsida 12 26.1 37 37.0 67 46.5 total (1–4) 46 100.0 100 100.0 144 100.0 magnoliophyta dominates among the rare fraction of �ora: they make up 84.8% of the total number of rare and endangered species, of which to magnoliopsida belong 58.7%, to liliopsida – 26.1% of species. spore vascular plants (lycopodiophyta, equisetophyta, polypodiophyta) include eight species. �e families orchidaceae (26 species), asteraceae bercht. & presl and cyperaceae juss. (11 species each), poaceae barnhart (10 species), rosaceae juss. (9 species), fabaceae lindl. (7 species), liliaceae juss. and lamiaceae martinov (6 species each), brassicaceae burnett (5 species) and juncaceae juss. (4 species) are the most numerous in the list. �e next 11 families comprise two rare and threatened speacies each r are and endangered vascular plant species of m ale o pillya (lviv region, u kraine) 55 (amaryllidaceae j.st.-hil., apiaceae lindl., boraginaceae juss., euphorbiaceae juss., gentianaceae juss., hydrocharitaceae juss., iridaceae juss., nymphaeaceae salisb., rubiaceae juss., ulmaceae mirb., violaceae batsch). half of the families (24), which include rare and endangered taxa are represented by one species (alismataceae vent., araceae juss., aspleniaceae newman, campanulaceae juss., caprifoliaceae juss., ceratophyllaceae gray, colchicaceae dc., cystopteridaceae (payer) shmakov, equisetaceae michx. ex dc., fagaceae dumort., hypericaceae juss., lycopodiaceae p.beauv. ex mirbel, lythraceae j.st.-hil., malvaceae juss., oleaceae ho�manns. & link, onocleaceae pichi sermolli, ophioglossaceae martinov, primulaceae batsch ex borkh., salicaceae mirb., salviniaceae martinov, saxifragaceae juss., solanaceae juss., �elypteridaceae pichi sermolli, typhaceae juss.). among the genera, the genus carex l. (10 species) is the most numerous in the list, which includes almost two times more species than the next in number – dactylorhiza l. (5 species) and epipactis l. (4 species). �e genera allium l., bromus l., cephalanthera rich., juncus l., pedicularis l., crepis l., contain three species each. another 91 genera are represented by 1–2 rare or endangered species. according to the data published almost 40 years ago (shelag-sosonko et al., 1982), 102 rare species of vascular plants were recorded in the �ora of the entire opillya. as of today, even in some parts of opillya, such as male opillya, a much larger number of plant species need protection. in addition to male opillya (with its 143 endangered and rare species), emergency measures to preserve phytodiversity require e.g. azonal loci of steppe vegetation in rohatyn opillya. in this area noticed 269 rare and endangered plants species which need to be preserved (dmytrash-vatseba, 2017). �e need to protect this kinds of species has already been emphasized by polish researchers in the 1930s (kozłowska, 1931; szafer, 1935). conclusion �e list of rare and endangered taxa of vascular plants of male opillya contains 143 species. of these, 55 species are listed in the red book of ukraine (en – 6 species, vu – 25, r – 9, ne – 15 species). from the list of regionally rare species in male opillya 127 species were recorded (vu – 52 species, nt – 22 species, lc – 12 species, r – 13 species, ne – 10 species, dd – 8 species, en – 8 species, and r – 2 species). from the iucn list 100 species are part of the rare and endangered species of �ora of male opillya: lc – 73 taxa, dd – 15 species, nt – 8 species, vu – 2 species, en – 1 species. from the cites appendix №2 the list includes 27 species. five species on this area are protected by the bern convention. li di a ta se nk ev ic h, m ar ya s en iv , k ry st yn a s kr yp ec 56 con�ict of interest �e autors declare no con�ict of interest related to the article. references besser, w. 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(1998). особливості рослинного покриву долини дністра (features of vegetation of the dnister valley). kyiv: politychna dumka. [in ukrainian] zhyzhyn, м.p., kagalo, a.a., shelyag-sosonko, yu.r. (1990). стан і перспективи охорони популяцій crambe tataria sebeók на опіллі (урср) (population state and conservation perspectives of crambe tataria sebeók on opillya (ukrainian rsr)). ukrainian botanical journal, 47(6), 77–79. [in ukrainian] r are and endangered vascular plant species of m ale o pillya (lviv region, u kraine) 59 abstract male opillya (586 km2) is a small part of the volhyn-podillya uplands, located southeast of lviv and is characterised by the presence of various forms of landscapes, which led to the diversity of its phytobiota. although located in the neighbourhood of lviv – one of the botanical centres, it was overlooked by botanists in the 19th and 20th centuries. �e aim of the study, results of which are presented, was to determine composition of rare and endangered species of male opillya’s �ora. �e article provides a list of these species according to their conservation status de�ned by the red book of ukraine, the list of regionally rare species in lviv region, the iucn list, the cites appendix №2, and the bern convention. based on the conducted analyses, it can be concluded that 143 species of �ora in male opillya may disappear due to increasing anthropopressure. key words: endangered species, rare species, threat categories, ukraine received: [2021.09.10] accepted: [2021.10.30] rzadkie i zagrożone gatunki roślin naczyniowych małego opola (obwód lwowski, ukraina) streszczenie małe opole (586 km2) to niewielka część wyżyny wołyńsko-podolskiej, położona na południowy zachód od lwowa. charakteryzuje się ona obecnością różnych form krajobrazowych, co doprowadziło do zróżnicowania jej �tobioty. choć teren ten położony jest w sąsiedztwie lwowa – jednego z centrów botanicznych, w xix i xx wieku został przeoczony przez botaników. celem badań, których wyniki zostały tu zaprezentowane, było określenie składu rzadkich i zagrożonych gatunków �ory tego obszaru. artykuł zawiera listę tych gatunków według ich stanu ochrony określonego przez czerwoną księgę ukrainy, listę regionalnie rzadkich gatunków obwodu lwowskiego, listę iucn, załącznik №2 cites i  konwencję berneńską. na podstawie przeprowadzonych analiz można stwierdzić, że 143 gatunki i jeden podgatunek �ory małego opola mogą zniknąć z powodu narastającej tu antropopresji. słowa kluczowe: gatunki zagrożone, gatunki rzadkie, kategorie zagrożeń, ukraina information about authors lidia tasenkevich https://orcid.org/0000-0001-9348-1218 she is professor at the department of botany at ivan franko national university of lviv. her interests are in �oristics, phytogeograpy, plant ecology, phytosociology and plant protection. marya seniv https://orcid.org/0000-0001-7365-7183 she currently works at the herbarium of ivan franko national university of lviv. she is interested in �oristics, �ora synanthropisation and plant protection. krystyna skrypec https://orcid.org/0000-0003-4833-1653 phd; she currently works at the herbarium of ivan franko national university of lviv. she is interested in plant morphology, reproductive biology in the family iridaceae and plant protection. 96 annales universitatis paedagogicae cracoviensis studia naturae, 5: 96–109, 2020, issn 2543-8832 doi: 10.24917/25438832.5.7 joanna biel-parzymięso independent research, institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland; jbp2@ onet.eu effect of morus alba l. leaf extracts on seeds germination and the seedlings growth of sinapis alba l. and cucumis sativus l. introduction hans molisch (1937) was the �rst to introduce and apply the concept of allelopathy to biological systems. he de�ned the mutual – both adverse and bene�cial, biochemical interactions of all plants, including soil microorganisms as allelopathy (wójcik-wojtkowiak et al., 1998). �e concept of allelopathy (harborne, 1997) was consolidated in the second half of the 20th century as a  result of pioneering research by muller and chou (1972), followed by rice (1984). currently, research on this phenomenon is growing because it is seen as an opportunity for more e�ective weed control; through learning about their allelopathic properties, natural compounds could be used to inhibit weed development (vyvyan, 2002; możdżeń et al., 2018). allelochemical substances that stimulate plant growth may actually act as inhibitors at high concentrations, and compounds considered to be inhibitors at low concentrations may stimulate some growth processes. �ere are more than 10,000 different allelopathic compounds, and the main sources of their mass release are from crop plants, weeds, and soil microorganisms (leather, einhellig, 1988; barazani, fredman, 2001; weston, 2005; kong et al., 2019). �ese compounds primarily belong to low-molecular-weight secondary metabolites (rice, 1984; einhelling, 1994). in the case of microorganisms, they can include enzymes involved in secondary metabolism pathways and antibiotics (sturz, christe, 2003). most allelopathic substances are highly soluble in water, hence they easily penetrate into the soil solution. �e most common allelo-inhibitors include: organic acids, including phenolic acids; �avonoids; tannins; glycosides; terpenoids; alkaloids; unsaturated lactones; coumarins; and quinones. allelopathins can be released into the immediate environment of plants through their various organs or parts: roots, seeds, fruits, �owers, and leaves. in the roots, inhibiting substances usually have weak 97 effect of m orus alba l. leaf extracts on seeds germ ination and the seedlings grow th of sinapis alba l. and cucum is sativus l. properties and occur in lower concentrations. one example of an exception to this is the root of alfalfa (medicago sativa l.), which is actually the primary source of saponin inhibitors. roots usually contain large amounts of allelopathic substances with a  wide spectrum of activity. in seeds, inhibitors prevent rotting and control germination by imposing absolute dormancy. fruits contain inhibitors that play a  role in regulating seed germination. some �owers also possess similar chemical substances (gniazdowska, bogatek, 2005). most researchers believe that, under balanced natural conditions, allelopathins from seeds, fruits, and �owers are not released in amounts that could pose a  threat to nearby plants (wójcik-wojtkowiak et al., 1998). �e concentration of allelopathic compounds depends on the season, the age of the plant, and ecological and habitat factors. allelopathins are released in the highest amounts by young plants in spring as compared to mature and aging plants in autumn (wardle et al., 1993; ahmed, wardle, 1994). �e biological activity of allelopathic compounds is assessed relatively easily using biotests. �e simplest biotest is seed germination. a more accurate method may be to measure plant growth parameters (oleszek, 1992). �e biotest that consists of measuring the mass of plants treated with allelopathic compounds is an order of magnitude more sensitive than the biotest based on seed germination, and the measurement of seedling growth is even �ve times more sensitive than that. �e aim of this study was to determine the e�ect of aqueous extracts of mulberry (morus alba l.) leaves, with di�erent percentage concentrations, on: (1) germination of mustard (sinapis alba l.) and cucumber (cucumis sativus l.) seeds – as crop plants, (2) their growth, and (3) the fresh and dry mass of underground and aboveground organs. material and methods plant material mulberry leaves (morus alba) were collected in southern poland, and then dried in laboratory conditions. mustard (sinapis alba) and cucumber (cucumis sativus) seeds were purchased from plantico zielonki spółka z.o.o., polan cultivation and seed plant. extracts preparation aqueous extracts of dry mulberry leaves were prepared at three di�erent percentage concentrations: 3, 5, and 10; for this purpose, 3 g of dry leaves were weighed and �ooded with 97 ml of distilled water, 5 g of leaves were �ooded with 95 ml of distilled water, and 10 g of leaves were �ooded with 90 ml of distilled water, respectively. a�er 24 hours of extraction at room temperature and in the dark, the extracts were �ltered jo an na b ie l-p ar zy m ię so 98 through whatman type �lter paper and stored in the 8°c temperature for the duration of the experiment. seeds preparations and germination conditions fi�y seeds, mustard or cucumber, were rinsed with running and distilled water and placed on sterilised petri dishes with �lter paper, moistened with prepared extracts. �e control consisted of seeds on petri dishes with distilled water. all seed dishes were placed in a  dark 25°c thermostat. �e percentages of germinated seeds were checked systematically a�er 24, 48, 72, and 96 h. to be considered a germinated seed, the sprout length was equal to or higher than 2 mm (możdżeń et al., 2018). plant growth proceeding analogously as in the �rst stage of the experiment (germination), petri dishes were prepared with �lter paper moistened with distilled water (control) and mulberry extracts. on each of the 7 petri dishes 50 mustard or cucumber seeds were placed, a�er washing under running and distilled water. �e petri dishes were placed in a dark, 25°c thermostat for 48 h. in the meantime, 70 pots were prepared with sand washed in running and distilled water. morphologically similar mustard and cucumber seedlings were planted into 35 pots, from controls and extracts, previously rinsed with distilled water. seedlings were watered alternately every 48 h with 15 ml distilled water per plant and 10 ml steiner medium per plant (steiner, 1961). �e mustard and cucumber seedlings that were germinated in distilled water were planted in the remaining 35 pots. seedlings were watered alternately every other day with distilled water (15 ml/plant) and mulberry extracts (5 ml/plant) and once a week with steiner medium (10 ml/plant). all pots were placed in a growth chamber, with a light intensity of 200 μmol × m–2 × s–1, in the photoperiod 12 h/12 h, day temperature 25°c, night temperature 20°c, and relative humidity (rh) 70–80%. biometric analysis biometric analysis of mustard and cucumber organs was carried out on day 21 since planting the seedlings in the pots with sand. plants were removed from pots and their roots were washed in water and dried with paper towel. using a ruler, with an accuracy of 1 mm, the length of the root, hypocotyl, petioles and remaining part of the shoot, among other parts, was measured. fresh and dry mass �e fresh mass of mustard and cucumber organs was determined using an electronic scale (radwag 120 wps, poland). to obtain the dry mass, open petri dishes with plants were placed in a dryer (wamed sup-100, poland) for 48 h at a temperature 99 of 105°c. �e dried plant material was placed in a desiccator for 1.5 h. a�er this time, the dry mass of the plants was determined on an electronic scale, with an accuracy of 0.1 mg. statistical analysis �e statistical analysis of signi�cance di�erences between the means ±se were made by tukey test at p ≤ 0.05 in statso�, inc. (2018). results seeds germination a  much higher percentage of mustard seeds germinated under control conditions than on aqueous mulberry leaf extract (tab. 1). 58% of seeds watered with distilled water began germination a�er 24 h. �is percentage subsequently increased and after 72 h reached 92%. germination of mustard seeds on 3% extract began a�er 48 h, when 4% of seeds had germinated; a�er 96 h, the seeds had germinated to a level of 40% less than control. petri dishes saturated with 5 and 10% extracts exhibited germination delayed by 2 and 3 days, respectively, from the time when the seeds were placed on the extracts. as a result, a�er 96 h, 14% of seeds germinated on the 5% extract and only 8% had germinated on the 10% extract. tab. 1. germination seeds capacity [%] of mustard (sinapis alba l.) – a and cucumber (cucumis sativus l.) – b, on the aqueous mulberry leaves extracts (morus alba l.) time [h] control concentration of morus alba extracts [%] 3 5 10 a b a b a b a b 24 58 78 0 72 0 74 0 22 48 87 94 4 90 0 94 0 72 72 92 94 28 92 8 96 0 80 96 92 94 52 92 14 96 8 82 cucumber seeds on petri dishes with aqueous mulberry extract began to germinate a�er the �rst day (tab. 1); a�er this time point, the percentage of germinated seeds was lower than in the control sample. starting from day 2, the percentage of germinated seeds on the 3 and 10% extracts decreased. however, the percentage of germinated seeds on the 5% extract a�er the second day was the same as in the control; a�er 3 and 4 days it remained at a level 2% higher than the control. �e use of an extract with a concentration of 10% most strongly inhibited the germination of cucumber seeds. effect of m orus alba l. leaf extracts on seeds germ ination and the seedlings grow th of sinapis alba l. and cucum is sativus l. jo an na b ie l-p ar zy m ię so 100 tab. 2. length of selected mustard organs (sinapis alba l.) for plants watered with extracts of mulberry leaves (morus alba l.): a – during germination phase, b – during growth phase; mean ±se values from 5 replicates marked with di�erent letters di�er signi�cantly according to tukey test at p ≤ 0.05 organ length [cm] control concentration of morus alba extracts [%] 3 5 10 a b a b a b root 5.6 a 4.6 ab 5.1 a 4.8 ab 3.5 b 0.0 cde 3.8 b hypocotyl 5.5 a 4.4 b 4.8 b 4.4 b 4.5 b 0.0 cde 4.5 b petioles of leaf 1.2 a 1.3 a 1.0 ab 1.0 ab 0.6 b 0.0 cde 0.3 b remaining part of shoot 1.7 b 2.3 a 0.5 c 2.2 a 0.0 d 0.0 cde 0.0 d biometric analysis biometric analysis of mustard organs revealed an adverse e�ect on root growth and hypocotyls for plants grown from seeds watered with extracts for 48 h. �ese organs were shorter than similar organs grown in control plants (tab. 2). slight changes in length were observed for leaf petioles. growth in 3 and 5% extracts resulted in increased lengths of the remaining parts of the shoot, relative to the control. �e 10% concentration extracts completely inhibited growth and development of the tested plants. compared to control, the length of mustard plant organs from seedlings watered with mulberry extracts during the growth period was signi�cantly inhibited. regardless of the time point of watering, 10% extracts exerted the most adverse e�ect on plant growth. tab. 3. length of selected cucumber organs (cucumis sativus l.) for plants watered with extracts of mulberry leaves (morus alba l.): a  – during germination phase, b – during growth phase; mean ±se values from 5 replicates marked with di�erent letters di�er signi�cantly according to tukey test at p ≤ 0.05 organ length [cm] control concentration of morus alba extracts [%] 3 5 10 a b a b a b root 15.8 b 23.8 a 7.6 c 21.1 a 6.1 cd 7.3 c 5.6 d hypocotyl 5.1 a 5.1 a 4.0 b 5.3 a 4.1 b 4.5 b 5.7 a petioles of leaf 2.1 a 2.2 a 1.4 b 2.3 a 1.4 b 2.1 a 1.5 b remaining part of shoot 1.8 ab 2.0 a 1.3 b 2.4 a 1.1 b 0.6 c 1.2 b fresh and dry mass measurement of fresh and dry mustard organ masses revealed that 3% extract as a seed germination medium resulted in an increase in the value of all tested parameters compared to control (tab. 4–5). �e 5% extract resulted in an increase in fresh mass only for cotyledons and the remaining part of shoot. �e dry mass of the cotyledons of plants watered with 3% and 5% extracts in germination phase, exceeded the dry mass of the control. dry mass values for other organs were signi�cantly lower 101 than in the control. �e percentage of water content in mustard organs was lower for each of the extract concentrations used (fig. 1a – appendix 1). tab. 4. fresh mass of selected mustard organs (sinapis alba l.) for plants watered with extracts of mulberry leaves (morus alba l.): a  – during germination phase, b – during growth phase; mean ±se values from 5 replicates marked with di�erent letters di�er signi�cantly according to tukey test at p ≤ 0.05 organ fresh mass [mg] control concentration of morus alba extracts [%] 3 5 10 a b a b a b root 496.6 ab 562.2 a 288.8 b 436.6 ab 174.4 c 0.0 e 120.0 cd hypocotyl 944.4 a 1006.6 a 598.8 c 768.6 b 338.8 d 0.0 e 344.4 d cotyledons 770.0 b 906.6 a 694.4 bc 1056.6 a 572.2 c 0.0 e 406.6 d petioles of leaf 132.2 b 164.4 a 84.4 c 98.6 bc 24.4 d 0.0 e 16.6 de leaf blades 740.0 a 762.2 a 446.6 b 468.8 b 138.8 c 0.0 e 114.4cd remaining part of shoot 116.6 bc 298.8 a 16.6 c 148.8 b 0.0d 0.0 e 0.0 d �e fresh and dry mass of mustard organs from plants watered during the growth phase with the extracts were signi�cantly lower than the control values (tab. 4–5). �e mass values varied depending on the concentration of the extract; as the concentration of allelopathins increased, a  decrease in the values of these parameters was observed. �e water content in plant organs decreased as the concentration of extracts increased (fig. 1b – appendix 1). �e fresh mass of cucumber organs grown from seeds germinating for 48 h on 3 and 5% extracts increased with an increase in the concentration of allelopathins in the extracts (tab. 6). �e exception were leaf petioles and the remaining part of the shoot, whose masses were less than the fresh mass of control plants. plants grown from seeds germinating on 10% mulberry extract had a  lower fresh mass for almost all organs, compared to the control. similar results were obtained for cucumber dry mass (tab. 7). �e water content of the cucumber organs was lower than the control. an increase in the water content value for organs watered with 10% mulberry extracts was observed (fig. 1c – appendix 1). �e fresh and dry mass of cucumber organs from plants watered with extracts during the growth phase was generally less than the masses of control plants (tab. 6–7). mulberry extracts with a concentration of 3 and 5% caused a signi�cant increase in fresh and dry mass of cotyledons and the 10% extract increased leaf mass. in the case of percentage water content, no statistically signi�cant di�erences were observed (fig. 1d – appendix 1). tab. 5. dry mass of selected mustard organs (sinapis alba l.) for plants watered with extracts of effect of m orus alba l. leaf extracts on seeds germ ination and the seedlings grow th of sinapis alba l. and cucum is sativus l. jo an na b ie l-p ar zy m ię so 102 mulberry leaves (morus alba l.): a  – during germination phase, b – during growth phase; mean ±se values from 5 replicates marked with di�erent letters di�er signi�cantly according to tukey test at p ≤ 0.05 organ dry mass [mg] control concentration of morus alba extracts [%] 3 5 10 a b a b a b root 180.0 b 234.4 a 74.4 c 176.6 b 65.5 c 0.0 d 50.0 c hypocotyl 40.0 ab 52.2 a 27.7 c 40.0 ab 25.5 c 0.0 d 24.4 c cotyledons 48.8 b 54.4 ab 41.1 bc 60.0 a 40.0 bc 0.0 d 40.0 bc petioles of leaf 6.0 b 14.4 a 4.4 b 6.6 b 2.2 c 0.0 d 0 d leaf blades 74.4 a 80.0 a 45.5 b 46.6 b 30.0 c 0.0 d 20.0 cd remaining part of shoot 16.6 b 34.4 a 2.2 c 16.6 b 0.0 d 0.0 d 0 d tab. 6. fresh mass of selected cucumber organs (cucumis sativus l.) for plants watered wit–h extracts of mulberry leaves (morus alba l.): a – during germination phase, b – during growth phase; mean ±se values from 5 replicates marked with di�erent letters di�er signi�cantly according to tukey test at p ≤ 0.05 organ fresh mass [mg] control concentration of morus alba extracts [%] 3 5 10 a b a b a b root 579.6 b 803.4 a 408.8 bc 846.2 a 315.8 c 219.5 d 255.8 cd hypocotyl 216.2 a 219.2 a 169.0 c 226.8 a 152.0 c 180.5 b 269.8 a cotyledons 335.6 b 389.2 b 496.6 a 421.0 a 448.2 a 352.5 b 291.8 c petioles of leaf 86.2 ab 70.4 b 35.4 d 101.0 a 39.0 c 45.0 c 72.4 b leaf blades 435.6 b 496.8 a 93.8 e 545.8 a 154.6 d 292.0 c 470.0 a remaining part of shoot 59.0 b 55.2 b 35.4 c 86.2 a 32.2 c 24.2 d 39.0 c tab. 7. dry mass of selected cucumber organs (cucumis sativus l.) for plants watered with extracts of mulberry leaves (morus alba l.): a – during germination phase, b – during growth phase; mean ±se values from 5 replicates marked with di�erent letters di�er signi�cantly according to tukey test at p ≤ 0.05 organ dry mass [mg] control concentration of morus alba extracts [%] 3 5 10 a b a b a b root 43.2 b 116.2 a 45.6 b 123.6 a 43.6 b 33.2 c 42.4 b hypocotyl 11.2 a 11.8 a 8.6 b 12.2 a 7.8 c 8.5 b 10.8 b cotyledons 25.4 c 29.2 b 48.0 a 30.8 b 37.8 ab 26.2 bc 22.0 c petioles of leaf 4.0 ab 3.6 b 1.8 d 4.8 a 1.8 d 2.5 c 3.8 b leaf blades 45.4 b 52.0 a 9.2 de 54 a 14.2 d 30.2 c 48.0 b remaining part of shoot 4.2 b 4.2 b 2.8 c 5.6 a 2.6 c 2.0 d 2.8 c 103 discussion �is experiment demonstrated the adverse e�ects of mulberry leaf extracts on the germination and growth of mustard and cucumber (tab. 1–7). considering the fact that most allelopathics are phenolic compounds that belong to secondary metabolites (grześkowiak, łochyńska, 2017), it can be assumed that, to a  large extent, they could be responsible for inhibiting the germination process of the analysed species. allelopathic substances already exert negative e�ects during the seed swelling process and then disrupt the metabolic processes that occur in the germinating seed. in this study, each concentration signi�cantly inhibited mustard seed germination (tab. 1). however, for cucumber the reactions were slightly di�erent and likely related to the size of seeds and other structures of the seed coat (możdżeń, rzepka, 2017; mazur, 2019). �e percentage of germinated cucumber seeds, for low concentrations of mulberry extracts (3 and 5%), did not di�er greatly from the percentage of germinated seeds for distilled water (tab. 1). only at a 10% concentration of mulberry extract was a signi�cant germination delay observed. zandi et al. (2018) showed that aqueous extracts from stellaria media l. (vill.) at low concentrations stimulated seed germination of raphanus sativus var. radicula, and extracts from helianthus annuus l., regardless of concentration, inhibited growth of sinapis alba l. cv. barka (puła et al., 2020). �e negative e�ect of 10% mulberry extract on seed germination was probably due to the higher content of allelochemical compounds in the extracts. �ese results suggest that mulberry extracts caused oxidative stress due to allelopathins. a�er use of allelopathic compounds, plant tissues increased the production of reactive oxygen species which caused lipid peroxidation and oxidative damage (ding et al., 2020). inhibition of germination is a secondary e�ect of allelopathics. before their effects become visible, allelopathic substances �rst a�ect metabolic changes and physiological processes (możdżeń et al., 2018; szafraniec et al., 2019), such as cell membrane permeability and water-ion balance (skrzypek et al., 2015). in this experiment, the action of 10% mulberry extract was the most apparent; watering mustard seeds with it for 48 h completely inhibited their development and similarly to cucumbers caused a reduction in the length of individual organs and their fresh and dry mass (tab. 2–7). �is e�ect was probably associated with a high concentration of allelo-inhibitors (możdżeń et al., 2020). inhibition of plant growth may result from the delay or incubation of mitotic divisions and reduction of cell elongation (vaughan, ord, 1991). rice (1984) believed that allelopathic substances inhibit cell division and elongation by deforming the nucleus and strong cell vacuolisation. inhibition of plant growth may also be associated with a reduction in nutrient uptake due to impaired membrane integrity (klein, blum, 1990; baziramakenga et al., 1995; możdżeń et al., 2016), inhibition of plasmolemic proton pump activity, which plays effect of m orus alba l. leaf extracts on seeds germ ination and the seedlings grow th of sinapis alba l. and cucum is sativus l. jo an na b ie l-p ar zy m ię so 104 a key role in the growth of plant cells (janicka-russak et al., 2004), or with changes in hormonal balance (rice, 1984; vaughan, ord, 1991; wójcik-wojtkowiak, 1998). �e e�ects of phenolic compounds can be observed in this study, as mentioned previously (grześkowiak, łochyńska, 2017). phenols reduce protein biosynthesis, disrupt lipid metabolism, and inhibit the synthesis of porphyrin compounds, including chlorophyll synthesis (wójcik-wojtkowiak, 1998). in this experiment, symptoms of chlorosis were noted in mustard plants watered with 10% extracts. bright patches on the leaf surface indicated a chlorophyll de�ciency and most likely a�ected photosynthesis and dark respiration (einhelling, 1994; hussain, reigosa, 2011; możdżeń, repka, 2014; skrzypek et al., 2015). inhibitory activity of allelopathic compounds includes disorders of oxidative phosphorylation, reduction of atp levels, and reduction of oxygen uptake. energy de�ciency interferes with active transport of substances within the plant and functioning of cytoplasmic membranes for which a constant supply of metabolic energy is necessary (barkosky, einhelling, 2003; einhelling, 1994). many studies (e.g. wójcik-wojtkowiak, 1998; hussain, reigosa, 2011; możdżeń et al., 2018, 2020) indicate that allelochemical compounds interfere with water intake, its transport, and cause its gradual loss. as a result of these phenomena, they reduce the intensity of transpiration, induce the closing of stomata, and reduce the overall and active sorption surface of the roots. in studies with mulberry leaf aqueous extracts, di�erences were found in the water content of mustard and cucumber organs treated with this type of extracts (fig. 1 – appendix 1). �e withering of mustard plants watered with 10% mulberry extract may have been a sign of irregularities in the uptake of water by the roots and reduction of their sorption surface. it is not known whether the production of plant allelopathic substances is a  deliberate strategy developed to counter competition or an accidental occurrence, preserved in subsequent generations, allowing the plant to synthesise an advantage over other plants in a  particular ecosystem (king, 2003). whittaker (1972) put forward the theory that allelopathic interactions of chemical compounds present in plants are created as a result of pressure from herbivores. it was intended to be a reaction that repelled herbivores by releasing plant secretions from leaves, stems, and roots into the environment. �ese types of substances may have accidentally played a  role in plant-plant interactions. because they gave the plant the bene�t of reduced competition, their synthesis was maintained. �e need to reduce the use of chemicals in horticulture and agriculture, due to the high costs of synthetic plant protection products and the emergence of herbicide-resistant weeds, provides an opportunity to use allelopathy as a source of safer substances that improve the quality of agricultural production (cheng, cheng, 2015). �is is why plant-based chemicals are constantly being sought as a basis for synthesising natural herbicides (duke et al., 2000; vyvyan, 2002; gniazdowska, bogatek, 2005). 105 conclusion (1) �e aqueous extracts of mulberry leaves (morus alba l.) inhibited the germination of mustard seeds (sinapis alba l.); as the concentration of extract increased, the time of seed germination was delayed and number of germinating seeds signi�cantly decreased; for cucumber (cucumis sativus l.) signi�cant inhibition of the seed germination process was only observed with 10% mulberry extract, as compared to control. (2) regardless of the concentration of extracts and the time point of watering, a negative e�ect of mulberry leaf extracts on mustard and cucumber growth was demonstrated; mustard plants were more sensitive to extracts than cucumber plants. (3) fresh and dry mass of organs grown from seeds germinated on substrates with mulberry extracts in low concentrations was higher than in the control; with increasing extract concentration, regardless of the time point of watering with extracts, the tested plants were characterised by a smaller increase in fresh and dry mass for almost every organ compared to control; di�erences in percentage water content depended on the plant organ, extract concentration, and watering time. con�ict of interest �e author declares no con�ict of interest related to this article. references ahmed, m., wardle, d.a. 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[in polish] wójcik-wojtkowiak, d., politycka, b., weyman-kaczmarkowa, w. (1998). allelopatia (allelopathy). poznań: wydawnictwo akademii rolniczej w poznaniu. [in polish] zandi, p., barabasz-krasny, b., stachurska-swakoń, a., puła j., możdżeń, k. (2018). allelopathic effects of stellaria media (l.) vill. on germination and early stages of growth of raphanus sativus var. radicula. annales universitatis paedagogicae cracoviensis studia naturae, 3, 90–99. https://doi. org/10.24917/25438832.3.7 effect of m orus alba l. leaf extracts on seeds germ ination and the seedlings grow th of sinapis alba l. and cucum is sativus l. jo an na b ie l-p ar zy m ię so 108 appendix 1 fi g. 1 . a  w at er c on te nt o f o rg an s i n m us ta rd (s in ap is al ba l .) – a , b a nd c uc um be r ( c uc um is sa tiv us l .) – c , d , f or p la nt s w at er ed w ith e xt ra ct s o f m ul be rr y le av es (m or us a lb a l. ): a , c – d ur in g ge rm in at io n ph as e, b , d – d ur in g gr ow th p ha se (m ea n va lu es fr om 5 re pl ic at es ) 109 abstract plant growth and development can be modi�ed, including modi�cation by chemical processes that result from neighbouring plants. if interactions in the natural environment between one plant and another are of a chemical nature, then this phenomenon is called allelopathy. �e aim of the study was to determine the e�ect of aqueous extracts of morus alba l., at concentrations of 3%, 5% and 10%, on the germination and growth of sinapis alba l. (mustard) and cucumis sativus l. (cucumber). it was found that allelopathins contained in the extracts slowed the germination of both species. �e highest, 10%, extracts signi�cantly inhibited germination. it was found that with an increase in allelopathin concentration, there was a significant inhibition of the growth of underground and above-ground plant organs. a complete lack of growth was observed for mustard plants grown from seeds watered with extracts during germination for 48 hours. compared to the control plants, a di�erences in the growth of fresh and dry mass in plants watered with extracts during the germination and growth phases were found. depending on the timing of treatment and the type of organ tested, aqueous mulberry leaf extracts at lower concentrations had a  positive e�ect on the growth and development of the analysed species. extracts with a higher concentration of chemical compounds had a negative impact on both mustard and cucumber. key words: aqueous extract, cucumis sativus l., fresh and dry mass, plants length, sinapis alba l. received: [2020.04.07] accepted: [2020.06.20] wpływ wyciągów z liści morus alba l. na kiełkowanie oraz wzrost sinapis alba l. i cucumis sativus l. streszczenie wzrost i rozwój roślin jest mody�kowany, m.in. przez procesy chemiczne, wynikające z sąsiedztwa innych roślin. jeśli oddziaływania w  środowisku naturalnym jednej rośliny na drugą mają charakter rywalizacji chemicznej, to zjawisko określa się mianem allelopatii. celem przeprowadzonych tu eksperymentów było zbadanie wpływu wodnych wyciągów z  liści morus alba l., o  stężeniach 3%, 5% i  10%, na kiełkowanie i  wzrost sinapis alba l. oraz cucumis sativus l. okazało się, że zawarte w  ekstraktach allelopatiny spowalniały kiełkowanie nasion obydwu gatunków. najwyższe, 10% ekstrakty, wyraźnie ograniczały zdolność kiełkowania. stwierdzono, że wraz ze wzrostem koncentracji allelopatin następowało istotne zahamowanie wzrostu organów podziemnych i nadziemnych badanych roślin. całkowity brak wzrostu wykazano dla roślin gorczycy wyrosłych z nasion podlewanych wyciągami w czasie kiełkowania przez 48 h. w porównaniu z roślinami z kontroli, wykazano zróżnicowanie przyrostu świeżej i suchej masy u roślin podlewanych ekstraktami w  fazach: kiełkowania oraz wzrostu. w  zależności od czasu traktowania i  od rodzaju badanego organu, wodne wyciągi z  liści morwy w  niższych stężeniach miały pozytywny wpływ na wzrost i  rozwój analizowanych gatunków. ekstrakty o większej koncentracji związków chemicznych wpływały negatywnie, zarówno na gorczycę, jak i na ogórka. słowa kluczowe: wyciągi wodne, cucumis sativus l., świeża i sucha masa, wzrost roślin, sinapis alba l. information on the author joanna biel-parzymięso she is interested in an allelopathic interaction between weeds and crop plants. effect of m orus alba l. leaf extracts on seeds germ ination and the seedlings grow th of sinapis alba l. and cucum is sativus l. 129 annales universitatis paedagogicae cracoviensis studia naturae, 5: 129–141, 2020, issn 2543-8832 doi: 10.24917/25438832.5.9 małgorzata romańska institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland; *malgorzata_romanska@onet.pl impact of water stress on physiological processes of moss polytrichum piliferum hedw. introduction plant responses to water de�cits depend not only on their age and development phase but also on the extent and rate of water stress. a  slow pace of water loss allows air conditioning and reduces damage caused by a water de�cit, while fast-pace water loss can block this process. �ough, a similar water de�cit can cause di�ering responses in sensitive plants compared to plants resistant to water de�ciency (bray, 1997). in addition to water, oxygen availability is also an important element in plant biochemical processes. improper oxygen conditions of the soil adversely a�ect plants, causing root hypoxia and, consequently, inhibition of respiration. in hypoxic roots, growth is �rst restricted, followed by reduced water permeability and nutrient uptake. maintenance of hypoxia and drought leads to irreversible physiological processes in both seed and spore plants, e.g. mosses (kozlowski, 1984; kozlowski, pallardy, 1997; rzepka et al., 2003; rzepka, 2008). mosses (bryophyta) are a group of plants present in various relatively humid habitats around the world (proctor, 2000, 2001). in general, their life requirements are small, which is why they o�en resettle in di�cult conditions, as pioneer plants. in some plant communities they cover huge areas, constituting the dominant �oristic group. �ey are commonly found on meadows, peat bogs, in brush, set-aside, rocks, stones, trees, in ditches, �res, debris and on walls. mosses are characterised by high resistance to variability of habitat edaphic factors, temperature and humidity (karczmarz, 2000; fojcik, 2011; możdżeń, skoczowski, 2016; kula et al., 2018; sołtys-lelek et al., 2018; możdżeń, 2019). �ey collect water thanks to the imbibition forces located in the plasma of cells, membranes and mucous substances that are secreted on the surface of the stalks or gametophyte leaves. �e role of rhizoids is limited to attaching the plant to the ground and is not important during intake of water. �erefore, for m ał go rz at a r om ań sk a 130 many species of mosses, rainwater and dew are a  direct source of moisture. water conduction in mosses occurs due to the presence of layers of highly hygroscopic mucilaginous substances that draw in water and distribute over the entire surface of the turf. water migration also takes place over the surface of leafy stalks, due to capillary forces present in the spaces between the leaves, or through primitive conductive tissue developed in the middle of the stem (pressel et al., 2006). bristly haircap moss polytrichum piliferum hedw. from the polytrichaceae schwägr. family belongs to cosmopolitan spore plants but is more commonly found in the northern hemisphere. in poland, it occurs both in the mountains and in the lowlands. it grows in sunny, sandy, dry and acidic places and creates brown-green, loose turf (szafran, 1948). in its life cycle, like other mosses, the dominant generation is haploid gametophyte (fig. 1a–b). �e sporophyte has the form of a simple telome ending with a sporangial (fig. 1c–d). �e underground part of the gametophores are made of rhizoids up to 5 cm long, and the above-ground part of the stalk has leaves that reach 4.5 to 6 mm in length and 0.5 mm in width. �ey are lanceolate, whole leaf edges, extended at the bottom in a single-layer, non-green sheath. �is moss has a double-layered leaf blade with a single-layer edge; a rib is present in the middle of the leaf (fig. 1b). �e ventral side of the leaf has lamellas, whose apical cells are bottle-shaped and papillary (wójciak, 2003). fig. 1. polytrichum piliferum hedw.: a – gametophores, b – fragment of gametophyte leaf with rib, c – sporophytes, d – fragment of capsule with peristome (source: https://atlas.roslin.pl/plant/9498) 131 im pact of w ater stress on physiological processes of m oss polytrichum piliferum h edw . mosses are convenient organisms for studying the reaction of plants to water stress because they are relatively primitive for terrestrial plants. �ey do not have an epidermis, which is why they are more sensitive to moisture changes than most other plants (harmens et al., 2011; schröder et al., 2014). �erefore, they are o�en a research object for exploring plant hydration. �e aim of the study was to determine the e�ect of water stress on the activity of photosynthesis (i) and dark respiration (ii) in bristly haircap moss p. piliferum. material and methods plant material �e turf of polytrichum piliferum mosses collected in situ in spring 2010, from the natural forest habitat surrounding rybna (50°03ʹ04ʺn 19°38ʹ50ʺe – southern poland) site was used for the research. conditions plant material was acclimated for 2 weeks in a  growth chamber. conditions for mosses during the acclimatisation were a 12 h/12 h photoperiod, density of quantum stream in the par range of 70 μmol × m–2 × s–1 – with the �uorescent light source fluora-osram (poland), a temperature of 15°c (±2°c) and a relative humidity that oscillated around 70–100%. mosses were regularly watered with distilled water. a�er the acclimatisation period, similar morphological p. piliferum gametophores ± 1.5 cm long were selected for the study. plants were rinsed with distilled water and dried with �lter paper. �en they were placed into holes of plexiglas plates in glass vessels with a volume of 1 dm3 with 10 ml of distilled water and stored in a growth chamber. gas exchange an infrared gas analyser adc–225 mk3 (uk) operating in a closed system was used to measure photosynthesis and respiration of gametophores. �e entire system consisted of an assimilation chamber with an air humidifying system and a water jacket, and its volume was 0.664 dm3. measurements were carried out in air with an oxygen content of 21%. �e temperature inside the assimilation chamber was 25°c throughout the measurements. during photosynthesis measurements, the intensity of light reaching the gametophores was 100 μmol × m–2 × s–1. �e concentration of carbon dioxide was 300–400 μmol co2 in 1 litre of air in a  closed system, and the relative humidity of the air was approximately 75%, which allowed air to pass through the scrubber with distilled water. m ał go rz at a r om ań sk a 132 experimental groups �e control group for photosynthesis and dark respiration were p. piliferum gametophores arranged on plexiglass plates, immediately a�er acclimatisation. a�er the fresh mass determination and gas exchange measurements, the mosses were dried for 1 hour at room temperature, and then their mass, photosynthesis and dark respiration were measured. a�er this treatment, the gametophores were sprayed with distilled water and allowed to rehydrate in air for 15 h. �en their mass was determined, photosynthesis and dark respiration were measured. a�er measurement, the gametophores were placed in a dryer (wamed sup-100, poland) until completely dry to determine their dry mass (dm). �e second experiment consisted of periodic �ooding of gametophores with distilled water for 1 h, a�er which photosynthesis and respiration parameters were determined, and then the plant material was transferred to the atmosphere for 15 h. a�er this time, the gas exchange parameters were determined. a�er completing these measurements, the plant material was placed in the dryer to completely determine the dm. statistical analysis statistical analysis was performed using one-way analysis of variance, anova/ manova, tests. �e signi�cance of di�erence between means (± se), n = 5, was determined with tukey test at p = 0.05. �e data was analysed with statistica data analysis so�ware system (statso�, inc., 2018, version 13.1, www.statso�.com). results drying and rehydration in the control group, photosynthesis intensity for polytrichym piliferum gametophores was 61.3 μmol co2 gdm –1 × h–1. drying the plant material for one hour reduced the intensity of photosynthesis to 0 μmol co2 gdm –1 × h–1. rehydrating the same material for 15 hours resulted in the restoration of photosynthetic activity to 32 μmol gdm –1 × h–1, which constituted 51% of the initial value (fig. 2a; tab. 1). �e dark respiration rate for the control plants was 35.5 μmol co2 gdm –1 × h–1. a�er drying the mosses for 1 hour, respiration decreased to 34.0 μmol co2 gdm –1 × h–1, compared to the control. a�er subjecting the same gametophores to 15 h of rehydration, the intensity of dark respiration increased signi�cantly to 45.6 μmol co2 gdm –1 × h–1 and was 28.5% higher than the control value (fig. 2a; tab. 1). 133 flooding and reoxygenation before �ooding p. piliferum gametophores, photosynthesis intensity was 78 μmol co2 gdm –1 × h–1 (control). hourly (1 h) �ooding of plant material reduced photosynthesis to 61.8 μmol co2 gdm –1 × h–1, i.e. 21% relative to the control. in addition, subjecting the plant material to 15 hours of reoxygenation in the air resulted in an increase in photosynthesis intensity to 66.3 μmol co2 gdm –1 × h–1, i.e. its intensity was 15% lower than the initial value (fig. 2a; tab. 1). �e intensity of dark respiration in the control sample was 41 μmol co2 gdm –1 × h–1. a�er 1 h of �ooding the plants, respiration decreased to 37.2 μmol co2 gdm –1 × h–1 (by 9% compared to the control). a�er subjecting the same gametophores to air reoxygenation for 15 hours, the intensity of this process increased to 38.0 μmol co2 gdm –1 × h–1 (by 7% of the initial value) (fig. 2b; tab. 1). fig. 2. changes in photosynthesis intensity (p) in polytrichum piliferum hedw. gametophores: a: p1 – a�er 1 h drying at room temperature, p2 – a�er 1 h drying and 15 h rehydration; b: p3 – a�er 1 hour of �ooding, p4 – a�er 1 hour of �ooding and 15 hours of reoxygenation in the air; mean values (± se) marked with di�erent letters di�er signi�cantly according to tukey test p = 0.05 im pact of w ater stress on physiological processes of m oss polytrichum piliferum h edw . m ał go rz at a r om ań sk a 134 tab. 1. changes in photosynthesis and respiration processes in polytrichum piliferum hedw. gametophores during drought and water stress; values expressed in % in relation to the control percentage of control mosses treatment a�er 1 hour of drying a�er 1 hour of drying and 15 h rehydration a�er 1 h �ooding a�er 1 hour of �ooding and 15 h reoxygenation p 0.00 51.02 79.12 85.44 r 96.01 128.50 91.14 93.25 p – photosynthesis, r – respiration; control – measurement taken at the beginning of the experiment, p1, r1 – a�er 1 h drying at room temperature, p2, r2 – a�er 1 h drying and 15 h rehydration, p3, r3 – a�er 1 hour of �ooding, p4, r4 – a�er 1 hour of �ooding and 15 hours of reoxygenation in the air fig. 3. changes in dark respiration intensity (r) in polytrichum piliferum hedw. gametophores: a: r1 – a�er 1 h drying at room temperature, r2 – a�er 1 h drying and 15 h rehydration; b: r3 – a�er 1 hour of �ooding, r4 – a�er 1 hour of �ooding and 15 hours of reoxygenation in the air; mean values (± se) marked with di�erent letters di�er signi�cantly according to tukey test p = 0.05 135 discussion as a solvent, water is an irreplaceable environment for biochemical reactions. it is a substrate for many reactions and a factor that keeps the protoplast in proper physical condition. with only a 10% water de�cit, the photosynthetic process was already decreasing due to the reduction of turgor and the automatic closing of stomata. water is also associated with the transport of organic substances. it is an important temperature regulator in plant tissues, and in the process of photosynthesis it is a source of hydrogen for assimilation force. dehydration of the green crumb inhibits chlorophyll synthesis and accelerates its degradation. however, plasma hydration has a signi�cant impact on the entire course of respiration (martim et al., 2009; mcelrone et al., 2013; silva et al., 2014). most mosses are very sensitive to changes in water conditions (rzepka et al., 2003, 2005), dependent on surface water and capillary forces action, which are o�en supported by setting gametophore leaves (wójciak, 2003). mosses belong to poikilohydric plants, i.e. they are characterised by variable hydration. in drought conditions, these plants can dry out completely, but their structure is not completely destroyed (lou, 2006; ruibal et al., 2012; zhou et al., 2011). increasing the availability of water, e.g. as a  result of atmospheric precipitation, causes rapid hydration of their tissues and activates metabolic processes that gradually dried up during drying. poikilohydric plants are hydrolabile, which means that their water balance can be negative for a long period. �is is the result of the low sensitivity of the stomata to dehydration, as only very low environmental humidity causes their gradual closing and limiting of transpiration. �e protoplasm of these plants is resistant to signi�cant and rapid �uctuations in water potential (rock et al., 2009). experiments carried out on polytrichum piliferum gametophores have shown that mosses adapt relatively quickly to changing water conditions (fig. 2–3; tab. 1). �e rate of photosynthesis of gametophores dried for 1 h was 0 μmol co 2 gdm –1 × h–1, and a�er 15 h of rehydration it constituted 51% of the control sample. �e dark respiration rate measured in p. piliferum, a�er 1 h of drying, decreased by 4% of the original value. a�er 15 h of rehydration, this process increased by 28.5% relative to the control sample. �is clearly illustrates a lower sensitivity of the dark respiration process than photosynthesis to a periodic lack of water (rzepka, 1990). �e process of drying and then rehydrating mosses can be repeated several times, without lethal changes to the body (proctor et al., 2007). one of the properties of mosses that allows them to survive di�cult periods of drought is the ability to preserve undamaged ribosomes. �is provides the opportunity to quickly regain the ability to synthesise proteins when the cells are rehydrated. it is an ecological adaptive feature not found in other plants because irreversible destruction of the protein synthesis apparatus occurs during drying (krupa, 1974; zeng et al., 2002). another feature of mosses is their relatively high level im pact of w ater stress on physiological processes of m oss polytrichum piliferum h edw . m ał go rz at a r om ań sk a 136 of sucrose (~10% of dry mass), which remains constant during dehydration and rehydration (bewley et al., 1978; alpert, oliver, 2002). during rehydration, the moss cells quickly return to homeostasis and become convex, and the cell organelles gradually return to their normal shape and function (pressel et al., 2006; proctor et al., 2007). dehydration causes general condensation of cell content, fragmentation of the central vacuole system, increased viscosity of the cytoplasm, condensation of chromatin and dense packing of ribosomes (pressel et al., 2006; proctor et al., 2007). �e plasma membranes of dried cells, e.g. syntrichia ruralis (hedw.) f. weber & d. mohr (= tortula ruralis), look like typical lipid bilayer membranes, with scattered endothelial particles. nuclei, chloroplasts and mitochondria lose their elongated shape and become round or ovate, probably due to the loss of the depolymerised microtubule cytoskeleton (platt et al., 1993; ligrone, duckett, 1996, 1998; hoekstra et al., 2001). �e e�ects of water de�cit are revealed in almost every cellular process. one such response may be the appearance of oxidative stress. reactive oxygen species are formed, which can trigger reactions leading to damage to cellular structures (rzepka, 2008). �is stress leads, e.g. for lipid peroxidation, to changes in thylakoid structure, inhibition of photosystem activity of psi and psii and electron transport chain and inhibition of photosynthesis due to the inactivation of thiol groups of the rubisco enzyme. reactive oxygen forms are removed using oxidants, e.g. catalase, peroxidase and superoxide dismutase (hoekstra et al., 2001; franca et al., 2007). under conditions of water de�cit, the processes of incorporating carbon are clearly modi�ed, rubp carboxylase activity is reduced and the incorporation of carbon into organic compounds is decreased. inhibition of photosynthesis activity is also caused by partially closed stomata. �e dark respiration process is less sensitive to periodic lack of water than photosynthesis and continues even when the potential values are low. experiments carried out on various species of bryophytes capable of withstanding periods of drought prove that the respiration rate, as these organisms are dried, clearly decreases (krupa, 1974; rzepka, krupa, 1996; rzepka et al., 2001). in the experiment conducted with p. piliferum, �ooding with water followed by reoxygenation in the air caused changes in the photosynthesis and the dark respiration processes of gametophores. a�er 1 hour of �ooding, photosynthesis activity decreased by 21% and dark respiration by 9%, compared to the control. a�er 15 h of reoxygenation in the air, the reduction of photosynthesis had reached approximately 15% and respiration 7%, compared to these processes in the control (tab. 1). hypoxia can induce various plant responses (rzepka, 2008; rzepka et al., 2003, 2005). many changes that adapt plants to oxygen de�ciency are preceded by the activation or repression of speci�c genes (mcdaniel et al., 2007; trouiller et al., 2007). according to recent hypotheses, morphological adjustment consists of the development of aerenchyma, whose large intercellular spaces cause the movement of oxygen from shoots to 137 underground organs and these �ooded organs (kopcewicz, lewak, 2005). osakabe et al. (2014) wrote that water stress is an important factor limiting plant growth and productivity. longer exposure to the stress factor causes weakness and loss of vigour of the body’s cells, which in turn increases susceptibility to diseases caused by pests and pathogens. �is leads to disturbances in the functioning of the plant and a decrease in its biological value. �e plant cell regains full physiological activity a�er being saturated with water. �e speed of recovery is generally inversely proportional to the rate and intensity of stress, but most mosses regain their normal morphology in a  few minutes and rebuild their architecture in 1–2 days (proctor, 2001; pressel et al., 2006; proctor et al., 2007). �e experiments carried out here showed that subjecting p. piliferum gametophores to water stress causes an imbalance of physiological processes such as photosynthesis and dark respiration. it can be argued that the plant’s ability to maintain homeostasis in the circumstances of the stressor or change homeostasis through adaptation determines the resistance of plants to stress and allows them to survive or overcome prolonged stress. conclusion mosses are poikilohydric organisms and resistant to various environmental stress factors. tolerance to drying or �ooding is a  relatively common phenomenon in moss plants. it helps them survive adverse environmental conditions. (i) drought stress caused complete inhibition of photosynthetic activity of polytrichum piliferum gametophores. however, rehydration for 15 h resulted in the restoration of the photosynthetic activity of the plants. �is is in contrast to hypoxia stress, which did not significantly a�ect photosynthesis or drying. (ii) dark respiration in the tested mosses was clearly less sensitive to drought stress or hypoxia, compared to photosynthesis. �ere were slight di�erences in the activity of dark respiration, compared to the controls. perhaps, the ability to change homeostasis by adapting, surviving or overcoming adverse living conditions plays an important role. global climate change causes a  rise in temperature, carbon dioxide and variable distribution of atmospheric precipitation. it is these factors that increase the demand for the monitoring and testing, including photosynthesis and dark respiration, of mosses and other plant species in the eco-physiology �eld. acknowledgements i would like to thank prof. jan krupa for valuable advice and help in editing the paper. i also thank phd grzegorz rut and phd katarzyna możdżeń for help in carrying out the experiments. con�ict of interest �e author declares no con�ict of interest related to this article. im pact of w ater stress on physiological processes of m oss polytrichum piliferum h edw . m ał go rz at a r om ań sk a 138 references alpert, p., oliver, m.j. 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(2011). combined e�ects of nitrogen deposition and water stress on growth and physiological responses of two annual desert plants in northwestern china. environmental and experimental botany, 74, 1–8. doi: 10.1016/j.envexpbot.2010.12.005 abstract mosses are convenient organisms for studying the reaction to water stress because they do not have an epidermis, which makes them more sensitive to changes in humidity than most other plants. �e aim of the study was to determine the e�ect of water stress on the course of physiological processes of mosses using polytrichum piliferum hedw. �e present study showed that the action of the abiotic stressor, which is water, adversely a�ects the photosynthesis and dark respiration processes by reducing their intensity. however, it is worth noting that the respiration process is less dependent on tissue hydration than the photosynthesis, which is clearly demonstrated by the study results. �e bryophytes’ resistance to stress factors is responsible for the plant’s ability to maintain homeostasis under stress conditions. �e ability to change homeostasis by adapting, surviving or overcoming adverse living conditions also plays an important role. key words: dark respiration, hypoxia, photosynthesis, polytrichum piliferum, rehydration received: [2020.03.31] accepted: [2020.06.16] 141 wpływ stresu wodnego na przebieg procesów fizjologicznych u mchów streszczenie mchy są organizmami dogodnymi do badania reakcji na stres wodny, ponieważ nie posiadają epidermy, przez co odznaczają się większą wrażliwością na zmiany wilgotności niż większość innych roślin. celem pracy było określenie wpływu stresu wodnego na przebieg procesów �zjologicznych mchów na przykładzie polytrichum piliferum hedw. przeprowadzone badania pokazały, że działanie abiotycznego stresora, jakim jest woda, wpływa niekorzystnie na przebieg procesów fotosyntezy i oddychania, poprzez zmniejszenie ich natężenia. jednak warto zaznaczyć, że proces oddychania jest w mniejszym stopniu uzależniony od uwodnienia tkanek niż proces fotosyntezy, co wyraźnie widać w przeprowadzonych tu badaniach. za odporność mszaków na czynniki stresowe odpowiada zdolność rośliny do utrzymania homeostazy w czasie działania stresora. również ważną rolę odgrywa zdolność zmiany homeostazy przez adaptację, przetrwanie albo pokonanie niekorzystnych warunków życiowych. słowa kluczowe: oddychanie ciemniowe, hipoksja, fotosynteza, polytrichum piliferum, rehydratacja information on the authors małgorzata romańska she is interested in mosses physiology in di�erent stress factors. im pact of w ater stress on physiological processes of m oss polytrichum piliferum h edw . 145 annales universitatis paedagogicae cracoviensis studia naturae, 5: 145–156, 2020, issn 2543-8832 doi: 10.24917/25438832.5.10 iveta marková*, mikuláš monoši department of fire engineering, faculty of security engineering, university of žilina 8215/1, 010 26 žilina, slovakia; *iveta. markova@çi.uniza expressions of climatic change in slovak republic introduction �e united nations framework convention on climate change (convention) (sažp, 1994, 2019a), kyoto protocol to the convention (sažp, 1999; adamišin et al., 2005; bédi, 2007) and the paris agreement (council eu, 2019) are the international legal instrument for the search for global solutions to climate change. slovakia is a party to the above conventions and is obliged to ful�l the resulting obligations. for example, slovakia has successfully reduced greenhouse gas emissions by 8% compared to the base year 1990 (sažp, 2019a). historically, the �rst universal agreement on climate change – the paris agreement, entered into force on 4 november 2016. �e aim of the agreement is to limit global temperature growth to a maximum 2°c by the end of the century and, if possible, to signi�cantly below 1.5°c. �is document includes reduction commitments not only for developed countries but for all countries that are parties to it. each country determines for itself how and in which sectors it will make an e�ort to reduce greenhouse gas emissions. �is agreement pays close attention to climate change and enshrines the obligation to prepare for the consequences of these changes. last but not least, this agreement introduced an obligation to monitor emissions and report on their quantities to all participating countries. finally, the paris agreement was rati�ed by 181 countries out of a total of 197 parties to the convention (in terms of developed countries, russia and turkey did not ratify this agreement) on 26 october 2018 (sažp, 2019a). �e climate-adapt project, led by the european environment agency with the support of the united nations framework convention on climate change, evaluates the so-called “indicators” of climate change (eea, 2019). �e group of climatic elements includes: greenhouse gas production as concentrations of pollutants, average annual air temperature, annual total atmospheric criteria, drought index and annual soil temiv et a m ar ko vá , m ik ul áš m on oš i 146 perature (soil index). �e selected indicators meet the following criteria: direct or indirect link to the climate system; consistent, homogenised and uninterrupted data over a longer period of time (minimum 30–50 years); guarantee of measuring, monitoring or recording data towards the future and good explanatory value (minďas et al., 2011). �e aim of this article is to describe the manifestations of climate change in slovakia (since its inception) according to selected indicators: (1) average annual air temperature, (2) soil temperature, (3) total atmospheric precipitation and (4) drought index over the last decade. material and methods on the slovakian territory, a selection of climate monitoring stations was chosen for the purposes of a representative evaluation of climatic indicators in relation to height above mean sea level (amsl) (e.g. bratislava-airport 48 10ʹ12ʺn, 17 12ʹ46ʺe, amsl 133 m; hurbanovo 47°52ʹ32ʺ n, 18 11ʹ36ʺ e, amsl 115 m; oravská lesná 49 22ʹ8ʺ n, 19°11ʹ1ʺ e, amsl 934 m; sliač 48°36ʹ43ʺ n, 19°8ʹ37ʺ e, amsl 305 m; lomnický štít 49 11ʹ45ʺ n, 20 12ʹ46ʺ e, amsl 2634.4 m; poprad 49°3ʹ34ʺ n, 20°17ʹ51ʺ e, amsl 672 m; košice airport 48°39ʹ47ʺ n, 21°14ʹ28ʺ e, amsl 230 m; liptovský hrádok 49 2ʹ15ʺn, 19 43ʹ33ʺ e amsl 637 m and others) (shmú, 2020a). monitoring of climate elements (air temperature, soil temperature, precipitation, air pressure, sunshine duration, number of ice days – maximum temperature lower than 0°c, number of freezing days – minimum temperature lower than 0°c, number of summer days – temperature higher than 25°c, number of tropical days – temperature higher than 30°c) allows for the quanti�cation of climate change. �e expression of climatic change is characterised by the chosen study parameters, of which only four were included in this paper (average annual air temperature, soil temperature as a soil dryness index (sdi), total atmospheric precipitation and drought index (di)). �e soil dryness index (sdi) is determined by the ratio of daily rainfall and the maximum temperature from local meteorological stations for estimating soil dryness, within tens of kilometres. it estimates the millimetres of rain needed to �ll the soil with water (mount, 1980; jamroz et al., 2014; hollá, 2016). �e current territorial average for total precipitation in slovakia is calculated in the slovak hydrometeorological institute (shmú) from the monthly totals of 203 stations. �is calculation was produced from the approximately 30–100 stations present before 1901 (lapin, 2020). �e drought index (di) is based on a comparison (ratio) of the annual potential evapotranspiration and the annual total atmospheric precipitation. alsumaiei (2020) and paulo et al. (2012) explained identi�cation and testing of the drought index. �ese indicators began to be monitored gradually. �ey were �rst published in the 2010 state of the environment report. expressions of clim atic change in s lovak r epublic 147 current data on climatic elements are published annually on the website of the slovak environment agency (sažp, 2019b); this data was used in these studies and discussed in terms of the �ndings and comments of slovak experts and climatologists. results and discussion air temperature climate change is most evident in air temperature, which is con�rmed by the readings of this parameter by many hydrometeorological stations. according to previous research, air temperature has been measured in hurbanovo at a professional level since 1872, with four shmú stations available since 1881, and one in bratislava since 1851 (lapin, 2020). �e average territorial deviation of temperature from normal, 1961–1990 ranges, in 2016 was incomparable with the year 2018. during the two years there was a 1.5°c di�erence in the deviation of the average annual air temperature from the normal 1961–1990 ranges. concerning means, the average the deviation ranges from 1°c, 1.0–1.5°c and 1.5°c in 2016 and in 2018, and the di�erence in the deviation of the average annual air temperature from the normal 1961–1990 ranges were increased (fig. 1). matejovič and libo (2020) reported record temperature values. �e absolute maximum reached (the highest measured air temperature) was 56.7°c, death valley july 10, 1913, california, usa and in slovakia: 40.3°c, hurbanovo, july 20, 2007. �ey reported the warmest summer (june–august) was in 2019, with an average air temperature of 23.2°c in hurbanovo. �is fact was con�rmed by bartošovičová (2019). during the period 1881–2016, an average annual air temperature increase of about 1.73°c (sažp, 2017a) was observed in slovakia. �e report on the state of the environment for 2015 (sažp, 2016) states: “warming has been most pronounced in the last twenty years”. �e year 2016 ended as extremely warm, in comparison with the climatic norm of 1961–1990; 2016 also ended as extremely warm, in comparison with the climatic norm 1961–1990 (sažp, 2017a), in most of slovakia. �is characteristic was maintained in the following years (sažp, 2018, 2019a) �e number of tropical days has been reported in the state of the environment report since 2012 (sažp, 2013). �e criterion for classifying days as tropical is higher than 30°c. �e normal summer day is day, when the maximum air temperature has increased to at least 25°c. based on study data, a growing trend in the number of tropical days was observed in the lowland and uplands of slovakia (hurbanovo increased by 20, liptovský hrádok increased by 10), for the period 1951–2016 (sažp, 2017b). in 2016, 30 tropical days in hurbanov and 5 days in liptovský hrádok were recorded. �is was also observed in 2018, where, in both places together, the number of tropical days increased by 16, compared to the period 1961–1990 (sažp, 2019a). an increase iv et a m ar ko vá , m ik ul áš m on oš i 148 fi g. 1 . d ev ia tio ns o f t he a ve ra ge a nn ua l a ir te m pe ra tu re (d t) fr om th e no rm al fr om 1 96 1– 19 90 in s lo va ki a fo r t he y ea rs 2 01 2, 2 01 4, 2 01 6 an d 20 18 (s ou rc e: s h m ú 2 02 0a , b ; s a žp 2 01 3– 20 19 a, b ) expressions of clim atic change in s lovak r epublic 149 fi g. 2 . a nn ua l o f c um ul at iv e pr ec ip ita tio n to ta ls in % fr om th e no rm al (r ) f ro m 1 96 1– 19 90 in s lo va ki a fo r t he y ea rs 2 01 3, 2 01 6, 2 01 7 an d 20 18 (s ou rc e: s h m ú 2 02 0a , b ; s a žp 2 01 2– 20 17 a, b ) iv et a m ar ko vá , m ik ul áš m on oš i 150 in warm days causes a decrease in the length of the heating season. for example, in hurbanovo a decrease of 21 days and in liptovský hrádok a decrease of 22 days was found during the analysed period (sažp, 2017b). based on the increase in the number of summer and tropical days, (45 more days compared to the period 1961–1990), the number of ice days, i.e. lower than 0°c, decreased (a decrease of 12 days compared to the period 1961–1990) (sažp, 2019a). a detailed explanation of the development of these trends and a forecast of their number through 2090 were reported by damborská et al. (2006). in the lowlands throughout central europe, due to the frequent alternation of periods with positive and negative average air temperature, snow cover occurrence is irregular (siman, slavková, 2019) in winter. �e report on the state of the environment for 2016 describes a decrease for all snow cover characteristics (mainly thickness) to a height of 1000 m in almost the entire territory of the slovak republic (an increase was recorded at higher altitudes). in the following years, reports on the state of the environment no longer specify the state of snow cover but only state its decrease. soil temperature (soil dryness index) based on this scienti�c analysis, the soil temperature at a depth of 10 cm in hurbanovo was 11.0°c and in liptovský hrádok was 9.2°c, in 2016. a trend of increasing average annual soil temperature at a depth of 10 cm was observed in lowlands and uplands of slovakia; in the mountains (hurbanovo 1.5°c, liptovský hrádok 2.1°c) for the period 1951–2016, the observed temperature increases were more signi�cant (sažp, 2018). a drought began to spread �rst in the northwest, later in the east, of slovakia in the second half of april 2018. at the beginning of may 2018, there was an extreme drought in 16% of the territory. �e worst situations were in the žilina, prešov and trenčín regions. extreme drought a�ected 7.5% of the country territory in june 2018 (sažp, 2019a). �is situation caused a de�cit of soil moisture and a loss of yields in eastern slovakia. cumulative precipitation totals �e annual precipitation totals began to be presented in the reports on the state of the environment in 2010. �e �rst map presented from 2012 employs a three-colour range with deep blue colours. in 2013, the colour of the map di�ers signi�cantly and areas are indicated with a signi�cant reduction in percentage cumulative precipitation, less than 60% of normal. �e map from 2015 revealed a declining trend in the annual precipitation totals (fig. 2) and a decrease in the relative humidity of the air (sažp, 2016). in 2016 (sažp, 2017a) there was a decrease in annual precipitation totals of 0.5%, on average, and 2018 precipitation was below normal �e upper limit of the percentexpressions of clim atic change in s lovak r epublic 151 age range of annual total atmospheric precipitation decreased from 140% to 120%. at the same time, there were areas with a 60% deviation from normal in annual precipitation totals during that period (sažp, 2019a). �e calculation methods for precipitation totals over a selected time period and for scenarios of precipitation totals during extreme precipitation situations in slovakia are explained by lapin et al. (2004). drought index �e drought index (hurbanovo station – 0.22) in the lowland areas of slovakia revealed an increasing trend for the period 1951–2016. for example, the drought indexes were 1.02 in hurbanovo and 0.41 in oravská lesná in 2016. detailed statistical highlights are found in the report on the state of the environment for 2016 (mžp, 2019; sažp, 2017a, b). on the other hand, local or widespread drought was much more common than before. �is phenomenon was caused mainly by long periods of relatively warm weather with small totals of precipitation in some parts of the growing season. in general, in this part of europe, the drought phenomenon was associated with relatively warmer winters (no snow cover) and warmer summers (greater evaporation). in some areas of slovakia, precipitation totals did not deviate from the long-term norms; however, intense, heavy rainfall in summer prevails that results in fast out�ow and low water retention. �e phenomenon of drought has become part of our lives. in 2018, documents were prepared which took a position and proposed speci�c measures to be taken to prevent water loss in our environment (sólymos, 2018); therefore an action plan to address the consequences of drought and water scarcity was created (mžp, 2018; sažp, 2018, 2019b). �ere is still debate in the scienti�c literature about which climatic parameters (e.g. precipitation, temperature, evapotranspiration, wind speed, relative humidity, solar radiation, etc.) are the most important in determining the severity of drought. �ere is general agreement on the importance of precipitation in explaining drought variability and the need to include this variable in the calculation of any drought index (vicente-serrano et al., 2009; jamroz et al., 2014; tigkas et al., 2014; amani et al., 2016). climate scenarios in the third national report on climate change (mžp, 2001) the climatic scenarios of temperature increase and decrease in annual total precipitation were presented. �e calculation was a regional modi�cation of the outputs from two related programs: gcms (cccm – the canadian centre for climate modelling) and giss (gaoldard’s institute for space studies in the usa). �e synthetic data for this topic is presented in table 1. iv et a m ar ko vá , m ik ul áš m on oš i 152 tab. 1. scenarios of changes in monthly averages of air temperature [°c] in 50–year horizons for the whole of slovakia in comparison with the normal 1951–1980 (lapin et al., 1996) horizont i ii iii iv v vi vii viii ix x xi xii cccm 1995 (30 year horizons compared to 1995–1980) 2010 1.2 1.4 1.4 1.0 0.9 0.9 1.1 1.0 1.1 1.1 0.9 0.9 2030 2.0 2.4 2.6 1.7 1.5 1.6 1.8 1.7 1.9 1.8 1.4 1.5 2075 3.7 4.5 4.3 3.2 2.9 3.0 3.3 3.2 3.6 3.4 2.7 2.8 cccm 1995 (50 year horizons compared to 1995–1980 from the modi�cation for 1901–1990) 2010 0.5 0.7 0.9 0.7 0.4 0.6 0.9 1.0 1.0 0.9 0.6 0.4 2030 0.9 1.2 1.4 1.1 0.8 1.1 1.4 1.5 1.6 1.2 0.7 0.7 2075 2.2 2.9 2.8 2.3 2.3 2.9 3.4 3.6 3.6 3.0 2.0 1.0 cccm – canadian centre for climate modelling from 1993, the report has developed and presented a series of di�erent climate scenarios for slovakia through the year 2100. �ese scenarios are now being evaluated through comparison with the actually recorded weather measurements (lapin et al., 1996, 2000). tab. 2. comparison of climatic elements as average deviations from the normal 1961–1990, based on the reports on the state of the environment prepared by the slovak environment agency in the years 2012 to 2018 (sažp, 2012–2019a, b) climate element 2012 2013 2014 2015 2016 2017 2018 average air temperature deviation dt [°c] 1.3 1.3 2.4 2.1 1.5 1.3 2.4 of cumulative precitipation totals on slovakia [%] 95.3 109.2 x 93.0 119.8 106.5 x x – not mentioned in the report by 2011 it was already clear that the forecasted goal of limiting the increase in global temperature to a maximum of 2°c by the end of the century was unrealistic (shmú, 2020a,b). at present, the growth trend is on average 1°c in a 100-year average. �e described 2011 analysis already forecasts the stated goal will be surpassed in 2075. �e real data from slovakia from 2012, for comparison (tab. 2), shows the di�erence as the average deviation of air temperature dt (increase) and total annual precipitation in slovakia as the % of normal, from the years 1961–1990 (decrease) (sažp, 2012–2019a, b). �e consequences of developing climate system changes are visible in the reactions of �ora and fauna. changing conditions, due to the e�ects of greenhouse gases, such as carbon dioxide concentration, increase the average air temperatures or reduce water availability thereby a�ecting the life cycle of plants and animals. in phenological phases, expressions of the life cycle of plants and animals, certain destabilising tendencies were registered, which may be related to the complex climatic-natural conditions in slovakia. changes in animal distribution areas as well as in changes in their behaviour expressions of clim atic change in s lovak r epublic 153 were also signi�cant (sažp, 2016). changes in the structure and composition of habitats and an exchange of species in the habitats have been observed. �ese factors will reduce the resilience of ecosystems, reduce their ability to provide ecosystem services or cause ecosystem disintegration (sažp, 2019a). conclusion slovakia, during the years 1881–2018, underwent signi�cant changes in all monitored climatic elements, e.g.: (1) an increase in the average cumulative air temperature by approximately 1.73°c, (2) a decrease in cumulative atmospheric precipitation totals by on average 0.5% (in the some parts of south slovakia the decrease was even greater than 10%, in the north and northeast it rarely increased to 3%) and (3) a decrease in soil moisture and an increase in the drought index. if we accept these changes, we must also accept the emergence of new adverse events, such as droughts, forest �res, �oods and torrential rains, for which rescue equipment must be 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(2009). a multiscalar drought index sensitive to global warming: �e standardized precipitation evapotranspiration index. journal of climate, 23, 1696–1718. https://doi.org/10.1175/2009jcli2909.1 abstract �e development of climate change is evaluated on the basis of trends in a long-term time series (1951– 2018) of individual climatic elements by comparing values from individual years with the normal period in climatology of 1961–1990. �e aim of the article is to present the manifestations of climate change in slovakia (since its inception) according to selected indicators: (1) average annual air temperature, (2) soil temperature, (3) total atmospheric precipitation and (4) drought index over the last decade. �e data presented in the article were obtained from public reports on the state of the environment in the slovakia and other related documents. slovakia, during the years 1881–2018, underwent signi�cant changes in all monitored climatic elements. �e most signi�cant changes were in 2017 and 2018. key words: annual air temperature, annual total precipitation, climate change, dryness index received: [2020.06.08] accepted: [2020.09.13] ekspresja zmian klimatu w republice słowackiej streszczenie postęp zmian klimatu ocenia się na podstawie trendów w długoterminowych szeregach czasowych (1951– 2018) poszczególnych elementów klimatu, na podstawie porównania wartości z wielu lat, z normalnym okresem w klimatologii 1961–1990. celem artykułu jest przedstawienie przejawów zmian klimatycznych na słowacji (od ich powstania) według wybranych wskaźników: (1) średnia roczna temperatura powietrza, (2) temperatura gleby, (3) suma opadów atmosferycznych, (4) susza indeks w ciągu ostatniej dekady. dane przedstawione w artykule pochodzą z publicznych raportów na temat stanu środowiska w republice słowackiej i innych powiązanych dokumentów. w słowacji w latach 1881–2018 znaczącym zmianom uległy wszystkie monitorowane elementy klimatyczne. największe zmiany zaobserwowano w latach 2017 i 2018. słowa kluczowe: roczna temperatura powietrza, roczne sumy opadów, zmiany klimatu, wskaźnik suszy information on the authors iveta marková https://orcid.org/0000-0001-9424-2024 she deals with simulate accidents (mainly �res) with the purpose of creation of experimental research database, creation of simulations and animations of chosen crises models for all �rst respondents (�re scenarios, evacuation models), on-site experimental monitoring activities (temperature, microclimatic conditions, �ames) and �re experiments. mikuláš monoši he deals with �re�ghting technics using by the fire and rescue corps in the slovak republic. �e importance of a criterion assessment of �re�ghting technics, climate changes and forest �res in the slovak republic. 95 annales universitatis paedagogicae cracoviensis studia naturae, 6: 95–108, 2021, issn 2543-8832 doi: 10.24917/25438832.6.6 alminda magbalot-fernandez1*, margie samones agan2 1college of agriculture, rizal memorial colleges inc., davao city 8000 philippines; *almindafernandez5@gmail.com 2college of agriculture and related sciences, university of southeastern philippines, tagum-mabini unit, 8100 philippines bio-forge promotes growth and yield performance of pechay (brassica rapa l. var. chinensis (l.) hanelt) introduction pechay brassica rapa l. var. chinensis (l.) hanelt (= b. chinensis l.) group cultivars ‘pak choi’, is an erect, biennial herb, cultivated as an annual about 15–30 cm tall in vegetative stage. ovate leaves are arranged spirally and spreading. �e petioles are enlarged and grow upright forming a subcylindrical bundle. in�orescence is a raceme with pale yellow �owers. seeds are 1 mm in diameter and are reddish to blackish brown in colour (gawryś, 2008). pechay is an important consitutent of filipino food such as “puchero” and “nilaga”. it is a green leafy vegetable rich in calcium and other essential nutrients. its nutritional values, in 100 g dry mass, are made up of 93.0 g water, 1.7 g protein, 0.2 g fat, 3.1 g carbohydrates, 0.7 g �ber, 0.8 g ash, 2.3 g β-carotene, 53.0 mg vitamin c, 102.0 mg calcium, 46.0 mg phosphorous, 2.6 mg iron, and an energy of 86.0 kj. pechay is used mainly for its immature, but fully expanded tender leaves (knot, deanon, 1967). �e succulent petioles are used as main ingredient for soup and stir-fried dishes. in chinese cuisine, its green petioles and leaves are also used as garnish (jim, tony, 2006; jimenez et al., 2021). �e food and agricultural organisation (fad) has identi�ed cabbage as one of the top twenty vegetables and an important source of food globally. �e country’s chinese pechay production slightly decreased by 0.95% in 2017. about 86.3% of the country’s total b. chinensis production come from the cordillera administrative region central visayas came next with 7.0% share. northern mindanao, davao region and the rest of the country had a combined share of 6.7% (philippine statistics authority, 2019). �erefore, the causes of this type of changes are sought. a lm in da m ag ba lo tfe rn an de z, m ar gi e s am on es a ga n 96 so far studies using various fertilisers and foliar supplements have been tested to maximize the growth and yield of various crops. for examples, for pummelo plants (citrus maxima merr.) fertiliser was used of fermented banana peel, nutrition by foliar or soil fertilisation with potassium (magbalot-fernandez, de guzman, 2019, 2021). for pechay (b. chinensis) was examined e�ect of organic foliar fertiliser or the stimulating e�ect of hormones (fernandez, miñoza, 2015; fernandez, andigan, 2017). for an abaca (musa textilis nee) and banana plants (m. paradisiaca l.) the e�ect of hormones and bio-forge fertiliser were analysed (fernandez, sabay, 2016; magbalot-fernandez et al., 2020). for rice (oryza l.) the e�ect of organic-based foliar fertiliser was checked (montifalcon, fernandez, 2017). bio-forge® (stoller usa) is a fertiliser use as a biological substance of plant growth. it is used to control stress processes caused by excessive ethylene production. bioforge improves the over expression of plant genes in terms of resistance to stress, facilitates nitrogen uptake and growth of the main root increasing its penetration and length, increases respiration and metabolism of the plants reactivating its growth, eliminates the e�ects of stress and blockage resulting from excess ethylene produced as a result of stress, delays aging of plants, helping them stay active, fresh and productive longer. it can be applied for seed treatment, soil, foliar or as a fertiliser to maximize potential bene�ts. its active ingredients are 3.0% total nitrogen, 1.25% urea nitrogen, 1.75% ammoniacal nitrogen, 1.0% soluble potash, 1.0% cobalt, 1.0% molybdenum (stoller, 2021). in the �ght against changing climatic conditions, soil contamination with heavy metals and pesticides, farmers are still looking for innovative solutions in crops. one of them seems to be the use of bio-forge as a plant protection agent against environmental stress, and its a positive e�ect on plant morphology and physiology. �erefore, a major aim of this study was (1) to evaluate the e�ects of bio-forge and bio-forge in combination with other fertilisers in the elongation growth of b. chinensis, (2) to determine the economic bene�ts of using bio-forge in pechay yield production, and (3) to determine the best treatment combination of fertilisers that will increase the yield of chinese pechay. material and methods location and duration of the study �is study was conducted at the research area of university of southeastern philippines, tagum-mabini campus, philippines at 7º28’8”n 125º85’63”e from december 2014 to january 2015 (fig. 1). 97 experimental design �e study was carried out in randomised complete block design (rcbd). field experiments were composed of seven treatments replicated three times. �ere were 64 pechay plants in a 0.25 × 0.25 planting distance with a plot size of 4 m2 per replication for a total area of 128 m2 with a total of 1,344 pechay plants. each plot was provided with a 0.5 m alleyway. �e climatic conditions throughout the duration of the study in reference to the pagasa agromet station have rainy days and temperatures which were favourable to the growth and development of pechay (pagasa, 2021). soil analysis soil analysis was done to determine the nutrient need for pechay. before the conduct of the experiment, soil samples were collected at random in the area. soil samples were air-dried, pulverised and sieved and were brought to the department of agriculture, regional soil laboratory, davao city for analysis. based on the result of the soil analysis (tab. 1), it was recommended to apply 10 bags of 50 kg / ha compost, 2–4 bags of 50 kg / ha ammonium phosphate, 0.90–1.75 bags of 50 kg / ha ammonium sulphate and 2.2–4.4 bags of 50 kg / ha urea. fig. 1. �e experimental area at the university of south-eastern philippines, tagum-mab in campus; a, b – row cultivation, c – leaf width measurements, d – plant height measurements (photo. m. agan) b io-forge prom otes grow th and yield perform ance of pechay (brassica rapa l. var. chinensis (l.) h anelt) a lm in da m ag ba lo tfe rn an de z, m ar gi e s am on es a ga n 98 tab. 1. soil analysis and requirement for pechay (brassica rapa l. var. chinensis (l.) hanelt) at the experimental area ph soil organic matter [%] 1p [ppm] 2k [ppm] nutrient requirement [kg / ha] n p2o3 k2o 6.2 – slightly acidic 1.3 – very low 8 – very low 955 – very high 150 40 0 st an da rd v al ue s 6.1–6.6 5.6–6.0 < 3.44 < 10 > 750 note: 1olsen methods (horta, torrent, 2007), 2h2so4 extraction method (hunter, pratt, 1957) preparation of the �eld for cultivation �e area was ploughed and harrowed thoroughly. plot lay-outing and drainage canal were constructed a�er ploughing and harrowing. two to three seedlings were planted per hill. one seedling per hill was maintained one week a�er transplanting. �is was done early in the morning or late in the a�ernoon or as needed to maintain soil moisture. �e di�erent fertiliser treatments were applied based on soil analysis and the manufacturer’s recommendation. weeding was done regularly to prevent weeds from competing with nutrients and water with the experimental plants. �e plants were sprayed with appropriate pesticides and fungicides such as dithiane to control the occurrence of insect pests and diseases following the manufacturer’s recommendations. pechay was harvested at maturity, 45 days a�er sowing. tested fertiliser modi�cations �e experiment investigated the e�ects of 7 di�erent combinations of fertilisers. t1 – was control group consisted of plots fed with distilled water and precipitation. group of t2 was consisted of recommended rate (rr) of inorganic npk fertiliser according to soil analysis (tab. 1). �e test of t3 was available on the fertiliser market – bio-forge fertiliser. �e t4 group consisted of t2 + s (simulate as a yield enhancer which contained a combination of 3 di�erent phytohormones including cytokinin (0.009% / 1l), gibberellic acid (0.005% / 1l), and indole-3-butyric acid (0.005% / 1l). treatment of t5 was consisted of t2 + bio-forge. �e treatment 6 – t6 was s + bio-forge and t7 = t2 + t6. �e recommended rate of stimulate was applied at 10 ml / 1 l of water. bio-forge (contains micronutrients that stimulate the production of auxin, a hormone generated by the plant to trigger vegetative growth) and other fertilisers were applied at 5 ml / 1 l of water based on manufacturer’s recommendation. 99 morphometric analysis �e plant height was measured by the ruler from 10 samples of plant per plot. leaves were taken from the base of the standing plant. measurements were done at 15 days intervals up to harvesting. �e length of the third leaves was measured by the ruler from the base of the leaf up to its tip. �e width of the leaves was examined from the one leaf in each of the 10 sample plants per plot. �e number of the leaves was determined by counting all fully expanded leaves of the plants at the end of experiment. �e mean values are presented in the table in the form of [cm] values and in the �gures in the percentage values using the inhibition percentage index according to mominul islam et al. (2012). fresh weight and yield of pechay plants fresh weight [g] was determined by weighing 10 sample plants per plot immediately a�er harvest using an electronic scale (rs pro, no. 111–3673, china). �e yield of pechay [kg / ha] was taken by weighing all the plants from each plot a�er harvest. it was determined on a hectare basis following the formula: yield [kg / ha] = plot yield [kg] × 10,000 sqm / plot size and expressed in percentage values. statistical analysis statistical analysis of the di�erent data gathered was analysed through the analysis of variance (anova) and the di�erences among treatments were compared using honest signi�cant di�erence (hsd). �e results were additionally compiled into ms excel in the form of percentage values. �ere were three replications per treatment for a total of 21 repetitions and the standard deviation of every replication means were taken. results plant height biometric analysis of pechay plants a�er 15 days showed no signi�cant di�erences between the applied fertiliser modi�cations with the addition of bio-forge, compared to the value in the control sample. a�er 30 days, slight plant growth was observed, however, no statistically signi�cant di�erences were found. a�er 45 days of pechay cultivation, clear di�erences were observed in the elongation growth of the plants compared to the control (t1). each of the applied fertilisers had a positive e�ect on the growth of the tested plants. �e highest plants were found in plots where plants were treated with fertilisers in the form of t2 (recommended rate (rr) of inorganic npk fertiliser according to soil analysis (tab. 1)) mixtures, t4 (t2 + s (simulate by phytohormones)), t5 (t2 + bioforge) and t7 (t2 + t6) (tab. 2, fig; 2a, d). b io-forge prom otes grow th and yield perform ance of pechay (brassica rapa l. var. chinensis (l.) h anelt) a lm in da m ag ba lo tfe rn an de z, m ar gi e s am on es a ga n 100 length of leaves �e length of the pechay leaves, at each of the three measurement dates, was the highest in the plants treated with the t7 fertiliser. compared to the control, all fertiliser mixtures had a positive e�ect on pechay leaf length. �e smallest di�erences in the values of this parameter were found between t1 (control group) and t3 (bio-forge fertiliser) (tab. 2; fig. 2b, e). fig. 2. plant height, length plant and leaves width of pechay (brassica rapa l. var. chinensis (l.) hanelt) a�er of application various fertilisers expressed in percentage values; t1 – control; t2 – recommended rate of inorganic fertiliser; t3 – bio-forge; t4 – t2 + s (simulate); t5 – t2 + bio-forge; t6 – s + bio-forge; t7 – t2 + s + bio-forge; ip – inhibition percentage expressed as % of control plant, negative values indicate a positive e�ect of fertilisers, positive values indicate plant growth inhibition; a red line represents 100% (control group), 15, 30, 45 – number of days of pechay cultivation 101 width of leaves results revealed no signi�cant di�erence was observed at 15 days of application of all t2–t7 fertilisers (tab. 2). while signi�cant di�erences in the e�ect on pechay leaf width were shown in plants a�er 30 and 45 days of cultivation. compared to the control and other fertiliser mixtures, the widest leaves of pechay were found in plants treated with t7 fertilisers (tab. 2; fig. 2c, f). number of leaves �e amount of leaves of brassica rapa l. var. chinensis (l.) hanelt was similar between the plants from the control and those grown on soils with the addition of t3, t4 and t6 fertilisers. in other cases, each fertiliser caused a signi�cant increase in the number of pechay leaves, compared to the number of plants in the control plant (t1) (fig. 3). fig. 3. �e amount of leaves of pechay (brassica rapa l. var. chinensis (l.) hanelt) a�er of application various fertilisers: t1 – control; t2 – recommended rate of inorganic fertiliser; t3 – bio-forge; t4 – t2 + s (simulate); t5 – t2 + bio-forge; t6 – s + bio-forge; t7 – t2 + s + bio-forge; means (± sd) with di�erent letters are signi�cantly di�erent at p < 0.01 according to the hsd test b io-forge prom otes grow th and yield perform ance of pechay (brassica rapa l. var. chinensis (l.) h anelt) tab. 2. plant height, length and width of leaves of pechay (brassica rapa l. var. chinensis (l.) hanelt) treated with various fertilisers a�er 15, 30 and 45 days of planting treatments plant height [cm] length of leaves [cm] width of leaves [cm] 15 ns 30 ns 45 ** 15 ** 30 ** 45 ** 15 ns 30 ** 45 ** t1 3.47±0.33 5.59 ±0.32 9.57 c ±0.29 11.97 d ±0.48 13.90 d ±0.56 20.35 c ±0.44 2.70 ±0.42 8.90 e ±0.11 13.16 d ±0.51 t2 3.41±0.48 5.78 ±0.06 13.09 a ±0.36 15.23 b ±0.17 16.55 b ± 1.08 23.68 b ±0.10 2.82 ±0.25 11.25 bc ±0.21 15.09 ab ±0.69 a lm in da m ag ba lo tfe rn an de z, m ar gi e s am on es a ga n 102 t3 3.76±0.10 5.31 ±0.21 11.32 b ± 0.69 12.98 cd ±0.81 14.92 cd ±0.30 20.57 c ±1.03 2.63 ±0.43 10.80 cd ±0.05 13.42 cd ±0.46 t4 3.53±0.06 5.54 ±0.46 13.46 a ± 0.20 14.56 bc ± 0.67 15.90 bc ±0.31 23.27 b ±0.27 2.93 ±0.20 11.81 ab ±0.33 14.69 bc ±0.57 t5 3.62±0.23 5.49 ±0.45 13.35 a ± 0.33 14.22 bc ±0.20 16.93 b ±0.80 23.56 b ±0.44 2.67 ±0.06 11.67 ab ±0.27 15.01 b ±0.79 t6 3.64±0.23 5.61 ±0.14 12.08 b ± 0.14 14.42 bc ± 0.70 14.77 cd ± 0.65 21.39 c ±0.92 2.90 ±0.08 10.26 d ±0.27 13.44 cd ±0.54 t7 3.39±0.14 5.67 ±0.03 13.54 a ± 0.27 17.24 a ±0.37 18.46 a ±0.17 25.66 a ±0.14 2.93 ±0.33 12.16 a ±0.23 16.50 a ±0.22 c.v.(%) 3.31 7.66 2.79 4.28 3.16 2.78 10.61 2.08 3.49 note: t1 – control; t2 – recommended rate of inorganic fertiliser; t3 – bio-forge; t4 – t2 + s (simulate); t5 – t2 + bio-forge; t6 – s + bio-forge; t7 – t2 + s + bio-forge; c.v. – coe�cient of variance; ns – not signi�cant; ** – highly signi�cant at p < 0.01; means (± sd) with di�erent letters in each column are signi�cantly di�erent at p < 0.01 according to the hsd test fresh weight �e fresh weight of pechay plants grown in soils with the addition of various fertilisers was greater than in the control. signi�cantly the highest values were recorded in plants treated with fertiliser in compositions t2, t3, t5 and t7. no statistically signi�cant di�erences in the fresh weight values, compared to the control were found in plants grown on soils with the addition of t3 and t6 fertiliser modi�cations (fig. 4). fig. 4. fresh weight of pechay (brassica rapa l. var. chinensis (l.) hanelt) a�er of application various fertilisers: t1 – control; t2 – recommended rate of inorganic fertiliser; t3 – bio-forge; t4 – t2 + s (simulate); t5 – t2 + bio-forge; t6 – s + bio-forge; t7 – t2 + s + bio-forge; means (± sd) with di�erent letters are signi�cantly di�erent at p < 0.01 according to the hsd test 103 yield compared to the control, the yield of pechay per plot was the highest in plants treated with fertilisers in the t7 modi�cation. no statistical di�erences were observed between control (t1) and t3 and t6 fertilisers. in other cases, each fertiliser had a stimulating e�ect on the values of this parameter. similar values and di�erences were found for the yield per hectare (tab. 3). tab. 3. �e yield of pechay (brassica rapa l. var. chinensis (l.) hanelt) a�er of application various fertilisers treatments yield / plot [kg / ha] [%] yield / ha [kg / ha] [%] t1 1.83 c±0.28 100.00 4583.3 c ±721 100.00 t2 3.50 ab± 0.50 192.26 8750.0 ab ±1250 190.91 t3 2.50 bc±0.66 136.61 6250.0 bc ±1653 136.36 t4 3.42 ab±0.38 186.89 8541.7 b ±954 186.37 t5 3.56 ab±0.32 194.54 8833.3 ab ±877 192.72 t6 2.89 bc±0.16 157.92 7208.3 bc ±401 157.27 t7 4.50 a± 0.50 245.90 11458.3 a ± 954 250.00 c.v. (%) 13.22 – 12.75 – note: t1 – control; t2 – recommended rate of inorganic fertiliser; t3 – bio-forge; t4 – t2 + s (simulate); t5 – t2 + bio-forge; t6 – s + bio-forge; t7 – t2 + s + bio-forge; c.v. – coe�cient of variance; ns – not signi�cant; ** – highly signi�cant at p < 0.01; means (± sd) with di�erent letters in columns are signi�cantly di�erent at p < 0.01 according to the hsd test discussion �e fertilisation e�ciency depends on many factors. it is related to the quality of the soil (agronomic category, content of humus and nutrients, ph values), the course of the weather during the growing season (amount of precipitation, air temperature) and the type of fertilisers used. only a regular supply of nutrients is the basis for proper plant growth (kruczek, 2005). �e substances contained in fertilisers improve the absorption of nutrients from the soil, increase resistance to low and high temperatures, strengthen the plants’ ability to defend themselves against pests and diseases (olaniyi, ojetayo, 2011). plants develop properly when grown in fertile and regulated of ph soil. fertile soil should not only contain a su�cient amount of minerals, but also provide them to the plant in a way that allows stress-free growth and development throughout the growing b io-forge prom otes grow th and yield perform ance of pechay (brassica rapa l. var. chinensis (l.) h anelt) a lm in da m ag ba lo tfe rn an de z, m ar gi e s am on es a ga n 104 season. in own research, the content of minerals in the soil was not high, as was the soil ph (tab. 1). �e acidic reaction of the soil limits the growth and development of plants by adversely a�ecting the soil structure and the biological activity of its microorganisms. in conditions of low soil ph, humus loses its sticking properties, air-water relations deteriorate and the activity of soil organisms is reduced (maziarek, krawczyk, 2015). �erefore, in this study, various combinations of fertilisers were used together with the bio-forge fertiliser in order to verify which form had the most bene�cial e�ect on the growth and development of the studied cabbage (brassica rapa l.). growth parameters monitored at di�erent time intervals con�rmed the positive e�ect of each nutrient mixture. compared to the control, plants fertilised simultaneously with fertiliser minerals with the addition of plant hormones and bio-forge showed the highest values of plant growth, fresh weight, number of developed leaves and, consequently, yield production (fig. 3, 4; tab. 2, 3). �e lowest e�ect of fertiliser mixtures was found a�er 15 days, compared to the control. with the extension of the treatment time with fertilisers, 30 and 45 days, a signi�cantly positive e�ect on the growth and development of pechay plants was observed (fig. 1–4). most likely, properly selected doses allowed for proper rooting and plant growth, leaf colour and resistance to external factors. �e di�erences in the response of plants to fertilisers depended on the form in which their nutrients were available in the soil (mozafar et al., 1993). �e uptake of certain nutrients depends on the uptake of inorganic nitrogen compounds, as a result of the mutual physiological relationships in the plant’s metabolism. �e meteorological conditions, mainly thermal ones, have the greatest impact on the e�ectiveness of fertilisation. �ey a�ect the availability of water in the soil and depend on the location of the fertiliser in its layers (kruczek, sulewska, 2005). �e use of bio stimulants can be an e�ective way to reduce the negative impact of environmental stresses on plants and to prevent them (arif et al., 2006; ali et al., 2014; gugała et al., 2017). bio stimulants increase plant resistance and the e�ectiveness of traditional soil fertilization (calvo et al., 2014). �ey are a component of proteins, the synthesis of which in living organisms requires very large amounts of energy. providing plants with additional amino acids reduces the energy input necessary for nitrogen absorption. under unfavourable environmental conditions, plants focus mainly on defence against stress, not on crop production. �e e�ect of bio stimulants on plants is to accelerate and improve the development of leaves and stems and improve their appearance by reducing the de�ciency of ingredients. �e modern market o�ers a fairly wide range of products containing amino acids, but not all of them work equally e�ectively on plants, which is mainly due to their acquisition in the production process (trawczyński, 2014). one of the fertilisers available on the market is bio-forge, which, as a plant biostimulator, can be one of the many solutions sought in plant cultivation. its use may increase the 105 e�ciency of fertiliser use in the �eld, along with an increase in plant productivity under abiotic stress conditions (du jardin, 2015, colla et al., 2017). bio-forge a�ects the hormonal balance, ensures e�ective absorption of microelements by plant tissues (fernandez, sabay, 2016; fernandez, andigan, 2017). it is especially e�ective when used simultaneously with foliar fertilisers (stollerusa, 2012). �e bene�cial e�ect of bio-forge was con�rmed by the analyses of pechay growth and development carried out in this experiment. �ese preliminary studies demonstrated the stimulating e�ect of this fertiliser not only on plant growth, but also on the yields obtained (tab. 3). in addition to high agricultural production, the main task currently facing agriculture is to protect the environment and natural resources. proper fertilisation consists in providing plants with nutrients in the right proportions and amounts, enabling them to obtain maximum yields of the desired consumption quality. �e unbalanced level of nutrients, as well as the high spatial variability of the content of assimilable nutrients in the soil, prompts the search for new and e�ective fertilisation technologies that will meet the production, economic and ecological goals. conclusion (1) �e use of bio-forge fertiliser had a positive e�ect on the growth of brassica rapa l. var. chinensis (l.) hanelt. however, the use of bio-forge alone has a less stimulating e�ect than its combination with mineral fertilisers or other stimulants of plant growth and development. (2) bio-forge can have a positive e�ect not only on yield, but also can reduce soil contamination. (3) in the studies carried out with the use of various mixtures of fertilisers with the addition of bio-forge, the most favourable e�ect of multi-component fertilisers containing the appropriate proportions of mineral fertilisers, growth stimulants and bio-forge was found. �is type of fertiliser mixture stimulated plant growth, fresh weight and overall pechay yield to the greatest extent. con�ict of interest �e authors declare no con�ict of interest related to this article. references ali, a., perveen, s., shah, s.n.m., zhang, z., wahid, f., shah, m., bibi, s., majid, a. (2014). e�ect of foliar application of micronutrients on fruit quality of peach. american journal of plant sciences, 5(9), 1258–1264. http://doi.org./10.4236/ajps.2014.59138 arif, m., chohan, m.a., ali, s., gul, r., khan, s. 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(2014). wpływ biostymulatorów aminokwasowych tecamin na plon i jakość ziemniaków (e�ect of amino acid biostimulators tecamin on the yield and quality of potatoes). ziemniak polski, 3, 29–34. [in polish] abstract �e aim of the study was to test the e�ectiveness of the fertiliser available on the market – bio-forge® on the growth and yielding of chinese cabbage (brassica rapa l. var. chinensis (l.) hanelt). in the experiment, not only the bio-forge fertiliser was used, but its action in combination with other fertilisers was tested. t1 – was control group that consisted of plots fed with distilled water and precipitation. group of t2 consisted of recommended rate (rr) of inorganic npk fertiliser according to soil analysis used in the experiment. �e test of t3 was available on the fertiliser market – bio-forge fertiliser. �e t4 group consisted of t2 + s (simulate as a yield enhancer which contained a combination of 3 di�erent phytohormones including cytokinin (0.009% / 1l), gibberellic acid (0.005% / 1l), and indole-3-butyric acid (0.005% / 1l). treatment of t5 consisted of t2 + bio-forge. �e treatment 6 – t6 was s + bio-forge and t7 = t2 + t6. based on the observations, it was found that the bio-forge fertiliser alone had a positive e�ect on the growth and yielding of pechay, but to a lesser extent than in combination with other growth stimulants and mineral fertilisers. compound fertilisers signi�cantly stimulated plant growth, increased fresh weight and contributed to better yielding. key words: fertiliser, �eld experience, growth, yield received: [2021.07.05] accepted: [2021.10.05] b io-forge prom otes grow th and yield perform ance of pechay (brassica rapa l. var. chinensis (l.) h anelt) a lm in da m ag ba lo tfe rn an de z, m ar gi e s am on es a ga n 108 bio-forge wspomaga wzrost i wydajność plonowania kapusty chińskiej (brassica rapa l. var. chinensis (l.) hanelt) streszczenie celem pracy było sprawdzenie skuteczności dostępnego na rynku nawozu – bio-forge® na wzrost i plonowanie kapusty chińskiej (brassica rapa l. var. chinensis (l.) hanelt). w doświadczeniu zastosowano nie tylko sam nawóz bio-forge, ale zbadano jego działanie w połączeniu z innymi nawozami. t1 – była grupą kontrolną, składającą się z poletek nawadnianych wodą destylowaną i opadami atmosferycznymi. grupa t2 składała się z poletek podlewanych zalecaną dawką (rr) nieorganicznego nawozu npk, zgodnie z analizą gleby wykorzystanej w doświadczeniu. test t3 obejmował dostępny na rynku nawozowym – nawóz bio-forge. grupa t4 składała się z t2 + s (wzmacniacz wydajności, który zawierał kombinację 3 różnych �tohormonów, w tym: cytokininę (0,009% / 1 l), kwas giberelinowy (0,005% / 1 l) i kwas indolo-3-masłowy (0,005% / 1 l). próba t5 składała się z t2 + bio-forge. próba 6 – t6 to s + bio-forge i t7 = t2 + t6. na podstawie przeprowadzonych obserwacji stwierdzono, że sam nawóz bio-forge wpływał pozytywnie na wzrost i plonowanie kapusty chińskiej, ale w mniejszym stopniu niż w połączeniu z innymi stymulatorami wzrostu oraz nawozami mineralnymi. nawozy wieloskładnikowe istotnie stymulowały wzrost roślin, przyrost ich świeżej masy, a także przyczyniały się do lepszego plonowania. słowa kluczowe: nawóz, doświadczenie polowe, wzrost, plon information on the authors alminda magbalot-fernandez https://orcid.org/0000-0001-5930-0671 she specialises in horticulture, crop protection and food science. margie samones agan she is a graduate of bachelor of science in agriculture majoring in crop science. 149 annales universitatis paedagogicae cracoviensis studia naturae, 6: 149–156, 2021, issn 2543-8832 doi: 10.24917/25438832.6.9 sylwia koczanowicz*, magdalena nowak-chmura department of zoology, institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland; *sylwia.koczanowicz@doktorant.up.krakow.pl exposition to ticks in the poprad landscape park – short faunistic note introduction �e poprad landscape park is one of the biggest landscape parks in poland. it covers the large part of beskid sądecki and the polish part of �e czerchów mountains. it is the place of unmatched natural, landscape and historical values. �e park has the area of 53 419.3 ha, where 70% is the forest with the majority of �r, beech, linden and alder trees. within the park, there are 13 nature reserves with a total area 600.6 ha. it has got a lot of educational-environmental paths, tourists trails and cycle lanes. �ere are also resorts famous for medicinal water. it is willingly visited by tourists (on average – 65 thousand tourists each year – data from 23 sensors located in the park) and its hiking trails are very popular (zawartka, 2013; boguś et al., 2020). in lesser poland, the occurrence and the number of ticks, especially ixodes ricinus l. have been examined for years (siuda, 1993). nevertheless, there are still a lot of places that have not been examined yet. �e poprad landscape park is one of them. ticks are vectors of many bacteria, viruses, protozoa. �ese pathogens are an etiological agents of various dangerous human and animal diseases. �erefore, it is very important to conduct research in tourist areas to assessment of the human potential risk of exposure to ticks and tick borne diseases (nowak-chmura, 2013). due to this knowledge we can improve people’s awareness of ticks and the epidemiological threat they represent. �e main aim of the study is to con�rm the occurrence of ticks i. ricinus in the poprad landscape park and to describe the potential risk of tick attacks on locals, tourists and park rangers. materials and methods ticks (species of ixodes ricinus l.) were collected on two educational-environmental paths and one tourist trail in the poprad landscape park (fig. 1). �e �rst collection s yl w ia k oc za no w ic z, m ag da le na n ow ak -c hm ur a 150 was conducted on 9th may 2020 on the educational-environmental path “rogasiowy szlak” in rytro (from 49°29ʹ05.4ʺn 20°38ʹ51.1ʺe to 49°28ʹ24.5ʺn 20°38ʹ04.3ʺe). it is a 12 km educational-environmental path and has 14 theme stands. it is mostly visited by children, school trips and families. �ere are two drinking water springs and a  quarry on this area. �e area is covered by beech-�r-pine forest. 4 stands (i–iv) were marked on the path. collecting started at 10.00 a.m., the temperature was 19°c, and it was sunny. fig. 1. �e location of study area in the poprad landscape park �e next collection was conducted on 6th june 2021, along the tourist trail “barani szlak” (from 49°25ʹ04.5ʺn 20°53ʹ49.7ʺe to 49°24ʹ41.4ʺn 20°55ʹ19.5ʺe). ticks were collected from the area between the ruins of rytro castle and the shelter “chata górska cyrla”. �e trail is very popular with tourists. �e shelter is usually full of locals and tourists. ticks were collected from 4 stands (i–iv). collecting started at 3 p.m.; the temperature was 23°c; it was sunny. �e last collection was conducted on 19th june 2021 in krynica-zdrój, on the educational-environmental path “na stoku jaworzyny krynickiej” (from 49°25ʹ04.5ʺn 20°53ʹ49.7ʺe to 49°24ʹ41.4ʺn 20°55ʹ19.5ʺe). it is a  4.5 km path, really picturesque and very touristy. it contains 14 theme information tables. ticks were collected from 5 stands (i  – v). collecting started at 3 p.m., the temperature was 28°c, it was hot and sunny. exposition to ticks in the p oprad landscape p ark – short faunistic note 151 ticks were collected using the �agging method (siuda, 1993). a  �annel �ag 60 cm × 40 cm was attached to a stick. �en, this stick was used to sweep low vegetation up to 1 m high. each time the �ag was checked for ticks. �e ticks were carried out by tweezers to the test-tubes �lled 1/3 with 70% ethanol (fig. 2). �e test-tubes were described: the place of collection, date, time, weather. ticks were designated according to the species and developmental stage using a stereoscopic microscope. to identify of ticks, guides by siuda (1993) and nowak-chmura (2013) were used. fig. 2. �e collecting of ticks: a–b – transfer of the ticks using tweezers to test tubes 1/3 �lled with 70% ethanol, c – ticks secured in a test-tube (photo. s. koczanowicz) results a total of 213 specimen of i. ricinus were found at all analysed research stands including: 59 ♀, 62 ♂, 91 nymphs and 1 larva. �e percentage comparison of these results is shown in �gure 3. �e highest number of ticks (111) was collected on 6 june 2021 on the tourist trail “barani szlak” in rytro. �e lowest number – only 32 ticks were collected on 19 june 2021 on the educational-environmental path “na stoku jaworzyny krynickiej”. in may, the highest number of nymph forms was recorded at the stands on “rogasiowy szlak”. on the other hand, in june at the stands on “barani szlak” – most adult males and females were found. at the stands of “na stoku jaworzyny krynickiej”, the structure of the share of individual forms of i. ricinus was similar to that recorded in may at “rogasiowy szlak”. s yl w ia k oc za no w ic z, m ag da le na n ow ak -c hm ur a 152 below is a list of stands and a numerical list of the collected on study area specimens of i. ricinus l. 1) �e educational-environmental path “rogasiowy szlak”: in total on this area, 70 individuals of i. ricinus were collected: 7 ♀ (10%), 6 ♂ (9%), 57 nymphs (81%). no larval forms were found. stands: i – 1 ♀; 3 ♂; 13 nymphs. ii – 1 ♀; 2 ♂; 4 nymphs. iii – 4 ♀; 1 ♂; 22 nymphs. iv – 1 ♀; 18 nymphs. 2) �e tourist trail “barani szlak”: in total on this area, 111 individuals of i. ricinus were collected: 41 ♀ (37%), 50 ♂ (45%), 19 nymphs (17%) and 1 larva (1%). stands: i – 1 ♂; 7 nymphs, 1 larva. ii – 10 ♀; 25 ♂; 3 nymphs. iii – 10 ♀; 6 ♂; 4 nymphs. iv – 21 ♀; 18 ♂; 5 nymphs. 3) �e educational-environmental path “na stoku jaworzyny krynickiej”: in total, on this area 32 individuals of i. ricinus were collected: 11 ♀ (34%), 6 ♂ males (19%), 15 nymphs (47%). no larval forms were found. stands: i – 1 ♂. ii – 1 ♂; 3 nymphs. fig. 3. percentage comparison of the total number of collected ixodes ricinus l. specimens by sex and developmental stage: a – “rogasiowy szlak”, b – “barani szlak”, c – “na stoku jaworzyny krynickiej” exposition to ticks in the p oprad landscape p ark – short faunistic note 153 iii – 2 ♀. iv – 5 ♀; 2 ♂; 12 nymphs. v – 4 ♀; 2 ♂. short discussion ixodes ricinus l. is the most common tick in poland and europe (cuber, 2009; nowak, siuda, 2006). it is the main vector of transmitted pathogens. it occurs unevenly, mainly in leafy and mixed forests, shrubs places and on wet pastures. optimal humidity for this species is 80–100%. �is tick occurs o�en along the trails frequented by wild animals, paths and trails for tourists. it is getting more common in wooded urban areas such as parks and allotments (cisak, zwoliński, 2010; nowak, 2013; nowak, siuda, 2012). in poland, i. ricinus is active from early spring to late autumn. we can talk about two peaks of activity: a spring peak and an autumn peak in late summer and autumn (siuda, 1991, 1993). while piercing the skin ticks inject the anesthetic substance so the victim does not feel pain or irritation. mature stages of i. ricinus feed on blood of big and medium size mammals (cattle, dogs, cats, wild animals, humans). younger ticks attack mainly small mammals, birds and reptiles. all developmental stages of mostly females and nymphs can feed on humans (wilhelmsson et al., 2013). before attacking, ticks look for the best place to take blood from. �ey tend to choose areas where epidermis is thin and wet: areas behind ears, armpits, knees, shanks and groins (bartosik et al., 2011). with children, they prefer areas of face or head (keklikçi et al., 2009). �e małopolska voivodship is constantly monitored for the presence of ticks (siuda et al., 2001; nowak et al., 2009). one of them is the bu�er zone of the poprad landscape park, where �eld research was conducted in barcice and barcice dolne in 2018 (janiczek, 2019). in the area designated for this study, there are favourable conditions for the development and spread of di�erent tick species. in the vicinity of the designated stands, within all three trails, there are areas potentially favoured by i. ricinus and migration routes of wild animals, o�en visited by them. together with the presence of educational and hiking trails, all these factors favour the presence and easy spread of this species. �is is con�rmed by the relatively large number of ticks (213 individuals) found in total in all 13 examined stands. �e preliminary research carried out here has shown that in the spring period, nymphs, i.e. juvenile developmental stages, dominate among the collected forms of this species (fig. 3), while in the early summer period – mature males and females. �is is consistent with the information about this species known from the literature (siuda, 1991, 1993, 2008). �e path “na stoku jaworzyny krynickiej” has conditions favoured by ticks: it is touristy, near meadows, mixed forest, with traces of wild animals; not very s yl w ia k oc za no w ic z, m ag da le na n ow ak -c hm ur a 154 high number individuals occur here was probably due to the high temperature – when the collecting started it was very hot (28°c), and the humidity was over 41%. on the other hand, the negligible share of larval forms can be explained by a di�erent time of their appearance in the analysed area (weather and vegetation season), which certainly requires further, more detailed exploration. scienti�c research conducted recently in poland con�rms that the most dangerous vector of tick-borne diseases for humans and animals is i. ricinus. in the słowiński national park registered the presence of i. ricinus on tourist trails and the threat to park rangers and tourists. ticks carried pathogens of anaplasma phagocytophilum dumler et al., babesia microti frança, borrelia burgdorferi s.l. johnson et al. emend. baranton et al. and toxoplasma gondii nicolle & manceaux (semla et al., 2014; asman et al., 2017). in the bieszczady national park and neighbouring areas, among 148 examined individuals of i. ricinus 25.7% carried a similar pathogens (nowak-chmura, siuda, 2016). �e research conducted in the polish jurassic highland indicated that the foresters who spend a lot of time in natural tick habitat are more likely to get infected with spirochaete of b. burgdorferi (nowak-chmura, 2013). �e risk of being attacked by ticks depends on many factors, i.e. geographical area, climate, temperature, humidity and how long one is exposed to ticks. another factor is the method of prevention – the basic method is to wear proper clothes, long trousers, long-sleeved shirts, high socks and boots. it also helps to use repellents and check the body for ticks each time one comes back from a walk (dutkiewicz et al., 2014). conclusions �ere are favourable habitats for ixodes ricinus l. in the poprad landscape park. a total of 213 specimens of this species were found at 13 sites, located within 3 educational and tourist routes. �ere are more nymphs in the spring, and adult males and females in the early summer. �e presence of larvae is likely to depend on weather conditions, season or the places where the collections were carried out. however, these issues require more careful research. local people, tourists and park rangers are in danger of tick-borne diseases that can threaten their health and lives. con�ict of interest �e authors declare no con�ict of interest related to this article. references asman, m., nowak-chmura, m., solarz, k., szilman, e., semla, m., zyśk, b. 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(2010). pierwsze stwierdzenia obecności wybranych patogenów chorób transmisyjnych w kleszczach ixodes ricinus (acari: ixodidae) zebranych w  okolicach zbiorników wodnych w  rogoźniku (województwo śląskie) (�e �rst observations of the presence of selected pathogens of transmission diseases in ixodes ricinus (acari: ixodidae) ticks collected in the vicinity of water reservoirs in rogoźnik (śląskie voivodeship)). in: a. buczek, c. błaszak (eds.), stawonogi. ekologiczne i patologiczne aspekty układu pasożyt – żywiciel. lublin: akapit, p. 155–164. [in polish] dutkiewicz, j., cisak, e., wójcik-fatla, a., zając, w., srok, j. (2014). pro�laktyka chorób odkleszczowych (tick-borne disease prevention). czynniki biologiczne, 21–23. [in polish] janiczek, s. (2019). ryzyko ekspozycji na kleszcze (acari: ixodida) i  choroby odkleszczowe w  barcicach i  barcicach dolnych (woj. małopolskie) (risk of exposure to ticks (acari: ixodida) and tick-borne diseases in barcice and barcice dolne (małopolskie voivodeship)). praca magisterska, zakład zoologii bezkręgowców i parazytologii, uniwersytet pedagogiczny, kraków. [in polish] keklikçi, u., unlü, k., cakmak, a., akdeniz, s., akpolat, n. (2009). tick infestation of the eyelid: a case report in a child. turkish journal of pediatrics, 251(2), 172–173. nowak, m., siuda, k. (2006). przegląd badań nad fauną kleszczy (acari: ixodida) wyżyny krakowsko-częstochowskiej (review of research on the fauna of ticks (acari: ixodida) of the kraków-częstochowa upland). prądnik. prace i materiały muzeum im. prof. władysława szafera, 16, 173–178. [in polish] nowak, m., siuda, k., solarz, k., góra, a., cuber, p. (2009). a risk of infection with ticks of ixodes ricinus (linnaeus, 1758) species (acari: ixodidae) in south-eastern poland depending on the daily and seasonal rhythm. in: a. buczek a., c. błaszak (eds.), arthropods. invasions and their control. lublin: akapit, p. 31–44. nowak-chmura, m. (2013). fauna kleszczy (ixodida) europy środkowej (�e fauna of ticks (ixodida) of central europe). kraków: wydawnictwo naukowe uniwersytetu pedagogicznego. nowak-chmura, m., siuda, k. (2012). ticks of poland. review of contemporary issues and latest research. annals of parasitology, 58, 125–155. nowak-chmura, m., siuda, k. (2016). kleszcze (ticks). in: a. górecki, b. zemanek (eds.), bieszczadzki park narodowy – 40 lat ochrony. ustrzyki górne, p. 256–260. [in polish] semla, m., nowak-chmura, m., zyśk, b., wojtaś, w., wilk, m. (2014). �e risk of attacks ticks (acari: ixodida) on the territory of slovinsky national park-prelimenary research. scienti�c review of physical culture, 4(2), 152–158. siuda, k. (1991). kleszcze (acari: ixodida) polski. i. zagadnienia ogólne. monogra�e parazytologiczne (ticks (acari: ixodida) of poland. i. general issues. parasitological monographs). warszawa-wrocław: wydawnictwo naukowe pwn. [in polish] s yl w ia k oc za no w ic z, m ag da le na n ow ak -c hm ur a 156 siuda, k. (1993). polish ticks (acari: ixodida). ii. systematics and arrangement (kleszcze polski (acari: ixodida). ii. systematyka i rozmieszczenie). polish parasitological society. siuda, k. (2008). kleszcze (ixodida) (ticks (ixodida)). in: w. bogdanowicz, e. chudzicka, i. pilipiuk, e. skibińska fauna polski. charakterystyka i  wykaz gatunków. tom 3. warszawa: muzeum i  instytut zoologii pan, p. 39–44. [in polish] siuda, k., nowak, m., urbanowicz, a. (2001). rytm sezonowej aktywności kleszcza pospolitego ixodes ricinus (linnaeus, 1758) (acar: ixodida) w  okolicy skały kmity koło krakowa (�e rhythm of the seasonal activity of the common tick ixodes ricinus (linnaeus, 1758) (acar: ixodida) near the kmita rock near kraków). in: j. partyka (ed.), materiały konferencji badania naukowe w południowej części wyżyny krakowsko-częstochowskiej, ojców, 10–11 maja 2001 ojcowski park narodowy, p. 299–301. wilhelmsson, p., lindblom, p., fryland, l., nyman, d., jaenson, t.g.t., forsberg, p., lindgren, p.e. (2013). ixodes ricinus ticks removed from humans in northern europe: seasonal pattern of infestation, attachment sites and duration of feeding. parasites vectors, 6, 362. https://doi.org/10.1186/1756-3305-6-362 zawartka, j. (2013). poprad landscape park (popradzki park krajobrazowy). kraków: zespół parków krajobrazowych województwa małopolskiego, p. 3–9. [in polish] ekspozycja na kleszcze w popradzkim parku krajobrazowym – krótka notatka faunistyczna streszczenie w popradzkim parku krajobrazowym występują dogodne siedliska dla ixodes ricinus l. łącznie na 13 stanowiskach, zlokalizowanych w obrębie dwóch szlaków edukacyjno-turystycznych i jednej ścieżki turystycznej, znaleziono 213 osobników tego gatunku. w okresie wiosennym jest więcej nimf, a wczesnym latem dorosłych samców i samic. obecność larw może zależeć od warunków pogodowych, sezonu lub charakterystyki miejsc, w których został przeprowadzony zbiór. zagadnienia te wymagają jednak dokładniejszych badań. miejscowa ludność, turyści i strażnicy parków są narażeni na choroby odkleszczowe, które mogą zagrażać ich zdrowiu i życiu. key words: ixodes ricinus, protected area, ticks received: [2021.10.25] accepted: [2021.11.17] 7 annales universitatis paedagogicae cracoviensis studia naturae, 4: 7–30, 2019, issn 2543-8832 doi: 10.24917/25438832.4.1 adriana brišová department of botany, institute of biology, pedagogical university of krakow, podchorążych 2, 30-084 kraków, poland, adriana.brisova@gmail.com inspirations by plant in the decorative motifs of st. mary’s basilica in kraków (poland) introduction plants, especially �owers, have been used as an element of various types of decorations since the beginning of civilisation (schwarz, szober, 1974). plant motifs from ancient times appeared in art, especially related to sacrum (latin) – the sphere of holiness. religious symbols of various artists have abundant symbolism, which appeal to a speci�c audience in a speci�c and intended way. at present, many monographic studies relating to the religious symbolism of plants can be found in the literature. examples include the “atlas of biblical plants” by b. szczepanowicz (2003) or “in the world of the bible flora” by j. picka (1998). �ese studies contain a list of species mentioned in the bible and they are particularly useful in explaining the meaning of plant religious symbols. for believers, god was the �rst being who cared for the earthly garden. �erefore, throughout the bible, illustrating the history of the world, there are plants that have a speci�c symbolism describing the relationship between man and god. human is also compared to the plant: (...) “planted in the house of the lord, they will �ourish in the courtyards of our god. �ey will bear fruit even in old age, full of juices and always alive (ps. 92.14)”. as many as eighty species of plants are listed in the bible that have not survived to their full extent. plants are part of the panorama of biblical events, from the old testament to the history of christ. biblical plant species are fascinating and attractive to many people, precisely because of the symbolism they carry. hence, they were o�en an inspiration in various artists studies (szczepanowicz, 2003; włodarczyk, 2004; lengiewicz, 2008). �e arch-presbytery church of the assumption of the blessed virgin mary in kraków, also called the st. mary’s church, the basilica of the assumption of the blessed virgin mary or st. mary’s basilica, is the subject of many studies and publications, a dr ia na b riš ov á 8 as it belongs to the “treasury of polish culture”. it is one of the most important and one of the greatest architectural monuments of kraków. for centuries it was under the care of wealthy middle-class families, thanks to which today it is included in poland in the group of buildings of masterful sacred composition (bujak, rożek, 1987). �e beginnings of the creation of this building, its reconstruction and other history were described in the �rst volume of the monograph “sacred art of krakow in the 19th century” by bałus et al. (2004). �e aim of this study is to identify plant motifs found in the interior decorations of the archpriestal church of the assumption of the virgin mary in kraków and to analyse their symbolism. characteristics of the object location and architecture st. mary’s basilica is located in the north-east corner of the main square in kraków, on the spacious st. mary’s square (50°03′42″ n, 19°56′21″ e). �e sanctuary is located on the route of the lesser poland road of saint james leading to tyniec. �e church is located obliquely in relation to the axis of the main square. �is is due to its location before establishment of the city. st. mary’s square was established only in 1257, when the city was located under magdeburg law. �e foundations of st. mary’s church date back to the years 1221–1222 (adamczewski, 1986; bałus et al., 2004; komorowski, sudacka, 2008). external architecture (appendix 1: fig. 1) �e facade of the temple shows two towers di�ering in height and a church porch (fig. 1a–c). �e higher tower „ekxcubarium” reaches 81 meters. it is crowned with a gothic helmet from 1478, whose author is maciej heringk. in 1666 year a golden crown was erected on the spire, which was funded by the italian merchant piotr antoni pestaloci from vicency. in the middle ages it had a defensive function, i.e. it was a guardroom. it was also called ‘hejnalica’. �e lower tower (fig. 1b) reaches a height of 69 meters and has been used as a belfry for centuries. �e mannerist helmet of the tower was covered in the second half of the 16th century. it is covered by an elliptical dome placed on an octagonal drum. in its corners there are four smaller domes, located on four-sided bases. in the belfry tower, �ve bells are hung (fig. 1b) – including four liturgical ones, which are the largest and oldest bells set in medieval poland. �e ��h bell is a clock dulcimer, because it does not work with the clock located on the higher tower. �e belfry of the tower is occupied by a renaissance chapel dedicated to st. paweł, which was funded by the kau�mann family. from the outside, right next to the chapel window, under the roof there is a bell ‘for those dying’ (fig. 1g) – it was used to ring when a human died (rożek, 1974, 1994, 2012). inspirations by plant in the decorative m otifs of s t. m ary’s b asilica in k raków (p oland) 9 �e walls of the temple are decorated with ogival windows with numerous �oral motifs, and the gables of the arches are �gural sculptures with symbolic themes. �e outer cornice of the temple surrounds 21 �gures. walls of the chapel st. jan nepomucen is decorated with a sundial made in 1954 by tadeusz przypkowski. on the eastern wall of the chapel there is a sculptural composition designed by czesław dźwigaj. it depicts the cruci�ed christ with the mother of god and saint john the evangelist, as well as smaller statues of saints (rożek, 2012). to the inside of the temple from the front side leads a baroque polygonal church porch (fig. 1c). it was created in 1750–1752, according to a design by the italian architect and sculptor franciszek placidi. from the north side of the church there is a small annex of the ecclesiastic treasury from the end of the 16th century. next to it is a gothic sacristy. �e northern entrance to the church is opened by a huge gate with bas-reliefs depicting the history of christ and mary (rożek, gondkowa, 2003; rożek, 2012). internal architecture �e interior of the st. mary’s church built on a cross plan (fig. 2) from the �oor to the vault is tightly covered with matejko’s paintings. among them we �nd various ornamental motifs, angel �gures, marian prayers texts and other religious inscriptions. �e vault throughout the church is covered with intensely blue pigment with golden stars. �e whole gives the impression of a starry sky (rożek, 1997; bałus, 2007). �e main altarpiece was the largest cosmetic and artistic undertaking in st. mary’s basilica. it was commissioned by city councillors to be implemented by master wit stwosz who arrived from nuremberg. �e altar was made in the years 1477–1489. it is considered a top-class work of sacred art from the middle ages. �e gothic pentaptic altar consists of a central wardrobe, two �xed wings and two movable wings and a crown. �e scene of the dormition of the virgin mary surrounded by the apostles is the main idea and artistic accent of the altar. above the central scene of falling asleep, stwosz placed the assumption, and at the peak – mary’s coronation. �e author of the altar accurately recreated the smallest elements and details, revealed in the life of mary and christ scenes, placed in the quarters of the wings. �e wit stwosz altar is the greatest achievement of european sculpture of the middle ages (dobrowolski, 1980). �e presbytery is also covered with paintings by jan matejko. on the walls are various �oral and heraldic motifs: loreto litanies placed on bands, marian prayers and angels in dynamic poses. �e wall paintings are in very intense and vivid colours. most of them are red, gold, purple, and blue is on the vaults. four carved keystones were made according to matejko’s design and they symbolise: the coat of arms of odrowąż, the piast eagle, the monogram of the blessed virgin mary, and the coat of arms of the city of kraków. �e windows in the nave and in the presbytery are three-parted (bałus, 2007). a dr ia na b riš ov á 10 �e cyborium is authored by jan maria padovan. its execution was completed in 1536. �e print consists of three �oors. �e central part of the cyborium is divided by four pilasters with corinthian capitals into three niches, based on the theme of the triumphal arch. �e cyborium contains the tempietta and tabernacle in which the blessed sacrament is. �e top is �nished with a dome. �e golden door on the tabernacle was decorated with a stylised rose and olive tree. it was reconstructed in 1745. vases, �nials and cartouches from the baroque era were placed here (wolańska, bałus, 2010). �ere are 11 chapels funded by rich kraków families such as: bonerowie, montelupi and salomonowie. on the north side of the nave there are the following chapels: trans�guration, loreto, st. lawrence’s, st. anthony’s, st. john the baptist’s and st. michael archangel’s. on the south side of the nave there are chapels: guardian angels, st. valentine’s, st. lazaurus’s, st. john nepomucene’s and our lady of częstochowa (wolańska, bałus, 2010). the analysed material �e material used to analyse plant inspirations in the interior decorative motifs of st. mary’s basilica in kraków, in the form of 100 photographs in the years 2017/2018 was collected and developed. in this study some photographs were included as documentation. in the basilica, 18 sectors (positions) were designated and numbered to simplify the description of the location of plant motifs (fig. 2). �e study omitted the wardrobe and wings of the main altar, made by wit stwosz, because they had already been the subject of botanical research (szafer, 1934, 1958), and the focus was on other parts of the church. �e documentation of the study refers to decorative motifs in the following forms: wall painting, sculpture (including scagiola and decorating the dress in paintings) with polychrome, stained glass. �e study identi�es plant taxa whose mappings were found in the above-mentioned ornamental forms. to identify species and genera the following studies, among others, were used: szafer et al. (1986), macků and krejča (1989), červenka et al. (1990), krzyściak-kosińska and kosiński (2007), halarewicz (2014). in the case of ambiguous identi�cation of the genus or species of the plant, an analysis was made of the entire context of the decoration in which the motif was considered. �e context was helpful when the symbolic meaning of the plants used in decorating was taken into account (wolańska, bałus, 2010). in the interpretation of christian symbolism were used, among others, studies of szczepanowicz (2003), włodarczyk (2004, 2011) and others. all plants identi�ed in ornamental motifs were catalogued alphabetically according to systematics with belonging to families. nomenclature of genera and species, as well as belonging to families was adopted according to the study by mirek et al. (2002) inspirations by plant in the decorative m otifs of s t. m ary’s b asilica in k raków (p oland) 11 and polish �ora (www.atlas-roslin.pl). at each identi�ed taxa, the following information is summarised: name of the genus or species (if possible), location of the motif in st. mary’s church according to the numbering of positions in the diagram (fig. 2), artistic form in which the motif with the given plant appeared (sg – stained glass, wp – wall painting, sc – sculpture, p – polychrome), plant parts (f – leaf, c – �ower, p – fruit, b – branch) and presentation (st. – stylised, r. – realistic, sy. – symbolic). for example, the abbreviated entry: 1 – wp st. f means that at stand no. 1 (presbytery) in wall painting a stylised leaf motif of a given plant appears. fig. 2. diagram of the distribution of the analysed sectors (stands) on the st. mary’s church (photo. adriana brišová); 1 – presbytery, 2 – cyborium, 3 – altar with the cruci�x of wit stwosz, 4 – chapel of st. john nepomucene, 5 – chapel of st. valentine, 6 – chapel of st. lazaurus, 7 – chapel of our lady of częstochowa, 8 – chapel of st. anthony, 9 – chapel of our lady of loreto, 10 – chapel of st. john the baptist, 11 – chapel of st. lawrence, 12 – chapel of the trans�guration, 13 – altar of st. stanislaw bishop, 14 – altar with the painting of the annunciation 15 – epitaph of prelate james januszowicz, 16 – altar of st. sebastian, 17 – fogelweder stalls, 18 – altar of st. simon and st. jude �addaeus a dr ia na b riš ov á 12 results alphabetical list includes 43 plants found in the ornamental motifs of the st. mary’s basilica in kraków, including 29 taxa of genus and 14 taxa of species (tab. 1 – appendix 2). most plant motifs concerned taxa from the following families: rosaceae (9), asteraceae (5), liliaceae (3), papaveraceae (2), malvaceae (2). �e most common motif appearing is the acanthus (acanthum sp.), which was found in 15 positions. �e next most frequent depictions are lilies (lilium sp.) which occur in 11 positions, then roses (rosa sp.) appearing at 9 points of the examined object (fig. 3). �e unique motifs are hibiscus (hibiscus sp.), gloriosis (gloriosa sp.) and lingonberry (vaccinium vitis-idaea l.) (tab. 1 – appendix 2). as for the manner of presentation, in all of the analysed artistic forms (wp, sc, sg) stylised performances dominate (fig. 4). selected, more interesting decorative motifs are presented in �gures 5–8 – appendix 1. many �oral motifs refer to the symbolism of mary, due to the fact that the basilica is for mary’s call (tab. 2 – appendix 2). fig. 3. comparison of the number of plant motifs in individual research sectors of st. mary’s basilica; 1 – presbytery, 2 – cyborium, 3 – altar with the cruci�x of wit stwosz, 4 – chapel of st. john nepomucene, 5 – chapel of st. valentine, 6 – chapel of st. lazaurus, 7 – chapel of our lady of częstochowa, 8 – chapel of st. anthony, 9 – chapel of our lady of loreto, 10 – chapel of st. john the baptist, 11 – chapel of st. lawrence, 12 – chapel of the trans�guration, 13 – altar of st. stanislaw bishop, 14 – altar with the painting of the annunciation 15 – epitaph of prelate james januszowicz, 16 – altar of st. sebastian, 17 – fogelweder stalls, 18 – altar of st. simon and st. jude �addaeus inspirations by plant in the decorative m otifs of s t. m ary’s b asilica in k raków (p oland) 13 discussion plants presented in decorative motifs were very o�en the result of a kind of fashion. sometimes, the artists created using patterns directly from the nature, but this required some botanical knowledge. �at is why they more o�en used templates or herbaria, i.e. medical books showing plant pictures. �ey even sometimes copied from other creators and this phenomenon was very common. in various artistic forms created around 1500, such unusual and unreal plants appear, and at the same time so similar to each other that they were certainly copied. however, it can be said that, more or less since the 1660s, the artists tried to faithfully and realistically render the appearance of plants (włodarczyk, 2011). �e �oral motifs in art have always been accompanied by various types of stylisations, which in some cases led to partially or completely unrealistic representation a given ornamental motif. in st. mary’s basilica, many decorative motifs were found, both in stained glass, wall painting and sculpture, which are di�cult to attribute to any plant, although the inspirations of the world of plants are clearly noticeable here. �ese types of motifs include: branch, thorns (fig. 5 a–b), pods (fig. 5c), triple leaves (fig. 8a), whether rosettes formed from them (fig. 5d) or stylised �owers (fig. 5e) – appendix 1. o�en, these motifs have a clear symbolic meaning, and it was probably the main artistic intention (tab. 2 – appendix 2). for example, thorns are associated with the crown of thorns and the passion of christ. according to f. n. hepper, fig. 4. comparison of the number of performances (st. – stylised, sy. – symbolic, r. – realistic) of �oral motifs in the analysed artistic forms a dr ia na b riš ov á 14 j. maillat and s. maillat (botanists dealing with biblical plants), there are several species from which branches a crown of thorns could be made, e.g. the thorn of christ (ziziphus spina-christi (l.) willd.), jerusalem thorn (paliurus spina-christi mill.), or prickly (sarcopoterium spinosum (l.) spach). however, there is no agreement about this (włodarczyk, 2011). in religion, the signi�cance of thorns is associated with unhappiness, pain and su�ering. �orns also symbolise the curse that resulted from adam’s disobedience to god. a crown of thorns placed on the head of christ (fig. 5a – appendix 1), means not only torture, but also redemption from the sin of adam’s entire family (szczepanowicz, 2003; lengiewicz, 2008). in the analysed sacred object, the most common appearing method of presentation is a stylised motif. only in a few cases, the performances are symbolic or realistic – this applies to all three examined artistic forms (tab. 1 – appendix 2; fig. 4). �ese styles are very di�erent – from small, almost real representations, to signi�cantly changed motifs. �eir identi�cation was possible only through the analysis of the whole decorative context. a good example of this are daisies placed in wall painting around the �gure of the mother of god in the chapel of st. jan nepomucen (fig. 6c – appendix 1). �eir stylisation is far advanced and they also look di�erent from real plants. however, the context of mary, accompanied by lilies, a symbol of purity, roses and dahlias as an attribute of virginity, peonies symbolising innocent embarrassment. white and pink daisies are a symbol of eternal youth and �t into marian context (szczepanowicz, 2003; lengiewicz, 2008). similarly, the �gures of angels in the presbytery placed in di�erent poses with instruments and with the text of the lorean litanies, set on lily �owers, which can be identi�ed only from the context of the presence of angels as the personi�cation of purity, expressed by the presence of these extremely stylised, fancy and colourful lilies. right next to it, as if for comparison, the artist posted white lilies that are easy to identify because the stylisation is insigni�cant here. among the 18 analysed sectors of st. mary’s basilica (fig. 2), the most identi�ed plant motifs were found in the presbytery: 21 in wall painting, 8 in sculpture and 2 in stained glass, and in the chapel of st. john nepomucene: 9 in wall painting, 2 in sculpture, 2 in stained glass (tab. 1 – appendix 2; fig. 3). �e most common �oral motifs appearing in the basilica decorations are: acanthus (acanthum sp.) – 15 positions, lilies (lilium sp.) – 11 positions, roses (rosa sp.) – 9 positions, cinquefoils (potentilla sp.) and chrysanthemums (dendranthema sp.) – 8 positions each. unique motifs are hibiscus (hibiscus sp. fig. 6f–g – appendix 1) and gloriosis (gloriosa sp.), placed in the wall painting of the presbytery and lingonberry (vaccinium vitis-idaea l.), which is part of the decorating the vault of the chapel of st. john the baptist. �e decorative element, in the form of a stylised acanthus leaf, was already used in architecture in ancient greece and rome. �is motif was also popular in the renaissance, baroque and classicism periods. its symbolic meaning is not so important, but inspirations by plant in the decorative m otifs of s t. m ary’s b asilica in k raków (p oland) 15 above all it was a favourite decorative element in various artistic forms. in st. mary’s basilica, it was found in as many as 16 examined sectors of the church (tab. 1–2 – appendix 2; fig. 7a–d – appendix 1). it is considered a symbol of the tree of life, the impermanence and fragility of life, the passing of the world and human life. on the other hand, as a symbol of su�ering and pain, while the leaves themselves as a symbol of moral virtues (szczepanowicz, 2003; lengiewicz, 2008; będkowska, zemanek, 2016). in the studied basilica a very common plant decoration are lily and rose. it is probably associated with the virgin mary, who is the patron of this church (tab. 2 – appendix 2). its attributes are, among others, �owers. �e lilies appear here in real, stylised and symbolic form. most o�en the lily is presented in a stylised form, but also in the form of the so-called bourbon lily as a symbol of power and majesty. from the 11th century, this �ower was part of the bourbon dynasty coat of arms. it is a kind of stylised lily, heraldic �gure, being the symbol of mary in the catholic church (szczepanowicz, 2003). for example, in the st. jan nepomucene’s chapel mary and the child, depicted in wall painting, are surrounded by small, golden bourbon lilies, decorating a navy blue background. �is motif also appears in their crowns (fig. 6c – appendix 1). lilies also occur in the st. anthony’s chapel for whom lily is also an attribute. in turn, the rose indicates the love of the mother of god, both for people and for god (tab. 2 – appendix 2). it is also a symbol of her beauty: physical and spiritual. in the past, theologians wrote that roses in paradise had no thorns. our lady is to be this rose without thorns, which refers to her immaculate conception. �e rose kept in the hands of the saints, in turn, can be a symbol of martyrdom. in the scriptures, roses appear several times, but they do not actually refer to the genus rosa l., but to completely di�erent plants, which do not even belong to the rosa family (rosaceae). many local biblical plants named ‘jericho rose’ or ‘sharon rose’ are the work of medieval monks who loved bible stories and created gardens with plants they gave biblical names to. �ey did not have basic botanical knowledge about biblical plants. �ey themselves identi�ed the �owers and gave them names that they thought were adequate or similar to some biblical species. to this day, many of these names have become part of botanical knowledge (garrett, 2008). hence, species that have nothing to do with the rose, e.g. hibiscus (hibiscus sp., fig. 6f – appendix 1) or peony (paeonia sp.), may have similar christian symbolism as a rose. �is is analogous to the lily described above, which has been confused with the tulip (tulipa sp.) and, such as: diced chessboard (fritillaria meleagris l.). jesse’s tree is an interesting plant motif occurring in the presbytery of st. mary’s basilica. it was a frequent motif used in art from the 11th to the 17th century. it is an artistic presentation of the family tree of christ, which in the analysed object resembles a vine (vitis vinifera l.). jesse was the father of david, king of israel. in prophecy from the isaiah book are mentions about the arrival of the messiah, a descendant of jesse a dr ia na b riš ov á 16 or jesus christ. in the presentations of this tree, jesse rests on the ground, from which a trunk with numerous branches springs; on them are images of the jewish kings and other ancestors of christ (śliwa, 2017). �e vine motif can also be seen here in several other decorations (fig. 8g–h – appendix 1). �ere were many reasons for placing speci�c plant motifs in decorations. certainly religious symbolism played an important role in this respect, which was to be legible to everyone. it was supposed to raise the majesty and dignity of the temple, which was mentioned earlier. �e placement of intricate �oral motifs also played an important aesthetic role, which is also very important for religious sites. �us, decorative motifs with lilies, roses, dahlias and other beautiful �owers, in addition to the symbolic and religious meaning, also ful�lled the decorative functions of this basilica. conclusion in the analysed object it is not possible to identify all representations of decorative motifs inspired by the world of plants. it is also di�cult to determine the exact functions of all depictions. one can only consider suppositions on this topic and formulate hypotheses. sometimes, however, it is worth considering the mapped species or type of plant, because it may carry some symbolic content. it happens above all on artistic forms depicting the mother of god, as well as on those on which maria, christ or the saints hold �owers or fruits in their hands. �e inspiration for many of these appearances was certainly the well-known symbolism of certain species. some of these species are recognised only because of their symbolic signi�cance in religious context, because they are not a natural component of our region’s �ora. an example would be the well-known olive, acanthus or pomegranate. �erefore, such representations also have a certain cognitive aspect. con�ict of interest �e author declares no con�ict of interest related to this article. references adamczewski, j. 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(2010). sztuka sakralna krakowa w wieku xix: część trzecia. kraków: wydawnictwo universitas, ss. 368. [in polish] a dr ia na b riš ov á 18 appendix 1 fig. 1. selected elements of the external architecture of st. mary’s basilica; a – main facade, b – lower tower (belfry), c – porch, d – board dedicated to saint. john paul ii, e – epitaph of stanislaw chudzicz, f – renaissance porch, g – bell ‘for dying’, h – southern entrance for visitors (photo. a. brišová, 2017) a ppendix 1 19 fig. 5. plant motifs in the decorations of st. mary’s basilica; artistic forms: sc – sculpture, p – polychrome, wp – wall painting, sg – stained glass; way of presenting the plant motif: r – realistic, sy. – symbolic, st. – stylised; 1 – 18 sector numbers as per �gure 2 thorns: a – 6 sc (sy.), b – 12 wp (sy.); pods: c – 2 sc (st.); rosettes with trifoliate elements: d – 1 wp (sy.) �owers: e – 1 wp (st.) (photo. a. brišová, 2018) a dr ia na b riš ov á 20 fig. 6. plant motifs in the decorations of st. mary’s basilica (cont.); artistic forms: sc – sculpture, p – polychrome, wp − wall painting, sg − stained glass window; way of presenting the plant motif: r. – realistic, sy. – symbolic, st. – stylised; 1–18 sector numbers as per �gure 2 oak (quercus sp.): a – 1 wp (st.); carnation (dianthus sp.): b – 6 sc (r.); peony (paeonia sp.): c – 4 wp (st.), other: roses, dahlias, lilies and bourbon lilies, daisies; mallow (alcea sp.): d, e – 17 sc / p (st.), other: bells, apple tree, peony; hibiscus (hibiscus sp.): f – 1 wp (st.), other: acanthus leaves; g – real plant (photo. a. brišová, 2018) a ppendix 1 21 fig. 7. plant motifs in the decorations of st. mary’s basilica (cont.); artistic forms: sc − sculpture, p − polychrome, wp − wall painting, sg − stained glass; way of presenting the plant motif: r. − realistic, sy. − symbolic, st. − stylised; 1−18 numbers sectors as per �gure 2 acanthus (acanthus sp.): a – 5 sc (st.), b – 18 sc (st.), c – 12 wp (st.), d – real plant; bell (campanula sp.): e – 13 sc (st.), f – 1 wp (st.); chrysanthemum (dendranthema sp.): g – 1 sg (st.) (photo. a. brišová, 2018) a dr ia na b riš ov á 22 fig. 8. plant motifs in the decorations of st. mary’s basilica (cont.); sc − sculpture, p − polychrome, wp − wall painting, sg − stained glass; way of presenting the plant motif: r. − realistic, sy. − symbolic, st. − stylised; 1−18 sector numbers as per �gure 2 clover (trifolium sp.): a – 1 wp (st.); linum (linum sp.): b – realistic plant, c – 16 sc (st.); geranium (pelargonium sp.): d – 1 wp (st.); proper grenade (punica granatum): e – 1 wp (st.); f – real plant; proper vine (vitis vinifera): g – 9 sc (st.), h – 9 wp (st.) (photo. a. brišová, 2018) a ppendix 1 23 a pp en di x 2 ta b. 1 . l is t o f a ll pl an t t ax a re co rd ed in th e de co ra tio ns o f t he se ct or s o f s t. m ar y’s b as ili ca ; 1– 18 – s ec to rs / po si tio ns n um be rs a cc or di ng to � gu re 2 ; a rt is tic fo rm s: sc – s cu lp tu re , p – p ol yc hr om e, w p – w al l p ai nt in g, s g – st ai ne d gl as s; w ay o f p re se nt in g th e �o ra l m ot if: r. – re al is tic , s y. – sy m bo lic , s t. – st yl is ed ; p la nt p ar ts : f – le af , c – � ow er , p – fr ui t, b – br an ch es pl an t m ot ifs presbitery cyborium altar with a cruci�x chapel of st. john nepomucene chapel of st. valentine chapel of st. lazaurus chapel of our lady of częstochowa chapel of st. anthony chapel of our lady of loreto chapel of st. john the baptist chapel of st. lawrence chapel of the trans�guration altar of st. stanislaw altar with a painting of the annunciation epitaph of bishop james januszowicz altar of st. sebastian fogelweder stalls altars of st. simon and st. jude �addaeus 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 c up re ss ac ea e 1. � uj a sp . sc st. b – – – – – – – – – w p st . b – – – – – – – pa pa ve ra ce ae 2. c he lid on iu m m aj us l. sg st. f – – – – – – – – – – – – – – – – – 3. pa pa ve r s p. w p st . c – sc st. c w p st . c sc r. c – – – – – w p st . c sc st . c – – – sc st. c – – la ur ac ea e 4. la ur us n ob ili s l . – – – – sc st . f – – – sc st. f w p sy . f sc st. f – – – – – – – fa ga ce ae 5. q ue rc us sp . w p st . b , f ; sg st. f – – – – – – – – – – – – – – – – – a dr ia na b riš ov á 24 c ar yo ph yl la ce ae 6. d ia nt hu s s p. – – – – – sc r. c – – – – – – sc r. c – – – – sc r. c pa eo ni ac ea e 7. pa eo ni a sp . – – – w p st . c – – – – – – – – – – – – sc /p st . c – m al va ce ae 8. a lc ea sp . – – – – – – – – – – – – – – – – sc /p st . c – 9. h ib is cu s s p. w p st . c – – – – – – – – – – – – – – – – – b ux ac ea e 10 . bu xu s s p. – – – w p st . b – – – – – – – – – – – – – – er ic ac ea e 11 . va cc in iu m v iti s– id ae a l. – – – – – – – – – w p st . p – – – – – – – – r os ac ea e 12 . a rm en ia ca v ul ga ri s l. – – – – – – – – – – – – sc st . p – – – – – 13 . c er as us sp . – – – – – sc st. p – – – – – – sc st. p – – – – sc st. p 14 . c ra ta eg us sp . w p st . f – – – – – – – – – – – – – – – – – 15 . m al us sp . w p st . p – – – – sc st. p – – – – – – sc st. p – – – sc /p st . p – 16 . pe rs ic a vu lg ar is m ill . – – – – – sc st. p – – – – – – sc st. p – – – – – 17 . po te nt ill a sp . w p st . f . – sc st . c w p st . c – – – sc st. c sc st. c w p st . c – sc st. c – sc st. c – – – – a ppendix 1 25 18 . pr un us sp . – – – – – – – – – – – – sc st. p – – – – – 19 . py ru s s p. – – – – – – – – – – – – sc st. p – – – – – 20 . ro sa sp . sg st. c ; sc st . c – sc st. c w p st . c sc r. c sc st. c – – – w p st . c sc st. c – – – – sc st. c – sc st. c 21 . ru bu s i da eu s l . – – – – – – – – – w p st . p – – – – – – – – fa ba ce ae 22 . tr ifo liu m sp . w p st . f – – – – – – – – – – – – – – – – – li na ce ae 23 . li nu m sp . – – sc st. c – – sc st. c – – – – – – – – – sc st. c – – g er an ia ce ae 24 . pe la rg on iu m sp . w p st . f , c ; sc st. f – – – – – – – – – – – – – – – – – ly th ra ce ae 25 . pu ni ca g ra na tu m l . w p st . p – – – – sc st. p – – – – – – sc st. p – – – – – v it ac ea e 26 . v iti s v in ife ra l . w p, s c st . f , p – sc st . f , p – – – – – sc st . f , p – – – – – – – – – o le ac ea e 27 . o le a eu ro pa ea l . – sc r. f, p – – – sc st . f , p – – – – – – – – – – – – a dr ia na b riš ov á 26 g en ti an ac ea e 28 . g en tia na sp . sc st. c – – – – – – – – – – – – – – – – – c ap ri fo lia ce ae 29 . v ib ur nu m sp . w p st . p – – – – – – – – – – – – – – – – – b or ag in ac ea e 30 . m yo so tis sp . w p st . c – – w p st . c – – – – – w p st . c w p st . c – – – – – – – a ca nt ha ce ae 31 . a ca nt hu s s p. w p, r z st . f sc st. f sc st. f sc st. f sc st. f sc st. f sc st. f sc st. f sc st. f – sc , w st. f sc st. f sc st. f sc st. f sc st. f sc st. f – sc st. f c am pa nu la ce ae 32 . c am pa nu la sp . w p st . c – – – – sc st. c – – – – – – sc st. c – – – sc /p st . c sc st. c a st er ac ea e 33 . be lli s p er en ni s l . – – – w p st . c – – – – – – – – – – – – – – 34 . d ah lia sp . – – – w p st . c – – – – – – – – sc st . c – – – – – 35 . d en dr an th em a sp . w p st . f , c – – sg st . f , c ; sc st. f – – sc st. c – – – – – – – – – 36 . h el ic hr ys um br ac te at um (v en t.) a nd re w s – – – – – sc st. c – – – – – – – – – – – – 37 . le uc an th em um vu lg ar e la m . s . s tr . – – – – sc r. c – sc st. c – sc st. c – – – – – – – – – c ol ch ic ac ea e 38 . g lo ri oz a sp . w p st . c – – – – – – – – – – – – – – – – – a ppendix 1 27 li lia ce ae 39 . fr iti lla ri a m el ea gr is l. – – – – – – sc st . c – – – – – – – – – – – 40 . li liu m sp . w p sy ., st . c ; sc st . c – – sg st . c ; w p sy ., st . c sc sy . c sc st. c sc st. c sc r. c sc st. c w p sy . c w p st . c – – sc st. c – – – sc st. c 41 . tu lip a sp . w p st . c ) – – – – – – – sc st. c – – – sc r. c – – – – sc st. c po ac ea e 42 . tr iti cu m sp . – – – – – – – – – – – – – – sc r. o – – – a re ca ce ae 43 . ph oe ni x da ct yl ife ra l. sc st. b – – – – – – – – – – – – – – – – – m ot ifs o f d i� er en t p ar ts o f p la nt s n r s ek to ró w / st an ow is k 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1. th or ns – – sc sy . – – sc sy . – – – – w p sy . w p sy . – – – – – – 2. tr ee sc st . – – – – – – – – w p st . – – – – – – – – 3. �o w er w p st . – – w p st . sc st . sc st . – – – – – – – – – – – – 4. po ds – sc r. – – – – – – – – – – – – – – – – 5. tr ip le le af w p sy . – – w p sy . – – – – – – – – – sc sy . – – – – a dr ia na b riš ov á 28 tab. 2. christian symbolism of plants noticed in the ornamental motifs of st. mary’s basilica in kraków successive number plants christian symbolism 1. �uja sp. branches are considered a symbol of happiness and longevity. 2. chelidonium majus l. it is not known. 3. papaver sp. symbol of sleep, death, shed blood. 4. laurus nobilis l. �e laurel wreath is a victory and a dignity of power; the wreath with the monogram of christ means triumph over death. 5. quercus sp. �e habit of the tree and the durability of wood is a symbol of strength; it also means a place sancti�ed by god’s presence and god’s great power – it is a tree of life and immortality. 6. dianthus sp. carnations mean purity and motherly love as well as the spiritual relationship between christ and mary; the red �ower is a symbol of christ’s passion and pure and true love. 7. paeonia sp. �e �ower is a symbol of mary or st. jan; considered a symbol of embarrassment and innocence. 8. alcea sp. it means pain, mercy, a request for forgiveness. 9. hibiscus sp. in christian symbolism, it has a similar meaning as a rose. 10. buxus sp. in christianity, it symbolises the hope of salvation and immortality. 11. vaccinum vitis-idaea l. fruits are a re�ection of o�spring, the virtues of interior life. 12. persica vulgaris mill. �e fruit symbolises the virtues of the interior life, christ, the artistic work of man, the o�spring of mankind. 13. cerasus sp. like the fruit of blueberry, raspberry, hawthorn, peach and apricot. 14. crataegus sp. like the fruit of blueberry, raspberry, cherry, peach and apricot. 15. pyrus sp. like the fruit of blueberry, raspberry, cherry, peach and apricot. 16. malus sp. an apple fruit symbolises christ, overcoming sin, eternity, marriage, and in the mouth of the snake or the hands of the devil means sin and even death. 17. rubus idaeus l. like the fruit of blueberry, cherry, peach and apricot, hawthorn. 18. armeniaca vulgaris lam. like the fruit of blueberry, cherry, peach and apricot, hawthorn. 19. potentilla sp. a �ower with 5 petals of the crown is �ve wounds of christ; occurs in the descriptions of the cruci�xion of christ. 20. rosa sp. flower means �ery love; a white rose hung from the ceiling meant silence; is the attribute of the virgin mary; is an image of the shed blood of christ; 5 rose petals symbolise (like a cinquefoil) the 5 wounds of christ. 21. prunus sp. �e branches of plum are made of thorns crowns, which are a symbol of the passion of christ, as well as su�ering and hope. 22. trifolium sp. �e leaf is a symbol of the holy trinity; it also means happiness and life. 23. linum sp. flower is patience, purity, innocence, steadfastness of customs, simplicity and even holiness. inspirations by plant in the decorative m otifs of s t. m ary’s b asilica in k raków (p oland) 29 24. pelargonium sp. it is not known. 25. punica granatum l. �e pomegranate is synonymous with resurrection, purity and compassionate love; red means – �aming love, a sign of blood and death, cracked fruit – emblem of “bonus frater” monks, clay fruit on the tombs – this is the hope of future life, the fruit crowning the column – it is rebirth, power and life. 26. vitis vinifera l. vine shoots are a symbol of heavenly bliss, leaves are life, eucharist grapes, paradise; grape juice is the blood of christ, and the grape on the bush means christ on the cross. 27. olea europaea l. mother of god attribute and in the scenes of the annunciation; means blessing and harvesting a time of peace, a just man, a holy tree, eternal wisdom, the chosen people, reconciliation, peace and forgiveness; the olive branch is a symbol of christ’s merciful love, reconciliation and peace. 28. gentiana sp. it is a symbol of bitterness and regret. 29. viburnum sp. �e viburnum shrub is a symbol of rebirth, fruit like blueberry hawthorn and others. 30. myosotis sp. sincere love, faithful memory and naive simplicity. 31. acanthus sp. acanthus thorns is a symbol of su�ering and pain, and the leaves themselves for moral virtues, impermanence and fragility of life, the passing of the world and human life. 32. campanula sp. bell �owers are called the hats or thimbles of the mother of god; combined with the mother of god, and mean persistence. 33. dendranthema sp. flower for the ‘dead’; symbol of faith and hope in the resurrection. 34. dahlia sp. �e red �ower is the equivalent of a rose, hence it has a similar symbolism as a rose. 35. helichrysum bracteatum (vent.) andrews it means immortality and perseverance. 36. bellis perennis l. eternal youth. 37. leucanthemum vulgare lam. a symbol of great joy, but also the su�ering and death of christ and martyrs. 38. gloriosa sp. it means wonderful, full of glory, hence the symbolism similar to that of lilies. 39. lilium sp. mary’s most important attribute; emphasizes virtues and innocence, means the body of christ, purity, soul striving for god, faithful soul, virginity, royal dignity, church of the blessed virgin mary, transience, the word of god, light. 40. fritillaria meleagris l. similar like a lily. 41. tulipa sp. similar like a lily; also means good name. 42. triticum sp. �e ear of grain is god’s kingdom, the mysteries of jesus and the resurrection; christ is likened to an ear; threshing and sieving grain symbolises the last judgment; the seed means christ and the resurrection, and the ear of the blessed virgin mary. 43. phoenix dactylifera l. �e palm branch symbolizes the tree of life, eternal award, triumph, sign of adoration, victory, christ’s victory over hell, a life that does not die, and the branch on the tombstone means martyrdom. a dr ia na b riš ov á 30 abstract �e aim behind this analysis was to identify the plants which are the �oristic motifs of ornaments inside of church of our lady assumed into heaven in kraków. �e art techniques used in ornaments submitted to the analysis were: paintings, sculptures and stained glass-works. �e analysis was based on the photographs of various parts of the st. mary’s basilica taken between 2017 and 2018. identi�ed species of plants were presented in the table. in the analysis performed here, the author was focused on the symbolism of chosen �oral art motifs, since the artist’s botanical knowledge was not the only inspiration for utilising such decorations. due to its symbolism, the motif of �owers was commonly used in the religious art, highlighting the dignity and majesty of this beautiful place of worship. even though the meaning of plants as a symbol o�en depends on the cultural basis in which the work is done, there are some examples of plants that are widely known as religious symbols by everyone. key words: �oral motifs, wall painting, stained glass, sculpture, plant symbols received: [2019.05.20] accepted: [2019.11.10] inspiracje roślinne w motywach zdobniczych bazyliki mariackiej w krakowie (polska) streszczenie celem niniejszej analizy była identy�kacja motywów roślinnych występujących w wewnętrznych zdobieniach kościoła archiprezbiterialnego wniebowzięcia najświętszej marii panny w krakowie. ujęto tu następujące formy artystyczne: malowidła, rzeźby, witraże. przegląd przeprowadzono w oparciu o fotogra�e wykonane w różnych częściach bazyliki mariackiej w latach 2017/2018. zidenty�kowane gatunki roślin zostały zestawione tabelarycznie. w przeprowadzonej tu analizie skupiono się również na znaczeniu symbolicznym wybranych motywów roślinnych, ponieważ nie tylko wiedza botaniczna artystów była inspiracją w umieszczaniu tego rodzaju zdobień. częstym bodźcem była powszechnie stosowana w religii symbolika roślin, co jeszcze bardziej podkreślało powagę i majestat tego pięknego miejsca kultu religijnego. mimo, iż symbolika roślin zależy ogólnie od podstaw kulturowych, w których dzieło powstało, to niektóre rośliny są powszechnie znanymi i czytelnymi dla wszystkich symbolami religijnymi. słowa kluczowe: motywy roślinne, malarstwo ścienne, witraże, rzeźba, symbole roślin information on the author adriana brišová she was a student at the department of botany of the pedagogical university. he is interested in �oral motifs in di�erent decorations. he is an avid biology teacher. annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832 doi: 10.24917/25438832.7.x wojciech w. a. kowalski department of botany and nature conservation, west pomeranian university of technology in szczecin, j. słowackiego 17 st., 71-434 szczecin, poland; e-mails: wojciech.wakowalski@wp.pl, botanika@zut.edu.pl, desmidia from engravings made by halina ryppowa in the archival collection of the department of plant systematics and geography of the university of warsaw (poland) the archival collection of iconographic materials from the interwar period, mostly by alicja lauer-jeziorańska (1939) and helena humblet-pawłowska (1939), also includes original drawings illustrating the results of research carried out in the peat bog ecosystems of wigry. information on a part of the collection developed by lauer-jeziorańska and humbletpawłowska was published by kowalski (2017), and the entire valuable collection was handed over to prof. waldemar surosz. currently, it is at the disposal of the department of marine biology and ecology, institute of oceanography, faculty of oceanography and geography, university of gdańsk (poland). figures of desmidia constitute a separate part of the preserved illustrative materials. these illustrations were not made by any of the above-mentioned authors who studied the plankton flora of the jeziorka river (lauer-jeziorańska, 1939) and the variability of phytoplankton in the settler at the river pump station in warsaw (humbelt-pawlowska, 1939). this is indicated by the different character of the handwriting presented next to the individual figures, the technique of their production, and the fact that they illustrate the cells of a specific systematic group, related primarily to the habitats of bog ecosystems. at that time, such type of research was conducted by halina ryppowa. partial results of these studies were published in the article “algae of peat ponds, the so-called "sucharów" near wigry“ (ryppowa 1927). the authorship of halina ryppowa, these extremely interesting figures, are indicated, among others, by her visiting card (fig. 1) and a valuable drawing, concerning the euastrum virgense rypp. cell, described for the first time for science by this author (fig. 2). this drawing was included in all major world editions of monographs devoted to this systematic group. fig. 1. the business card of the author of the drawings preserved among the figures fig. 2. the cell of euastrum virgense rypp., a new species for the world flora, described by halina ryppowa (published) the authorship of halina ryppowa’s figures is also confirmed by the handwritten description for table iv (fig. 3) preserved in the materials, which is an integral part of the article published by her and devoted to the phycoflora of peat bogs in the vicinity of wigry (ryppowa 1927). fig. 3. the original handwritten description to table iv with figures included in the work entitled “algae of peat bogs, the so-called “sucharów” near wigry”, by halina ryppowa (1927) the preserved collection includes a total of 56 drawings which are illustrations of various taxa of desmidia. illustration boards a-c (appendix 1) contain figures, the state of preservation of which allowed for their relatively legible scanning. illegible captions next to individual figures required additional numbers to be attached to the taxa. descriptions of the numbered specimens were made based on the original nomenclature introduced in the figures of the presented species, as well as on the basis of their illustrations in the author's article (ryppowa 1927). taxa, in relation to which the current author had no identification doubts, and whose species signatures did not survive in the presented illustrative material, were given an appropriate name, with the reference number in parentheses (wk). only some of the drawings can be found on the boards in the author's work as illustrative material – euastrum virgense rypp., e. crassum kütz., e. affine ralfs, and e. arctiscon ehren. this extremely interesting and valuable iconographic documentation was made mainly on small scraps of thin cardboard, less often on tracing paper. figures were made with black ink or a pencil. in good condition, the documentation illustrating individual taxa has been preserved only in the figures made with ink (illustration board a, c). the remaining part has become brightened with time (illustration board b: 5-6, 8-9); hence the contours of some cells and their descriptions are poorly visible, especially on yellowed cardboard pages (illustration board c:9-10). it is a natural consequence of the influence of light, but also of the aging process of the drawings. this archival collection is almost 100 years old, and for its further preservation it will be necessary to record in digital technology and not to expose it to the effects of light. a surprising fact is the very precise technique of the figures, reflecting the features of the cells that enable them to be recognized (e.g. fig. 4: cosmarium portianum arch.). the lack of dimensions of the cells of staurastrum sp. (fig. 5), although with very characteristic ornamentation, does not allow the identification of this species. fig. 4. cosmarium portianum arch. cell the author’s work on the flora of peat bogs (ryppowa 1927) contains the information that the figures are approximately 270 times magnified and were made with reichert's drawing apparatus. however, this does not allow for a precise determination of this taxon (fig. 5). the analysis of the available, basic bibliography and keys concerning this systematic group as well as the figures and descriptions presented in them (west, west, 1912; hirano, 1959; kosinskaja, 1960; palamar-mordvinceva, 1982; croasdale, et al. 1994; lenzenweger, 1997, coesel, 1997; coesel, meesters, 2007) do not allow for including it among the species, varieties, and forms described so far. in terms of some features of the sculpt, the top surface of the half-cell, and its side view, the undescribed drawing is the closest to the species staurastrum vestitum ralfs. fig. 5. sculptura of the unsigned and possibly unidentified by h. ryppowa species of the genus staurastrum sp. however, the ornamentation of its structure of the top cell surface does not correspond to any of the previously described varieties and forms of this taxon. it is possible that the cell found by the author belongs to a new, hitherto unknown taxon for science. the species with this kind of morphological features of cells was also not found in algological materials during later studies of the “suchary” near wigry (tomaszewicz et al. 1996), as well as monographic studies on the desmidia of bory tucholskie lakes (oleksowicz, 1988) and the area of mazovia (tomaszewicz, 1988). most of the surviving figures have latin taxa names. this mainly concerns the figures used in the published article by the author. in total, the entire archival collection includes 56 figures of desmidia cells, 2 drawings of micocystis aeruginosa kützing colonies, one of the ceratium hirundinella cell (o. f. müll.) bergh and one of the cenobium pediastrum tetras (ehrenb.) ralfs. (tablica b:6,8 – appendix 1). the preserved drawings illustrate cells belonging to such genera as micrasterias sp. div. closterium sp. div., cosmarium sp. div., tetmemorus sp., staurastrum sp. div., desmidium sp. div., pleurotaenium sp., netrium sp., hyalotheca sp. not all of the illustrated taxa are provided with detailed information. only for some genus are given, incl. the size of the cells or their species names. these data are included in relation to the species that the author used as illustrative material, or their detailed descriptions are provided in the text of the work. after set in order of all the materials, they were transferred to the icon library, which is located in the department of algology of the institute of botany of the polish academy of sciences, władysław szafer in krakow. conflict of interest the author declare no conflict of interest related to this article. references coesel p.f.m. 1997. de desmidiaceeën van nederland 6. fam. desmidiaceae (4). wetensch. meed. knnv. 220, 1–93. utrecht. [in dutch] coesel p.f.m., meesters k. j. 2007. desmids of the lowlands. 207. mesotaeniaceae and desmidiaceae of the european lowelands. stowa. knnv publishing. croasdale h., flint e.a., racine m.m. 1994. flora of new zeland desmids. 3. manaaki whenua press, lincoln, canterbury, new zeland. 1–218. hirano m. 1959. flora desmidiarum japonicarum vi. – contr. biol. lab. kyoto univ. 9, 302–386. humblet-pawłowska h. 1939. roczna zmienność fitoplanktonu w osadniku na stacji pomp rzecznych w warszawie. la varialion annuelle du phytoplancton dans le bassin de sedimentation a la station des pompes fluviales a varsovie. [in polish] kosinskaja e.k. 1960. flora sporovych rastenij sssr. 5. conjugatae (2). desmidievye vodorosli. akad. nauk ssr, moskva-leningrad. [in russion] kowalski w.w.a. 2017. archiwalna kolekcja rycin z badań fykologicznych prowadzonych w okresie międzywojennym w zakładzie systematyki i geografii roślin uniwersytetu warszawskiego. wiad. bot., 61. https://doi.org/10.5586/wb.2017.017 [in polish] lauer-jeziorańska a. 1939. materiały do flory planktonu rzeki jeziorki. recherches sur le phytoplancton de la riviere jeziorka. warszawa. nakładem towarzystwa naukowego warszawskiego z zasiłku przedsiębiorstwa wodociągów i kanalizacji m. st. warszawy 193. [in polish] lenzenweger r. 1997. desmidiaceenflora von österreich. 2. bibl. phycol., j. cramer, berlinstuttgart, 102: 1– 216. [in german] oleksowicz a., s. 1988. desmidie jezior borów tucholskich. msk. [in polish] palamar-mordvinceva g.m. 1982. opredelitel presnovodnych vodoroslej sssr. 11, 2. chlorophyta: conjugatophyceae, desmidiales (2). nauka, leningrad. [in russion] ryppowa h. 1927. glony jeziorek torfowcowych tzw. „sucharów” w okolicach wigier. [in polish] tomaszewicz g.h. 1988. desmids of the transitional bogs of the middle mazowsze lowland. monogr. bot., 70, 1–86. tomaszewicz g.h., kowalski w.w.a., lesiak t. 1996. flora desmidii kilku sucharów wigierskiego parku narodowego. fragm. flor. geobot. ser. polonica, 3, 261–276. [in polish] west w., west g. s. 1912. a monograph of the british desmidiaceae. 4. ray soc., london. appendix 1 illustration board a. 1 – cosmarium pyramidatum bréb., 2 – c. reniforme (ralfs) arch., 3 – euastrum binale (turp.) ehenb., 4 – euastrum denticulatum (kirchn.) gay, 5 – cosmarium venustum f. hypohexagonum west, 6 – euastrum sublobatum bréb., 7 – cosmarium retangulare grund., 8 – staurastrum sp., 9 – cosmarium conspersum ralfs, 10 – euastrum virgense rypp.(wk), 11 – xanthidium armatum (bréb.) rabenh. (wk), 12 – staurastrum muricatum bréb.; (wk) – identified by wojciech kowalski illustration board b. 1 – pleurotaenium nodosum (bail.) lund. (wk), 2 – penium spirostriolatum barker. cf. f. amplicatum (schmidt) (wk), 3 – euastrum affine ralfs,(wk), 4 – cosmarium ovale ralfs, (wk), euastrum sp., 5, 7 – staurastrum arctison ehrenb. (wk), 6 – pediastrum tetras (ehenb.) ralfs, (wk), euastrum cf. binale (turp.) ehrenc., 8 – ceratium hirundinnella (o.f.m.) bergh. (wk), 9 – micrasterias americana ehrenb. (wk); (wk) – identified by wojciech kowalski illustration board c. 1 – staurastrum arctison ehrenb. (wk), 2 – euastrum crassum kütz. (wk), 3 – cosmarium cf. ornatum ralfs, 4 – desmidium quadrangulatum (ralfs) roy., 5 – spirotaenia condensata bréb., 6 – penium minutum (ralfs) cleve. f. minor racib., 7 – gymnozyga moliformis ehrenb., 8 – micrasterias papilliferra bréb. (wk). 9 – staurastrum aranchn ralfs, 10 – euastrum intermedium cleve.; (wk) – identified by wojciech kowalski zbiór archiwalny rycin z badań fykologicznych prowadzonych w okresie międzywojennym w katedrze systematyki i geografii roślin uw (polska) streszczenie w pracy przedstawiono archiwalne ryciny desmidii, autorstwa haliny ryppowej z kowalski. są to oryginalne rysunki wykonane przez autorkę, ilustrujące fykoflore ekosystemów mszarnych z rejonu wigier. kolekcja obejmuje 56 rycin wykonanych na skrawkach kartonu, bądź kalce technicznej. znaczna część tych rycin wykorzystana została jako materiał ilustracyjny w opublikowanej pracy pt. „glony jeziorek torfowcowych tzw. „sucharów” w okolicach wigier” (ryppowa 1927). na podstawie zawartych w materiałach notatek i rysunków udokumentowano wykonanie ich przez autorke. ilustracje zestawiono na tablicach zbiorczych (a-c) oraz jako samodzielne rysunki w tekście. słowa kluczowe: desmidie, fykoflora, ikonografia, zbiory archiwalne received: [2022.09.10] accepted: [2022.09.29] 7 annales universitatis paedagogicae cracoviensis studia naturae, 5: 7–24, 2020, issn 2543-8832 doi: 10.24917/25438832.5.1 yura drach*, zvenysvala mamchur ivan franko national university of lviv, 4, hrushevskyi st, lviv 79005, ukraine; *yuriy.drach@lnu.edu.ua bryophytes of the upper reaches of the western bug river (lviv region, ukraine) introduction �e upper reaches of the western bug river (lviv region) are physically and geographically located within the boundaries of male polissya, partly roztochia, and to a minor extent in the gologoro-voronyatsky denudo-structural hills. on the territory of lviv region, the natural area of male polissya lies within the central part of the western bug; its geological composition is dominated by marls covered with alluvial sand deposits. only on the territory of the batyatytsia remains, neogene sandstones have been preserved, indicating the presence of a tertiary stratum in this area in the past (tsys, 1962). geographers distinguish �ve main geomorphological regions within male polissya in lviv region: the ratyn gently undulating aquaglacial alluvial denudation plain, the bug-styr gently undulating aquaglacial denudation alluvial plain, the radekhiv undulating denudation plain, the pidpodilska undulating-residual denudation plain, and the upper pobuzhia with aeolian loess ridges and wide valleys between the ridges (nazaruk et al., 2018). �e area is represented by pine, oak and pine, and occasionally hornbeam and oak forests, as well as �oodplain meadows and swamps (eutrophic and less frequently mesoand oligotrophic). �e latter are represented by a wide range of aquatic vegetation (didukh, 2003). �e bryophytes of the above-mentioned area have not been studied well. �e �rst scienti�c records of the bryophytes in lviv region can be found in the works of j.h. lobarzewski, a botanist and a professor of the lviv university who was the �rst to publish data on mosses of galicia based on his own collection (lobarzewski, 1847, 1849). most data on bryophytes were published by j. krupa: 71 species of liverworts and 225 species of mosses, some of which were collected outside the city of lviv (krupa, 1885). we can �nd some data in the work of bryologists such as a. geheeb y ur a d ra ch , z ve ny sv al a m am ch ur 8 (1899), f. lilienfeldówna (1910, 1911, 1914), a.j. żmuda (1911), t. wilczyński in 1912 (tasenkevich et al., 2013), who completed “zielnik wilczyńskiego” – a  collection of bryophytes including those from the outskirts of lviv and roztochia, a.t. wisniewski (1923) and others. however, this information concerns only individual collections from the outskirts of lviv. in the second half of the twentieth century, zerov (1964), slobodyan (1967), ulychna (1978, 1979), bradis (1969), and bryologists of the institute of ecology of the carpathians (danylkiv et al., 2002) studied bryophytes of this area. among modern researchers, the most valuable contributions were made by a  lviv botanist a. kuzyarin, who studied the �oodplain vegetation of the upper reaches of the western bug basin and, in addition to vascular plants, collected mosses (kuzyarin, 2010, 2012, 2013), and m. ragulina, who studied bryophytes of quarry outcrops of lviv region (ragulina et al., 2012; ragulina, kuzyarin, 2014). �e bryo�ora of the chervonohrad coal basin was studied by kuzyarin (2013) and karpinets et al. (2017). i. danylkiv compiled the �rst annotated list of bryophytes for the chronicle of nature of the national park “northern podillya”. however, this information was not published. some data on mosses from the upper reaches of the western bug river are presented in the collections of �e moss flora of the ukrainian ssr and �e moss flora of ukraine (bachuryna, melnychuk, 1987, 1988, 1989, 2003). �is article reports new �ndings on the species composition and distribution of bryophytes, as well as analyses of substrate preferences, ecological groups and life forms of bryophyte species from the upper reaches of the western bug river. materials and methods �e �eld survey was carried out by the route method (lazarenko, 1955; ignatov, ignatova, 2003) in the period from 2017 to 2019 in the upper reaches of the western bug river within male polissya and partly on the territory of the gologoro-voronyatsky denudo-structural hills (fig. 1). in the course of the study, 14 sites were surveyed: the volytsky botanical reserve of national importance, the romaniv landscape reserve of local importance, the storonybaby nature reserve of local importance, the pidlyska hill (shashkevych hill) complex natural monument of local importance, the zhovkivska botanical natural monument of local importance, the ivan franko park in busk, a railway track outside the village of zakomarya, a �oodplain of the solotvyna river, the outskirts of the villages of kulychkiv, zarvanytsia, poltva, zavady, pidlyssia, romaniv and shopky. various types of ecotopes were examined: swamps, swampy forests, �oodplains of rivers, oak, pine, beech and mixed forests, urban ecosystems, railway tracks, etc. b ryophytes of the upper reaches of the w estern b ug r iver (lviv r egion, u kraine) 9 tab. 1. information on localities of the study area within the upper reaches of the western bug river no. place coordinates date 1. the volytsky botanical reserve 50°17ʹ25.2ʺn, 23°57ʹ57.6ʺe 16.05.2018 29.04.2018 2. swampy mixed forests near the village of kulychkiv 50°16ʹ50.3ʺn, 24°06ʹ33.3ʺe 22.07.2018 3. ivan franko park in busk 49°57ʹ59.9ʺn, 24°36ʹ30.3ʺe 28.03.2018 4. �oodplain of the solotvyna river 49°56ʹ55.0ʺn, 24°47u48.2ʺe 25.03.2017 5. railway track near the village of zakomarya 49°57ʹ00.0ʺn, 24°45ʹ30.2ʺe 28.03.2018 6. the storonybaby nature reserve 49°56ʹ37.6ʺn, 24°39ʹ44.9ʺe 03.09.2017 7. swampy meadow near the village of zarvanytsia 49°46ʹ16.9ʺn, 24°59ʹ36.0ʺe 17.07.2018 8. mixed forest near the village of zarvanytsia 49°45ʹ21.8ʺn, 24°58ʹ26.8ʺe 19.07.2018 9. the outskirts of the village of poltva 49°52ʹ34.0ʺn, 24°30ʹ47.4ʺe 24.09.2019 10. the outskirts of the village of romaniv and the romaniv landscape reserve 49°41ʹ57.4ʺn, 24°21ʹ26.6ʺe 23.03.2018 11. the outskirts of the village of zavady and the zhovkivska botanical natural monument of local importance 50°03ʹ00.5ʺn, 23°54ʹ25.2ʺe 28.08.2017 12. the outskirts of the village of pidlyssia and the pidlyska hill (shashkevych hill) complex natural monument of local importance 49°55ʹ58.4ʺn, 24°51ʹ06.0ʺe 16.04.2018 13. the outskirts of the village of shopky 49°44ʹ51.7ʺn, 24°31ʹ41.7ʺe 09.06.2018 14. the outskirts of the village of novyi yarychiv 49°52ʹ43.9ʺn 24°17ʹ44.8ʺe 29.09.2019 fig. 1. map of the study area within the upper reaches of the western bug river (numbers of research areas correspond to those in table 1 and table 2 – appendix 1) y ur a d ra ch , z ve ny sv al a m am ch ur 10 sample collection and identi�cation was conducted by methods of bright-�eld microscopy. latin names of taxa are given according to the checklists by boіko (2014) and mosyakin, fedoronchuk (1999). sozological assessment is given according to boiko (2010) and hodgetts et al. (2019). �e following substrate types on which the species grew were noted: ro – rocks; sta – arti�cial stone; sg – soil with gravel or sand; so – soil; sv – soil among grass or leaf litter; wr – rotten wood; wl – living wood; am – aqueous medium. ecological groups and life forms of are given according to hill et al. (2007) and ellenberg, leuschner (2010). results based on our survey, 165 species belonging to 92 genera, 41 families, and 18 orders were identi�ed (tab. 2 – appendix 1). out of the recorded taxa, 151 species were mosses (bryophyta), 13 species were liverworts (marchantiophyta) and one was a hornwort (anthocerotophyta). families with the highest numbers of recorded species, which comprise 66.7% of all identi�ed species, are: brachytheciaceae (12.1%), pottiaceae (11.5%), amblystegiaceae (8.5%), mniaceae (7.3%), orthotrichaceae (6.1%), hypnaceae (4.8%), bryaceae (4.8%), sphagnaceae (4.2%), polytrichaceae (3.6%), dicranaceae (3.6%). among the found species, three are on the list of species o�cially recognised as rare – alleniella besseri, campyliadelphus elodes, tomenthypnum nitens and 16 are recognised as regionally rare – aloina rigida, aulacomnium androgynum, calliergon giganteum, dicranum �agellare, didymodon tophaceus, encalypta streptocarpa, porella platyphylla, rhodobryum ontariense, sciurohypnum plumosum, s. re�exum, sphagnum cuspidatum, s. fallax, s. �mbriatum, straminergon stramineum, syntrichia papillosa, taxiphyllum wissgrillii. substrate preferences by substrate preference, most of the identi�ed species were epigean (116 species, 70.3%). �ey are dominated by bryophytes that grow on bare soil or with minor inclusion of grasses (so – 98 species, 59.4%). species that grow on soil among grass (sv – 33 species, 20%) and on sandy or gravelly soil (sg – 31 species, 18.8%) also constitute a  signi�cant share. �e most frequent are typical forest species: atrichum undulatum, dicranella heteromalla, fissidens taxifolius, polytrichum commune, pseudoscleropodium purum, rhytidiadelphus squarrosus, eurhynchium angustirete. species cratoneuron �licinum, drepanocladus aduncus, kindbergia praelonga, leptodictyum riparium, oxyrrhynchium speciosum and sciurohypnum plumosum grow along the banks of rivers. b ryophytes of the upper reaches of the w estern b ug r iver (lviv r egion, u kraine) 11 a group of epiphytes (46 species, 27.9%) is signi�cantly represented by forest species: anomodon attenuatus, homalia trichomanoides, leucodon sciuroides, metzgeria furcata, porella platyphylla, radula complanate. �ese species are most commonly found in mixed forests on the bark of trees such as acer platanoides, a. pseudoplatanus, alnus glutinosa, fraxinus excelsior, quercus robur, occasionally fagus sylvatica and others. due to the signi�cant presence of wood at di�erent stages of decomposition in moist fairly intact mixed forests, epixile species are quite common in the study area (wr – 56 species, 33.9%). among the obligate epixiles, relatively rare species were found: dicranum �agellare, lepidozia reptans, nowellia curvifolia, ptilidium pulcherrimum, riccardia latifrons and tetraphis pellucida. polysubstrate species also inhabit decaying wood; they include: brachythecium salebrosum, callicladium haldanianum, dicranum montanum, d. polysetum, herzogiella seligeri, hypnum cupressiforme, platygyrium repens, sciurohypnum populeum and others. �e fourth most numerous bryophyte group is composed of epilithic species – 43 species (29.6%). among them, 28 species (17.0%) grow on arti�cial rocky substrate (sta) and 23 species (13.9%) on natural stone or rock outcrops (ro). seven species (4.2%) were found on both types of rocky substrates. �e group is represented by both typical obligate epiliths (alleniella besseri, didymodon rigidulus, homalothecium lutescens, orthotrichum anomalum, rhynchostegium murale, tortula muralis) and facultative ones (amblystegium serpens, ceratodon purpureus, brachythecium rutabulum, leskea polycarpa, plagiomnium cuspidatum). �e lowest number of identi�ed species belong to a group of bryophytes that are completely or partially submerged in water (22 species, 13.3%). �ese are species that grow in swamps, ditches and other wetland areas – brachythecium mildeanum, calliergon cordifolium, c. giganteum, calliergonella cuspidata, campyliadelphus elodes, campylium stellatum, cratoneuron �licinum, leptodictyum riparium, drepanocladus aduncus, hygroamblystegium tenax, fissidens adianthoides, ptychostomum pseudotriquetrum, scorpidium cossonii and warnstor�a �uitans. ecological groups although the substrate plays an important role in the life of the bryophytes, environmental conditions are no less important, since factors such as humidity, light and temperature play a decisive role for them. �us, within the spectrum of heliomorphs, dominate the species that inhabit both moderately shaded and illuminated places – subheliophytes (51 species, 30.9%) and hemisciophytes (51 species, 30.9%). slightly fewer of the identi�ed species belong to the group of bryophytes adapted to a habitat with a very intensive insolation – heliophytes (24 species, 14.5%) and ultraheliophytes (14 species, 8.5%). a small number (19 species, 11.5%) of the identi�ed bryophytes – y ur a d ra ch , z ve ny sv al a m am ch ur 12 sciophytes, prefer well-shaded ecotypes. six species (3.6%) are indi�erent to light or the indicator has not been established. �e distribution of hydromorphs indicates the predominance of species with intermediate characteristics in relation to moisture among the studied bryophytes – mesophytes (49 species, 29.7%), hygromesophytes (35 species, 21.2%) and xeromesophytes (32 species, 19.4%). signi�cantly fewer species represent the groups of drought-tolerant species (xerophytes – 20 species, 12.1% and ultraxerophytes – 4 species, 2.4%) and species that inhabit wet and humid places (hygrophytes – 25 species, 15.2%). in terms of temperature regime, more than half of the bryophytes are cold-tolerant (98 species, 59.4%). signi�cantly fewer moderately heat-tolerant (29 species, 17.6%) and heat-tolerant (6 species, 3.6%) species were found. almost one-��h of the species are indi�erent or demonstrate no speci�c characteristics in relation to temperature. by ph preference dominate neutrophils (83 species, 50.3 %) followed by acidophiles – species growing on acidic substrates (34 species, 20.6 %) and extreme acidophiles (2 species, 1.2 %). �e smallest number of the detected species (19 species, 11.5 %) belong to sub-basophils and basophils (3 species, 1.8 %) that grow on substrates with free calcium carbonate, mainly chalk and limestone. life forms in terms of the structure of life forms of the bryophytes distinguishes two groups of bryophytes: shoots that are not part of an organised colony and shoots that form a part fig. 2. �e range of life forms of bryophytes in the upper reaches of the western bug river: fa – fan, we – we�, mt – thalloid mat, ms – smooth mat, mr – rough mat, de – dendroid, cu – cushion, tu� – loose cushion, tf – turf, ts – scattered turf, st – solitary thalloid, sc – solitary creeping shoots b ryophytes of the upper reaches of the w estern b ug r iver (lviv r egion, u kraine) 13 of an organised colony. in terms of the structure of life forms of the bryophytes (fig. 2), in the study area, the �rst group was represented by only 5 species (2.9%): straminergon stramineum – solitary creeping shoots (sc), riccia glauca and anthoceros agrestis – solitary thalloid (st), aloina rigida and phascum piliferum – scattered turf (ts). �e second group is dominated by species forming turfs (tf ) – 50 species (30.3%), loose cushions (tu�) – 17 species (10.3%). next come species with a mat life form: rough mat (mr – 30 species, 18.2%), smooth mat (ms – 16 species, 9.7%), thalloid mat (mt – 4 species, 2.4%). �e third place is occupied by species that form tightly intertwined turf – we� (we) – 25 species (15.2%). �e cushion life form (cu) is characteristic of 13 species (7.9%) – these are mainly epiliths and epiphytes (orthotrichum spp., dicranum montanum, grimmia pulvinata and others). �e least numerous is dendroid (de) – 2 species (1.2%) and fans (fa) – 3 species (1.8%). discussion �e upper reaches of the western bug river (lviv region) are distinguished by rich species diversity, a wide range of ecomorphs and substrate groups of the bryophytes due to heterogeneity of the relief, a  considerable diversity of ecotopes (pine, oak and mixed forests, open and forest bogs, river �oodplains, areas a�ected by anthropogenic transformations, etc.), as well as a  signi�cant number of protected areas (the roztochia nature reserve, the yavorivsky national park, the northern podillya national park, the volytsky botanical reserve of national importance, the romaniv landscape reserve of local importance, the storonybaby nature reserve, the pidlyska hill (shashkevych hill) complex natural monument of local importance, the zhovkivska botanical natural monument of local importance, the hryada forest reserve and others). �e following nine species are reported for the �rst time for lviv region: orthotrichum patens, palustriella falcata, pedinophyllum interruptum, ptychostomum torquescens, rhodobryum ontariense, sphagnum angustifolium, s. inundatum, taxiphyllum wissgrillii, tortella humilis (zerov, 1964; bachuryna, melnychuk, 1987, 1988, 1989, 2003; boiko, 2014). we have also con�rmed the presence of campyliadelphus elodes on the outskirts of the village of zarvanytsia (kuzarin, 2012), which was found in a swampy reclamation canal. �is is the third locality of the distribution of this species in ukraine (bachuryna, melnychuk, 2003). pedinophyllum interruptum, which was found on soil in the wood on the outskirts of the village of zarvanytsia, is reported for the �rst time not only for lviv region but also for the whole plain territory of ukraine. previous �ndings of this species were reported from the territory of the ukrainian carpathians (zerov, 1964; boiko, 2014). �e families brachytheciaceae and pottiaceae typically dominate on most of the plains y ur a d ra ch , z ve ny sv al a m am ch ur 14 of lviv region as well as the upper reaches of the western bug river (kuzyarin, 2010, 2013; mamchur et al., 2017a, b, 2018; rahulina, kuzyarin, 2014). �e bryo�ora of this territory is represented by a  large number of boreal and nemoral species as well as typical arid species characterised by a  signi�cant level of synanthropisation (lazarenko, 1956; boiko, 1999). relative to light tolerance, the spectrum of ecomorphs is dominated by hemisciophytes and subheliophytes, in relation to humidity – by mesophytes, and in terms of temperature regime – by cold-tolerant species, by substrate preference – by neutrophils. a wide diversity in the life forms of bryophytes in the study area is also due to the variety of growth conditions. however, turf, rough mat, we�, tu� and smooth mat forms prevail. apparently, the dominance of turfs is due the signi�cant environmental tolerance of this life form, in particular, the species that represent it are capable of withstanding high levels of solar radiation. such life forms as mat and we� are characteristic of the species with perennial life strategies and consequently are more widely distributed (during, 1979, 1992; batista et. al., 2018). �e bryophytes grow on a variety of substrates of both natural and anthropogenic origin. among the variety of substrates, the bryophytes most o�en give preference to bare soil, or soil with a slight inclusion of grasses. in terms of moisture conditions, epigean bryophytes usually occur in turfs, mats or even we�s, which is characteristic of species of both natural and anthropogenically a�ected ecosystems (mamchur et al., 2017b). �e epixile group is well represented due to the availability of wood at di�erent stages of decomposition in the natural ecosystems. �e most common life forms are rough mat, smooth mat, we�, tu� and turf. �e epilithic species grow on di�erent types of rocky substrates, of both natural and arti�cial origin. an increase in the number of anthropogenic rocky ecotopes contributes to an increase in the proportion of epilithic species (mamchur et al., 2018). �e predominant life forms of epiliths are rough mats, turfs, tu�s and cushions. �e species that grow on natural rocks are dominated by rough mat, turf and we�, while those growing on arti�cial rock are mainly represented by rough mat, turf, tu� and cushion. among the bryophytes that are partially or completely submerged in water, the most common life forms are turf, we� and rough mat. �e life form of rough mat is the most common in the group of polysubstrate species, as it is suitable for bryophytes with various substrate preferences. in general, the species composition of the bryophytes occurring in habitats under anthropogenic impact is relatively insigni�cant. it is mainly represented by synanthropic species distinguished by a  considerable environmental plasticity: amblystegium serpens, brachytheciastrum velutinum, brachythecium glareosum, b. rutabulum, bryoerythrophyllum recurvirostrum, bryum argenteum, b. caespiticium, ceratodon b ryophytes of the upper reaches of the w estern b ug r iver (lviv r egion, u kraine) 15 purpureus, fissidens taxifolius, funaria hygrometrica, hypnum cupressiforme, plagiomnium cuspidatum, p. rostratum, pohlia nutans, syntrichia ruralis (ochyra,1983; wolski, fudali, 2013; maslovsky, 2012; godovičová, 2019; szűcs, 2020). anthropotolerant epiphytes in the upper reaches of the western bug river are dominated by leskea polycarpa, platygyrium repens, pseudoleskeella nervosa, ptychostomum moravicum, pylaisia polyantha, syntrichia papillosa (fudali, 2012; marka, 2017; mamchur et al., 2018; szűcs, 2020). in terms of life forms, epiphytes are mainly represented by rough and smooth mats, much less frequently by cushions or turfs. on the contrary, a considerable diversity of species is typical of various types of areas under protection. it was on those sites that the o�cially and regionally recognised rare species of the bryophytes were found. in particular, such species as alleniella besseri, aloina rigida, encalypta streptocarpa, porella platyphylla, taxiphyllum wissgrillii were found on the territory of the romaniv landscape reserve of local importance; tomenthypnum nitens, calliergon giganteum, sphagnum cuspidatum, s. fallaх, s. �mbriatum, straminergon stramineum, syntrichia papillosa – in the volytsky botanical reserve of national importance; didymodon tophaceus, rhodobryum ontariense – on the territory of the pidlyska hill (shashkevych hill) complex natural monument of local importance; aulacomnium androgynum, dicranum �agellare, calliergon giganteum, sphagnum cuspidatum, s. fallax and s. �mbriatum – in the mixed forest on the outskirts of the village of kulychkiv. �is fact proves the signi�cant role of protected areas for the preservation of the biodiversity of species, particularly the bryophytes (virchenko, orlov, 2009; virchenko, 2014; barsukov, 2015). �e study area is also distinguished by a  signi�cant number of species growing in wetlands. given the large areas of drained land in lviv region, rare species are of particular value. swamps and water bodies are among the most endangered habitats of the bryophytes in europe, primarily due to drainage of wetlands (hodgetts et al., 2019). �erefore, the preservation of the o�cially and regionally recognised rare species of the bryophytes in the upper reaches of the western bug river (19 species altogether) is particularly important, especially wetland species such as aulacomnium androgynum, calliergon giganteum, campyliadelphus elodes, sphagnum cuspidatum, s. fallax, s. �mbriatum, straminergon stramineum and tomenthypnum nitens. in phytocenoses, the bryophytes are particularly sensitive components that respond to minor changes in the environmental conditions. in ecosystems, the bryophyte component plays a signi�cant role in the nitrogen, carbon, biomass, and water balance cycles (turetsky, 2003). in accordance with the development strategy of lviv region by 2027, the preservation of biodiversity, the protection of valuable natural areas and expansion of nature reserves are top priority issues (development strategy of lviv region, 2019). given that, inventory and further study of the bryo�ora are highly important. y ur a d ra ch , z ve ny sv al a m am ch ur 16 acknowledgements �e authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. con�ict of interest �e author declares no con�ict of interest related to this article. references bachuryna, h.f., melnychuk, v.m. 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[in ukrainian] y ur a d ra ch , z ve ny sv al a m am ch ur 20 appendix 1 tab. 2. recorded species and substrate preferences of bryophytes in the upper reaches of the western bug river; explanation of abbreviations in the text (see "materials and methods") no. species substrate research area anthocerotophyta 1. anthoceros agrestis so 6 marchantiophyta 2. aneura pinguis am 7 3. conocephalum conicum ro 9, 10 4. lepidozia reptans wr 2 5. lophocolea heterophylla sg, wr, wl, am 1, 2, 6, 8 6. metzgeria furcata so, wl 8, 10 7. nowellia curvifolia wr 2 8. pedinophyllum interruptum so 8 9. plagiochila asplenioides so 10 10. porella platyphylla ro, wl 3, 10 11. ptilidium pulcherrimum wr 2 12. radula complanata wr, wl 3, 4, 6, 8, 14 13. riccardia latifrons wr 2 14. riccia glauca so 14 bryophyta 15. abietinella abietina ro, sg, so, sv 1, 2, 8, 9, 10, 11, 12 16. alleniella besseri ro 10 17. aloina rigida sg 10 18. amblystegium serpens ro, sta, sg, so, sv, wr, wl 1, 2, 3, 4, 5, 6, 8, 10, 11, 12, 14 19. anomodon attenuatus so, wr, wl 4, 8 20. anomodon longifolius wr, wl 4, 8 21. anomodon viticulosus ro 10 22. atrichum undulatum sg, so, wr 6, 10, 11 23. aulacomnium androgynum so 2 24. aulacomnium palustre so, sv, am 1, 2 25. barbula convoluta ro, sg 5, 12 26. barbula unguiculata ro, sta, sg 2, 4, 8, 10, 11 27. brachytheciastrum velutinum ro, so, wr, wl 2, 6, 8, 10, 11, 12 28. brachythecium albicans sta, sg 1, 5, 11 29. brachythecium campestre so, wr 1, 2, 6 30. brachythecium glareosum ro, sta, sg, so, sv, wr, wl 4, 5, 8, 9, 10, 11, 12 31. brachythecium mildeanum am 1 32. brachythecium rutabulum sta, sg, so, sv, wr, wl 1, 2, 3, 4, 5, 6, 8, 9, 11, 14 33. brachythecium salebrosum ro, sta, sg, so, wr, wl 3, 5, 6, 8, 10, 14 34. bryoerythrophyllum recurvirostrum so 2 35. bryum argenteum sta, sg 1, 2, 3, 4, 8, 10, 11 b ryophytes of the upper reaches of the w estern b ug r iver (lviv r egion, u kraine) 21 36. bryum caespiticium ro, sta, sg 1, 5, 11, 12, 13, 14, 37. bryum elegans so 10 38. callicladium haldanianum wr, wl 2, 6, 9 39. calliergon cordifolium sg, so, wr, am 1, 2 40. calliergon giganteum sv, am 1, 2, 7 41. calliergonella cuspidata sg, so, sv, wr, am 1, 2, 8, 9, 11 42. campyliadelphus chrysophyllus ro, so, sv 1, 10, 12, 13 43. campyliadelphus elodes am 7 44. campylium sommerfeltii sg, so 8 45. campylium stellatum so, sv, am 1, 2, 7 46. ceratodon purpureus sta, sg, so, sv, wr, wl 1, 2, 3, 4, 5, 7, 9, 10, 11 47. climacium dendroides so, sv, am 1, 10 48. cratoneuron �licinum ro, am 7, 12 49. ctenidium molluscum ro, am 7, 12 50. dicranella heteromalla so, sv 1, 6, 9, 11 51. dicranella varia so 4 52. dicranum bonjeanii so 1 53. dicranum �agellare wr 2 54. dicranum montanum so, wr, wl 1, 2, 9, 10 55. dicranum polysetum so, wr 1, 9 56. dicranum scoparium so, wr 1, 2 57. didymodon acutus so 13 58. didymodon fallax so, wl 4, 10 59. didymodon rigidulus sta, sg, sv 1, 2, 10 60. didymodon tophaceus ro 12 61. drepanocladus aduncus sv 1, 9 62. drepanocladus polygamus so, sv, wr 1 63. encalypta streptocarpa so, sv 7, 10, 13 64. eurhynchium angustirete so, wr, wl 2, 4, 6, 8, 65. eurhynchium striatum so, sv, wl 1, 6, 8 66. fissidens adianthoides sta, sv, am 1, 2, 7 67. fissidens bryoides so 6 68. fissidens dubius so 13 69. fissidens taxifolius so, am 1, 3, 4, 6, 14 70. funaria hygrometrica sta, sg 12, 13 71. grimmia pulvinata so 10 72. herzogiella seligeri so, wr 1, 9, 10 73. homalia trichomanoides wr, wl 3, 4, 6, 8, 74. homalothecium lutescens ro, sg 10 75. hygroamblystegium tenax am 7 76. hygroamblystegium varium wl 6, 8 77. hylocomium splendens so 2 78. hypnum cupressiforme sg, so, sv, wr, wl 1, 2, 3, 4, 5, 6, 8, 9, 10, 12 79. hypnum fertile wr 12 80. hypnum pallescens wr, wl 1, 2, 11 y ur a d ra ch , z ve ny sv al a m am ch ur 22 81. isothecium alopecuroides wl 8, 10 82. kindbergia praelonga so 4 83. leptodictyum riparium sta, sg, sv, wr, am 1, 3, 4, 11 84. leskea polycarpa sta, so, wr, wl 3, 4, 6, 8, 10 85. leucobryum glaucum so 2 86. leucodon sciuroides wr, wl 12 87. mnium marginatum so 8 88. mnium spinosum wl 4 89. mnium stellare ro 10 90. nyholmiella obtusifolia wr, wl 3, 6, 8 91. orthotrichum a�ne wl 4, 8 92. orthotrichum anomalum ro, sta, sg 2, 10, 12 93. orthotrichum cupulatum wr 12 94. orthotrichum diaphanum sta, wl 1, 3, 4 95. orthotrichum pallens wl 4, 6, 8 96. orthotrichum patens wl 8 97. orthotrichum pumilum sta, wl 1, 14 98. orthotrichum speciosum sta, wl 3, 4, 6, 8, 11, 14 99. orthotrichum stramineum wr 4 100. oxyrrhynchium hians so, sv, wr, wl 3, 4, 6, 8, 101. oxyrrhynchium speciosum so, wr 4 102. palustriella falcata am 7 103. phascum piliferum so 4 104. plagiomnium a�ne so 1, 8 105. plagiomnium cuspidatum ro, sta, sg, so, sv, wr, wl 1, 2, 4, 5, 6, 8, 10, 14 106. plagiomnium elatum so, am 1, 2, 8, 14 107. plagiomnium ellipticum so, sv 1, 2, 9 108. plagiomnium rostratum ro, sg, so, wr 4, 10, 11, 12 109. plagiomnium undulatum so, sv 4, 6, 8, 12 110. plagiothecium cavifolium so, wl 8 111. plagiothecium denticulatum so, wl 1, 8 112. plagiothecium laetum so, wr, wl 1, 8, 10 113. plagiothecium nemorale so, wr, wl 4, 6, 10 114. platygyrium repens sv, wr, wl 1, 2, 3, 4, 6 115. pleurozium schreberi so, sv, wr 2, 8, 9 116. pohlia nutans sta, so, wr, wl 1, 2, 4, 8, 9 117. pohlia wahlenbergii so 10 118. polytrichum commune so, sv, am 1, 2, 11 119. polytrichum formosum so 1, 10 120. polytrichum juniperinum sg, so 1, 2, 9 121. polytrichum longisetum so, wl 1 122. polytrichum piliferum sg, so 1, 11 123. pseudoamblystegium subtile wr 8 124. pseudoleskeella nervosa wr, wl 3, 4, 8, 10 125. pseudoscleropodium purum so 2, 9, 11 b ryophytes of the upper reaches of the w estern b ug r iver (lviv r egion, u kraine) 23 126. ptilium crista-castrensis wr 2 127. ptychostomum moravicum sta, sg, wr, wl 1, 2, 3, 6, 8, 12 128. ptychostomum pallens so 4 129. ptychostomum pseudotriquetrum sg, sv, wr, am 1, 7, 11 130. ptychostomum torquescens so 4 131. pylaisia polyantha wr, wl 2, 3, 4, 6, 8, 9, 12, 14 132. rhizomnium punctatum so 4 133. rhodobryum ontariense so 12 134. rhynchostegium murale sta 3 135. rhytidiadelphus squarrosus so 2, 9 136. rhytidiadelphus triquetrus so, sv 8 137. schistidium crassipilum sta, sg 1, 10 138. sciurohypnum plumosum sta 3 139. sciurohypnum populeum so, sv, wr 4, 6, 12 140. sciurohypnum re�exum so 4 141. scorpidium cossonii am 1, 7 142. sphagnum angustifolium so 2 143. sphagnum cuspidatum so 1, 2 144. sphagnum fallax so, sv 1, 2 145. sphagnum. �mbriatum so 1, 2 146. sphagnum girgensohnii so 1 147. sphagnum inundatum so, am 11 148. sphagnum palustre so, sv 1, 2 149. straminergon stramineum so 1 150. syntrichia papillosa sta, wr, wl 1, 3, 6 151. syntrichia ruraliformis so 4 152. syntrichia ruralis sta, sg, so, wl 1, 2, 12, 13 153. taxiphyllum wissgrillii ro 10 154. tetraphis pellucida wr 1, 2, 4 155. �uidium assimile. so, sv 2, 4, 8, 10, 12 156. �uidium delicatulum so 1 157. tomenthypnum nitens so 1 158. tortella humilis so 12 159. tortella inclinata ro 12, 13 160. tortula aestiva sta 1 161. tortula caucasica so 4 162. tortula muralis sta, sg 1, 3, 10, 12 163. tortula subulata wr 8 164. tortula truncata so 4 165. warnstor�a �uitans sv 1 y ur a d ra ch , z ve ny sv al a m am ch ur 24 abstract in the article, the bryophytes of the upper reaches of the western bug river (ukraine), which is physically and geographically located within male polissya, partly roztochia, and to a minor extent in the gologoro-voronyatsky denudo-structural hills, have been studied. based on our survey, a list of the bryophytes has been compiled for the �rst time. ecological features, substrate preferences and life forms of the bryophytes have been analysed. according to the ecological features, subheliophytes (30.9%) and hemisciophytes (30.9%) predominate in the spectrum of heliomorphs; mesophytes (29.7%), hygromesophytes (21.2%) and xeromesophytes – in the spectrum of hydromorphs (19.4%); cold-tolerant species (59.4%) – in the spectrum of thermomorphs. based on the analysis of the substrate preferences of the bryophytes, the following groups were identi�ed: epigeans (116 species), epixils (56 species), epiphytes (46 species), epiliths (43 species), aquatic (22 species). �e prevailing life forms are turf (30.3%), rough mat (18.2%), we� (15.2%), tu� (10.3%) and smooth mat (9.7%). 3 species that are o�cially recognised as rare and 16 species that are recognised as regionally rare have been found. in the group of bryophytes associated with wetland ecosystems, 2 o�cially rare and 6 regionally rare species were found in the study area. given the large areas of drained land in lviv region, these species are of particular value, especially in the context of conservation of the biodiversity and protection of the valuable natural areas in accordance with the development strategy of lviv region by 2027. key words: rare bryophytes, ecomorphs, life forms, ecological groups received: [2020.08.06] accepted: [2020.10.13] mszaki górnego biegu zachodniego bugu (obwód lwowski, ukraina) streszczenie w artykule przedstawiono wyniki badań, dotyczących mszaków górnego biegu zachodniego bugu (ukraina). teren ten �zycznie i geogra�cznie położony jest w obrębie małego polesia, częściowo roztochii oraz w niewielkim stopniu na wzgórzach gołogóro-woronieckich. na podstawie badań terenowych, po raz pierwszy zestawiono listę mszaków tego obszaru. przeanalizowano cechy ekologiczne, preferencje podłoża i formy życiowe stwierdzonych tu mszaków. według cech ekologicznych w spektrum heliomorfów dominują subhelio�ty (30,9%) i  hemiscio�ty (30,9%); mezo�ty (29,7%), higromezo�ty (21,2%) i kseromezo�ty – w spektrum hydromorfów (19,4%); gatunki tolerujące zimno (59,4%) – w spektrum termomorfów. na podstawie analizy preferencji substratowych, zidenty�kowano następujące grupy mszaków: epigeity (116 gatunków), epiksylity (56 gatunków), epi�ty (46 gatunków), epility (43 gatunki), hydro�ty (całkowicie lub częściowo zanurzone) (22 gatunki). dominującymi formami życiowymi są formy darniowe (30,3%), tworzące szorstkie maty (18,2%), wełniste (15,2%), kępkowe (10,3%) oraz tworzące gładkie maty (9,7%). znaleziono 3 gatunki w  skali ogólnej uznane za rzadkie oraz 16 gatunków uznanych za rzadkie regionalnie. w  grupie mszaków związanych z ekosystemami wodno-błotnymi, stwierdzono 2 gatunki ogólnie rzadkie i 6 gatunków rzadkich regionalnie. ze względu na duże powierzchnie odwodnionych terenów w obwodzie lwowskim, gatunki te są szczególnie cenne, zwłaszcza w kontekście zachowania różnorodności biologicznej i ochrony ważnych przyrodniczo obszarów, zgodnie ze strategią rozwoju obwodu lwowskiego do 2027 roku. słowa kluczowe: mszaki rzadkie, ekomorfy, formy życiowe, grupy ekologiczne information about authors yura drach https://orcid.org/0000-0002-6497-5638 his currently works at the department of ecology (head of department), ivan franko national university of lviv research is connected with ecology and biodiversity of bryophytes of the western bug river and the ukrainian carpathians. zvenysvala mamchur https://orcid.org/0000-0003-0527-5639 zvenyslava mamchur currently works at the department of ecology (head of department), of ivan franko national university of lviv. she does research in ecology and botany. her current projects are “ecological features of synanthropic �ora of lviv”, “bryophyta”. 66 annales universitatis paedagogicae cracoviensis studia naturae, 5: 66–80, 2020, issn 2543-8832 doi: 10.24917/25438832.5.5 beata jakubik*, krzysztof lewandowski, aleksandra biernat siedlce university of natural sciences and humanities, institute of biological sciences, b. prusa 14 st., 08-110 siedlce, poland; *beata.jakubik@uph.edu.pl changes of malacofauna in a small lowland river in eastern poland introduction molluscs are an important component of the biodiversity of freshwater invertebrate fauna (e.g. jurkiewicz-karnkowska, 2004; królak, korycińska, 2008; piechocki, 2008). �e malacofauna of large polish rivers is well recognised (e.g. piechocki, 1996; jurkiewicz-karnkowska, 2004; lewandowski, 2004; jurkiewicz-karnkowska, karnkowski, 2013; piechocki, szlauer-łukaszewska, 2013; lewin, 2014). studies on the occurrence of molluscs in small lowland rivers in our climate zone, despite their long tradition, are fragmentary (e.g. piechocki, 1969, 1972; kasprzak, 1975; kołodziejczyk, 1994; jakubik, 2003, 2008; jurkiewicz-karnkowska, 2019). areas less recognised for malacofauna include the siedlce upland – part of the south podlasie lowland (kondracki, 2002). malacofauna of small watercourses in the siedlce upland is poorly understood. some remarks on species present in the region were provided by królak (1998), who described the results of her studies on the content of heavy metals in molluscs of the siedlce upland. apart from papers by jakubik (2003), korycińska (2002) and jurkiewicz-karnkowska (2016, 2019), no detailed data on molluscs from this region can be found. �e muchawka river is a le�-bank tributary of the liwiec river – the main river of the siedlce upland. �erefore, species composition of molluscs in the former may markedly a�ect taxonomic diversity in the latter. �e muchawka river drains mostly agricultural grounds and, thus, is not signi�cantly polluted by domestic and/or industrial wastewaters. until 1999, data about the river were limited to general faunistic and �oristic characteristics and to annual assessment of water quality performed by the environmental protection inspectorate based on physical, chemical and biological parameters (bakiera et al., 1993). �at is why detailed faunistic studies on molluscs in the river were undertaken in the years 1999–2000 (jakubik, 2003). sixteen years later, the assessment of taxonomic composition of molluscs in the muchawka river, in relation to certain environmental factors, was repeated. c hanges of m alacofauna in a sm all low land river in eastern p oland 67 material and methods study area studies were carried out in the muchawka river (52°12ʹ35ʺn, 22°13ʹ10ʹʹe) – a  le�bank tributary of the liwiec river. it springs about 2 km south of daćbogi village and �ows across the siedlce upland. �e river is about 30 km long and its catchment area is 292 km2. �e river forms a  valley overgrown by periodically �ooded meadows. �e mu chawka river valley is �oristically rich, especially in vascular plants. �ere are about 400 species of vascular plants dominated by the families asteraceae, poaceae and fabaceae from meadow, aquatic and rush communities (kot, dombrowski, 2001). waters of the muchawka river feed a  recreational dam reservoir in siedlce. eight sampling sites, the same as in jakubik (2003), were selected along the course of the muchawka river (fig. 1, 2). characteristics of the sampling sites are given in table 1. fig.1. location of the sampling sites (jakubik, 2003) b ea ta j ak ub ik , k rz ys zt of l ew an do w sk i, a le ks an dr a b ie rn at 68 tab. 1. characteristics of study sites; plant nomenclature according to podbielkowski, tomaszewicz (1996) no. site type width the river [m] sampling depth [cm] vegetation bottom river bank 1 muddy high, enforced with wooden stakes 2–3 50 acorus calamus l., elodea canadensis michx, phragmites australis (cav.) trin. ex steud, sparganium ramosum curtis 2 sandy-stony natural 2 20 sparganium ramosum curtis 3 stony-muddy natural, gentle slope 3 20 elodea canadensis michx, juncus e�usus l., nuphar lutea l. sibth & sm, sagittaria sagittifolia l. 4 sandy-stony 5 30 elodea canadensis michx, nuphar lutea l. sibth & sm, phragmites australis (cav.) trin.ex steud sagittaria sagittifolia l. 5 6 80 elodea canadensis michx, nuphar lutea l. sibth & sm, sagittaria sagittifolia l., sparganium ramosum curtis 6 sandy-muddy 6 30 elodea canadensis michx, nuphar lutea l. sibth & sm, sagittaria sagittifolia l. 7 6 30 nuphar lutea l. sibth & sm, sagittaria sagittifolia l., sparganium ramosum curtis, typha latifolia l. 8 muddy natural, gently sloping, steep in some places 8 30 nuphar lutea l. sibth & sm, phragmites australis (cav.) trin.ex steud, sagittaria sagittifolia l., typha latifolia l. methods water and molluscs were sampled in late spring (may to june) and in summer (july to the middle of september) 2016 in triplicate from every sampling site. water for chemical analyses was collected in polyethylene containers and preserved with 2–3 cm3 of chloroform per 1 dm3 of water (hermanowicz et al., 1999). dissolved oxygen (measured with the oxygen probe eot 196), temperature and electrolytic conductivity of water (conductivity meter cc-317) were determined in the �eld at each sampling site. water ph (digital ph-meter cp-215), water hardness and concentrations of ammonium-nitrogen, nitrate-nitrogen, phosphates (�eld photometer lf-205) and chlorides (with the argentometric method as in hermanowicz et al., 1999) were determined in the laboratory. based on physical and chemical parameters, water quality of the muchawka river was estimated and compared with earlier data (tab. 2). molluscs were sampled with a 20-cm-wide grab sampler (jakubik, 2003). �e sample was material collected from an area of one square meter. bivalves of the family c hanges of m alacofauna in a sm all low land river in eastern p oland 69 ta b. 2 . w at er q ua lit y in th e m uc ha w ka r iv er : a – d at a fr om 1 99 9– 20 00 (j ak ub ik , 2 00 3) , b – d at a fr om 2 01 6 pa ra m et er es si te s m ea n fi na l w at er qu al ity 1 2 3 4 5 6 7 8 a b a b a b a b a b a b a b a b a b a b te m pe ra tu re [° c ] 15 .7 3 17 .2 0 19 .7 0 17 .0 0 16 .7 3 18 .3 0 20 .3 3 21 .0 0 21 .0 0 21 .3 0 20 .1 0 19 .9 0 14 .2 0 19 .1 0 18 .1 0 18 .7 0 18 .2 3 19 .0 6 i i a ci di ty [p h ] 7. 53 7. 61 7. 66 7. 90 7. 66 7. 58 7. 86 7. 54 7. 86 7. 65 7. 53 7. 71 7. 40 7. 63 7. 37 7. 57 7. 61 7. 64 i ii c on du ct iv ity [m s/ cm ] 0. 51 0. 31 0. 61 0. 29 0. 67 0. 39 0. 66 0. 42 0. 64 0. 49 0. 64 0. 42 0. 69 0. 43 0. 70 0. 44 0. 64 0. 40 i ii o 2 [ m g/ dm 3 ] 8. 63 4. 00 7. 40 6. 40 7. 76 4. 00 7. 90 4. 60 6. 43 6. 40 5. 86 6. 40 6. 16 5. 40 7. 03 4. 40 7. 15 5. 20 i * n – n o 3– [m g/ dm 3 ] 1. 53 1. 46 1. 46 1. 15 0. 67 0. 62 0. 52 5. 58 0. 49 5. 05 0. 56 3. 14 0. 75 3. 45 0. 65 3. 45 0. 83 2. 98 i * n – n h 4 [m g/ dm 3 ] 0. 62 0. 03 0. 28 0. 04 0. 34 0. 10 0. 20 0. 26 0. 25 1. 08 0. 19 0. 03 0. 20 0. 12 0. 12 0. 03 0. 28 0. 21 i ii po 43 – [m g/ dm 3 ] 0. 43 0. 82 0. 43 0. 87 0. 36 0. 51 0. 33 0. 30 0. 43 0. 16 0. 46 0. 23 0. 40 0. 23 0. 30 0. 25 0. 39 0. 42 ii * c l– [m g/ dm 3 ] 4. 99 16 .0 0 5. 94 16 .0 0 5. 77 14 .0 0 5. 77 21 .0 0 5. 66 30 .0 0 5. 66 21 6. 27 22 6. 22 25 5. 78 20 .6 0 i ii h ar dn es s [m g c ac o 3/ dm 3 ] 25 5 17 6 33 1 18 0 36 4 24 2 36 2 22 8 34 1 24 4 34 7 23 4 37 0 24 2 37 3 24 2 34 5 22 4 i i n ot e: * d oe s n ot � t i n ii b ea ta j ak ub ik , k rz ys zt of l ew an do w sk i, a le ks an dr a b ie rn at 70 fig. 2. a, b – sites (1–2) in the upper section of the river; c, d – sites (4–5) in the middle stretch; e, f – sites (7–8) in the lower river section (photo. k. lewandowski) unionidae were determined to the species level and then released back into the water. in the �eld, collected material was washed on a  sieve of 0.5 mm mesh. in the laboratory, molluscs were preserved in 70% ethanol. molluscs were identi�ed to the species level based on their morphological and anatomical features according to piechocki and dyduch-falniowska (1993), jackiewicz (2000), piechocki and wawrzyniak-wydrowska (2016). �e species nomenclature follows glöer (2002). c hanges of m alacofauna in a sm all low land river in eastern p oland 71 statistical analysis �e following parameters were calculated (górny, grüm, 1981): (1) the total number of species (s); (2) density − expressed as the number of individuals per square metre; (3) domination according to the formula d = 100 × ni /n where ni is the number of individuals of the i-th species and n is the number of individuals of all species; the value of the domination, d, was divided into 5 classes: eudominants > 10.0%, dominants 5.1–10.0%, subdominants 2.1–5.0%, recedents 1.1–2.0% and subrecedents ≤ 1.0% of the sample; (4) the shannon-wiener index (h’) (hauer, lamberti, 2007): h’ = −s pi ln pi where pi = ni/n − the share of individuals of the i-th species. �e signi�cance of di�erences between the number of species and densities at particular sites visited in 1999–2000 and in 2016 were evaluated with the tukey one-way anova test (statistica v. 10). �e compared parameters had a normal distribution. results temperature and water hardness did not exceed standards established for the �rst class of water quality (rozporządzenie ministra środowiska..., 2016). water ph, electrolytic conductivity and concentrations of ammonium-nitrogen and chlorides corresponded to the second class of water quality. only the concentrations of nitrate-nitrogen, phosphates and oxygen were higher than those typical for the second class of water quality (tab. 2). because of agricultural management of surrounding lands, the main source of phosphates delivered to the river is surface runo� from �elds and meadows. studies on malacofauna of the muchawka river carried out in the years 1999–2000 revealed the presence of 12 mollusc species: �ve species of bivalves and seven species of snails (tab. 3). tab. 3. species composition of molluscs in the muchawka river 1999–2000 (jakubik, 2003) and the present study no. species 1999–2000 2016 1. anisus vortex linnaeus × 2. anodonta anatina linnaeus × × 3. a. cygnea linnaeus × 4. bathyomphalus contortus linnaeus × 5. bithynia tentaculata linnaeus × × 6. lymnaea stagnalis linnaeus × × 7. physa acuta draparnaud × 8. p. fontinalis linnaeus × × 9. planorbarius corneus linnaeus × 10. planorbis carinatus o.f.müller × b ea ta j ak ub ik , k rz ys zt of l ew an do w sk i, a le ks an dr a b ie rn at 72 11. p. planorbis linnaeus × × 12. pisidium amnicum o.f.müller × × 13. p. casertanum poli × 14. p. henslowanum sheppard × 15. p. nitidum jenyns × 16. p. subtruncatum malm × 17. radix ampla hartmann × 18. r. auricularia linnaeus × 19. sphaerium corneum linnaeus × 20. s. rivicola lamarck × 21. unio crassus philipsson × 22. u. pictorum linnaeus × 23. valvata cristata o.f.müller × 24. viviparus contectus millet × × total 12 19 in 2016, there were 19 species of molluscs in the river including 11 species of snails and 8 species of bivalves. seven species of molluscs (5 species of snails and 2 species of bivalves) were recorded in both periods. species invariably present in the river since 1999 were: anodonta anatina, bithynia tentaculata, lymnaea stagnalis, physa fontinalis, pisidium amnicum, planorbis planorbis, viviparus contectus. malacofauna showed both qualitative and quantitative diversity. in total, 390 individuals of molluscs were noted at all sites. pisidium casertanum – one of eudominants – was present only at the last two sites (with densities of 153 individual × m–2 and 15 individual × m–2, respectively) and constituted 43.1% of all molluscs (tab. 4). tab. 4. density (individual × m–2) and dominance [d%] of the molluscs at the sampling sites in the muchawka river (2016) species sites n d 1 2 3 4 5 6 7 8 anisus vortex 1 1 0.3 anodonta anatina 1 1 0.3 bathyomphalus contortus 2 9 11 2.8 bithynia tentaculata 2 12 2 1 4 21 5.4 lymnaea stagnalis 3 3 0.8 physa acuta 2 2 0.5 p. fontinalis 8 8 2.0 pisidium amnicum 9 5 4 18 4.6 p. casertanum 153 15 168 43.1 p. henslowanum 1 1 0.3 c hanges of m alacofauna in a sm all low land river in eastern p oland 73 p. nitidum 9 9 2.3 p. subtruncatum 75 12 2 89 22.8 planorbarius corneus 1 2 1 1 5 1.3 planorbis carinatus 2 2 0.5 p. planorbis 2 2 0.5 sphaerium corneum 29 1 1 5 4 40 10.2 unio crassus 2 2 0.5 valvata cristata 1 1 0.3 viviparus contectus 6 6 1.5 total 90 25 72 2 3 11 159 28 390.0 100.0 number of species 6 4 9 1 2 6 3 5 – – fig. 3. comparison of the domination [d%] of molluscs in the muchawka river between 1999–2000 (jakubik, 2003) and 2016 b ea ta j ak ub ik , k rz ys zt of l ew an do w sk i, a le ks an dr a b ie rn at 74 fig. 4. comparison of the density (individual × m–2) of molluscs in the muchawka river between 1999– 2000 (jakubik, 2003) – green and 2016 – red fig. 5. comparison of shannon-wiener indices h’ of the molluscs in the muchawka river between 1999–2000 – (a) (jakubik, 2003) and 2016 – (b) two other species of bivalves of the family sphaeriidae identi�ed were also eudominants – pisidium subtruncatum (22.8%) and sphaerium corneum (10.3%) (fig. 3). bivalves of the family sphaeriidae constituted as many as 83.3% of all collected molluscs and, together with other bivalves (unio crassus and anodonta anatina), comprised 84.1%. snails were mostly represented by the dominant bithynia tentaculata (5.4% of all molluscs) and by subdominant bathyomphalus contortus (2.8%). c hanges of m alacofauna in a sm all low land river in eastern p oland 75 out of eight sites analysed, sphaerium corneum and the snail bithynia tentaculata were recorded in �ve sites. most of the noted species were present in small densities in single sites. �e highest number of mollusc species (9) was found at site 3 and the lowest numbers of species (1–2) were noted at sites within the city limits of siedlce (sites 4 and 5). noteworthy was the presence of a protected species, unio crassus, at a density of 2 individual × m–2 in site 3. apart from living molluscs, the empty shells of other species, such as valvata piscinalis müll., radix balthica l. and acroloxus lacustris l., were recorded. densities of molluscs in the year 2016 were markedly higher compared to those in the years 1999–2000 (fig. 4). signi�cant di�erences were found in sites in the upper course of the river (sites 1, 2 and 3) and in sites (7 and 8) near its outlet (tukey test, p < 0.05). �e shannon-wiener index was lower in the year 2016 (fig. 5). discussion during the sixteen-year period since the �rst malacological analysis, the number of mollusc species in the muchawka river increased from 12 to 19 and was comparable with that in the skierniewka river (21 species) (jurkiewicz-karnkowska, 1989). �e number of taxa was, however, lower than that found in the rivers grabia and pasłęka (piechocki, 1969, 1972), krutynia (lewandowski, 1996; jakubik et al., 2014) and raba, łubrzanka, łośna, biała nida, czarna nida (piechocki, 1981), koprzywianka (piechocki, 1987), szeszupa (lewandowski, 1990) and wieprz (piechocki, łuczak, 1989; piechocki, 1992). in each of the described habitats about 40 species of molluscs were noted. out of 19 species of molluscs found in the muchawka river, 11 species were snails. �e number is comparable with the 12 snail species recorded by korycińska (2002) in the liwiec river – the recipient of waters of the muchawka. later studies of molluscs carried out in the years 2013-2015 showed 26 species in the liwiec river (jurkiewicz-karnkowska 2016, 2019). in the muchawka river, among the taxa found by jurkiewicz-karnkowska in 2016, were noted all species with the exception of pisidium carinatus and bathyomphalus contortus. grużewski (2000) noted as many as 25 mollusc species including 15 species of snails and 10 species of bivalves in the pęza river (right-bank tributary of the middle narew river) which is much smaller than the muchawka river. studies from the same author (1996) in the kamionka river in the wigry national park revealed 13 species of molluscs (10 of which were bivalves) – a  number similar to that recorded in the muchawka river in the �rst study period. malacofauna of the muchawka river was dominated by bivalves of the genera pisidium and sphaerium – pisidium amnicum, sphaerium rivicola, in the �rst study peb ea ta j ak ub ik , k rz ys zt of l ew an do w sk i, a le ks an dr a b ie rn at 76 riod and by pisidium casertanum and p. subtruncatum in 2016. �is phenomenon was con�rmed by studies on molluscs by jurkiewicz-karnkowska (2016, 2019) who found a high frequency of bivalves of both genera in the middle stretch of the liwiec river, i.e. near the inlet of the muchawka river. according to piechocki and dyduch-falniowska (1993), these bivalves mainly inhabit lowland rivers and prefer sandy or sandy-muddy largely overgrown substrata and clean or only slightly polluted water (stadničenko, 1984). pisidium amnicum is particularly sensitive to water pollution by domestic and industrial wastewaters and does not tolerate intensive eutrophication. �e presence of bivalves of the family unionidae in waters of the muchawka river was evidence of high water quality, con�rmed by physical and chemical analyses in the �rst study period. unio pictorum was found in the years 1999–2000 at four sites (1, 2, 7 and 8) of the upper and outlet section of the river (jakubik, 2003). in view of drastically decreasing numbers of unionids in lakes and rivers of poland, the presence of anodonta anatina, unio pictorum, u. tumidus, u. crassus (królak, korycińska, 2001) in the liwiec river in the years 1996–1997 is noteworthy. in 2016, the situation for protected species changed, which might be associated with worsening water quality. increased concentrations of nitrates and phosphates originated from agricultural management of the river drainage basin. anodonta cygnea, sphaerium rivicola and unio pictorum disappeared from the river, while another protected species, unio crassus, was noted. studies by jurkiewicz-karnkowska (2016, 2019) also showed a  lack of anodonta cygnea and unio pictorum in the middle stretch of the liwiec river. of particular importance was the presence of unio crassus – a species whose occurrence in poland’s waters regularly diminishes due to proceeding eutrophication and degradation of river valleys (abraszewska-kowalczyk, 2002; zając, 2004). regarding snails in the muchawka river, a remarkable share were bithynia tentaculata, while in the previous study period it was lymnaea stagnalis. a similar change was reported by jurkiewicz-karnkowska (2016, 2019) for malacofauna of the liwiec river. both species prefer sandy-muddy substrates with some stones, which is characteristic of lowland rivers (jackiewicz, 2000; lewin, 2014). �e shannon-wiener index for data from 2016 was lower than in the years 1999– 2000. �e index increases with increasing number of species in a  given community and with the evenness of their densities (głowaciński, 1996). despite an increased number of species in 2016, their densities were quite variable – from 2 individual × m–2 in the middle course of the river to 159 individuals × m–2 in the outlet. malacofauna of the muchawka river re�ects the natural character of a watercourse like this small lowland river (wiśniewski et al., 1985; jurkiewicz-karnkowska, 1989, 2016; raczyńska, 1999; grużewski, 2000; jakubik, 2008; lewin, 2014). �e present study is part of a larger project, which is scheduled to end in 2021. c hanges of m alacofauna in a sm all low land river in eastern p oland 77 acknowledgements �e authors wish to express their gratitude to ewa jurkiewicz-karnkowska for help in determining bivalves of the genus pisidium, to elżbieta biardzka for help in chemical analyses and to lech kufel for translating this text. we are very grateful to the anonymous reviewers for their valuable comments which help us to improve manuscript. con�ict of interest �e author declares no con�ict of interest related to this article. references abraszewska-kowalczyk, a. 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(1985). �e e�ect of impoundments on the quality of water and biocoenosis in the lowland streams rawka and skierniewka. ekologia polska, 33, 511–536. zając, k. (2004). unio crassus philipsson, 1788. in: z. głowaciński, j. nowacki (eds.), polska czerwona księga zwierząt. bezkręgowce (polish red data book of animals. invertebrates). p. 353–355. kraków–poznań: institute of nature conservation pas and august cieszkowski agricultural university of poznań. b ea ta j ak ub ik , k rz ys zt of l ew an do w sk i, a le ks an dr a b ie rn at 80 abstract in 2016, the species composition and the structure of the dominance of molluscs in the muchawka river (le�-bank tributary of the liwiec river) were assessed. �e occurrence of 19 species of molluscs were recorded, including 11 species of snails and 8 species of mussels. �e eudominant was pisidium casertanum, which, only in the last two sites, constituted 43.1% of all molluscs. two other species of molluscs from the sphaeriidae family were also eudominants – pisidium subtruncatum (22.8%) and sphaerium corneum (10.3%). snails were most frequently represented by the dominant bithynia tentaculata, constituting 5.4% of all molluscs, and the subdominant bathyomphalus contortus at 2.8%. sixteen years a�er the �rst malacological analysis, an increase in species richness and di�erences in the dominance of molluscs were found in the muchawka river. clams from the sphaeriidae family invariably dominated but with a di�erent species composition. �e disappearance of the protected anodonta cygnea and sphaerium rivicola has been noted, and the occurrence of the protected unio crassus has also been noted. key words: lowland river, molluscs, the muchawka river received: [2020.07.29] accepted: [2020.09.24] zmiany malakofauny w małej rzece nizinnej wschodniej polski streszczenie w 2016 roku dokonano oceny składu gatunkowego oraz struktury dominacji mięczaków rzeki muchawki (lewobrzeżny dopływ liwca). odnotowano występowanie 19 gatunków mięczaków, a w tym 11 gatunków ślimaków i  8 gatunków małży. eudominantem był małż pisidium casertanum, który występując tylko na dwóch stanowiskach stanowił 43,1% wszystkich mięczaków. eudominantami były także dwa inne gatunki małży z rodziny sphaeriidae – pisidium subtruncatum (22,8%) i sphaerium corneum (10,3%). ślimaki najliczniej reprezentował dominant bithynia tentaculata, stanowiąc 5,4% wszystkich mięczaków oraz subdominant bathyomphalus contortus – 2,8%. po szesnastu latach od pierwszej analizy malakologicznej w rzece muchawka stwierdzono wzrost bogactwa gatunkowego oraz różnice w dominacji mięczaków. niezmiennie dominowały małże z rodziny sphaeriidae, ale przy innym składzie gatunkowym. odnotowano zanik chronionej anodonta cygnea i sphaerium rivicola, a pojawienie się również chronionej unio crassus. słowa kluczowe: rzeka nizinna, mięczaki, rzeka muchawka information on the authors beata jakubik https://orcid.org/0000-0002-1367-2805 ecology of molluscs in various types of aquatic habitats, studies on life strategies of molluscs in response to di�erent habitat conditions using family viviparidae as an example. krzysztof lewandowski he is a professor emeritus of the siedlce university of natural sciences and humanities). ecology of freshwater molluscs with special focus on abundant species (like e.g. zebra mussel dreissena polymorpha, bivalves of the family unionidae, the river snail viviparus viviparus), that play an important role in the functioning of freshwater ecosystems; lake eutrophication and protection. aleksandra biernat graduate student of the institute of biology, siedlce university of natural sciences and humanities. annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x the checklist of impatiens riv. ex l. (balsaminaceae) in nilgiri and palani hills of southern western ghats (india) s. jeevith 1* , ravi kiran arigela 2 , rajeev kumar singh 3 , k. althaf ahamed kabeer 4 , c. rajasekar 5 , c. kunhikannan 6 1 division of forest ecology and climate change, institute of forest genetics and tree breeding, coimbatore – 641 002, tamil nadu, india. 2 botanical survey of india, deccan regional centre, rooms 228–238, kendriya sadan, sultan bazar, hyderabad – 500 095, telangana, india. 3 industrial section and indian museum, botanical survey of india, 1, sudder street, kolkata– 700 016, west bengal, india. 4 central botanical laboratory, botanical survey of india, acharya jagadish chandra bose indian botanic garden, howrah – 711 103, west bengal, india. 5 department of botany, alagappa university, karaikudi – 630 003, tamil nadu, india. 6 institute of forest genetics and tree breeding, coimbatore – 641 002, tamil nadu, india. *e-mail of corresponding author: jeevithbotany@gmail.com introduction the genus impatiens riv. ex l. consists of more than 1050 species, distributed across the old world tropics and subtropics as well as in the north and central america (ruchisansakun et al., 2018; arigela et al., 2019a; singh et al., 2021; borah et al., 2022). most of the impatiens species occurs in five centres of diversity of the old world, namely madagascar, tropical africa, sinohimalayan region, western ghats region of india and southeast asia. in india, there are about 242 species of impatiens recorded so far, of which 152 species are endemic, and the highest diversity occurs in two regions, the eastern himalayas and the western ghats (singh, 2016a, 2016b; singh & garg, 2016; singh, 2017; arigela, 2019a; singh et al., 2021). western ghats holds about 116 endemic species of impatiens (bhaskar, 2012; arigela et al., 2019a). the species of impatiens are generally known as balsam or jewel weed, and several species have ornamental potential. in western ghats, the floristic studies and inventories of the genus impatiens were early reported in floras by gamble (1915), fyson (1932) and matthew (1983, 1999). recently, bhaskar (2012) provided a comprehensive account of 106 species from western ghats, and dessai and janarthanam (2011) reported 26 species and 2 varieties from the northern and central western ghats. singh (2016) typified thirty six names of thirty five recognised taxa of impatiens, which are endemic to the western ghats. in nilgiri and palani, the genus is widespread in montane wet temperate (shola) forests and high altitude grassland to semi-evergreen forests. to effectively protect the endemic vascular plants, a comprehensive document of endemic plants of these ecosystems, is prerequisite. the present study on the genus impatiens brought out an updated checklist of balsams in the nilgiri and palani hills of western ghats, india (fig. 1 – appendix 1). study area the nilgiri hills of tamil nadu is located between 11°10’–11°43’ n and 76°14’–77°00’ e, in nilgiris district with an area of 2,452.50 km 2 . the nilgiri is one of the high mountain ranges of the western ghats, at an elevation of 900 to 2,636 m a. s. l. and it is surrounded by the plains of coimbatore in the southeast, bhavani in the northeast, moyar valley in the north and gudalur plateau in the northwest (fig. 1c – appendix 1). in the nilgiri, underlying rocks are crystalline and mostly gneisses in the entire plateau. moist evergreen forests at 900–1300 m and montane wet temperate (shola) forests combined with high altitude grasslands at 1300–2600 m are the major vegetation types. the entire district soil is classified under ‘humic-ferritic soil’ on account of topsoil, as per profile characteristics. the nilgiris district lies in the tropical zone with subtropical to temperate climate. the western regions of the plateau receive about 3000 mm of rainfall which exceeds 5000 mm in certain sites. the temperature ranged from a minimum of 2°c and a maximum of 25°c, with a layer of mist and fog covering the south eastern slopes. the palani (palni) hills lie between 10°07’–10°25’ n and 77°15’–77°43’ e in dindigul and theni districts of tamil nadu. the palani hills are situated in south central tamil nadu, an east-west spur of the western ghats, extending roughly from the kerala border near munnar, past the town of kodaikanal as far as dindigul in the east (fig. 1b – appendix 1). the maximum length is 65 km and width is 40 km, with an area of about 2000 km 2 . these hills in turn are divided into the upper palani (west) and the lower palani (east). the upper palani have an average elevation of 2,200 m and the lower palani have elevation less than 1700 m. the upper palani (1700–2100 m) are predominantly grasslands interspersed with tropical montane shola forests which tend to cluster along the streams and rivers. below this, from 1300 to 1700 m is dry evergreen forest. from 800 to 1300 m is dry deciduous forest, from 300 to 800 m is savannah interspersed with dry deciduous trees and from the plains to approximately 300 m elevation is scrub forest (matthew, 1999, arigela et al., 2019b). the upper palani hills have a quasitemperate climate with temperature ranges from 2ºc to 17ºc in the winter 12ºc to 21ºc in the summer. the average year round temperature is 16°c. the lower hills become progressively warmer and temperature ranges from 25°c to 40°c. the average annual rainfall of kodaikanal is approximately 1700 mm. materials and methods based on literature and phenology, the specimens were sampled in different sites of nilgiri and palani from 2012 to 2020 in different seasons. exploration sites include shola forests, grasslands and transitional zones between different vegetation types for ecotype variants. photographs of habitat, habit, inflorescence and close up of flowers of each species were taken to prepare the digital data base and for identification. height, inflorescence size and colour, wildlife signs and threat factors were recorded for all plant species. gps coordinates with altitude were collected for geo-tagging and for further studies. the species were identified with the help of literature (roxburgh, 1824; wight, arnott, 1834; wight, 1837, 1838–1853, 1839; hooker, 1875; gamble, 1924; matthew, 1983, 1999; dessai, janarthanam, 2011; bhaskar, 2012; singh et al., 2015; singh, 2016a, 2016b, 2017) and specimens available with central national herbarium (cal), howrah, fischer herbarium (frc), ifgtb, coimbatore and madras herbarium (mh), bsi, coimbatore. the plant specimens of present study have been deposited at mh and frc for future reference. results in the present study, documented 38 species of impatiens from nilgiri and palani hills. among balsams, 36 species were recorded from nilgiri (figs. 2–4 – appendix 1), 13 species were recorded from palani (fig. 5 – appendix 1), 11 species occur in both nilgiri and palani hills. out of 38 taxa, 32 are endemic to india (6 species are endemic to western ghats and 26 species are endemic to southern part of western ghats) and 6 taxa have wider distribution. photo plates of the impatiens species from both the localities are presented separately to show the flower colour variations. few species like impatiens fasciculata (figs. 3c and 5c – appendix 1) or i. tomentosa (figs. 4h and 5k – appendix 1) produce both white and pink colour flowers. the habitat, occurrence in the study areas and distribution of these 38 impatiens species are given in table (1) – appendix 2. short discussion presently, madagascar has the highest impatiens species richness with about 260 species (rahelivololona et al., 2018) and india is at the second position with about 242 species. india nurtures a wide range of impatiens species due to its diverse topography, vegetation and forest types. the diversity of climate types and habitats of indian himalaya and western ghats is also associated with impatiens species diversity and richness. phylogenetic studies of the genus impatiens show the centre of origin in south china (janssens et al., 2006), but bhaskar (2012) claimed that it lies in the western ghats, as the primitive species of impatiens with lowest chromosome numbers (n=3 in impatiens latifolia – fig. 2b and i. leschenaultii – fig. 2a–5g – appendix 1) and the largest (n=20 and 25 in i. grandis – fig. 3k) are recorded from the western ghats. detailed ecological study of impatiens species in different habitats of nilgiri and palani hills showed that some species show variation in the vegetative morphological traits (i. inconspicua and i. minor – fig. 3f) some in the reproductive (floral or fruiting or both) traits (i. fasciculata, i. goughii, i. tomentosa and i. scapiflora), and some in both vegetative and reproductive traits (i. acaulis, i. balsamina, i. chinensis, i. diversifolia, i. oppositifolia and i. scabriuscula – figs. 2–5 – appendix 1). it is observed that abiotic ecological factors (climatic, edaphic and physiographic) play a major role in the morphological variation of some taxa, while biotic factors play a minor role. impatiens balsamina (figs. 4i and 5a – appendix 1) is native to south india and sri lanka, but widely cultivated as garden ornamental plant throughout india and many parts of globe (bhaskar, 2012). during the present study we found that 10 endemic species of balsam (impatiens clavicornu, i. campanulata, i. dasysperma, i. gardneriana, i. grandis, i. henslowiana, i. latifolia, i. leschenaultii, i. maculata and i. minor) and 8 threatened species (i. fruticosa, i. elegans, i. parasitica, i. phoenicea, i. tangachee, i. tanyae, i. viscida and i. viscosa) – tab. 1 – appendix 2 – have great horticultural potential. the promising potentials of utilisation of wild genes during breeding experiments and cultivar development cannot be ruled out of all these beautiful balsam. these endemic and threatened species of balsam have beautiful foliage and flower (often of different colours within the same species), and can be introduced as garden ornamental plants, especially in south and central india. multiplication through culture techniques and regeneration of individuals in similar habitats to their natural habitat is the most probable method to increase population number. by this way, we can conserve (ex-situ) these endemic and threatened species of balsam. acknowledgements the authors are thankful to the director of botanical survey of india, kolkata and the head of office, botanical survey of india, deccan regional centre, hyderabad. authors also thank dr. shrishali kulloli, spices board of india and dr. prasad kothareddy, yogi vemana university for sharing their knowledge to fix the identity of few species. special thanks to mr. amruth k, sacon, for his support in developing gis-based maps. thanks are due to the forest department, tamil nadu state and its staffs for all the help during field explorations. conflict of interest the authors declare no conflict of interest related to this article. references arigela, r.k., singh, r.k., kabeer, k.a.a. (2019a). impatiens tanyae (balsaminaceae), a new species from western ghats, india. kew bulletin, 74(3), 48. https://doi.org/10.1007/s12225-019-9831-4 arigela, r.k., singh, r.k., kabeer, k.a.a., murthy, g.v.s., robin, v.v. (2019b). phytodiversity inventorisation and mapping of shola grasslands of kodaikanal wildlife sanctuary, western ghats, india. indian forester, 145(3), 214–226. bhaskar, v. (2012). taxonomic monograph on impatiens l. (balsaminaceae) of western ghats, south india. bangalore: centre for plant taxonomic studies. borah, d., singh, r.k., taram, m. (2022). 118. impatiens pasighatensis (balsaminaceae), a new species from arunachal pradesh, northeast india. indian forester, 148(2), 233–235. dessai, j.r.n., janarthanam, m.k. (2011). the genus impatiens (balsaminaceae) in the northern and parts of central western ghats. rheedea, 21, 23–80. fyson, p.f. (1932). the flora of south india hill stations. vol. 1–2. madras: govt. press. gamble, j.s. (1915). flora of the presidency of madras. vol.1. london: adlard & sons. hooker, j.d. (1875). flora of british india. vol. 1. london: l. reeve & co. matthew, k.m. (1983). the flora of tamil nadu carnatic. vol. 1. tiruchirapalli: the rapinat herbarium. matthew, k.m. (1999). flora of palni hills, south india. vol. 1. tiruchirapalli: the rapinat herbarium. rahelivololona, e.m., fischer, e., janssens, s.b., razafimandimbison, s.g. 2018. phylogeny, infrageneric classification and species delimitation in the malagasy impatiens (balsaminaceae). phytokeys, 110, 51–67. https://doi.org/10.3897/phytokeys.110.28216 roxburgh, w. (1824). flora indica; or descriptions of indian plants. vol. 2. 452–465. serampore: the mission press. ruchisansakun, s., suksathan p., van der niet, t., smets, e.f., saw-lwin, janssens, s.b. (2018). balsaminaceae of myanmar. blumea, 63, 199–267. https://doi.org/10.3767/blumea.2018.63.03.01 singh, p., karthigeyan, k., lakshminarasimhan, p., dash, s.s. (2015). endemic vascular plants of india. kolkata: botanical survey of india. singh, r.k. (2016a). notes on author attribution and typification of two names of impatiens (balsaminaceae). journal of japanese botany, 91(4), 237–241. singh, r.k. (2016b). typification of thirty-six names of thirty five recognized taxa in impatiens (balsaminaceae), endemic to western ghats. phytotaxa, 268, 167–180. https://doi.org/10.11646/phytotaxa.268.3.1 singh, r.k. (2017). lectotypification of three linnaean names in impatiens (balsaminaceae). phytotaxa, 321, 299– 300. https://doi.org/10.11646/phytotaxa.321.3.8 singh, r.k., garg, a. (2016). call for conservation of the critically endangered saffron balsam of karnataka – impatiens raziana bhaskar & razi (balsaminaceae). indian forester, 142, 803–805. singh, r.k., borah, d., taram, m. (2021). typifications, new combinations and new synonyms in indian impatiens (balsaminaceae). biodiversity research and conservation, 61, 1–27. https://doi.org/10.2478/biorc-2021-0001 wight, r., arnott, g.a.w. (1834). prodromus florae peninsulae indiae orientalis. vol. 1. london: purbury, allen & co. wight, r. (1837). contributions to indian botany, no.1. on the genus impatiens. madras journal of literature and science, 5, 1–15. wight, r. (1839). illustrations of indian botany. vol. 1. madras: j.b. pharoah. wight, r. (1838–1853). icones plantarum indiae orientalis: or figures of indian plants. vol. 1–6. madras: j.b. pharoah. appendix 1 fig. 1. study area: a – india with tamil nadu districts, b – palani hills, c – nilgiri hills fig. 2. species of impatiens from nilgiri: a – impatiens leschenaultii (dc.) wall. ex wight & arn. (white flower), b – i. latifolia l., c – i. cuspidata wight & arn., d – i. parasitica bedd., e – i. fruticoasa lesch. ex dc., f – i. henslowiana arn. (light pink flower), g – i. modesta wight, h – i. viscosa bedd., i – i. clavicornu turcz., j – i. goughii wight (pink flower), k – i. scapiflora b.heyne ex wall., l – i. inconspicua (photo: s. jeevith and c. rajasekar) fig. 3. species of impatiens from nilgiri: a – impatiens dasysperma wight, b – i. tenella b.heyne ex wight & arn., c – i. fasciculata lam. (white flower), d – i. rufescens benth., e – i. denisonii bedd., f – i. minor (dc.) bennet, g – i. cordata wight, h – i. acualis arn., i – i. oppositifolia l., j – i. scabriuscula b.heyne ex wall., k – i. grandis b.heyne ex wall., l – i. tangachee bedd. (photo: s. jeevith and c. rajasekar) fig. 4. species of impatiens from nilgiri: a – impatiens nilgirica c.e.c.fisch., b – i. elegans bedd., c – i. levingei gamble ex hook.f., d – i. maculata wight, e – i. phoenicea bedd., f – i. herbicola hook.f., g – i. campanulata wight, h – i. tomentosa b.heyne ex wight & arn. (white flower), i – i. balsamina l. j – i. gardneriana wight, k – i. chinensis l., l – i. diversifolia wall. ex wight & arn. (photo: s. jeevith and c. rajasekar) fig. 5. species of impatiens from palani: a – impatiens balsamina l. (pink flower), b – i. campanulata wight, c – i. fasciculata lam. (pink flower), d – i. goughii wight (red flower), e – i. henslowiana arn. (white flower), f – i. dasysperma wight, g – i. leschenaultii (dc.) wall. ex wight & arn. (light orange flower), h – i. phoenicea bedd., i – i. tangachee bedd., j – i. tanyae r.kr. singh, arigela & kabeer, k – i. tomentosa b.heyne ex wight & arn. (pink flower), l – i. viscida wight (photo: ravi kiran arigela) appendix 2 tab. 1. species list of impatiens in nilgiri and palani hills (southern india) no. species distribution habitat occurrence in the study area 1. impatiens acaulis arn. southern india and sri lanka wet rocky slopes and moist soil walls of evergreen forests nilgiri 2. impatiens balsamina l. native to india and sri lanka. introduced in new world countries on open dry, wet rocky slopes and plains from foothills to 2500 m elevation nilgiri and palani 3. impatiens clavicornu turcz. endemic to southern western ghats (karnataka, kerala and tamil nadu) montane (shola) grasslands and wet rock slopes nilgiri 4. impatiens campanulata wight endemic to southern western ghats (kerala and tamil nadu) dripping rocky slopes and on tree trunks in shola forests and shola forest borders nilgiri and palani 5. impatiens chinensis l. native to asia and introduced in africa on open dry, wet rocky slopes, swamps, marshes and plains from foothills to 2500 m elevation nilgiri and palani 6. impatiens cordata wight endemic to southern western ghats (kerala and tamil nadu) evergreen forests along the streams and shola grasslands nilgiri 7. impatiens cuspidata wight & arn. endemic to southern western ghats (karnataka, kerala and tamil nadu) fringes of shola forests nilgiri 8. impatiens dasysperma wight endemic to southern western ghats (karnataka, kerala and tamil nadu) dripping rocky slopes in shola forests and wet evergreen forests borders nilgiri and palani 9. impatiens denisonii bedd. endemic to southern western ghats (kerala and tamil nadu) wet rocks and moist grasslands slopes in evergreen and shola forests nilgiri 10. impatiens diversifolia wall. ex wight & arn. endemic to southern western ghats (karnataka, kerala and tamil nadu) evergreen forests and moist grasslands, also found along the streams from 700–1200 m elevation nilgiri 11. impatiens elegans bedd. endemic to southern western ghats (kerala and tamil nadu) moist evergreen and shady places, also prefers dripping rocks in sholas and shola grasslands nilgiri 12. impatiens fasciculata lam. endemic to southern western ghats (karnataka, kerala and tamil nadu) dripping rocky slopes in shola forests, wet evergreen forests borders and swamps in grasslands nilgiri and palani 13. impatiens fruticosa lesch. ex dc. endemic to southern western ghats (kerala and tamil nadu) near streams of shola forests nilgiri 14. impatiens gardneriana wight endemic to southern western ghats (karnataka, kerala and tamil nadu) wet evergreen forests and found along the margins of shola forests nilgiri 15. impatiens goughii wight endemic to southern dripping rocky slopes at shola nilgiri and palani western ghats (karnataka, kerala and tamil nadu) forest borders and shola grasslands from 1800–2500 m elevation 16. impatiens grandis b.heyne ex wall. karnataka, kerala, tamil nadu and sri lanka moist shady places in shola forests nilgiri 17. impatiens henslowiana arn. karnataka, kerala, tamil nadu and sri lanka dripping rocky slopes in shola forests and wet evergreen forests borders nilgiri and palani 18. impatiens herbicola hook.f. endemic to southern western ghats (kerala and tamil nadu) moist grasslands, swampy areas and wet rocky slopes from 1700–2200 m elevation nilgiri 19. impatiens inconspicua benth. ex wight & arn. endemic to western ghats (goa, karnataka, kerala, maharashtra and tamil nadu) dripping rocky slopes of shola forest borders and shola grasslands from 1800–2500 m elevation nilgiri 20. impatiens latifolia l. endemic to western ghats (karnataka, kerala, maharashtra and tamil nadu) fringes of shola forests nilgiri 21. impatiens leschenaultii (dc.) wall. ex wight & arn. endemic to southern western ghats (kerala and tamil nadu) rocky slopes of shola forest borders from 1800–2500 m elevation nilgiri and palani 22. impatiens levingei gamble ex hook.f. endemic to southern western ghats (kerala and tamil nadu) wet rocky slopes of shola forests and shola grasslands nilgiri 23. impatiens maculata wight endemic to southern western ghats (kerala and tamil nadu) moist evergreen and shola forests along the streams nilgiri 24. impatiens minor (dc.) bennet endemic to western ghats (goa, karnataka, kerala, maharashtra and tamil nadu) semi-evergreen forests and wet evergreen forests from 1600–2000 m elevation nilgiri 25. impatiens modesta wight endemic to southern western ghats (kerala and tamil nadu) grasslands and rocky slopes of moist evergreen forests nilgiri 26. impatiens nilgirica c.e.c.fisch. endemic to southern western ghats (kerala and tamil nadu) dripping rocks of shola grasslands nilgiri 27. impatiens oppositifolia l. india, myanmar, sri lanka and thailand dripping rocky slopes of shola forest borders and shola grasslands from 1800–2500 m elevation nilgiri 28. impatiens parasitica bedd. endemic to southern western ghats (kerala and tamil nadu) mostly epiphytic on shola trees, also found on wet rocks nilgiri 29. impatiens phoenicea bedd. endemic to southern western ghats (kerala and tamil nadu) along the stream of shola forests and fringes of moist evergreen forests nilgiri and palani 30. impatiens rufescens benth. endemic to southern western ghats (kerala and tamil nadu) mostly in swampy areas and shola grasslands at 2100–2300 m elevation nilgiri 31. impatiens scabriuscula b.heyne ex wall. endemic to western ghats (karnataka, kerala, maharashtra and tamil nadu) wet slopes and dripping rocks of shola forest and grasslands nilgiri 32. impatiens scapiflora b.heyne ex wall. endemic to southern western ghats (karnataka, kerala and tamil nadu) dripping rocky slopes of evergreen forests and grasslands nilgiri 33. impatiens tangachee bedd. endemic to southern western ghats (kerala and tamil nadu) dripping rocky slopes in shola forests nilgiri and palani 34. impatiens tanyae r.kr. singh, arigela & kabeer endemic to southern western ghats (kerala and tamil nadu) swampy areas and along the streams of shola grasslands palani 35. impatiens tenella b.heyne ex wight & arn. endemic to western ghats (karnataka, kerala, maharashtra and tamil nadu) shola forests and shola grasslands nilgiri 36. impatiens tomentosa b.heyne ex wight & arn. endemic to western ghats (goa, karnataka, kerala, maharashtra and tamil nadu) dripping rocky slopes of shola forest borders and shola grasslands from 1800–2500 m elevation nilgiri and palani 37. impatiens viscida wight endemic to southern western ghats (kerala and tamil nadu) dripping rocky slopes of shola forests and shola forest borders palani 38. impatiens viscosa bedd. endemic to southern western ghats (kerala and tamil nadu) dripping rocky slopes of shola forest borders and shola grasslands 1800–2500 m elevation nilgiri abstract a comprehensive checklist of the genus impatiens riv. ex l. (balsaminaceae) in nilgiri and palani of southern western ghats, india is presented. after thorough field explorations in these areas from 2012 to 2020 in different seasons, 38 species of impatiens are recorded, in which 6 species are endemic to western ghats and 26 species are endemic to southern western ghats of india. horticultural potential of 18 endemic species of balsam is also discussed. key words: balsams, endemic, grasslands, shola forests, western ghats received: [2022.05.08] accepted: [2022.07.26] wykaz gatunków z rodzaju impatiens riv. ex l. (balsaminaceae) z nilgiri i wzgórz palani, w południowej części ghatów zachodnich (indie) w artykule zamieszczono listę gatunków z rodzaju impatiens riv. ex l. (balsaminaceae), odnotowanych w nilgiri i na wzgórzach palani (południowa część ghatów zachodnich w indiach). po dokładnych badaniach terenowych, przeprowadzonych w różnych porach roku w latach 2012–2020, zarejestrowano na analizowanym obszarze 38 gatunków niecierpka, w tym 6 gatunków endemicznych dla całych ghatów zachodnich i 26 gatunków dla południowej części ghatów zachodnich. krótko omówiono również potencjał ogrodniczy 18 endemicznych gatunków niecierpków. słowa kluczowe: niecierpki, endemiczne, lasy shola, łąki, ghaty zachodnie information on the authors s. jeevith https://orcid.org/0000-0002-1003-8016 he is biologist and interested in forest ecology and plant taxonomy, ethnobotany, flora fauna interactions, wildlife explorations in different landscapes of western ghats. ravi kiran arigela http://orcid.org/0000-0001-5804-3423 he is interested in plant taxonomy, ecology, plant – bird and plant – animal interactions. his study deals with the ecosystems and biodiversity of them; in particular endemic and threatened species. rajeev kumar singh https://orcid.org/0000-0002-0136-9243 his special interests are plant taxonomy, plant nomenclature and biodiversity. he has worked on tiger reserves and protected areas in india. k. althaf ahamed kabeer https://orcid.org/0000-0002-7547-1363 he is an agrostologist and working on the grasses of india. he has investigated and documented the grasses of tamil nadu state of india. c. rajasekar https://orcid.org/0000-0002-9134-2828 he is interested in plant taxonomy, molecular biology and biodiversity conservation. he is working as teaching faculty in botany and passionate on research work. he has done dna sequencing on several plant species in western ghats. c. kunhikannan he is well known specialist in plant taxonomy, ecology and biodiversity assessment. he has worked on different plant taxa in various landscapes and completed many projects across the country. 34 annales universitatis paedagogicae cracoviensis studia naturae, 5: 34–57, 2020, issn 2543-8832 doi: 10.24917/25438832.5.3 ksenia strzeżoń institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland; ksenia1922@gmail.com evaluation of the grassland sward floristic composition of the wadowice commune introduction pastures and meadows are mainly anthropogenic (semi-natural) communities, which were created thanks to man. �eir existence is conditioned by appropriate management methods, such as mowing, grazing or fertilisation (kornaś, 1990; kornaś, dubiel, 1991; barabasz, 1994, 2011). deforested areas, which were not suitable for arable �elds due to steep location, high humidity or shallow and relatively infertile soil, were selected for pastures. meadows were established in places that were relatively easily accessible, although not too distant from the settlement, so that they could be mowed, dried, and conveniently brought hay, which in winter was the only source of feed for farm animals (kocan, jacniacki, 1980; falkowski, 1983). proper care of meadows and pastures is necessary because it prevents, for example, the occurrence of undesirable species (włodarczyk, 1983). high concentrations of organic compounds have a negative e�ect on many meadow perennials, even those relatively resistant to large amounts of nitrogen, e.g. species from the fabaceae lindl. family. �ese species are eagerly eaten by cattle because they have high nutritional value. too much manure provides an opportunity for the expansion of nitrophilous species not belonging to the fodder plants, which also adversely a�ects the usage value of a sward (kornaś, dubiel, 1991; bobbink et al., 1998; radkowski, barabasz-krasny, 2008). additionally, the lack of light and oxygen can cause damage and rot in many fodder plants. in general, weed infestation of meadow sward reduces its productivity. �e most common causes of this phenomenon are improper usage, excess humidity or soil drought, or incorrect nutrient proportions in soil (haghighi et al., 2010; teague et al., 2011). care treatments and a rationally used mowing and grazing system contribute to improving the �oristic composition of green utility areas (włodarczyk, 1983; baraevaluation of the grassland sw ard floristic com position of the w adow ice com m une 35 basz, 1994; bakker, berendse, 1999; díaz et al., 2007). moderate grazing has a positive e�ect on the quality of meadows and pastures because weeds omitted by cattle become visible, and they can easily be mechanically removed (grynia, 1974). additionally, soil kneading by animals helps to get rid of weed roots that are not resistant to pressure. nibbling by animals deprives plants of the aerial parts responsible for photosynthesis, which weakens the growth of many non-resistant weeds (nowiński, 1970; pauler et al., 2019). mowing can contribute to the growth of tall grasses and reduce the number of perennials. during the �owering phase and in�orescence emergence, or earing, tall grasses drown out lower growing weeds and less e�cient low grasses. it is best to utilize mowing throughout the season on a separate part of the meadow, collecting up to three swaths (falkowski, 1983). however, it should be remembered that intensive use of meadows can lead to species poorness (kornaś, 1990; sienkiewicz, 2010). �e aim of this study was to (1) analyse the �oristic composition, (2) the characteristics and (3) the agricultural suitability of meadows and pastures of the wadowice commune. to date, no analyses related to this topic have been carried out in this area even though agriculture in the commune of wadowice is of great importance. characteristic of the study area �e urban and rural commune of wadowice, together with the city of wadowice (49°52ʹ57ʺ n 19°29ʹ40ʺ e), is located in southern poland in the lesser poland voivodeship. �e area of the community is 113 km², of which 102 km² is rural and the remainder is city – wadowice (local development strategy..., 2014). kondracki (2011) classi�ed the analysed area as extending to the carpathian and subcarpathian provinces, the macroregions of the western beskids and the west beskidian foothills, as well as to the mesoregions of the little beskids and the wieliczka foothills. �e commune consists of the following land usage elements: agricultural lands (include arable areas, grasslands, orchards) and fallows, forest land, urbanised areas, as well as places under water (fig. 1). �e little beskids can be described as a plateau approximately 35 km long and 12 km wide, with an average height of 800 m a.s.l. from the west, it is limited by the wilkowicka gate. �e northern border with the silesian beskids is demarcated by the bielsko-biała-wadowice road, while the southern and eastern extremities are river valleys, including łękawka, kocońka, tarnawka, krzeszowski potok, and skawa. �e wieliczka foothills, which are also associated with the study area, cover an area extending east of the skawa river to the regions of wieliczka and gdów. �e western part of the wieliczka foothills is important for the wadowice region. altitudes are approximately 200–550 m a.s.l. �e peaks do not have an island character and are connected with each other by short ridges, forming latitudinal bands (siemionow, 1984). k se ni a s tr ze żo ń 36 soils found in the area of the wadowice commune are mainly shallow, acidic and stony. most are pseudo-podzols and brown soils. skeletal and stony soils are also present. erosion in this area occurs faster than in the lowlands, due to the greater slope and signi�cant rainfall. many river valleys, as well as the sources and streams present here, favour the development of fertile valley soils. �ere are fertile soils in the skawa valley, and the rest of the commune is covered by podzol and brown soils, as well as rusty soils and aerosols (bednarek, skiba, 2015). �e area of the wadowice commune, located in the little beskids zone, has a moderate-cold, warm and slightly cold climate (hess, 1965); respectively, the �rst is characterised by temperatures from 4°c to 6°c and the second from 6°c to 8°c. only in the highest parts can you �nd the features of a cold climate, with temperatures up to 2°c. part of the wieliczka foothills has a milder climate, which is illustrated by a higher average temperature of up to 9°c. an important climatic aspect of the area is the phenomenon of levelling summer-winter di�erences; the summers are relatively cold and the winters mild, with little snow cover (woś, 1999). material and methods �e study was conducted in june 2016 in the area of the wadowice commune. a total of 51 plots with an area of 5 m × 5 m (25 m2 each) were designated on the meadows and pastures in all municipalities’ villages (fig. 2). a botanical evaluation of meadow sward was performed on each plot using the klapp (1962) estimation method. using this method, the share of species in the square was estimated, with an accuracy of 1%, fig. 1. �e structure of land use in the wadowice commune (source: local development strategy..., 2014) evaluation of the grassland sw ard floristic com position of the w adow ice com m une 37 recording all species in the order of grasses, legumes, and others. �en the percentage share of individual species within each group was estimated, starting from the most abundant. species with an occurrence of less than 1% received the designation “+”. plants that were not directly identi�ed in an area were collected and then identi�ed in laboratory conditions. szafer et al. (1986) and nawara (2012) were used as resources to recognise plants. �e nomenclature of vascular plants and ferns used in the study was in line with mirek et al. (2002). analyses were carried out using the turboveg program, which is a standard botanical database in europe. it allows, among other things, for the collection and processing of information about the composition and the share of species in plots (zarzycki, 2009). floristic lists of all plots were entered into the database and subjected to numerical classi�cation based on the percent coverage of species. for classi�cation, similarities between plots were calculated using the van der maarel coe�cient (westho�, van der maarel, 1978), according to the formula: fig. 2. distribution of studied plots (1–51) in the wadowice commune area k se ni a s tr ze żo ń 38 where r is the similarity between the plots and x, y are vectors of the species percentage occurrence in the plots. for grouping, the ward method (minimum variance clustering) was used – a value of 0.5 was assumed as a “+” (dzwonko, 2007). for classi�cation, the mulva-5 program was used (wildi, orlóci, 1996). based on the resulting dendrogram, tables were constructed that were used for detailed analysis of the botanical composition of similar groups of plots. to characterise the habitat of similar plots, indicator numbers were used (ellenberg et al., 1992), which is the recommended method of indirect inference (roo-zielińska, 2014). weighted average ellenberg’s indicators values were calculated, in relation to the percentage share of plants for: light (l), humidity (f), soil reaction (r), and nitrogen content in soil (n). �en arithmetic means were calculated for groups of similar plots. in order to determine the utility values of the studied area, the usage value of the meadow (uvm) was calculated for all plots, and then individual plots were assigned to the utility scale according to filipek (1973). �e calculations were made on the basis of the use value index (uvi), which is assigned to each species ful�lling a utility role, especially forage plants. �e uvm index was calculated as the sum of the products of the percentage share of each species and its uvi divided by 100 (łyszczarz, 2014). results in the dendrogram of similarity (fig. 3) two main groups were separated, with one including three smaller subgroups of plots. in total, four groups of similar plots were distinguished, which were marked with numbers i–iv. �ese groups were named depending on the species that gave the plot appearance: i – plots with phleum pratense and vicia cracca, ii – plots with dactylis glomerata and trifolium repens, iii – plots with holcus lanatus, and iv – plots with arrhenatherum elatius (appendix 1 – tab. 1–4). comparing the average values of ellenberg’s indicators calculated for groups of similar plots, it was found that group i is characterised by the highest humidity (f) and nitrogen content (n) among all analysed groups, while in group iv indicators for light (l) and soil ph (r) showed the highest values. all of the analysed groups were in the same range of the ellenberg’s indicators values: moderately sunny places, with fresh soils and moderately moist, with a fairly high nitrogen content and a ph corresponding to slightly acidic soil (appendix 1 – tab. 5). evaluation of the grassland sw ard floristic com position of the w adow ice com m une 39 �e studies of the botanical composition of the plots, in terms of use, revealed that most had a good and average uvm. from group iv, only one plot showed very good usage value, and from group i only one plot had poor usage value (appendix 1 – tab. 6). most plots of good usage quality were found in group iv, and the fewest were found in group iii. overall, more than 50% of the plots had good usage value, and about 40% were characterised by the average value of the analysed indicator (fig. 4). characteristic of distinguished groups of plots group i – plots with phleum pratense and vicia cracca (appendix 1 – tab. 1) 16 plots were included in the group. among the grasses, phleum pratense had the largest share in the plots – from 1 to 40% coverage per 12 occurrences. other species, such as poa pratensis, anthoxanthum odoratum, arrhenatherum elatius, dactylis glomerata, and alopecurus pratensis, had coverage of up to 60% in plots, but showed lower numbers of occurrences – from 5 to 9. among legumes vicia cracca was found most o�en in the plots but with a relatively low coverage – from 1 to 30%. less o�en but with larger coverage, trifolium pratense and t. repens occurred here – from 5 to 50% coverage per 4–8 occurrences. among other species, the highest number of occurrences was recorded for taraxacum o�cinale – 8, with coverage up to 10%. certain crops’ weeds occur in some plots (e.g. papaver rhoeas – approx. 1%) with some having high coverage, e.g. matricaria maritima ssp. inodora (up to 45%). �e average uvm of these plots was a medium level (appendix 1 – tab. 6; fig. 4). fig. 3. classi�cation of the studied plots based on the percentage scale of species coverage: i – plots with phleum pratense and vicia cracca, ii – plots with dactylis glomerata and trifolium repens, iii – plots with holcus lanatus, iv – plots with arrhenatherum elatius k se ni a s tr ze żo ń 40 group ii – plots with dactylis glomerata and trifolium pratense (appendix 1 – tab. 2) 12 plots were in the group. of the grasses, the most occurrences were of dactylis glomerata – 12, reaching 10 to 40% coverage in plots. �e second most common grass was poa pratensis, which was recorded in 10 plots at 5 to 60% coverage. among the legume plants, trifolium repens deserves attention, which grew on all studied plots with a coverage of 2 to 30%. t. pratense (7 plots) with up to 15% coverage was also common. legume species, such as vicia cracca, medicago lupulina, and lathyrus pratensis, which occurred in 2 to 5 plots with a coverage of up to 5%, had a smaller share. of the other plants growing on the group ii plots, the following grazing species were notable: ranunculus acris, taraxacum o�cinale, plantago lanceolata, and alchemilla monticola, occurring from + to 10–15% in plots. plantago major, typical for trampled sites, was also present in 4 plots – with coverage up to 10%. in terms of average uvm, these plots also were classi�ed at a medium level (appendix 1 – tab. 6; fig. 4). group iii – plots with holcus lanatus (appendix 1 – tab. 3) 7 plots were included in this group. holcus lanatus exhibited the largest share in this group, growing in all 7 plots, with coverage from 10 to 35%. �e second most common grass, present in 6 plots, was poa pratensis, with coverage from 2 to 35%. of the legumes, the following species were notable: trifolium repens, t. pratense, and lathyrus pratensis, which, although not present in all plots (from 3 to 5), their coverage was fig. 4. comparison of the percentage participation of plots from six usage categories (according to filipek, 1973) between the groups of meadow distinguished on the study area: i – plots with phleum pratense and vicia cracca, ii – plots with dactylis glomerata and trifolium repens, iii – plots with holcus lanatus, iv – plots with arrhenatherum elatius evaluation of the grassland sw ard floristic com position of the w adow ice com m une 41 relatively large – up to 30%. among other species, taraxacum o�cinale had the largest share (in 5 plots, with coverage up to 20%) and plantago major, p. lanceolata, and leontodon hispidus all reached a maximum of 10% coverage in plots. �ey are grazing species, resistant to biting and trampling. in some plots, weeds from arable �elds were recorded: chamomilla recutita and matricaria maritima ssp. inodora (in 3 and 2 plots, respectively), with coverage of 1%. as in previous groups, the average uvm of these plots was a medium level (appendix 1 – tab. 6; fig. 4). group iv – plots with arrhenatherum elatius (appendix 1 – tab. 4) �ere were 16 plots in this group. grasses were the most common in this group: arrhenatherum elatius, phleum pratense, dactylis glomerata, and poa pratensis. a. elatius was recorded in all 16 plots, with coverage of 10 to 50%, with the other grasses noted above being found in 13, 12, and 10 plots, respectively, with coverage of up to 60% (d. glomerata). of the legume plants, vicia cracca (11 plots) and trifolium repens (7 plots) had the largest share. �eir coverage was up to a maximum of 10–30% but usually 1–10%. of the other plants, achillea millefolium was recorded the most (in 15 plots, with coverage 2 to 15%). other noticed plants included ranunculus acris, taraxacum o�cinale, stellaria graminea and plantago major, associated with pastures. although buttercup was present in 12 plots, its coverage was small – from >1 to 2%. �e average uvm of this group of plots was at the level of good (appendix 1 – tab. 6; fig. 4). discussion in meadows and pastures you �nd not only species that have a positive e�ect on the quality of fodder but also those that can reduce the utility value and above all threaten the health and life of animals, as they are o�en poisonous. �ey can also be harmful to the vegetation of meadows and pastures and hinder hay harvesting or animal grazing (włodarczyk, 1983; dobrzański, 2009). among the poisonous species commonly found in meadows and pastures are representatives of the ranunculaceae juss. family. buttercups contain poisonous glycosides and alkaloids, which are dangerous to animals. �ere are relatively many on pastures, as they are le� by grazing cattle (nowiński, 1970). for the plots in the wadowice commune grassland that was analysed, ranunculus acris occurred in all four groups of meadows (appendix 1 – tab. 1–4). in group i with phleum pratense and vicia cracca, buttercup was present in only 3 plots, and in the other groups it occurred frequently but with low coverage – usually from + to 2% (sporadically 5–10%). of note, buttercups are dangerous for animals only when fresh. most o�en poisoning occurs in the spring, when only its leaves are present in the sward. when �owers appear, k se ni a s tr ze żo ń 42 the animals bypass this plant. in hay, buttercups lose their toxic properties and are not dangerous (nawara, 2012). stellaria graminea is another toxic species. chickweed belongs to the caryophyllaceae juss. family, and its herb is particularly dangerous for animals (grynia, 1974). however, the share of chickweed found in the analysed plots was insigni�cant – in groups ii and iii it sporadically covers up to 5% and in other groups up to 2% (appendix 1 – tab. 1–4). other undesirable species in the meadows include rumex acetosa (polygonaceae juss.), veronica chamaedrys (scrophulariaceae juss.), equisetum arvense, and e. palustre (equisetaceae michx. ex dc.) (nowiński, 1970; nawara, 2012). �eir share in the studied area was relatively small (up to 2%), thus not of great importance for the overall usage value of the analysed plots. only sorrel sporadically reached coverage of up to 5% in the group ii plots with holcus lanatus (appendix 1 – tab. 2). it is characterised by a high content of calcium oxalate, which is deposited in the organs. horsetails, in addition to being hard due to the presence of silica, also contain substances that have a toxic e�ect on the functioning of organisms (grynia, 1974). �e larger share of these plants in the plots may be the result of being bypassed by grazing animals. similarly, the rushes juncus e�usus or j. in�exus (juncaceae juss.) are species clearly bypassed by animals due to the hardness of the stems. in the presence of intensive defoliation of fodder plants and trampling, to which the rushes are quite resistant, they can colonise the habitat without major barriers (barabasz, 1994). strong growth of rhizomes and high durability of seeds (even up to 60 years) makes competition of other plants with rushes very di�cult (nowiński, 1970). however, this unfavourable phenomenon was not observed in the study area, as rushes occurred here relatively rarely and with low coverage up to 5% (appendix 1 – tab. 1–4). it is worth noting that rushes belong to the group of weeds that make mowing di�cult (włodarczyk, 1983). in some of the study plots, mainly in the �rst group, weeds characteristic of arable �elds also occurred: papaver rhoeas, matricaria maritima ssp. inodora, or agrostemma githago (appendix 1 – tab. 1). �eir presence indicates the close proximity of the studied meadows to arable �elds, or they may be a remnant of former crops that have since been converted to grassland (barabasz-krasny, 2011). cultivated weeds can easily in�ltrate neighbouring areas and, although they are quite sensitive to other habitat conditions, they persist quite well on the outskirts of meadows, where the sward is not so dense (dubiel, 1984). according to picket and cadenasso (2005), the remainder of the last crop (some weeds) can be observed in plant communities for about 10 years a�er the crop has been abandoned. for proper management of grasslands, the care of meadows and pastures is just as important as rational usage (falkowski, 1983; aldrich, 1984; teague et al., 2011). for example, a weakly compact sward is bad for young grasses and plants that are poorly evaluation of the grassland sw ard floristic com position of the w adow ice com m une 43 rooted in soil. meanwhile, if a sward is too dense it degrades and reduces the yield of the meadow (adler et al., 2001). both mounds of moles and the accumulation of animal droppings cause gaps in the continuity of meadow sward. �erefore, plants in these types of areas, although they grow luxuriantly, are bypassed and reluctantly eaten by animals due to the unpleasant smell (kocan, jacniacki, 1980). as a result, weeds of the meadow sward are favoured and species such as thistle (cirsium sp. asteraceae dum.) and nettle urtica dioica (urticaceae juss.) begin to appear (nowiński, 1970; dobrzański, 2009). in this study area, the share of these types of weeds was relatively small (appendix 1 – tab. 1–4). however, in the analysed meadows, a fairly large group of plants were weeds, the addition of which is bene�cial for animals. for example, the presence of up to 5% coverage of species such as achillea millefolium, alchemilla monticola, taraxacum of�cinale, plantago major, or p. lanceolata increases the usage value of meadows and pastures. yarrow improves the digestion of animals, and lady’s mantle has many nutrients and positive e�ects on milk quality. dandelion is also a fodder herb and has medicinal properties. however, in large quantities, just like plantains, lady’s mantle, and yarrow, it can be poisonous to animals (nowiński, 1970; nawara, 2012). a. monticola occurred at an appropriate level in the study area in group ii plots, some of which are used as pastures. �e exception was 1 plot of this group, where it reached 10% coverage. similarly, p. major occurred at a level above 5% only in 1 plot of each group, and p. lanceolata occurred in group ii and iii in 2 plots and 1 plot, respectively. t. o�cinale, found in 15 plots from all four groups, also had a larger share, 10–20% of coverage (appendix 1 – tab. 1–4). similarly, a. millefolium occurred in as many as 10 plots, also with high coverage. hence, it would be worth undertaking actions to balance the share of these plants in some plots. research indicates that direct weed control with chemicals has a worse e�ect on meadows and pastures than slower but thoughtful and rational prevention of their spread (falkowski, 1983; heap, 2014). incorrect usage promotes larger concentrations of weeds and weakens meadows sward. both an excess or a de�ciency in grazing and mowing adversely a�ect the meadows (kornaś, 1990; barabasz, 1994; załuski, 2002; bator, 2005; kompała-bąba, bąba, 2007; díaz et al., 2007; pauler et al., 2019). �e improvement of species composition can be achieved by proper mowing, which prevents the �owering phase of some undesirable species. moderate grazing can also contribute to improving the quality of the meadow, as it helps in the mechanical removal of harmful plants, as mentioned previously. �e most favourable composition of meadow sward in the study area occurred in plots included in group iv with arrhenatherum elatius (appendix 1 – tab. 4, fig. 4). �ere was a fairly good proportion of legumes, which was also re�ected in the usage values of those plots, whose average uvm was at a good level (appendix 1 – k se ni a s tr ze żo ń 44 tab. 6). in addition to false oat-grass, other forage grasses were abundant here: phleum pratense, dactylis glomerata and poa pratensis. �ese species are characterised by high usage value and fertility. �ey are the most desirable grass species in grassland (jagła, 2001; nawara, 2012). tall grasses, such as cocksfoot, timothy, or false oat-grass, are more common in meadow areas because they tolerate mowing better than being trampled and bitten by grazing animals (barabasz, 1994). �ey are usually associated with relatively fertile places, which can also be seen in the study area (appendix 1 – tab. 5). �ey are excellent for fodder, both fresh and in the form of hay. bluegrass is even more valuable, additionally resistant to various unfavourable habitat factors and giving high yields. it prefers medium or high humidity and fertile environments (falkowski, 1983). on the other hand, group iii with holcus lanatus had the relatively weakest usage value in the analysed commune – the fewest number of plots of good usable quality were noted here (appendix 1 – tab. 6; fig. 4). �e velvet grass that dominates here has a fairly low fodder value as it is not suitable for animal feed because of its hairy stalks. its hay is also too tomentose and slight (nawara, 2012). however, in this group there are also species such as poa pratensis or arrhenatheretum elatius, as well as legume species that help to maintain the usage value at an average level (appendix 1 – tab. 3). observations carried out during �eld studies con�rm the fact that in the analysed area, the meadow management was rather balanced, although care treatments would certainly improve the usage values of many meadow plots. to improve the quality of meadows and pastures, care should be taken to ensure proper conditions for high-quality plants and to reduce undesirable species. most o�en, these type of management methods include maintaining the compactness of the turf (so that it is not too loose or compact); cleaning works in sward – clearing mounds of mole; removing animal droppings; removing weeds; as well as rational mowing and grazing. properly fertilised soil can also contribute to an increase in crops – fertilisation with nitrogen, phosphorus, and potassium, especially in the case of grasses, and fertilisation with magnesium, in the case of legumes. care should be spread over time – cleaning work in autumn and fertilisation and care for proper drainage management in early spring (falkowski, 1983; kocan, jacniacki, 1980). conclusions (1), (2) on the basis of �oristic analyses, it can be stated that the species composition was found to be quite diverse in utility meadows and pastures of wadowice commune; they can be classi�ed as moderately sunny meadows, with average soil moisture (fresh), slightly acidic, and quite rich in nitrogen. (3) �e studied grasslands had average (groups: i with phleum pratense and vicia cracca, ii with dactylis glomerata and evaluation of the grassland sw ard floristic com position of the w adow ice com m une 45 trifolium repens, iii with holcus lanatus) and good (group iv with arrhentherum elatius) usage values. balanced fertilisation could increase the usage value of the meadows and pastures examined. however, the improvement of the quality of the botanical composition of medium fodder value plots could be achieved through appropriate comprehensive care and sustainable use. con�ict of interest �e author declares no con�ict of interest related to this article. references adler, p., ra�, d., lauenroth, w. (2001). �e e�ect of grazing on the spatial heterogeneity of vegetation. oecologia, 128, 465–479. https://doi.org/10.1007/s004420100737 aldrich, r.j. 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[in polish] k se ni a s tr ze żo ń 48 appendix 1 tab. 1. group i – plots with phleum pratense and vicia cracca successive no. of plots 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 number of occurrenceno. plot in the area 34 27 33 49 43 46 32 41 18 13 4 19 42 14 35 8 number of species in plot 11 12 10 8 14 9 13 9 12 14 13 15 14 12 11 13 grass phleum pratense 10 35 30 2 10 10 35 5 40 . 20 . 1 10 . . 12 poa pratensis 20 10 . 2 10 10 10 2 10 . . . 10 . . . 9 anthoxanthum odoratum 8 . 10 . . . 10 1 10 5 5 20 . . . . 8 arrhenatherum elatius . . 5 2 . . . 10 . 20 . . . . 40 35 6 dactylis glomerata . 10 2 15 25 . 20 . . . . . . . . . 5 alopecurus pratensis . . . 60 10 40 . . . . . 35 . . 10 . 5 legumes vicia cracca 30 10 30 . 10 2 . 2 2 1 . 2 10 10 10 + 13 lathyrus pratensis 10 10 . 15 2 2 2 . . . 5 . 1 . . . 8 trifolium pratense . . . . . 25 . 10 20 30 40 10 25 . . 20 8 t. repens . . . . . . . 50 10 25 5 . . . . . 4 medicago lupulina . . . . . . . . . . 5 10 . 10 . . 3 others taraxacum o�cinale 5 . 10 . . 1 . . 2 5 5 10 . 1 . . 8 plantago major 5 3 . . 2 . . 10 . . 5 . . 1 . . 6 aegopodium podagraria . + . 2 + 10 8 . 2 . . . . . . . 6 papaver rhoeas . + . . . . . . . 1 + . . 1 + . 5 matricaria maritima ssp. inodora . . . . . . . . . 1 . . 40 45 2 + 5 symphytum o�cinale . . . . + 1 . . . . . 2 + . . 10 5 centaurea jacea . . + . . . . . . . . . + 10 + . 4 achillea millefolium 10 20 . . . . . . . . . . . 10 . 10 4 rumex acetosa + . + . + . . . . . . . . . + . 4 equisetum arvense . + . . . . . . + + . . . + . . 4 cirsium rivulare . . . . 2 . 2 . . . . . . . 1 + 4 stellaria graminea . . . . . . . . 2 + . 2 + . . . 4 hypericum perforatum + . . . . . . . 2 . . 2 . . . . 3 leontodon hispidus + . 10 . . . . . . . . . . . 1 . 3 ranunculus acris . + . . . . . . . . 2 . . . . + 3 urtica dioica . . . 2 10 . + . . . . . . . . . 3 cirsium arvense . . . . . . + . . . . + + . . . 3 capsella bursa-pastoris . . . . . . . . . 5 . . 10 . 10 . 3 convolvulus arvensis . . . . . . 2 . . . . 2 . . . . 2 potentilla anserina . . . . . . . 10 . . . 2 . . . . 2 plantago lanceolata . . . . . . . . . . 5 . . 1 . . 2 evaluation of the grassland sw ard floristic com position of the w adow ice com m une 49 lychnis �os-cuculi . . . . . . . . . . 1 . . . . 1 2 chamomilla recutita . . . . . . . . . + . . 1 . . . 2 myosotis arvensis . . . . . . . . . . . . 1 1 . . 2 sporadic species – grass: phragmites australis 32: 1; elymus repens 13: 5; trisetum �avescens 35: 25. others: campanula patula 27: +; geranium dissectum 33: +; galium aparine 43: 10; mentha longifolia 43: 10; oxalis acetosella 32: 2; solidago gigantea 32: 8; veronica chamaedrys 18: +; ajuga reptans 13: +; campanula trachelium 4: +; agrimonia eupatoria 19: +; armoratia rusticana 19: 2; artemisia vulgaris 19: 2; agrostemma githago 42: +; daucus carota 8: 2; galium mollugo 8: 10; juncus e�usus 8: 2; lysymachia vulgaris 8: 10. k se ni a s tr ze żo ń 50 tab. 2. group ii – plots with dactylis glomerata and trifolium repens successive no. of plots 1 2 3 4 5 6 7 8 9 10 11 12 number of occurrenceno. plot in the area 40 7 48 37 26 5 22 29 28 23 21 36 number of species in plot 12 12 14 16 14 17 13 13 17 13 19 12 grasses dactylis glomerata 15 10 30 10 40 30 15 40 40 20 15 10 12 poa pratensis 60 40 10 10 5 20 5 . . 10 5 10 10 alopecurus pratensis . . 10 20 . . 15 . . 2 1 . 5 phleum pratense . . . . . . . 10 2 5 5 25 5 festuca pratensis . . . . 10 . . . 10 10 5 . 4 holcus lanatus . 5 . . . . 1 . . . . . 2 anthoxanthum odoratum . . . 5 . 5 . . . . . . 2 legumes trifolium repens 2 10 5 30 10 8 5 10 10 20 15 10 12 t. pratense . . 5 5 10 5 15 . 10 . . 10 7 vicia cracca 2 . . . + 5 . 2 2 . . . 5 medicago lupulina 2 . . 1 + . . . . . . 2 4 lathyrus pratensis . . . . . . . 2 2 . . . 2 others ranunculus acris + 1 + 1 1 1 5 2 + . 10 + 11 taraxacum o�cinale 10 10 10 5 10 . . 2 2 2 15 10 10 plantago lanceolata . 10 1 5 1 5 . . . . 15 . 6 alchemilla monticola . . 10 + . . 1 2 2 . + . 6 achillea millefolium . . . 1 5 5 5 2 2 . . . 6 rumex acetosa . . . . . 5 + 1 2 5 1 . 6 equisetum arvense . + . . + 1 . . + . + . 5 aegopodium podagraria . . . 1 . 1 . . . 20 5 2 5 stellaria graminea 2 . 5 . . 1 . . . . + . 4 plantago major . 10 5 1 5 . . . . . . . 4 cirsium rivulare . . . . . . 5 25 10 . 5 . 4 galium mollugo . . . . . . . 1 2 2 . 10 4 capsella bursa-pastoris 2 . . . . . 1 . . 2 . . 3 veronica chamaedrys . . . + . + . . . 2 . . 3 urtica dioica 1 1 . . . . . . . . . . 2 matricaria maritima ssp. inodora 2 . . . . . 25 . . . . . 2 myosotis palustris + . . . . . . . . . + . 2 glechoma hederacea . . 5 1 . . . . . . . . 2 campanula patula . . 1 . . + . . . . . . 2 potentilla anserina . . . . . . . 2 2 . . . 2 lychnis �os-cuculi . . . . . . . . . 2 + . 2 sporadic species – grasses: arrhenatherum elatius 48: 1. others: ranunculus repens 7: +; symphytum o�cinale 7: 1; petasites albus 26: +; juncus in�exus 5: 5; pteridium aquilinum 5: +; oxalis acetosella 28: 2; galium aparine 21: +; dactylorhiza majalis 21: +; convolvulus arvensis 36: 2; armoratia rusticana 36: 10. evaluation of the grassland sw ard floristic com position of the w adow ice com m une 51 tab. 3. group iii – plots with holcus lanatus successive no. of plots 1 2 3 4 5 6 7 number of occurrenceno. plot in the area 31 30 11 6 25 50 24 number of species in plot 14 16 14 12 15 13 15 grasses holcus lanatus 30 30 30 20 10 30 35 7 poa pratensis 5 8 2 10 35 . 5 6 arrhenatherum elatius 10 10 5 30 . . . 4 dactylis glomerata . . 10 . . 2 . 2 legumes vicia cracca 1 1 . 5 2 1 + 6 trifolium pratense . . 20 1 5 10 5 5 t. repens 20 20 . . . 30 8 4 lathyrus pratensis . . 10 10 10 . . 3 medicago lupulina . . . 5 . . 2 2 others ranunculus acris 1 2 1 10 2 . . 5 taraxacum o�cinale 20 10 5 1 10 . . 5 plantago major 5 10 5 . 5 2 . 5 leontodon hispidus 1 1 . . + 1 10 5 plantago lanceolata . . 5 5 5 1 10 5 stellaria graminea + 5 . + 2 . 2 5 veronica chamaedrys + 1 . . 2 1 . 4 achillea millefolium . . 1 . 10 . 20 3 chamomilla recutita 1 1 . . . . 2 3 matricaria maritima ssp. inodora 1 1 . . . . . 2 viola arvensis . + . . . . + 2 rumex acetosa . . 2 . + . . 2 aegopodium podagraria . . 1 . . 10 . 2 equisetum arvense . . . . . + + 2 sporadic species – grasses: lolium perenne 31: 1; elymus repens 11: 1; phleum pratense 50: 10. others: centaurea jacea 30: +; oxalis fontana 30: +; senecio jacobaea 6: +; galium mollugo 25: 2; lychnis �os-cuculi 50: +; agrimonia eupatoria 24: +; juncus e�usus 24: +. k se ni a s tr ze żo ń 52 tab. 4. group iv – plots with arrhenatherum elatius successive no. of plots 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 n um be r o f oc cu rr en ce no. plot in the area 10 9 16 1 2 3 15 39 12 51 17 45 44 47 20 38 number of species in plot 15 13 15 18 17 19 9 16 12 12 18 17 13 15 13 17 grasses arrhenatherum elatius 10 30 30 20 40 10 10 30 50 10 20 10 10 10 15 20 16 phleum pratense 10 10 10 . 1 . . 10 10 2 10 10 10 10 10 10 13 dactylis glomerata 10 10 10 30 10 35 60 . . . . 20 10 40 10 10 12 poa pratensis . . . . . 20 . 20 10 50 10 10 50 2 50 10 10 anthoxanthum odoratum . . . 5 10 . 8 5 . . . . . . . . 4 holcus lanatus . . . 5 . 2 5 . . . . . . 1 . . 4 elymus repens . . 2 1 . 2 . . . . . . . . . . 3 lolium perenne 10 20 . . . . . . . . . . . . . . 2 festuca pratensis . . . . 5 . 10 . . . . . . . . . 2 alopecurus pratensis . . . . . . . 5 . . . . 2 . . . 2 legumes vicia cracca 10 1 2 1 1 10 . 1 10 . 2 . 2 1 . . 11 trifolium repens 30 . 5 20 10 . . 5 . 5 10 . . . . . 7 t. pratense . . . . 1 . . 5 . 10 . 25 10 . . . 5 medicago lupulina . . 5 . . . . . . 10 20 . . . . . 3 lathyrus pratensis . 10 . . . . . . . . . . 2 . . . 2 others achillea millefolium 5 15 10 5 1 5 2 5 2 . 2 2 2 10 2 10 15 ranunculus acris + + 2 + . 2 2 1 1 . . 2 1 2 2 . 12 taraxacum o�cinale . . 20 2 5 2 . 5 10 10 10 2 . 10 2 5 12 stellaria graminea 2 1 2 1 . + . + . + 2 1 + . . 1 11 lychnis �os-cuculi . + 2 . . 2 . + + + + 2 . + . . 9 veronica chamaedrys 2 + 2 + + + . . . . . . . + 2 2 9 rumex acetosa + . . . 1 + . + + 2 . . . . 2 + 8 plantago major . 3 . 5 5 2 2 . . . . 10 . . . . 6 aegopodium podagraria 5 . . . . . . . . . . + . 10 2 + 5 equisetum arvense + . . . . . . . + . + . . . . + 4 plantago lanceolata . + . . 1 5 2 . . . . . . . . . 4 campanula patula . . . + 1 2 . . . . . 3 . . . . 4 mentha longifolia . . . . . . . 5 2 . . 2 . . . 20 4 hypericum perforatum 2 . . . . . . . . . . . . . + + 3 equisetum palustre . . + . . . . . . . . + + . . . 3 convolvulus arvensis . . . 1 . . . . . + . . . + . . 3 potentilla anserina . . . . . 2 . . 2 . . . . 1 . . 3 myosotis palustris . . . . . . . . . . + . + . + . 3 senecio jacobaea . . + + . . . . . . . . . . . . 2 evaluation of the grassland sw ard floristic com position of the w adow ice com m une 53 viola arvensis . . . + . . . . . . + . . . . . 2 juncus e�usus . . . . 1 2 . . . . . . . . . . 2 alchemilla monticola . . . . . . . 1 . . . 2 . . . . 2 matricaria maritima ssp. inodora . . . . . . . . . . 2 1 . . . . 2 geranium dissectum . . . . . . . . . . 2 . . . . + 2 urtica dioica . . . . . . . . . . . . . 2 + . 2 sporadic species – others: daucus carota 10: 2; cirsium rivulare 1: 1; hieracium murorum 2: 1; luzula campestris 3: +; symphytum o�cinale 51: +; chamomilla recutita 17: +; centaurea jacea 17: +; conyza canadensis 17: 10; galium mollugo 38: 10; artemisia vulgaris 38: +; euphorbia cyparissias 38: 2. k se ni a s tr ze żo ń 54 tab. 5. comparison of average values of ellenberg’s indicators (l, f, r, n) calculated for groups of plots distinguished on the studied area of the wadowice commune; grey colour indicates the highest values group name n um be r o f p lo ts in th e gr ou p average number of species in plot ±sd ellenberg’s indicators (average values and ranges) li gh t l m oi st ur e f s oi l p h r n itr og en n i – plots with phleum pratense and vicia cracca 16 12 ±2.06 6.91(6.41-7.50) 5.40 (5.00-5.96) 6.38 (5.57-7.09) 6.49 (5.89-7.08) ii – plots with dactylis glomerata and trifolium repens 12 14 ±2.35 6.94(6.69-7.42) 5.31 (4.88-7.42) 6.41 (5.80-7.00) 6.19 (5.41-7.07) iii – plots with holcus lanatus 7 14 ±1.60 7.00 (6.77-7.21) 5.32 (5.24-5.41) 6.41 (5.81-7.06) 5.93 (5.15-6.50) iv – plots with arrhenatherum elatius 16 15 ±2.69 7.05 (6.70-7.40) 5.31 (4.91-5.68) 6.57 (5.50-7.40) 6.32 (5.94-6.87) evaluation of the grassland sw ard floristic com position of the w adow ice com m une 55 tab. 6. comparison of utility value of meadows (uvm) on the study area based on the use value index (uvi) numbers according to filipek (1973) plots group name no. plot in the area feeding value of plants very good 10-9 good 8-7 medium 6-4 small 3-1 none 0 poisonous plants (-1)-(-3) group i – plots with phleum pratense and vicia cracca 34 6.72 27 7.66 33 6.63 49 8.53 43 5.97 46 8.19 32 6.89 41 7.96 18 7.56 13 7.91 4 7.88 19 6.21 42 4.03 14 3.25 35 7.42 8 5.72 average uvm for all group i ±sd medium – 6.78 ±1.48 group ii – plots with dactylis glomerata and trifolium repens 40 8.13 7 7.26 48 6.69 37 7.93 26 7.68 5 7.12 22 5.17 29 6.62 28 7.52 23 6.02 21 5.97 36 7.06 average uvm for all group ii ±sd medium – 6.93 ±0.76 group iii – plots with holcus lanatus 31 6.13 30 6.12 11 6.67 6 7.10 25 6.78 50 6.84 24 5.38 k se ni a s tr ze żo ń 56 average uvm for all group iii ±sd medium – 6.43 ±0.53 group iv – plots with arrhenatherum elatius 10 7.86 9 8.07 16 7.08 1 7.62 2 7.25 3 7.82 15 8.19 39 8.04 12 7.65 51 8.78 17 7.06 45 7.65 44 9.25 47 6.64 20 8.65 38 5.75 average uvm for all group iv ±sd good – 7.71 ±1.49 evaluation of the grassland sw ard floristic com position of the w adow ice com m une 57 abstract �e quality of the botanical composition of meadows and pastures is decisive in the pro�tability of livestock farming. �e aim of this study was to examine the �oristic composition of meadows and pastures of the wadowice commune and to assess their feed value. field explorations were carried out in june 2016 throughout the commune. floristic lists were made on 51 designated plots using the estimated klapp method. four groups of similar plots with the following dominant species were distinguished in the analysed area: i – plots with phleum pratense and vicia cracca, ii – plots with dactylis glomerata and trifolium repens, iii – plots with holcus lanatus, and iv – plots with arrhenatherum elatius. �e results of the research showed that, in the area of this commune, meadows and pastures were characterised by average and good pasture values, which favours their use for agricultural purposes. improving the quality of the botanical composition of plots of medium fodder value could be achieved through proper care and sustainable use. key words: ellenberg’s indicators, grasslands, habitat, klapp method, utility value of meadows (uvm) received: [2020.03.20] accepted: [2020.05.15] ocena składu florystycznego runi użytków zielonych gminy wadowice streszczenie jakość składu botanicznego łąk i pastwisk ma kluczowe znaczenie w opłacalności hodowli zwierząt gospodarskich. celem niniejszej pracy było zbadanie składu �orystycznego łąk i pastwisk gminy wadowice oraz ocena ich wartości paszowej. eksploracje terenowe zostały przeprowadzone w czerwcu 2016 roku na obszarze całej gminy. na 51 wyznaczonych poletkach dokonano spisów �orystycznych, przy użyciu szacunkowej metody klappa. na analizowanym terenie stwierdzono obecność czterech grup poletek podobnych z następującymi gatunkami dominującymi: i – z phleum pratense i vicia cracca, ii – z dactylis glomerata i trifolium pratense, iii – z holcus lanatus, iv – z arrhenatherum elatius. rezultaty przeprowadzonych badań pokazały, że na obszarze tej gminy łąki i pastwiska odznaczają się średnią oraz dobrą wartością pastewną, co sprzyja ich wykorzystaniu do celów rolniczych. poprawę jakości składu botanicznego płatów o średniej wartości pastewnej można uzyskać poprzez odpowiednią pielęgnację oraz zrównoważone użytkowanie. słowa kluczowe: wskaźniki ellenberga, użytki zielone, siedlisko, metoda klappa, wartość użytkowa łąk (wuł) information on the author ksenia strzeżoń she was a student at the department of botany of the pedagogical university. she is interested in herbalism and �ora as well as the dynamics of plants communities in non-forest areas. 177 annales universitatis paedagogicae cracoviensis studia naturae, 5: 177–193, 2020, issn 2543-8832 doi: 10.24917/25438832.5.12 saikat kumar basu1, william cetzal-ix2*, alminda magbalot-fernandez3, peiman zandi4,5 1ps lethbridge, ab canada t1j 4b3 2tecnológico nacional de méxico, instituto tecnológico de chiná. calle 11 entre 22y 28, colonia centro chiná 24050. campeche, méxico; *rolito22@hotmail.com 3school of agriculture & food technology, �e university of the south paci�c, private mail bag, apia, samoa 4international faculty of applied technology, yibin university, yibin, sichuan, 644600, p. r. china 5institute of environment and sustainable development in agriculture, chinese academy of agricultural sciences, beijing 100081, p. r. china traditional and novel proposals for the protection of endangered pollinating insects an alarming decline has been observed in insect pollinator populations and subpopulations living in various ecosystems across the planet. among insect pollinators, bees have been the most impacted. native populations of several endemic (indigenous) bees are showing very serious decline, being pushed almost to the verge of extinction (tollefson, 2019). many countries around the world have documented elevated declines in bee populations, including china, brazil, north america and europe. �e reasons underlying such a worrying decline are not well-known. numerous factors, such as: excessive pollution, rapid rises in aggressive industrial agriculture, excess use of various toxic synthetic chemicals in agriculture, transformation in land use policies and practices, colony collapse disorder (ccd), poor immunity, loss of vigour in genetic strains, genetic bottlenecks, a rise in various parasitic diseases and a lack of suitable foraging plants (melliferous vegetation) and adequate nutrition (inadequate supply of nectar and pollen across di�erent seasons), are attributed to this rapid decline of pollinator insect populations, including bees and bumblebees (gallai et al., 2009; potts et al., 2010; henry et al., 2012). �e scenario is extremely disturbing as it is inter-connected with the future agricultural productivity, food security and stability of our natural or semi-natural global ecosystems. it is therefore essential to formulate a comprehensive, long-term, cost e�cient and sustainable model for conserving insect pollinators to secure the future of two important global areas, namely: agriculture (including apiculture) and forestry (basu, cetzal-ix, 2018 a, b, 2019; coh-martinez et al., 2019). �e wide diversity of �owering plants – both dicotyledonous and monocotyledons ai ka t k um ar b as u, w ill ia m c et za l-i x, a lm in da m ag ba lo tfe rn an de z, p ei m an z an di 178 ous plants (which include a vast majority of our food and industrial crops), are directly or indirectly dependent on natural pollinators through cross-pollination necessary for their generative reproduction. natural pollinators include not only insects like bees (such as honeybees and/or native bees), bumblebees, moths and butter�ies (fig. 1–2 – appendix 1) and some species of pollination-friendly ants, wasps, beetles and �ies but certain species of slugs and snails, birds (humming birds, sun birds, parakeets, etc.) and mammals (such as di�erent species of bats). several species other than insects, like reptiles (lizards) and amphibians, are also suspected of being active agents of cross-pollination. a rich diversity of animal species, not only insects, are involved in the process of cross-pollination (robinson et al., 2017). in fact, honey bees are responsible for only a third of the pollination of crops and a very small percentage of pollination of wild plants. �ere are many other insects that carry out this work and they are also in trouble – their population and subpopulations across the planet are decreasing alarmingly (kremen et al., 2007). however, the native bees, bumblebees (tab. 1 – appendix 2) and honey bees are the most endangered (garibaldi et al., 2013; kleijn et al., 2015). �e gradual reduction of insect pollinator populations is alarming since they have direct implications for our future and the stability of fragile global ecosystems (ripple et al., 2017). �ey play a key role in maintaining plant diversity. �e local extinction of even a single species of pollinator can lead to the disappearance of some plant species populations (lever et al., 2014; embry, 2018). �us, it is critical to focus on developing a comprehensive, but sustainable and a�ordable, conservation policy for insect pollinators for di�erent countries. if the human factor is not included in the conservation equation for protecting the insect pollinators the conservation policy cannot be successful in any part of the planet. hence, it is essential that we develop a low-cost and low-maintenance, farmerand environment-friendly simple “green approach” that can help save the endangered pollinator populations without stressing economies. �e aim of this study is to brie�y review traditional and novel methods for protecting pollinating insects and to propose the creation of special refuges – “pollinator sanctuaries”, that condition suitable habitats and an abundance of food for bees, bumblebees and other pollinators. traditional ways to protect pollinators traditional ways of protecting insects and other pollinators are classi�ed in terms of species protection or biotope protection. �e disappearance of biotopes of a given species is the most common reason for quantity decline and, consequently, extinction. inclusion of a species on a list prohibiting their destruction by individuals is usually not su�cient to safeguard its durability if the biotope in which it occurs is transformed. traditional and novel proposals for the protection of endangered pollinating insects 179 �erefore, modern species protection emphasizes the protection of habitats of species, not just the species themselves. it is currently thought that the most e�ective method is the preservation of all natural diversity in the context of preserving rich gene pools in changing environmental conditions (pickett, cadenasso, 2002; gastauera et al., 2013). in terms of risk to the richness of pollinating insects, the loss of habitats and �ower resources results from structural simpli�cation of the agricultural landscape and increased intensity of the use of arable land and grassland (kleijn et al., 2009; goulson et al., 2015). it is believed that the loss of uncultivated habitats (fallow lands) and increased land use are particularly harmful to rare pollinator species (kleijn et al., 2015). �e systematic loss of this type of species may contribute to the biotic homogenization of entomofauna groups (gámez-virués et al., 2015). studies generally show negative relationships between the local richness of wild bees and the decreasing complexity of the landscape in terms of the reduction of natural and semi-natural habitat areas (hendrickx et al., 2007; scheper et al., 2013; ekroos et al., 2020). in the second half of the last century, the approach to protective management also gradually changed. it was once thought that the most e�ective way of protection was the so-called passive (conservation) protection, which is e�ective, but more so in preserving the resources of diversity of forest communities (sołtys-lelek et al., 2014), including forest pollinators (robinson et al., 2017). in the case of semi-natural communities – xerothermic grasslands, meadows and pastures, which focus the greatest richness of pollinating insects, conservation protection leads to the overgrowth of these surfaces and the gradual elimination of light-sensitive species (drury, nisbet, 1973). as a result, it also signi�cantly reduces the diversity of entomofauna (maina et al., 2019). �erefore, it is currently believed that in the preservation of non-forest biotopes and all the diversity associated with these habitats balanced, extensive utilitarian managements, such as moderate grazing or mowing and extensive fertilisation, are necessary. in neighbouring arable �elds, it is recommended to limit the use of herbicides and other plant protection products that can harm pollinating insects (holzschuh et al., 2007; kleijn et al., 2009; robinson et al., 2017; ekroos et al., 2020). �ese treatments are not only important for maintaining the diversity of �owering plants but for maintaining the species richness of bees, bumblebees, butter�ies and other insects. meadows and other grasslands o�er both nests and �ower resources for insects for most of the growing season, unless they are used too intensively (gathmann et. al., 1994; albrecht et al., 2007; batáry et al., 2010; winfree, 2010). grasslands support both species important for plant pollination and species requiring protection, particularly if they o�er a variety of �ower resources (sutter et al., 2017). however, since many (immeasurable) factors can simultaneously a�ect biodiversity (cornell, harrison, 2014), it can be assumed that the high local intensity of land use will have an s ai ka t k um ar b as u, w ill ia m c et za l-i x, a lm in da m ag ba lo tfe rn an de z, p ei m an z an di 180 overriding negative impact on pollinating insects, thus limiting their diversity, despite the potential availability of resource habitat (ekroos, kuussaari, 2012; kennedy et. al., 2013; hopfenmüller et al., 2014; ekroos et al., 2020). innovative proposals in the protection of pollinating insects since bees and other pollinators are mobile organisms, they can survive even in intensively managed landscapes, as long as there are enough semi-natural habitats for their nesting and foraging (jabr, 2013). �is may suggest that the protection of common pollinating insects can be maintained in intensive farming systems, as long as a minimum number of semi-natural habitats are available to them (baldock et al., 2015; ekroos et al., 2016). �ese conclusions led to the emergence of new postulates important in the protection of pollinating insects in anthropogenic habitats, especially in areas of large urban agglomerations. considering how important numerous plants pollinated by insects are for the entire human economy (potts et al., 2016), the notion of pollinator protection in urban areas has begun to be promoted (garbuzov, ratnieks, 2014). over the last several years, building houses for insects has become popular in both city parks and green areas, in rural areas, in gardens and even on farms themselves fig. 3. an example of a house for insects of pollinating with wood (a) and mixed construction (b) (photo. s.k basu) traditional and novel proposals for the protection of endangered pollinating insects 181 (falk, 2015). �e construction of an insect house is very simple and can be a do-ityourself project (fig. 3). detailed instructions on this topic can be found in many available online sources (e.g. carlton, 2017). �e houses should be placed in a quiet, sunny and dry place. �ey can be made of wood or using elements of straw, reed or bricks. a “wild �ower meadow”, as well as a piece of un-mowed neglected lawn or planting nectaring plants, such as erica sp., rubus sp., salix sp., �ymus sp., tilia sp., attracts useful pollinating insects to them (carvell et al., 2007; salisbury et al., 2015). such insect houses also have a very important didactic function; we can better get to know and understand the world of nature, and some housing structures allow for the observing the stages of insect development. another interesting idea for the protection of pollinating insects is the establishment of “�ower meadows” in urban areas. �is idea has been popular in recent years, both in western europe, e.g. great britain, france and germany, and in central europe, e.g. in poland. �is type of meadow is an alternative to monotonous urban lawns, and can also be a feeding place and refuge for pollinating insects (baldock et al., 2015). �rough �owering plants, the meadow can have a positive e�ect on the aesthetics of the area, delighting with colours and smell (hoylea et al., 2017). residents can also use the meadow, and the installation of information boards about plants and insect species occurring there and the purpose of the entire undertaking can ful�l educational functions (lindemann-matthies, bose, 2007). urban “�ower meadows” can be made on virtually any well-sunned surface that is not dominated by large trees. �e species composition, properly selected for the type of soil, allows for the use of areas that are o�en degraded by humans, where it may be di�cult to maintain a nice lawn (roadsides, road lanes, neighbourhoods of public transport tracks, etc.). exemplary compositions of plant mixtures can easily be found on many online sources (tab. 2 – appendix 2), which strongly support the idea of protection, but they are not always satisfactory in the context of the utility quality for pollinators and the length of access (garbuzov, ratnieks, 2014; harmon-�reatt, hendrix, 2015; hicks et al., 2016). �e “�ower meadow” is also an economically viable alternative to city lawns. savings arise from the caretaking of the area by limiting mowing to one or two mowing operations in a season. however, there are also opponents of this type of project, who hold that urban “�ower meadows” are not ideal for protection of insects. �e fact that city meadows are o�en in the vicinity of streets may have unintended but harmful consequences: insects attracted by plants may be more vulnerable to being hit by a car. in addition, the creation of small isolated meadows �lled with “potential prey” is a sure way to attract predators (such as birds) that very quickly reduce the number of insects. even the best mix of meadow plants selected to a given area should be cared (weeded) for the �rst several years to prevent expansive weeds from dominating and displacing the nectar plants (hoylea et al., 2017); this obviously requires �nancial expenditure. s ai ka t k um ar b as u, w ill ia m c et za l-i x, a lm in da m ag ba lo tfe rn an de z, p ei m an z an di 182 also, the use of alien species in decorative plants mixtures should be avoided as they can quickly become a threat to the native �ora (salisbury et al., 2015; williams et al., 2011). certainly, the idea of urban “�ower meadows” is very interesting and worthy of further scienti�c investigation. another idea for protecting pollinating insects is to leave small set-aside fragments in agricultural areas where crop and meadow weeds can survive. �ey can be �eld edges or mid�eld balks. �ey can provide refuging, nesting (svensson et al., 2000) and food for pollinators, especially when they are additionally sown with native mixtures of nectaring plants (carvell et al., 2007; kuussaari et al., 2011; williams et al., 2011). studies conducted in many countries, in canada, the usa, germany, and great britain, con�rm the fact that these habitats are usually rich in �owering weed species, therefore they are a natural refuge for many pollinating insects. �e richness of �ower species translates directly into the richness of insects found in these areas (carvell et al., 2007; maina et al., 2019). of course, it is very important that the neighbouring �elds are cultivated organically, without intensive use of pesticides, as this has a direct impact on the number of insects. if agricultural lands are to be managed to preserve the diversity of species, including wild bees, maintaining diverse communities close to arable �elds and/or modifying agricultural production practices in these areas can signi�cantly improve the protection of local bee communities (holzschuh et al., 2007; winfree, 2010; kennedy et. al., 2013; maina et al., 2019). “pollinator sanctuaries” – our novel proposeof pollinating insects to protect our proposal is a combination of the concept of �ower meadows and the creation of areas separated from agricultural use in order to increase the resource of wild nectar �owers. we propose the creation of “pollinator sanctuaries”, which will be simultaneously gardens and habitats for pollinating insects. “pollinator sanctuaries” can occur in the form of small plots, placed in various, normally unused places, e.g. along highways, boulevards, avenues, parks, gardens and lawns, wetland areas and around perimeters of large and small golf courses (basu, cetzal-ix, 2017; martínez-puc et al., 2018). �ose places will be sown with locally adapted “plant mixes” consisting of local honey-bearing plants (such as: fabaceae: lotus corniculatus l. fig. 1a, medicago sativa l. fig. 1c, vicia villosa roth fig. 2f; asteraceae: tripleurospermum inodorum (l.) sch. bip. fig. 2d – appendix 1) and pollinator-friendly acclimatised annual/biennial/perennial forage crop species (such as asteraceae: gaillardia sp., fig. 1b, echinops sphaerocephalus l., fig. 1d, echinacea purpurea l., fig. 2e; brassicaceae: brassica napus l. fig. 2c; and hydrophyllaceae: phacelia sp., fig. 2a, b – appendix 1), in various proportions used in agriculture. an important step in achieving that goal will be to establish “pollinator sanctuaries” across di�erent agro-climatic zones. traditional and novel proposals for the protection of endangered pollinating insects 183 plant species selected for the mix must be �owering in sequence, one a�er another, to extend the pollinator (bees) foraging period and provide them with an adequate supply of nectar and pollen. plants mixes need to be developed based on appropriate agronomic parameters of the growing region and on local agro-climatic conditions, keeping in mind the local pollinator diversity and their preferences. plant mixes constituting only native wild�owers, currently available commercially, are not a viable option due to their poor adaptability to local agronomic conditions, high yield �uctuations (based on locality and annual production variation) and high production cost. development of suitable, environment-friendly plant mixes for various agro-climatic regions could therefore provide a long-term, low-cost and sustainable measure for conserving endangered pollinator insects (basu, cetzal-ix, 2017; basu, 2019). �is model targets several trophic levels within a natural or arti�cial ecosystem to conserve multiple species simultaneously in addition to local pollinator insects thereby e�ciently protecting and enriching local biodiversity (robinson et al., 2017; basu, cetzal-ix, 2018 a, b). in this way, a “pollinator sanctuary” would not only attract pollinator insects but other species of insects and small passerine birds and then raptors that survive on other birds and rodents that take refuge in such ecosystems. di�erent birds, small or medium sized mammals, amphibians and reptiles are all drawn to such natural ecosystems, providing a multiple tier or multiple trophic level dynamic ecosystem operating within just a few years of establishment at an extremely low and a�ordable cost requiring only simple management (basu, 2019). �e proposed model has been in use in canada for the enhancement of local biodiversity. �e model not only helps pollinator insect conservation but at the same time the can provide cover crops, used in promoting soil health, in phytoremediation, in transformation of agriculturally non-suitable areas into ecologically productive natural ecosystems units, as grazing area for pasture animals, in maintenance of pasture and rangelands, in crop rotation, as organic agriculture, in biomass generation and, last but not least, in local biodiversity enrichment. our proposed model is simple and nature-based and does not interfere or put any negative pressure on the local economy (basu, cetzal-ix, 2017). �us, it has the potential for integrating economy with ecology while protecting species and enhancing biodiversity of small island nations through a simple nature-based approach. conclusions in face of mass extinction of pollinating insects in many regions of the world, all proposals for their protection, both the older, classic as well as the new, should be comprehensively used. numerous agri-environmental programs that directly or indirectly s ai ka t k um ar b as u, w ill ia m c et za l-i x, a lm in da m ag ba lo tfe rn an de z, p ei m an z an di 184 protect pollinating insects seem to help in this regard. of course, attention should be paid to the local conditions and an e�ort made to try to adapt the selection of appropriate methods and protective strategies to them as much as possible. universal social education is also important in this respect, which should emphasise, especially in isolated (island) areas, the absolute need to protect the biological resources of local �ora and thus local entomofauna. �is is an essential element in preserving 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(photo. s.k basu) s ai ka t k um ar b as u, w ill ia m c et za l-i x, a lm in da m ag ba lo tfe rn an de z, p ei m an z an di 190 fig. 2. insect pollinators cont.: a – a large nevada bumblebee (bombus nevadensis cresson) foraging on scorpion weed (phacelia sp.); b – hunt’s bumblebees (b. huntii greene) foraging on scorpion weed (phacelia sp.); c – a western checkered butter�y (pontia protodice boisduval & le conte) foraging on canola (brassica napus l.); d – a stratiomyid �y visiting scentless chamomile (tripleurospermum inodorum (l.) sch. bip.); e – a honey bee (apis mellifera l.) foraging on purple cone �ower (echinacea purpurea l.); f – a bumblebee (bombus sp.) foraging on hairy vetch (vicia villosa roth) �ower (photo. s.k basu) traditional and novel proposals for the protection of endangered pollinating insects 191 appendix 2 tab. 1. endangered or threatened bumblebees species in north america according to iucn (2020); rlc = iucn – red list of �reatened species, cr – critically endangered, lc – least concern, en – endangered, vu – vulnerable, dd – data de�cient; nomenclature of animal species added according to animal diversity web… taxa population trend rlc geographic range elevation [m] bombus a�nis cresson decreasing cr canada – b. appositus cresson unknown lc canada, united states – b. bimaculatus cresson stable lc canada, united states – b. borealis kirby stable lc canada, united states – b. brachycephalus handlirsch decreasing en mexico, el salvador, guatemala, honduras 300–2800 b. caliginosus frison decreasing vu canada, united states – b. centralis cresson stable lc canada, united states – b. citrinus smith stable lc canada, united states – b. ephippiatus say stable lc mexico, el salvador, guatemala, honduras, nicaragua, costa rica, panama 0–3595 b. fervidus fabricius decreasing vu canada, united states, mexico – b. �avifrons cresson stable lc canada, united states – b. franklin frison decreasing cr united states – b. fraternus smith decreasing en united states – b. frigidus smith stable lc canada, united states – b. griseocollis de geer stable lc canada, united states – b. haueri handlirsch decreasing en mexico 1025–2700 b. huntii greene decreasing lc canada, united states, mexico – b. macgregori labougle & ayala stable lc mexico, guatemala 1250–3260 b. medius cresson decreasing vu mexico, el salvador, nicaragua 700–2500 b. melanopygus nylander stable lc canada, united states – b. mexicanus cresson decreasing vu mexico, el salvador, guatemala, honduras, nicaragua, costa rica 0–2693 b. morrisoni cresson decreasing vu canada, united states – b. nevadensis cresson stable lc canada, united states, mexico 0–2741 b. occidentalis greene decreasing vu canada, united states – b. pensylvanicus de geer decreasing vu canada, united states, mexico – b. perplexus cresson stable lc canada, united states – b. pullatus franklin unknown dd mexico, guatemala, honduras, nicaragua, costa rica, panama 0–3400 b. rufocinctus cresson stable lc canada, united states – b. sandersoni franklin stable lc canada, united states – b. sitkensis nylander stable lc canada, united states – b. steindachneri handlirsch decreasing en mexico 0–2600 s ai ka t k um ar b as u, w ill ia m c et za l-i x, a lm in da m ag ba lo tfe rn an de z, p ei m an z an di 192 b. suckleyi greene decreasing cr canada, united states – b. sylvicola kirby stable lc canada, united states – b. ternarius say stable lc canada, united states – b. terricola kirby decreasing vu canada – b. trinominatus dalla torre stable lc mexico, guatemala 2527–3500 b. vagans smith stable lc canada, united states – b. vandykei frison stable lc canada, united states 0–2200 b. variabilis cresson decreasing cr canada, united states, mexico, guatemala – b. vosnesenskii radoszkowski stable lc canada, united states 0 b. weisi friese stable lc mexico, el salvador, honduras, nicaragua, costa rica 1000–3200 tab. 2. examples of seed mixtures recommended for establishing �ower meadows in central europe (sources: dąbrowska, kulik, 2020); nomenclature of plants according flora polski atlas… family dry habitats, neutral or alkaline soils (grassland species) fresh habitats, neutral soils, wide ph range (fresh meadows species) fresh habitats of di�erent fertility (�eld �owers, usually 1–year’s) asteraceae achillea millefolium achillea millefolium centaurea cyanus centaurea scabiosa centaurea jacea chamomilla recutita cichorium intybus leucanthemum vulgare matricaria perforata – tragopogon pratensis – apiaceae daucus carota daucus carota – – pastinaca sativa – boraginaceae echium vulgare – – campanulaceae campanula patula – caryophyllaceae saponaria o�cinalis – – anthemis tinctoria – – dipsacaceae – knautia arvensis – fabaceae – lotus corniculatus vicia villosa – trifolium pratense – – vicia cracca – geraniaceae – geranium pratense – lamiaceae salvia pratensis – – scrophulariaceae verbascum nigrum – – v. densi�orum – – papaveraceae – – papaver rhoeas poaceae brachypodium pinnatum arrhenatherum elatius – – festuca rubra – – poa pratensis – rosaceae agrimonia eupatora sanguisroba o�cinalis – rubiaceae – galium verum – traditional and novel proposals for the protection of endangered pollinating insects 193 tradycyjne i nowe propozycje ochrony zagrożonych owadów zapylających streszczenie obecnie wymieranie owadów zapylających staje się problemem ogólnoświatowym. jest to istotne nie tylko z punktu widzenia utraty bioróżnorodności, ale ma ogromne znaczenie dla rolnictwa i gospodarki żywnościowej świata. tradycyjne sposoby ochrony owadów i innych zapylaczy postrzega się, albo w kategoriach ochrony gatunkowej, albo ochrony biotopowej, w ich naturalnych lub półnaturalnych siedliskach. jednak coraz częściej podejmuje się próby ochrony tej grupy owadów w środowiskach silnie zmienionych przez człowieka, jak na przykład aglomeracje miejskie i towarzysząca im infrastruktura. istnieją też inne propozycje, np. zakładanie pewnego rodzaju upraw – „ostoje zapylaczy”, na niewielkich poletkach w sąsiedztwie pól uprawnych lub miejscach nie przydatnych gospodarczo. w obecnej sytuacji wszystkie propozycje ochrony, zarówno te starsze – klasyczne, jak i te nowe, powinny być stosowane w sposób kompleksowy, bo tylko to może przynieść poprawę sytuacji owadów zapylających. key words: bee, biodiversity, insects, methods of protection, pollination received: [2020.05.15] accepted: [2020.07.30] 110 annales universitatis paedagogicae cracoviensis studia naturae, 5: 110–128, 2020, issn 2543-8832 doi: 10.24917/25438832.5.8 katarzyna lipniak1, angelika kliszcz2* 1cracow institute of development and education, wielicka 42/105 st., 30-552 kraków, poland 2university of agriculture in krakow, department of agroecology and plant production, mickiewicz 21 ave, 30-120 kraków, poland; *angelika.kliszcz@student.urk.edu.pl allelopathic effect of goosefoot on germination and early stage growth of triticale and radish introduction weeds are plants that are very well adapted to growth and development in changing environmental conditions. �eir proliferation is a  signi�cant and growing problem for agriculture. �ey have the ability to produce an ample amount of seeds which are characterised by prominent vitality and easy germination. �ese plants demonstrate a wide range of temperature tolerance and variable soil conditions. �ey use nutrients in larger quantities than crop plants and they can appear in secondary weed infestation. one example is goosefoot (chenopodium album l.). it is an annual plant that grows to about one meter in height (paczyńska, 2016). in various classi�cation systems it belongs to the goosefoot family (chenopodiaceae vent.) or amaranthus (amaranthaceae juss.). c. album has been cultivated in europe since ancient times. in poland, it occurs in the lowlands as well as in lower mountain regions, and it o�en grows on the fallow, �elds or gardens (sudnik-wójcikowska, 2011). goosefoot has a  pile root and is branched. its stem also branches strongly and sometimes has red discoloration. �e whole plant is pubescent (czubiński, paradowski, 2014). �e leaves take on a diamond shape or are lanceolate, with a serrated edge and a wedge-shaped base. c. album begins �owering in july, which can last until november. its �owers are small and pale green. a�er �owering, it produces fruit in the form of a small nut with black seeds. goosefoot seeds can easily adapt to all weather conditions, even retaining their ability to germinate for several years (klaaβen, freitag, 2004). �is plant is wind-proof and reproduces and develops very quickly, thus taking valuable nutrients and water from crop plants (domagała-świątkiewicz, 2007). c. album contains vitamins a, b1, b2 and c and microelements. it is also rich in �avonoids, essential a llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 111 oils, carbohydrates and proteins (byłka, kowalewski, 1997; dutt et al., 2003; jardim et al., 2008; usman et al., 2010; gęsiński, nowak, 2011). its seeds are abundant in valuable fats and albumins. shoots, older leaves and seeds contain oxalic acid, saponins, phenols, lignans and alkaloids (cutillo et al., 2004, 2006; lavaud et al., 2007; laghari et al., 2011) crop plants and weeds are a community with an additive system of components in which individuals compete for limited resources of the habitat, especially for water, nutrients and light. competition results in the adjustment and weakening of competing organisms, which manifests as a  reduction in the amount of biomass produced, in the size of individual organs and in seed yield. in cases of strong competitive impacts, individuals may even die and be eliminated from the community (begon et al., 1999). in the literature, the high allelopathic potential of c. album is widely discussed (batish et al., 2006; jafari, kholdebarin, 2002; laghari et al., 2011). although, reinhardt et al. (1994) pointed out, this plant did not exert an inhibitory e�ect on radish germination when milled dry shoot biomass was added to the soil (1% w/w) or when radish seeds were treated with aqueous soil extracts from pots that previously contained mature c. album plants. in natural conditions, the allelopathic actions of c. album are not only limited to plant-plant interactions. c. album plants strongly propagate soil microorganism growth (like anabena genus, a nitrogen-�xing cyanobacteria) and their aqueous extracts can enhance the growth of bradyrhizobium japonicum kirchner (vokou et al., 2006). �e aim of the study was to examine the e�ect of aqueous extracts from dry goosefoot (chenopodium album l.) plants on germination and early stage growth of triticale grains (×triticosecale wittm. ex a.camus) and radish seeds (raphanus sativus l.). �e e�ect of di�erent percent concentrations of c. album extracts on (1) germination index values, (2) percent inhibition of seedling growth, (3) fresh and dry seedling mass, relative water content and percent water content in 7-day triticale and radish seedlings and (4) the degree of cell membrane destabilisation by measuring the electrolyte leakage were determined. material and methods plant material triticale (×triticosecale) grains and radish (raphanus sativus) seeds were used in the experiment. goosefoot (chenopodium album) shoots in the vegetative phase were collected in spring 2019, from the southern poland stand (49°59ʹ10ʺn 20°03ʹ42ʺe) and dried in laboratory conditions. plant material was stored in the dark to avoid photochemical destruction of allelopathic compounds. k at ar zy na l ip ni ak , a ng el ik a k lis zc z 112 preparation of extracts dry material (goosefoot shoots) was ground in a mortar and distilled water was added, in the following amounts: (i) 0.5% extract: 0.5 g dry material + 95.5 ml distilled water, (ii) 1.0% extract: 1 g dry material + 99 ml distilled water, (iii) 1.5% extract: 1.5 g dry material + 98.5 ml distilled water and (iv) 2.0% extract: 2 g dry material + 98 ml distilled water. �e preparations were le� for 24 hours in the dark at approximately 25°c to allow for extraction of the compounds. a�er one day, the extracts from dry c. album plants were strained and stored in a refrigerator at 8°c ± 2°c for the duration of the experiment. seed germination 25 pieces of triticale grains and radish seeds (a�er washing with running water for 30 minutes, and then 3 times with distilled water) were placed with a sterile tweezer on petri dishes (ø 9 cm) covered with a triple layer of �lter paper, moistened with 5 ml of the appropriate aqueous extract of c. album plants in the concentrations: 0.5%, 1.0%, 1.5% or 2.0%. for the control, distilled water was used. during the experiment, all grains and seeds were placed in the dark, at room temperature. every 24 h for 7 days the number of germinated grains and seeds was counted. grains and seeds were considered germinated when their germinal root was equal to half the size of a grain or seed. germination parameters a�er 7 days of the experiment, the e�ect of c. album aqueous extracts on the germination capacity of triticale grains and radish seeds was evaluated. a germination index – gi (aosa, 1983), speed of emergence – se, seedling vigour index – svi (islam et al., 2009), coe�cient of the rate of germination – crg (chiapusio et al., 1997) and time required for 50% germination – t50 (farooq et al., 2006) were assessed. biometric analysis triticale and radish seedling length was measured using a calliper with an accuracy of 1 mm. �e e�ect of goosefoot extracts on seedling growth was determined according to islam and kato-noguchi (2012). fresh and dry mass, relative water content and percentage water content fresh mass (fm) of triticale and radish seedlings was determined with a balance (ohaus adventurer pro, usa). to obtain the dry mass (dm), the plant material was dried for 48 h at 105°c in a dryer (wamed sup 100, poland) and then weighed. �e relative water content (rwc) and the percentage content were determined based on the mass values. a llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 113 �e rwc was determined according to the method described by mullan and pietragalla (2012). brie�y, an individual seedling was weighed for fm and incubated for 24 h at 25°c in vials with 10 ml distilled water for saturation of plant tissues with water. a�er the incubation time, the turgor seedling mass (tm) was determined. �en, each was dried at 105°c for 48 h in the laboratory oven (wamed sup 100, poland) and dm weighed. �e rwc parameter for every seedling was calculated according to the formula: rwc = [(fm − dm) / (tm − dm)] × 100. �e percentage of water content (% h2o) was determined based on the mass values according to the formula % h2o = 100 − [(dm × 100) / fm]. electrolyte leakage cell membrane permeability was measured by electrolytze leakage in triticale and radish seedlings according to the method used by możdżeń et al. (2018). statistical analysis experimental results were compiled in microso� excel. additionally, statistical analysis was performed using one-way anova/manova. to assess the signi�cance of di�erences between the means ± sd (n = 3), duncan’s test at p ≤ 0.05 was used. �e data was analysed with the statistica program (statso�, inc. 2018, data analysis so�ware system, version 13.1). results germination indexes �e germination capacity of triticale grains (×triticosecale) from the control group (distilled water) a�er 2 days was 100%. a similar result was obtained for seeds germinating on the aqueous extracts of dry chenopodium album at a concentration of 0.5%, where the germination capacity a�er 3 days was 90%. higher concentrations of goosefoot extracts (1.0%, 1.5% and 2.0%) inhibited the germination of grains. regardless of the concentration of extracts, the largest number of newly germinated triticale grains was observed on the third day of the experiment. similar results were observed for radish (raphanus sativus) seeds. �e highest percentage of germinated seeds was recorded a�er 4 days for the control sample and a�er 6 days for the 0.5% aqueous extracts. with increasing concentrations, a clear reduction in the number of germinated seeds was observed. radish seeds watered with 2.0% extracts exhibited the lowest germination capacity (tab. 1 – appendix 1). �e coe�cient of rate of germination (crg) index for triticale grains slightly decreased compared to control with increasing aqueous goosefoot extract concentrations. �e only statistically signi�cant di�erences observed for this index, for grains, k at ar zy na l ip ni ak , a ng el ik a k lis zc z 114 were between the control and 2.0% extracts. compared to the control, the crg for radish seeds clearly decreased in each of the c. album extracts used. a  signi�cant decrease in the crg index was demonstrated even at a  concentration of 1.0% (tab. 2 – appendix 1). �e shortest time needed to reach 50% germinated seeds (t50) for triticale and radish seeds was in the control case. for aqueous c. album extracts, the t50 values for each of the extracts increased with the concentration of allelopathic compounds (tab. 2 – appendix 1). �e germination index (gi) reached higher values for radish seeds compared to triticale grains. regardless of the type of seeds studied, the gi values decreased with increasing concentrations of aqueous c. album extracts. compared to the control, the lowest gi was observed for grains and seeds germinating in petri dishes saturated with 2.0% goosefoot extracts. �e speed of emergence (se), regardless of the type of studied seeds, reached its highest values in the control sample (tab. 2 – appendix 1). for c. album extracts, the se index values decreased for both triticale and radish seeds with an increase in the concentration of extracts. for radish seeds treated with extracts (1.5% and 2.0% concentrations) a clear inhibition of germination was observed. compared to the control, the seed vigour index (svi), irrespective of the organ and type of seeds, signi�cantly decreased with increasing concentration of c. album extracts (tab. 3 – appendix 1). �e lowest values were observed in seedlings watered with 2.0% extracts. biometric analysis biometric analysis of whole triticale and radish seedlings revealed signi�cant inhibition of growth in the presence of aqueous extracts from c. album plants (tab. 4 – appendix 1; fig. 1–2 – appendix 2). stimulation of seedling growth in relation to control was observed only for radish seedlings watered with 0.5% goosefoot extract. �e root length of triticale and radish seedlings was similar in all studied samples. with increasing concentration of allelopathic compounds in aqueous extracts, inhibition of root growth was observed. �e exception was for r. sativus seedlings watered with 0.5% c. album extract, which resulted in a stimulatory e�ect on the root length of seedlings. �e inhibition percentage index of growth (ip), expressed as a percentage of the control value, reached higher positive values for the triticale and radish above-ground organs with increasing concentration of goosefoot extract (tab. 4 – appendix 1). �e only exception were radish seedlings watered with 0.5% extract; compared to controls, they reached negative values, which indicated stimulation of hypocotyl growth. a llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 115 fresh and dry mass, relative water content and percentage water content �e aqueous extracts from c. album plants had an inhibitory e�ect on the fm values of triticale and radish seedlings. compared to the control sample, when the concentration of the extracts increased, a signi�cant decrease in the mass values was observed. �e lowest values for this parameter were found in seedlings watered with 2.0% extract. �e exception was radish seedlings treated with 0.5% extract, which showed a signi�cant increase in fm, compared to control seedlings (tab. 5). �e dm of triticale seedlings reached di�erent values depending on the concentration of the extract. compared to the control, the highest values were found in seedlings watered with 0.5% extract and the lowest for seedlings grown in petri dishes with 1.0% and 2.0% goosefoot extract. in the case of radish seedlings, dm increased for each extract concentration, in relation to the control (tab. 5). tab. 5. fresh and dry masses, percentage water content and relative water content of triticale (×triticosecale wittm. ex a.camus) – (a) and radish (raphanus sativus l.) – (b) seedlings watered with the aqueous extracts of chenopodium album l. shoots in di�erent concentrations (0.5%, 1.0%, 1.5%, 2.0%) and control conditions pa ra m et er concentration of aqueous extracts [%] control 0.5 1.0 1.5 2.0 a b a b a b a b a b fm 0.2531a±0.03 0.0988b ±0.02 0.2196ab ±0.07 0.1587a ±0.01 0.1813b ±0.03 0.0673c ±0.03 0.1699c ±0.02 0.0283d ±0.02 0.1018d ±0.01 0.0197e ±0.005 dm 0.0251c±0.00 0.0028d ±0.001 0.0796a ±0.06 0.0050a ±0.00 0.0227c ±0.01 0.0043b ±0.00 0.0577b ±0.08 0.0025d ±0.00 0.0267c ±0.01 0.0039c ±0.002 wc 90.09a±0.29 97.10a ±0.47 57.88de ±39.07 96.83a ±0.66 87.52ab ±1.51 92.80a ±2.65 68.20d ±39.01 90.82a ±2.61 73.80c ±4.67 78.05b ±17.19 rwc 79.61a±8.89 58.50b ±15.00 60.73bc ±29.84 70.51a ±3.61 66.74b ±6.23 57.64b ±12.44 50.54c ±29.93 75.28a ±53.29 60.82bc ±14.87 51.77b ±35.39 mean values ±sd (n = 3) in row marked with letters a, b, c di�er signi�cantly according to duncan’s test at p ≤ 0.05; fm – fresh mass [g], dm – dry mass [g], wc – water content [%], rwc – relative water content [%] �e total water content of triticale seedlings decreased signi�cantly when extract concentration increased, compared to the control values (tab. 5). in the case of radish seedlings, the values for this parameter changed slightly. in seedlings watered with 2.0% extract, a  signi�cant decrease in the water content was found. �e rwc in the tested triticale seedlings decreased with the increasing extract concentration, compared to the control. for radish seedlings, the values for this parameter di�ered depending on the interaction with allelopathic compounds concentrated in aqueous extracts from c. album. k at ar zy na l ip ni ak , a ng el ik a k lis zc z 116 electrolyte leakage �e electrolyte leakage measurements revealed an increase in cell membrane destabilisation under the in�uence of goosefoot extracts (fig. 3). regardless of the type of seedling, a statistically signi�cant increase in electrolyte leakage occurred along with increasing extract concentration. �e highest degree of cell membrane disorganisation was found for triticale and radish seedlings treated with 2.0% c. album extract. fig. 3. electrolyte leakage from cell membranes of triticale (×triticosecale wittm. ex a.camus) and radish (raphanus sativus l.) seedlings watered with the aqueous extracts of chenopodium album l. shoots in concentrations 0.5%, 1.0%, 1.5%, 2.0% and control conditions; mean values ±sd (n = 3) marked with letters a, b, c di�er signi�cantly according to duncan’s test at p ≤ 0.05 discussion seed germination is a complex process that includes both catabolic and anabolic reactions and biochemical transformations. it consists of processes occurring inside the seed that lead to the activation of the embryo (nonogaki et al., 2010). �e germination is controlled by external (environmental) and internal (genetic and hormonal) factors realised as a function of time. �e exogenous factors include water, temperature and light (benech-arnold et al., 2000). �e endogenous factors are hormones, growth and development regulators, and reactive oxygen species content. in addition, various chemical substances that occur in nature, e.g. nitrogen oxides, butenolide derivatives (janas et al., 2010), glyceronitrile (downes et al., 2013) and allelopathic compounds secreted by neighbouring plants (zandi et al., 2018, 2019; puła et al., 2020) have impact on germination. a llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 117 �is research carried out on c. album con�rms that, during germination, grains and seeds become sensitive to allelopathic compounds (vokou et al., 2006; valizadeh, mirshekari, 2011; konieczna et al., 2018). �e germination capacity of triticale grains and radish seeds from the control sample was identical to the sample treated with 0.5% c. album aqueous extract. meanwhile, in other shoot extracts it turned out that the higher the concentration of allelopathic substances the smaller the number of germinated grains and seeds (tab. 1–2 – appendix 1). for gi values, a similar result was observed. it reached higher values for radish seeds, compared to triticale grains. in the control groups, the highest values of se index were observed. �e lowest se values were observed for grains and seeds treated with allelochemical compounds that had accumulated in the 1.5% and 2.0% extracts (tab. 2 – appendix 1). for crg, values clearly decreased with increasing concentration of aqueous c. album extract, compared to the control sample (tab. 2 – appendix 1). �e svi reached its lowest values for seedlings watered with 2.0% shoot extract, regardless of the type of organ and seeds (tab. 3 – appendix 1). similar observations in terms of inhibition of seed germination in the presence of aqueous extracts from c. album shoot for raphanus sativus l. were observed by e.g. mallik et al. (1994) and konieczna et al. (2019). di�erent degrees of seed sensitivity to allelopathic compounds may result from the size of diaspores and the thickness of seed coats (sołtys et al., 2012). according to vaughn and spencer (1993), large-sized seeds show higher stress tolerance. możdżeń and rzepka (2016) and mazur (2019) con�rmed the important protective role of seed coat in the germination and early stages of growth of vicia faba l. and phaseolus vulgaris l. �e mechanism of action of allelopathic compounds on seed germination is also associated with disturbances in the phytohormonal balance, slowing down the activation of spare materials, as well as the induction of oxidative stress (krasuska et al., 2014). biometric analysis of underground and above-ground organs of triticale and radish seedlings revealed that aqueous extracts from c. album shoots inhibited their growth, as the concentration of allelophatins in the extracts increased (tab. 4 – appendix 1). �e exception turned out to be radish seedlings watered with 0.5% extracts. �is extract stimulated their growth, compared to seedlings from the control. most likely contained in the leaves and stems of c. album, nitrates, among others phenols, saponins and alkaloids, negatively a�ected the germination and early stages of growth of triticale and radish seeds (cutillo et al., 2004, 2006; lavaud et al., 2007; czubiński, paradowski, 2014). allelopathic substances inhibit cell division and cell lengthening by limiting proton transport from the cytoplasm to the apoplast (burgos et al., 2004). �ey reduce the uptake of micro and macroelements by changing the hydraulic conductivity of cell membranes. one of the �rst e�ects of allelophatic compounds at a cellular level is to k at ar zy na l ip ni ak , a ng el ik a k lis zc z 118 reduce the transmembrane electrochemical potential of cell membranes. membrane depolarisation causes disturbances in the transport of anions and cations, which is associated with increased permeability of these structures. under stress conditions, (1) inhibition of phosphorus, potassium, magnesium and nitrate ion intake, (2) modi�cation of membrane proteins, (3) lipid oxidation by the presence of free radicals due to the reduction of catalase and peroxidase activity and (4) disorders in the functioning of channels, membrane conveyors and proton pumps occur. damage to the cell membranes depends on, among other things, the concentration and solubility of allelopathic substances and the ph of the environment (einhellig, 2004). in the experiment, di�erences in the electrolyte leakages from triticale and radish seedlings proved that allelopathic compounds released from goosefoot shoots clearly increased the degree of cell membrane destabilisation (fig. 3). studies on the allelopathic properties of c. album showed that the presence of goosefoot reduced the quantity and quality of various crop plant species (krop� et al., 1992; salam et al., 2014). for example, in zea mays l., glycine max (l.) merr., solanum l. section lycopersicon (mill.) wettst., avena l., hordeum l., medicago l., and beta vulgaris l. subsp. vulgaris, at a density of 172 to 300 plants per m2, goosefoot caused crop losses from 6 to 58% (staniforth, lovely, 1964; sibuga, bandeen, 1980; shurtle�, coble, 1985; torner et al., 1995; ngouajio et al., 1999). dyck and liebman (1995) attributed uncontrolled c. album populations to as much as 59% reduction in yields. in tomato cultivation, goosefoot caused a  36% reduction in the quality of marketable fruit (bhowmik, reddy, 1988). for barley yield, changes from 23 to 36% were attributed to its competition with c. album plants (conn, �omas, 1987). c. album caused an approximate 60% reduction in grain yield during the oat growing season and a 23% reduction in lucerne biomass yield (lapointe et al., 1985). �is study also con�rmed a negative e�ect of c. album extract on the masses and water content of triticale and radish seedlings (tab. 5). a decrease in mass accumulation was observed with increasing extract concentration, regardless of the type of seeds. only for radish seedlings was an increase in the dm value for each of the extract concentrations found. �e intense and negative allelopathic e�ect of goosefoot extracts on triticale grains and radish seeds most likely resulted from the type of extracts used in the experiment. leaves are organs that accumulate the largest amount of allelopathic compounds, and roots contain the lowest concentration (kryzeviciene, paplauskiene, 2004). goosefoot is a common weed in poland. in limiting c. album, regular care of arable land in connection with simultaneous prevention of weed control is important. otherwise, c. album plants le� without interference quickly cover large areas (even eliminating crop species), will use the nutrients intended for the crop plants and sharply increase their seed bank in the soil. �is problem especially a�ects plants cula llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 119 tivated in wide rows, such as maize, sugar beets and potatoes. �e weed economic thresholds determine the number of weed plants per unit area at which the crop yield loss will be greater than the total costs of the plant protection treatments. for maize, the threshold is 2 pcs of c. album per m2 and for sugar beet 5 plants per 30 m in a row (�e threshold of pests…, 2020). in cereal crops (both spring and winter forms), besides the negative potential from released allelochemicals and competition for nutrients, the timing of the appearance of goosefoot is also problematic because c. album’s ability to germinate occurs during the entire growing season and this weed may yield seeds several times. additionally, due to the fact that these seeds hold moisture well, even small numbers per 1 ha may disturb the long-term storage of grain. determination of such weed thresholds is quite di�cult due to many factors that occur (e.g. the moment of weed appearance in relation to the developmental stage of the crop, weather and soil conditions and the actual prices of plant protection treatments) (rola et al., 2013). however, research conducted by valizadeh and mirshekari (2011) suggests that the c. album threshold for rapeseed may be 4–8 pcs per m2 (coverage of rapeseed in the terms of lai drops from 3 to 1.5). �e lack of selective herbicides, which at the same time could control the occurrence of goosefoot and be harmless to plants (kucharski, sekutowski, 2007) means that only regular mechanical treatments help in reducing the occurrence of goosefoot in crop �elds. if necessary, chemical weed control is used along with areas adjacent to them, e.g. wasteland, ditches, paths and areas prepared for growing plants. it is also important to clean the equipment used for sowing seeds and harvesting plants, fertilise the soil only with well-spread manure and use good quality seed that is free from weed seed contamination (domagała-świątkiewicz, 2007). although the controversy around the phenomenon of allelopathy has not yet been unequivocally resolved, the negative e�ect of one plant on other neighbouring individuals remains an indisputable fact (gniazdowska, 2007). weston (2005) emphasised that all plants contain allelochemical substances with di�erent structures and functions. �e germination and early growth analyses carried out have revealed the allelopathic e�ects of c. album extracts at various concentrations, which similarly interfere with the metabolism of grains and seeds and indicate the need for further research. poonia et al. (2015) emphasized that attention should be paid to c. album not only as a bothersome weed but as a source of functional nutrients and useful medicinal properties. however, this species is a good host for many dangerous crop pests (agrotis segetum denis & schi�ermüller, aphis fabae scop., delia radicum l., heterodera schachtii schmidt, silphida latreille latreille) and the host of some pathogens that cause viral diseases (e.g. beet mosaic, hepatitis jaundice (abe, tamada, 1986). c. k at ar zy na l ip ni ak , a ng el ik a k lis zc z 120 album was also reported to be the host of a  new plant disease caused by the fungus stagonospora atriplicis (westend.) lind in new zealand (mckenzie, dingley, 1996). conclusion (1) �e results presented in this paper con�rm the allelopathic properties of aqueous extracts of dry goosefoot (chenopodium album). based on the observations, an inhibitory e�ect of allelopathic compounds released from c. album shoots on the gis of triticale grains (×triticosecale) and radish seeds (raphanus sativus) was found. (2) biometric analysis of triticale and radish seedlings demonstrated a signi�cant inhibition of growth in the presence of aqueous extracts from dry c. album shoots. (3) with an increasing concentration of c. album extracts, the fm values of 7-day triticale and radish seedlings were decreased. �e dm and the percentage of water content values changed, depending on the type of seed and the concentration of goosefoot shoot extract. (4) �e negative e�ect of c. album was con�rmed by the electrolyte leakage detected from triticale and radish seedlings cells. with increasing allelopathic substances in the shoot extracts, an increase in water-ion balance disorders in the studied seedlings was observed. con�ict of interest �e authors declare no con�ict of interest related to this article. references abe, h., tamada, t. 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(2019). �e in�uence of aqueous extracts from stellaria media l. on the growth of zea mays l. cultivars. notulae botanicae horti agrobotanici cluj-napoca, 47(3), 921–928. https://doi.org/10.15835/nbha47311597 k at ar zy na l ip ni ak , a ng el ik a k lis zc z 124 appendix 1 tab. 1. percentage of germinated triticale grains (×triticosecale wittm. ex a.camus) – (a) and radish seeds (raphanus sativus l.) – (b) watered with the aqueous extracts of chenopodium album l. shoots in di�erent concentrations (0.5%, 1.0%, 1.5%, 2.0%) and control conditions aqueous extracts [%] days [24h] 1 2 3 4 5 6 7 a b a b a b a b a b a b a b control 80 a ±1.58 80 a ±0.84 96 a ±0.89 84 a ±0.84 96 a ±0.71 96 a ±1.14 96 a ±0.55 100 a ±0.55 100 a ±0.00 100 a ±0.45 100 a ±0.00 100 a ±0.45 100 a ±0.00 100 a ±0.00 0.5 68 ab±0.71 60 ab ±1.14 80 ab ±0.89 64 ab ±1.30 96 a ±2.07 80 a ±2.28 96 a ±1.64 92 a ±1.92 100 a ±0.00 96 a ±1.30 100 a ±0.00 100 a ±0.45 100 a ±0.00 100 a ±0.45 1.0 40 b±1.14 8 c ±1.14 44 b ±0.84 12 b ±1.48 64 b ±2.97 16 b ±1.52 68 b ±2.07 20 b ±1.52 72 b ±1.52 28 b ±1.52 76 b ±1.22 44 b ±2.41 80 b ±1.30 44 b ±2.49 1.5 36 b±1.14 0 c ±0.00 40 b ±0.84 8 b ±0.71 56 b ±2.07 8 c ±1.14 60 b ±2.59 16 b ±0.84 64 b ±2.59 28 b ±1.14 68 b ±2.30 32 bc ±0.89 76 b ±2.17 40 b ±0.84 2.0 20 c±1.14 0 c ±0.00 28 c ±1.30 8 b ±1.30 52 b ±3.05 8 c ±1.14 56 bc ±2.68 16 b ±1.14 56 b ±2.97 20 bc ±1.48 60 bc ±3.85 24 c ±2.07 60 bc ±3.85 28 c ±2.07 mean values ±sd (n = 3) marked letter (a, b, c) in columns di�er signi�cantly according to duncan’s test at p < 0.05 tab. 2. �e germination parameters of triticale grains (×triticosecale wittm. ex a.camus) – (a) and radish seeds (raphanus sativus l.) – (b) watered with the aqueous extracts of chenopodium album l. shoots in di�erent concentrations (0.5%, 1.0%, 1.5%, 2.0%) and control conditions aqueous extracts [%] crg t50 gi se a b a b a b a b control 24.34 a 24.12 a 0.35 b 0.35 cd 58.74 a 59.42 a 80.00 a 80.00 a 0.5 23.70 a 22.98 ab 0.59 ab 0.67 a 53.74 a 55.42 ab 68.00 b 60.00 b 1.0 22.56 ab 19.72 b 0.67 a 0.59 ab 34.71 b 36.30 b 50.00 c 18.18 c 1.5 22.42 ab 18.44 bc 0.66 a 0.63 a 31.16 b 32.57 b 47.37 d 0.00 d 2.0 21.90 b 19.12 b 0.65 a 0.40 c 23.78 c 27.17 c 33.33 e 0.00 d the germination index (gi), speed emergence (se), the coe�cient rate germination (crg) index and the time needed to reach 50% of germinated seeds (t50); mean values (n = 3) marked letter (a, b, c) in columns di�er signi�cantly according to duncan’s test at p < 0.05 tab. 3. seed vigour index (svi) triticale (×triticosecale wittm. ex a.camus) – (a) and radish (raphanus sativus l.) – (b) seedlings a�er 7 days of germination on the aqueous extracts of chenopodium album l. shoots in di�erent concentrations (0.5%, 1.0%, 1.5%, 2.0%) and control conditions svi concentration of aqueous extracts [%] control 0.5 1.0 1.5 2.0 a b a b a b a b a b underground organ 98.40 a 96.00 a 90.00 ab 64.80 b 53.25 b 10.91 c 39.74 c 3.50 d 5.67 d 12.50 e aboveground organ 114.40 a 49.60 d 99.60 b 80.60 b 76.75 c 72.27 c 95.26 b 19.50 e 23.67 d 6.07 f whole seedlings 212.80 a 145.60 a 189.60 b 145.40 a 130.00 c 83.18 b 135.00 c 23.00 c 29.33 d 18.57 d mean values (n = 3) in row marked with letters a, b, c di�er signi�cantly according to duncan’s test at p ≤ 0.05 a llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 125 tab. 4. length and inhibition percentage (ip) of growth (expressed as a percentage of control) of triticale (×triticosecale wittm. ex a.camus) and radish (raphanus sativus l.) seedlings germinated on the aqueous extracts of chenopodium album l. shoots in di�erent concentrations (0.5%, 1.0%, 1.5%, 2.0%) and control conditions organs concentration of aqueous extracts [%] control 0.5 1.0 1.5 2.0 [mm] [mm] ip% [mm] ip% [mm] ip% [mm] ip% triticale underground parts 98.4 a ±1.90 90.0 a ±2.37 6.31 42.6 b ±0.52 55.89 30.2 c ±0.91 68.87 3.4 d ±0.15 96.64 aboveground parts 114.4 a ±1.21 99.6 a ±1.65 12.97 61.4 b ±2.09 46.79 72.4 c ±1.36 36.76 14.2 d ±0.80 87.35 whole seedlings 212.8 a ±2.52 189.6 a ±3.83 104.20 104.0 b ±2.10 51.15 102.6 b ±2.14 51.72 17.6 c ±0.79 91.57 radish underground parts 49.6 a ±4.15 6.48 b ±0.92 –9.06 4.8 c ±0.87 93.83 1.4 d ±0.30 98.14 3.5 c ±0.29 94.39 aboveground parts 96.0 a ±0.43 8.06 c ±2.03 –64.20 31.8 b ±0.30 35.85 7.8 c ±0.09 84.45 1.7 d ±0.17 96.51 whole seedlings 14.56 a ±4.43 14.54 a ±2.88 –19.25 3.66 b ±0.93 72.55 0.92 c ±0.38 93.21 0.52 c ±0.19 96.07 minus (–) values of ip indicates growth stimulation, and plus (+) values of ip indicates growth inhibition; mean values (n = 3) in row marked with letters a, b, c di�er signi�cantly according to duncan’s test at p ≤ 0.05 k at ar zy na l ip ni ak , a ng el ik a k lis zc z 126 appendix 2 fi g. 1 . r ad is h se ed lin gs (r ap ha nu s s at iv us l .) on th e 3r d ( a –e ), 5t h ( f– j) a nd 7 th (k –o ) d ay o f g er m in at io n w at er ed w ith a qu eo us ex tr ac ts o f g oo se fo ot (c he no po di um a lb um l .) sh oo ts a nd d is til le d w at er : a , f , k – c on tr ol (d is til le d w at er ); b, g , l – 0 .5 % e xt ra ct ; c , h , m – 1 % e xt ra ct , d , i , n – 1 .5 % e xt ra ct ; e , j , o – 2 % e xt ra ct (p ho to . k . l ip ni ak ) a llelopathic effect of goosefoot on germ ination and early stage grow th of triticale and radish 127 fi g. 2 . t ri tic al e se ed lin gs (× tr iti co se ca le w itt m . e x a .c am us ) o n th e 3r d ( a –e ), 5t h ( f– j) a nd 7 th (k –o ) d ay s o f g er m in at io n w at er ed w ith a qu eo us e xt ra ct s o f g oo se fo ot (c he no po di um a lb um l .) sh oo ts a nd d is til le d w at er : a , f , k – c on tr ol (d is til le d w at er ); b, g , l – 0. 5% e xt ra ct ; c , h , m – 1 % e xt ra ct ; d , i , n – 1 .5 % e xt ra ct ; e , j , o – 2 % e xt ra ct (p ho to . k . l ip ni ak ) k at ar zy na l ip ni ak , a ng el ik a k lis zc z 128 abstract �e aim of this study was to investigate the e�ect of aqueous extracts from chenopodium album l. on germination and early stages of triticale grains (×triticosecale wittm. ex a.camus) and radish seeds (raphanus sativus l.). germination indexes, fresh and dry mass, water content and electrolyte leakage were measured. studies revealed the di�erent germination capacity of triticale grains and radish seeds, where increased concentrations of allelopathins in aqueous c. album extracts signi�cantly inhibited seedling growth for both species. �e extracts had an inhibitory e�ect on the growth of seedling fresh mass. an increase in dry mass of radish seedlings was demonstrated for each of the extracts and, for triticale seedlings, only at concentrations of 0.5% and 1.5%. water content in triticale and radish seedlings varied depending on the concentration of allelopathins in the extract. with increasing concentrations of c. album extract, regardless of seedling type, a statistically signi�cant increase in electrolyte leakage was observed. key words: fresh and dry mass, seed germination indexes, electrolyte leakage, relative water content received: [2020.03.16] accepted: [2020.05.25] allelopatyczny wpływ komosy białej na kiełkowanie i wczesne stadia wzrostu pszenżyta i rzodkiewki streszczenie celem pracy było zbadanie wpływu wyciągów wodnych z chenopodium album l. na kiełkowanie oraz wczesne stadia wzrostu ziarniaków pszenżyta (×triticosecale wittm. ex a.camus) i nasion rzodkiewki (raphanus sativus l.). wyznaczono wskaźniki kiełkowania, świeżą i suchą masę, zawartość wody oraz wyciek elektrolitów. badania wykazały różną zdolność kiełkowania ziaren pszenżyta i  nasion rzodkiewki – wraz ze wzrostem stężeń substancji allelopatycznych zawartych w wodnych ekstraktach z  c. album obserwowano znaczne zahamowanie wzrostu siewek obydwu gatunków. wodne wyciągi hamowały przyrost świeżej masy siewek. natomiast przyrost suchej masy siewek rzodkiewki wykazano przy każdym z ekstraktów, a dla pszenżyta jedynie w stężeniach 0,5% i 1,5%. zawartość wody w siewkach pszenżyta i rzodkiewki była zróżnicowana, w zależności od stężenia allelopatin w ekstrakcie. wraz ze wzrostem stężenia ekstraktów c. album, niezależnie od rodzaju siewek, zaobserwowano statystycznie istotny wzrost wycieku elektrolitów. słowa kluczowe: świeża i sucha masa, wskaźniki kiełkowania nasion, wyciek elektrolitu, względna zawartość wody information on the authors katarzyna lipniak she is interested in weeds in�uence on germination and growth of seeds. angelika kliszcz https://orcid.org/0000-0002-1270-4414 she is focusing on enhancing the understanding of the in�uence of di�erent factors on soil structure and fertility. particularly, she is investigating the interaction of plants with the physical, chemical, and biological properties of the soil. she is also interested in earthworm ecology and mesofauna function in agroecosystems. 109 annales universitatis paedagogicae cracoviensis studia naturae, 6: 109–124, 2021, issn 2543-8832 doi: 10.24917/25438832.6.7 andrzej rzepka*, wioleta szarek institute of biology, pedagogical university of krakow, podchorążych 2 st., 32-084 kraków, poland; *andrzej.rzepka@up.krakow.pl resistance of mosses to drying, measured by the intensity of gas exchange and the content of malate and citrate introduction water stress can lead to plant dehydration, wilt, and in extreme cases to its complete death. known as drought, it is one of the many factors limiting the occurrence of plants in the environment. in contrast, drought is an abiotic condition that occurs when plant transpiration is greater than the amount of precipitation supplied. it is closely related to the loss of water from plant tissues and cells. climatology treats drought as a reduction in the amount of rainfall, which is signi�cantly di�erent from the norm. weather conditions, such as low air humidity, high temperatures, strong winds make rainfall less e�ective, resulting in drought (chojnacka-ożga, lorenc, 2019). drought causes a serious disturbance in the life of plants, especially in those whose capacity to retain water for physiological processes for a long time is limited. even if the water de�cit does not cause the plant’s death, it can cause signi�cant morphological and physiological dysfunctions (vitt et al., 2014). mosses are among the most primitive land plants. in contrast to vascular plants which have developed tissues for transportation and retention of water, mosses retained simple morphology and anatomy (charron, quatrano, 2009). leafy gametophytes have properties that indicate their adaptation to terrestrial living conditions by developing e�cient, capillary-driven water transport for maintenance of growth, e�cient photosynthesis and reproductive processes (oliver, 2009). however, poorly specialised moss leaves, o�en made of a single layer of cells, do not always allow to survive in conditions of water de�cit. �erefore, mosses generally only tolerate some degree of cellular dehydration. a�er removing the liquid water from the cell surface, they quickly lose turgor and balance with the water potential of the air. �is is common in many bryophytes and most species have developed di�erent ways of surviving. for example, mosses avoid water stress by colonising wet or heavily shaded ecological niches, growing in short, dense a nd rz ej r ze pk a, w io le ta s za re k 110 clusters that reduce water evaporation (proctor, 1980; rice et al., 2001). �e evolution of mechanisms to withstand water stress presumably arose from individuals uniquely suited to occupy the outer limits of mesic niches (olivier et al., 2005; rzepka, 2008). another example is the fact that many mosses have developed the ability to survive even completely drying out, returning to vital functions within a few hours of rehydration. as poikilohydric organisms, they are generally considered to be relatively resistant to desiccation (proctor, 1990; proctor, pence, 2002). �is allows them to survive, both short and long periods of dehydration to a level at which the entire mass of the water is lost from the cells (oliver, 2009). fig. 1. plagiomnium undulatum (hedw.) t.j.kop.: a – gametophyte appearance (photo. w. starmach), b – leaf fragment (source: https://commons.wikimedia.org/wiki/); polytrichum commune hedw.: c – gametophyte appearance (photo. w. szarek), d – leaf with lamellas (source: https://commons.wikimedia.org/wiki/) plagiomnium undulatum (hedw.) t.j.kop. = mnium undulatum hedw. (hart’s-tongue thyme-moss) plagiomniaceae t.j.kop. family, is orthotropic moss, with creeping stolons forming large, loose and vivid green turf (fig. 1a). it has stems up to 15 cm long, towards the top with increasing side branches. its leaves are longitudinally lingual, strongly folded transversely, long running along the stem, rounded at the top, with a short, sharp tip and with sharp single teeth along the entire edge (fig. 1b). �eir rib is r esistance of m osses to drying, m easured by the intensity of gas exchange and the content of m alate and citrate 111 thick, reaching the top of the leaf. �ere are several oval-shaped capsules on one stem, growing on reddish sets. �e operculum is with a short spout. �is species grows in wet deciduous forests – riparian forests and hornbeam forests – as well as on humus (szafran, 1961; düll, düll-wunder, 2012; plášek, 2013). polytrichum commune hedw. (common haircap) polytrichaceae schwägr. family, is an orthotropic moss that creates loose, dark green turf (fig. 1c). its stems are single and reach up to 20–30 cm in length (sometimes up to 40 cm). �e leaves (1–2 cm long) have a  sheath-like base and a  lanceolate leaf blade, protruding from the stem. �e edge of the leaves from the top almost to the base is sharply toothed. �e leaf blade is dark green with square cells. �e vaginal part of the base of leaf is white, with elongated cells (up to 10 times longer than wider). �e rib is wide, with sharp serrations on the dorsal side, and shortly protruding at the top as a  serrated, brown spike. up to 70 longitudinal lamellas appear on the ventral side of the rib (fig. 1d). �e upper lamella cell is larger than the others, with a deep crescent-shaped furrow. �e capsule is four, �ve or hexagonal, with a  well-developed neck, with yellow-red, long (up to 10 cm) seta. �e calyptra is dense, golden yellow hairy (remnants of the archegonium). most o�en, this moss grows in peat bogs, both in the lowlands and in the mountains. it also occurs in damp forests or along the banks of streams (szafran, 1961; ignatov, ignatova, 2003; plášek, 2013). bryophytes belong to the homoiochlorophyllous groups and cannot maintain constant internal water content by regulating water loss (tuba et al., 1998; proctor et al., 2007). previous studies on the physiological level do not fully explain the mechanisms of drought tolerance in bryophytes. �erefore, the aim of this experiment was to determine the relationship between the stressor (drought) and changes in the intensity of relative photosynthesis (1), respiration (2) and the content of malate and citrate (3) in two species of mosses – plagiomnium undulatum (hedw.) t.j.kop. and polytrichum commune hedw. material and methods plant material �e gametophores plagiomnium undulatum and polytrichum commune were used in the research. �e plant material was collected in southern poland, near krakow (rybna, 50°03′04″n 19°38′50″e) and tarnów (pleśna, 49°55′19″n 20°56′43″e). both moss stands were remote from communication routes. �e cultivate conditions �e collected moss turf was stored in 1 dm3 glass aquariums, in air-conditioned growth chambers (fig. 2a). a nd rz ej r ze pk a, w io le ta s za re k 112 �e density of the quantum stream used here in the par range was 70 μmol × m–2 × s–1 (fluora-osram, poland), with a photoperiod of 12 h day/12 h night, with temperature 15°c (±2°c) and relative air humidity from 70 to 100%. mosses were watered regularly with distilled water. �e digital hygrometer pwt-221 (elmetron, poland) was used to measure the relative air humidity. quantum �ux density in the par range was measured with a radiometer (li-cor model li-189, usa). course of experiments moss gametophores of equal length (about 4–5 cm in the case of plagiomnium undulatum and 1.5–2 cm in the case of polytrichum commune) were rinsed with distilled water fig. 2. gametophores of plagiomnium undulatum (hedw.) t.j.kop.: a – placed in a glass vessel, b – placed in a plexiglass plate, c – dried on plates with �lter paper (photo. w. szarek) r esistance of m osses to drying, m easured by the intensity of gas exchange and the content of m alate and citrate 113 and dried with �lter paper. �en they were placed evenly in the holes of the plexiglass plates, in glass vessels with a volume of 1 dm3 (fig. 2b). a�er the control measurement of photosynthesis and respiration, the gametophores of both species were subjected to the drought. for this aim, leafy moss stalks were placed in a petri dish lined with three layers of �lter paper discs, kept in air with a  relative humidity of 30–40% until a  loss of about 50% relative water content (fig. 2c). �e intensity of the luminous �ux reaching the moss leaves during drying was 80 μmol × m–2 × s–1, and the temperature was 20°c (±1°c). �en, the gametophores were rehydrated, placed in a plexiglass plate and kept in glass aquariums in a growth chamber for 24 h. a�er this time, further measurements were made. preparation of moss gametophores for malate and citrate measurements looked similar to photosynthesis and respiration measurements. additionally, before measuring the malate content, a�er a period of darkness, the vessel with the gametophores was darkened and placed in the growth chamber. gas exchange �e infrared gas analyser adc-225 mk-3 (great britain) was used to measure the intensity of photosynthesis and respiration of the moss gametophores in a closed system. �e entire closed system consisted of: an assimilation chamber, a water jacket and an air humidifying system, and its volume was 0.644 dm3. �e intensity of the light that reached the gametophores was 100 μmol × m–2 × s–1. �e temperature inside the assimilation chamber was 25ºc. measurements were carried out in air with 21% oxygen. koh scrubbers, together with the control kit, made it possible to obtain various concentrations of carbon dioxide. �e carbon dioxide concentration was controlled using an adc-225 mk-3 gas analyser. �e co2 concentration during the measurement was 300–500 μmol × mol–1 in a closed system. in contrast, the relative humidity was about 75%, thanks to the air being passed through a scrubber with distilled water. measurement of malate and citrate a speci�c sample of plant material (about 100–150 mg) was triturated with distilled water in a weight ratio of 1:4 for plagiomnium undulatum and 1:6 for polytrichum commune, and then centrifuged for 5 minutes at 5000 g at 4°c (hermle labortechnik z 36 hk, germany). �e content of malate and citrate in the test sample was measured at the end of the day (around 5:00 pm) and at the end of the night period (around 7:00 am). in order to determine the malate concentration, 500 μl of the bu�er (0.6 m glycylgycin, 0.1 m l-glucamate ph 10), 100 μl of nad+ (sigma), 5 μl of got (roché), 745 μl of redistilled water, 25 μl of the test sample and 5 μl of malate l-dehydrogenase, were mixed in the spectrophotometric cuvette (mdh, roché). for the determination a nd rz ej r ze pk a, w io le ta s za re k 114 of citrate, the following were mixed in the spectrophotometric cuvette: 180 μl of the bu�er (0.6 m glycylgycin, 0.6 mm zncl2 ph 7.8), 25 μl of nadh, 10 μl of mdh/ldh (sigma), 331 μl of redistilled water, 25 μl of the test sample and 5 μl of citrate lyase. �e concentration of malate and citrate was determined spectrophotometrically (cecil 9500, great britain), according to the möllering method (1985), at the wavelength λ = 340 nm, at room temperature. �e concentration of malate and citrate was expressed as the di�erence, according to the following formulas: (δ) malate (μm) = (malate)night – (malate)day (1) (δ) citrate (μm) = (citrate)night – (citrate)day (2) statistical analysis �e experiment was carried out in 3 repetitions. �e signi�cance of di�erences between mean values (± se) were analysed by the parametric test using the duncan’s test (p ≤ 0.05) in statistica 13.0 for windows. results changes in the intensity of gas exchange �e drying of the gametophores of both moss species to the loss of about 50% of water resulted in changes in the intensity of gas exchange (fig. 3, 4). in the case of polytrichum commune, an increase in the value of photosynthesis by 16.45 μmol co2 × gdm –1 × h–1 was observed, which was a di�erence of 21% compared to the control sample. conversely in plagiomnium undulatum – a reduction in the intensity of photosynthesis by 30.74 μmol co2 × gdm –1 × h–1 was shown here, which was a 25% di�erence in e�ciency compared to the control. �e respiration rate of gametophores subjected to the stress of drying and rehydration decreased in both moss species, compared to control. �is parameter in p. commune was 8.32 μmol co2 × gdm –1 × h–1, and in p. undulatum was 16.13 μmol co2 × gdm –1 × h–1, which constituted 18% and 15%, respectively, in relation to the control values. changes in the content of malate and citrate as a result of the conducted experiments, di�erences in the content of malate in the leaves of both species of mosses and between night and day, a�er drying and rehydration of the leaves, were found, compared to the control (tab. 1; fig. 5). in the case of the control sample in p. commune, the di�erence was 1.86 µm × gfm –1. in the test sample, this value was signi�cantly higher – 3.28 μm × gfm –1. in p. undulatum r esistance of m osses to drying, m easured by the intensity of gas exchange and the content of m alate and citrate 115 fig. 3. changes in the intensity of photosynthesis (a) and respiration (b) in gametophores of the plagiomnium undulatum (hedw.) t.j.kop. and polytrichum commune hedw., caused by dehydration and rehydration; mean values of 3 replicates (± se), data marked with di�erent letters di�er signi�cantly according to duncan’s test at p ≤ 0.05 there was clearly greater accumulation of malate in the dark period. �e di�erence between night and day for the control sample was 9.60 μm × gfm –1. when p. undulatum gametophores were dried to a  loss of 50% fresh mass and then rehydrated, the di�erence in malate concentration between night and day increased compared to the control sample (10.55 μm × gfm –1). in relation to the control, drought stress increased the malate concentration in p. commune by 1.42 μm × gfm –1, that is approximately 76%, while in p. undulatum – 0.95 μm × gfm –1, that is, approximately 10%. a nd rz ej r ze pk a, w io le ta s za re k 116 fig. 4. percentage changes in the intensity of photosynthesis (a) and respiration (b) in the gametophores of plagiomnium undulatum (hedw.) t.j.kop. and polytrichum commune hedw. caused by dehydration and rehydration (d/r); mean values of 3 replicates (± se), data marked with di�erent letters di�er signi�cantly according to duncan’s test at p ≤ 0.05 �e analysis of citrate content showed di�erences in its concentration in the leaves of p. commune and p. undulatum, between night and day, between the control and the experimental sample (tab. 1, fig. 5). in p. commune, in the control sample, this difference was 3.36 μm × gfm –1. �is value was higher in the test performed a�er drying and rehydration – 4.18 μm × gfm –1. in p. undulatum the di�erences in citrate content between night and day in the samples taken were smaller in comparison to p. commune. it reached the values of 1.75 μm × gfm –1 for the control sample and 1.45 μm × gfm –1 for the sample a�er drying and repeated rehydration. dehydration and rehydration increased the amount of citrate in p. commune by 0.81 µm × gfm –1, which was 25% of r esistance of m osses to drying, m easured by the intensity of gas exchange and the content of m alate and citrate 117 the control value. in p. undulatum there was a reduction in the amount of citrate by 0.32 µm × gfm –1, which was 21% compared to the control sample. tab. 1. changes in the concentration of malate and citrate in the leaves of plagiomnium undulatum (hedw.) t.j.kop. (a) and polytrichum commune hedw. (b), measured at the end of night and day, due to dehydration and rehydration; mean values of 3 replicates (± se), data marked with di�erent letters (within one species) di�er signi�cantly according to duncan’s test at p ≤ 0.05 treatment malate [μm × gfm–1] day (17:00) night (7:00) (δ) a b a b a b control 16.70 b±3.90 20.70 b ±0.70 26.30 a ±6.50 22.56 a ±0.47 9.60 b ±2.67 1.86 d ±0.23 dehydration/ rehydration 16.02 b ±7.71 20.18 b ±0.84 26.57 a ±9.82 23.46 a ±0.91 10.55 a ±2.67 3.28 c ±0.07 citrate [μm × gfm–1] control 10.55 b±1.27 11.85 b ±0.64 12.30 a ±1.67 15.21 a ±0.73 1.75 c ±0.41 3.36 b ±0.09 dehydration/ rehydration 10.98 b ±1.81 11.61 b ±0.78 12.41 a ±2.69 15.79 a ±1.36 1.43 cd ±0.87 4.17 a ±0.57 note: (δ) – malate or citrate content measured as the concentration di�erence between the end of night and day fig. 5. changes in the concentration of malate and citrate, expressed in% of control, in leaves of plagiomnium undulatum (hedw.) t.j.kop. and polytrichum commune hedw. measured at the end of the night and day, due to dehydration and rehydration a nd rz ej r ze pk a, w io le ta s za re k 118 discussion �e proper functioning of mosses in a natural environment depends on many external factors (rzepka, krupa, 2008; możdżeń, 2019). access to water is a particularly important parameter in land conditions. a slight water de�cit can cause signi�cant changes in metabolism and in the biochemical transformation occurring in moss cells, which were already mentioned in the introduction (green et al., 2011; de carvalho et al., 2014; romańska, 2020). mosses have the ability to completely dry out by balancing the internal potential of water with extremely dry air and resume normal functioning when rehydrated (alpert, 2000). desiccation tolerance has been experimentally con�rmed in over 200 species of mosses (wood, 2007). biological soil crusts are frequently subjected to cyclical desiccation-rehydration events and mosses have developed unusual constitutive and inducible desiccation tolerance mechanisms (wood, 2007; zheng et al., 2011; li et al., 2014). leafy stalks of mosses quickly balance with the potential of the surrounding water and can usually be fully hydrated or dry and metabolically inactive. when, the time required to recover from desiccation increases, the degree of regeneration decreases with the length of drying (proctor et al., 2007). in the present experiment it was shown that the plagiomnium undulatum and polytrichum commune gametophores reacted di�erently to dehydration and rehydration of cells. (fig. 3, 4; tab. 1). p. commune showed less changes in metabolic activity due to the stressor than p. undulatum (fig. 3, 4). both species regenerated very quickly upon rehydration. �e photosynthesis process was much more sensitive to slight disturbances in the water balance than dark respiration. �e reaction to partial dehydration is to reduce photosynthesis, while periodically increasing the respiration process (krupa, 1974). �is kind of response could be due to the fact that mosses are evergreen and perennial plants that show seasonal changes in metabolic activity related to the variation in temperature and water relations throughout the year (rzepka, 2008). in mosses, full photosynthesis, respiration and protein synthesis activity is restored within minutes and sometimes one or two hours; it may take 24 hours or more to activate the cell cycle, food transport and rebuild the cytoskeleton (pressel et al., 2006; proctor et al., 2007). �e time needed to reactivate the photosynthetic process depends largely on the amount of water lost, the dehydration time and the natural conditions in which the species lives. many mosses respiration intensity increases over time a�er rehydration, and eventually reaches a value similar to that prior to dehydration (krupa, 1974). in addition to the physiological tolerance to drying, mosses have anatomical and morphological features that allow you to e�ectively combat drought (vitt et al., 2014). �erefore, it can be assumed that the di�erences in the response of the studied mosses to the stress of drought could result from the di�erent shape, structure, arrangement r esistance of m osses to drying, m easured by the intensity of gas exchange and the content of m alate and citrate 119 of the lea�ets and the heterogeneous number and type of cells that build them (pressel et al., 2006). in some species of moss, the leaves are sharped and positioned laterally and overlap along the edges. �is may suggest that the capillary movement of water occurs from the base to the top of the plant (stark et al., 2017). at the narrower, more closed system leaves, reduces the rate of evaporation of water from the cells. �erefore, short periods of drought are better tolerated by mosses. other mosses have single-layer, symmetrically arranged leaves, growing from the stalks attached to the ground with a densely branched system of rhizoids. in this case, the water may be held in capillary spaces between individual gametophores or their elevations on the substrate. some mosses create tightly intertwined, while others form loosely layering turf, which also protects against water loss (vitt et al., 2014). anatomically, the lea�ets of p. commune are more diverse than p. undulatum. �e p. commune has lamellas, composed of a series of cells arranged along the rib of the leaf, with apical cells devoid of chloroplasts and with thickened walls (szafran, 1963) – fig. 1d. o�en the outer part of the thickened wall is warty, which protects against water loss from the remaining lamella cells or acts as a  light �lter. in this case, the presence of rows of lamella cells next to each other on the leaf surface determines the increase of the assimilation and the gas exchange areas (krupa, 1974; rzepka et al., 2005; rzepka, 2008). whereas p. undulatum has a single-layer leaf blade with a multi-layer rib, and the upper surface of the leaf is smooth (szafran, 1961, 1963) – fig. 1b. �e lack of anatomical structures in the leaves, protecting the assimilation organ against water loss, makes their metabolic activity dependent on water conditions in the environment. �is diversity in the anatomy of the lea�ets plays an essential role in photosynthetic production, closely related to water management. �e di�erences in the structure of the lea�ets are most o�en related to qualitative changes and are associated with a  di�erent assimilation surface and the size of the leaf blade. probably the structural diversity of moss leaves is an expression of evolutionary trends and their adaptation to terrestrial living conditions (krupa, 1974). �is is one of the many factors responsible for the di�erent reactions in photosynthetic and respiration activity in the analysed mosses. di�erences in the gas exchange of p. undulatum and p. commune gametophores subjected to dehydration and rehydration, could also result from changes in the activity of malate and citrate (table 1; fig. 5). malate is involved in processes related to the biosynthesis of organic compounds, energy transformation and mineral nutrition. it plays an important role as a factor regulating photosynthesis and glycolysis. �e concentration of citrate is related to the synthesis and accumulation of atp (rzepka, 2008). mosses have developed various systems to counteract water loss as previously mentioned (olivier et al., 2005). under stress, they show the ability of expression of lea proteins, high content of non-reducing sugars and e�ective antioxidant and photo-protection, a nd rz ej r ze pk a, w io le ta s za re k 120 are at least partly constitutive, allowing survival of rapid drying, but changes in gene expression resulting from mrna sequestration and alterations in translational controls elicited upon rehydration are also important to repair processes following re-wetting (bewley et al., 1978, 1993; proctor et al., 2007). �e results obtained in this experiment suggest that changes in the concentration of malate and citrate in mosses under stressful conditions could be an element of an adaptation strategy to water de�cit and allowed for high plasticity of changes in metabolism (rut et al., 2010). gabriel et al. (2005) described the tolerance to drying out in bryophytes as a reversible plastic feature. even drying and rehydrating a few times does not cause any serious changes in the functioning of the whole organism (krupa, 1974; mayaba et al., 2001). dehydration tolerant mosses can survive in dry atmospheres – with 50% and lower humidity values (alpert, oliver, 2002; wood, 2007). many species are able to withstand drying to a water content of 5–10% of their dry mass and return to normal metabolism and growth when rehydrated (alpert, 2006). such a way of surviving drought probably depends on: the speed of drying the plants, the relative water content of the surrounding environment and the drying time (green et al., 2011; stark, brinda, 2015; brinda et al., 2016). high resistance to drying of mosses and their ability to regenerate physiological processes a�er rehydration is a  characteristic feature that distinguishes them from vascular plants. �e lack of mechanisms protecting them against excessive water loss allows us to assume that there are physiological adaptations to the changing water content in cells (krupa, 1974). �e resistance of plants on the stress factor determines the plasticity of the cellular structures resulting from genetic conditions, the body’s ability to repair damage caused and the ability to adapt to the changing conditions of stress (levitt, 1980; bohnert et al., 1995). conclusion �e results of the experiment showed that drying to 50% of the relative water content and then rehydration of the plagiomnium undulatum and polytrichum commune gametophores a�ected changes in the daily activity of gas exchange and the content of malate and citrate. (1, 2) p. commune responded to the stressor to a lesser extent than p. undulatum. �e photosynthesis process was much more sensitive to changes in the amount of water than respiration. (3) changes in the content of malate and citrate indicated a  close relationship between cell hydration and their metabolic e�ciency. �is reaction was speci�c and depended on the species and anatomical and morphological structure of moss leaves. r esistance of m osses to drying, m easured by the intensity of gas exchange and the content of m alate and citrate 121 acknowledgments we would like to express my warm thanks to prof. jan krupa and phd grzegorz rut for invaluable substantive help, inspiring comments and suggestions, and help in planning the experiment. con�ict of interest �e authors declare no con�ict of interest related to this article. references alpert, p. 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(1998). ecophysiological responses of homoiochlorophyllous and poikilochlorophyllous desiccation tolerant plants: a comparison and an ecological perspective. plant growth regulation, 19(24), 211–217. https://doi.org/10.1023/a:1005951908229 vitt, d.h., crandall-stotler, b., wood, a. (2014). survival in a dry world through avoidance and tolerance. in: n. rajakaruna, r. boyd, t. harris (eds.), plant ecology and evolution in harsh environments, p. 267–295. new york: nova publishers. wood, a.j. (2007). �e nature and distribution of vegetative desiccation-tolerance in hornworts, liverworts and mosses. bryologist, 110, 163–177. https://doi.org/10.1639/0007-2745(2007)110[163:ienfib]2.0 .co;2 zheng, y., xu, m., zhao, j., zhang b., bei, s., hao, l. (2011). morphological adaptations to drought and reproductive strategy of the moss syntrichia caninervis in the gurbantunggut desert, china. arid land research management, 25, 116–127. https://doi.org/10.1080/15324982.2011.554956 abstract desiccation tolerance, the ability to lose virtually all of its free intracellular water and then restore normal function when rehydrated, is one of the most remarkable features of bryophytes. �e aim of the study was to determine the resistance of two species of plagiomnium undulatum (hedw.) t.j.kop. and polytrichum commune hedw. to drying to 50% relative water content of the air and rehydration. changes in the intensity of photosynthesis and respiration, as well as the content of malate and citrate in leafy moss stems were analysed. p. commune gametophores showed greater resistance to drought stress than p. undulatum. in both species, photosynthesis was much more sensitive to drought than respiration. changes in the content of malate and citrate indicated a high plasticity of moss metabolism in conditions of water shortage and may be one of many important elements of the adaptation strategy to water de�cit. �e reactions of the tested mosses to dehydration and rehydration con�rmed their adaptation to speci�c land conditions. key words: dehydration and rehydration, enzymatic activity, photosynthesis, respiration received: [2021.02.22] accepted: [2021.04.29] odporność mchów na wysychanie, mierzona intensywnością wymiany gazowej oraz zawartością jabłczanu i cytrynianu streszczenie tolerancja na wysychanie, czyli zdolność do utraty praktycznie całej wolnej wody wewnątrzkomórkowej, a następnie przywrócenia normalnego funkcjonowania po nawodnieniu, jest jedną z najbardziej niezwykłych cech mszaków. celem eksperymentu było określenie odporności dwóch gatunków plagiomnium undulatum (hedw.) t.j.kop. i polytrichum commune hedw. na wysuszanie do 50% względnej zawartości wody w powietrzu i ponownemu uwodnieniu. przeanalizowano zmiany intensywności fotosyntezy i oddychania oraz zawartości jabłczanu i cytrynianu w ulistnionych łodyżkach mchów. gametofory p. commune wykazały większą odporność na stres suszy niż p. undulatum. u obu gatunków proces fotosyntezy był znacznie bardziej wrażliwy na działanie suszy niż oddychanie. zmiany w zawartości jabłczanu i cytrynianu wskazywały na dużą plastyczność metabolizmu mchów w warunkach niedoboru wody. mogą one być jednym z wielu ważnych elementów strategii przystosowawczej do de�cytu wody. reakcje badanych mchów na wysuszanie i ponowne a nd rz ej r ze pk a, w io le ta s za re k 124 uwadnianie potwierdziły ich adaptację do specy�cznych warunków lądowych. słowa kluczowe: wysuszanie i uwadnianie, aktywność enzymatyczna, fotosynteza, oddychanie information on the authors andrzej rzepka https://orcid.org/0000-0003-3344-7768 his research interests are related to the response of plants, especially mosses, to abiotic and biotic stress factors. �e analyses include changes in selected metabolites, as well as the intensity of photosynthesis, respiration, transpiration, stomatal conductance and chlorophyll a �uorescence. wioleta szarek she is an msc student of biology. she is interested in mosses physiology. 220 annales universitatis paedagogicae cracoviensis studia naturae, 5: 220–237, 2020, issn 2543-8832 doi: 10.24917/25438832.5.14 sylwia śliwińska-wilczewska*, gracjana budzałek, zuzanna kowalska, marek klin, adam latała institute of oceanography, university of gdansk, av. piłsudskiego 46, 81-378 gdynia, poland; *ocessl@ug.edu.pl baltic macroalgae as a potential source for commercial applications – review introduction �e baltic sea is a well-known shallow inland sea that lies on the continental shelf in northern europe. �e baltic sea separates the scandinavian peninsula from the rest of continental europe and is surrounded by nine countries (i.e. sweden, finland, russia, estonia, latvia, lithuania, poland, germany, and denmark). �e baltic sea is an arm of the atlantic ocean, extending from northern germany and poland in the south almost to the arctic circle in the north. �e area of the baltic sea, together with the kattegat, covers about 415,000 km2. �e baltic sea is classi�ed as one of the world’s 66 “large marine ecosystems” (snoeijs-leijonmalm et al., 2017). �e marine in�uence on the baltic sea ecosystem is large but it is strongly in�uenced by freshwater as well. �e average salinity of the baltic sea is 7.5%, while the ocean’s salinity is 36.6%. salinity is also one of the main determinants of species diversity in macroalgae (rybak, 2018). �e �ora of the baltic sea is relatively poor, which means that there are few plant species in the baltic sea (deluga, 2018). however, despite the low salinity of baltic waters, there is a group of organisms that creates an interesting ecosystem. algae are considered one of the �rst forms of life that demonstrated the ability to photosynthesize, and they appeared on earth about one and a half billion years ago. �ese autotrophic non-tissue organisms are the most numerous group of self-nutrient aquatic organisms, and their growth is enabled by components such as phosphorus and nitrogen, as well as carbon dioxide and sunlight (buchholz et al., 2012). �ey do not have vascular systems, leaves, stems, and roots, but only elements that resemble them (bedoux et al., 2014). macroalgae are rich in various biologically active substances valued for their unique properties. currently, algae biomass is a  renewable source with many active substances that are widely used the chemical, food, agriculture, cosmetics, pharmacy, and medicine industries, among others (mclachlan, b altic m acroalgae as a potential source for com m ercial applications – review 221 1985). an example of the commercial application of algae mass resources from the polish part of the baltic sea is the use of furcellaria fastigiata for agar production (biłos et al., 2016). moreover, in poland, selected groups of macroalgae have been used as a raw material for industry and agriculture. �is group of algae includes ulva sp. and cladophora sp., which most o�en form high biomass in freshwater reservoirs (schroeder et al., 2013). macroalgae are classi�ed as so-called: green algae (chlorophyta reichenbach, emend. pascher, emend. lewis & mccourt), brown algae (phaeophyceae kjellman) and red algae (rhodophyta �uret, emend. rabenhorst, emend. adl et al.). macroalgae are a  source of mineral substances, polysaccharides, proteins, lipids, phytohormones, and pigments, as well as many di�erent secondary metabolites (phenolic compounds, terpenoids, halogenated compounds, sulfur derivatives, and nitrogen derivatives), and their biochemical composition depends on where the samples were taken, harvest season, and the environment (buchholz et al., 2012). hence, it is of important interest to present the possibilities for practical use of macroalgae derived from diverse ecosystems, such as the baltic sea (fig. 1). the potential use of bioactive compounds of baltic macroalgae in the medical and cosmetic industry macroalgae abound in various active substances used in medicine and the cosmetic industry (bedoux et al., 2014). macroalgae are considered a source of bioactive compounds because they can produce many di�erent secondary metabolites characterised by a broad spectrum of biological activity (gupta, abu-ghannam, 2011). fig. 1. examples of the possible use of baltic macroalgae as industrial resources s yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 222 baltic macroalgae used in the cosmetic industry �e use of macroalgae in the cosmetic industry is becoming increasingly desirable due to the natural ingredients’ broad spectrum and that they are easy to harvest, while at the same time they do not harm the environment. �e biomass of marine macroalgae is o�en used to produce ingredients in cosmetics. �ese ingredients can have one of three main functions: as additives that contribute to organoleptic properties, to stabilise and preserve a  product, or as bioactive compounds that perform the true function and cosmetic activity (bedoux et al., 2014). it has been shown that metabolites derived from macroalgae are useful in many aspects of anti-aging, anti-cellulite, and slimming care, as well as having antioxidant, photoprotective, moisturizing, and whitening properties (wang et al., 2014). �e diversity of macroalgae species and their broad biochemical composition means that they are a potentially a source of bioactive compounds (buchholz et al., 2012). macroalgae are rich in vitamins such as a, b1, b2, b3, b5, b6, b9, b12, c, d, e, and k, i.e. virtually all essential amino acids, macroelements and trace elements, such as iodine. �e skin aging process results from various factors inside and outside the body, which include diet, the passage of time and subsequent changes in the body, environmental pollution, and exposure to light or stress (bedoux et al., 2014). during skin aging, cells lose the ability to regulate generated reactive oxygen species and su�er from oxidative stress. in the epidermis and dermis, the number of melanocytes and keratinocytes is reduced, as well as the ability to multiply cells and migrate �broblasts, leading to a decrease in cellular activity and protein synthesis (wang et al., 2014). sulphated polysaccharides that are derived from marine algae may have various biological and biochemical e�ects, i.e. modulating proteolysis of connective tissue, anti-in�ammatory e�ects, adversely a�ecting the development of free radicals and antioxidants, as well as prevention of oxidative damage. �ese properties, among others, have been identi�ed for green algae, ulva lactuca l. and codium tomentosum stackhouse (bedoux et al., 2014). ulva populations originated mainly from shores and estuaries, with a minor number occurring in ocean lagoons, coastal lakes, canals, and rivers. ulva species were present in a  very wide range of saline waters (from fresh to hypersaline waters) (rybak, 2018). �erefore, the genus ulva is common in the baltic sea and may constitute 22–75% of biomass in the gulf of gdańsk (filipkowska et al., 2009). it is also worth mentioning that u. lactuca is one of the macroalga from the baltic sea described by linnaeus (dominguez, loret, 2019) and an organism that can be e�ciently used in the cosmetic industry. additionally, other species of ulva occur in the baltic sea – u. �exuosa wulfen, u. intestinalis l., and u. linza l., whose potential health properties have not yet been studied (fig. 2). on the other hand, codium tomentosum is not a  typical species b altic m acroalgae as a potential source for com m ercial applications – review 223 found in the baltic sea. however, the other species, c. fragile, can be found in the kattegat (nyberg, 2007). macroalgae are also an important source of amino acids, amino acid derivatives and peptides which can stimulate collagen production in the human skin. additionally, hydrolysates of algae proteins from macroalgae of the genus undaria, sargassum, porphyra, palmaria, and ulva show antioxidant activity, which is important in anti-aging care. �ese species: sargassum muticum (yendo) fensholt, porphyra umbilicalis kützing, palmaria palmata l., and ulva sp., also occur in the baltic sea (snoeijs-leijonmalm et al., 2017). it is worth noting that many marine macroalgae cannot survive the low salinity of the baltic sea; however, these species are still found in the lower sublittoral of the belt sea (helcom, 2012; snoeijs-leijonmalm et al., 2017). furthermore, a polypeptide from ulva sp., called aosa biopeptide, is used in cosmetic products and consists of glycine, arginine, lysine, valine, and aspartic acid. �e tripeptide consisting of arginine-glycine-aspartic acid was found to stimulate mechanisms underlying collagen production through tissue growth factor and cause an increase in collagen biosynthesis in human �broblasts. biopeptides from macroalgae can be used as a potential source of synthetic components and can contribute to the prevention of aging. enzymatic hydrolysis of algae proteins could allow for the isolation of peptides useful in the development of skin bioactive compounds. external aging causes exposure to uv radiation and associated irritation, characterized by a deterioration of the extracellular skin matrix and keratinocyte dysplasia in the epidermis, causing wrinkles, �abbiness, roughness, and spotty pigmentation. uv light is divided into uva, uvb, and uvc. uva can penetrate the dermis and not only causes wrinkles but also various skin-related ailments; uvb is more carcinogenic and causes the skin to turn red and burn 1,000 times faster than uva; while uvc is blocked by the ozone layer, and therefore does not pose a  risk to the skin (wells et al., 2017). macroalgae have mechanisms that counteract the harmful e�ects of uvb and uva, producing photosynthetic pigments such as carotenoids and phenolic compounds (e.g. phlorotannins) fig. 2. di�erent ulva species that occur in the baltic sea: ulva lactuca l. (a), ulva �exuosa wulfen (b), ulva intestinalis l. (c), and u. linza l. (d) (photo. s. śliwińska-wilczewska) s yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 224 or compounds that absorb uv radiation. uv absorbing compounds are bioactive and can protect human �broblast cells against cell death caused by uv radiation and inhibit the aging of human skin (bedoux et al., 2014). moisturizing is the most important step in preventing skin aging by helping to maintain a healthy appearance and elasticity while strengthening its protective barrier against harmful environmental factors. polysaccharides derived from macroalgae are environmentally friendly substances, relatively inexpensive, and can be used as substitutes for petrochemical products. polysaccharides and oligosaccharides provide a  high water storage capacity. macroalgae extracts contain �oridoside and iso�oridoside acids. �ese components have properties that enable them to increase water retention in the stratum corneum, and thus take part in moisturizing the skin. it was noted that the red algae chondrus crispus (l.) j.stackhouse are rich in polysaccharides and minerals that have a moisturizing, �rming, and soothing e�ect. moreover, extract from the green algae codium sp. can regulate the distribution of water in the skin, and thus protect the skin from dryness, especially in an arid environment (wells et al., 2017). �e described macroalgae species (chondrus crispus and codium fragile (suringar) hariot) can be found in the west area of the baltic sea (snoeijs-leijonmalm et al., 2017). c. crispus and c. fragile live permanently submerged in the kattegat at a water depth that ranges from 0.5–1 m to 4–5 m (nyberg, 2007). cellulite is a cosmetic problem that causes the appearance of hollows on the skin surface. �is problem is a combination of expanding fat cells that accumulate under the skin and �brous stripes that run perpendicular to the skin’s surface. cellulite includes changes in the biochemistry of the subcutaneous fat layer. �ese changes include increased lipogenesis, reduced lipolysis, and increased lipid storage in fat cells. energy homeostasis is regulated by the chronobiology of fat cells through the mechanisms of lipogenesis and lipolysis. some local cellulite treatments include macroalgae extracts, such as those used to increase microcirculation �ow, reduce lipogenesis, and improve lipolysis, which restore the normal structure of the dermis and subcutaneous tissue and remove or prevent free radicals. �e macroalgae most commonly used in anti-cellulite and slimming preparations are brown algae, fucus vesiculosus l., furcellaria lumbricalis (hudson) j.v.lamouroux, laminaria digitata (hudson) j.v.lamouroux, and pelvetia sp. (bedoux et al., 2014). �ey stimulate blood �ow and help the body excrete excess �uid. �us, the baltic macroalgae fucus vesiculosus and furcellaria lumbricalis can reduce or eliminate cellulite problems (fig. 3). the potential use of macroalgae in the medical industry macroalgae are also becoming increasingly popular due to properties that are useful in the medical industry (buchholz et al., 2012). �e most bioactive compounds in macb altic m acroalgae as a potential source for com m ercial applications – review 225 roalgae include sulfated polysaccharides, phlorotannins, and diterpenes. �ese compounds have strong antiviral, antibacterial, and anticancer properties. polysaccharides are a  class of macromolecules that are increasingly gaining importance in the �eld of biochemistry and medicine due to their immunomodulatory, anticancer, antiviral, and antithrombotic e�ects. �ey are mainly present in cell walls and their composition varies depending on the season, age, species, and geographical location of the macroalgae (gupta, abu-ghannam, 2011). in addition to con�rmed anticoagulant and antiviral activity, they act on the in�ammatory and immune systems. macroalgae produce various polysaccharides, such as alginic acid (also called algin), fucoidan, and laminarin (also known as laminaran) (milledge et al., 2015). alginate produced by brown algae, especially in the form of sodium and calcium alginate, is widely used in the pharmaceutical industry due to its ability to chelate metal ions and create very sticky solutions, while sulfated polysaccharides from macroalgae show various biological activities with potential medical value, including antitumor, antiviral, and antioxidant activity. fucoxanthin is another interesting compound. fucoxanthin is the main biofunctional pigment present in brown algae and is one of the most numerous carotenoids found in nature. fucoxanthin has antioxidant and anti-cancer properties. it has recently been found that fucoxanthin can help increase metabolism, thereby controlling weight gain (gupta, abu-ghannam, 2011). �ere are many di�erent species of brown algae in the baltic sea, including species of the genus fucus (fucus cottonii m. j. wynne & magne, f. evanescens c. agardh, f. radicans l. bergström & l. kautsky, f. serratus �unberg, f. spiralis l., f. vesiculosus), as well as ascophyllum nodosum l., ectocarpus siliculosus (dillwyn) lyngbye, scytosiphon lomentaria (lyngbye) link, chorda �lum l. and pylaiella littoralis l., hence, baltic brown algae could useful to both the pharmaceutical and the medical industries (snoeijs-leijonmalm et al., 2017). fig. 3. examples of the baltic macroalgae: brown algae sargassum muticum (yendo) fensholt (a), red algae chondrus crispus l. (b), and green algae codium fragile (suringar) hariot (c) that can be used in the cosmetic industry (photo. s. śliwińska-wilczewska) s yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 226 baltic macroalgae used in the food industry macroalgae, due to their richness in vitamins and macroelements, are o�en use in the food industry. from a  nutritional point of view, edible macroalgae are low-calorie foods with a high concentration of minerals, vitamins, and proteins, and low lipid content. currently, they are willingly consumed on a global scale. �anks to their interesting properties, they are also used in the production and processing of food products such as emulsifying, gelling, or water-retaining agents. macroalgae are an excellent source of vitamins a, c, d, and e and group b vitamins such as ribose, niacin, pantothenic acid, and folic acid (fitzgerald et al., 2011). �ey also derive an unusual wealth of minerals from the sea, which can constitute up to 36% of their dry matter. macronutrients include sodium, calcium, magnesium, potassium, chlorine, sulfur, and phosphorus as well as micronutrients such as iodine, iron, zinc, copper, selenium, molybdenum, �uorine, manganese, boron, nickel, and cobalt (kandale et al., 2011). it follows that the composition of macroalgae comprises almost all of the vitamins, the most important macroelements necessary for the proper functioning of the human body, as well as iodine which is responsible for a healthy thyroid (buchholz et al., 2012). as a result, algae are o�en used as components of a healthy diet, in the form of salads, soups, various types of snacks, and also as widely distributed dietary supplements in well-developed countries to help maintain health. macroalgae are a  rich source of iodine. �e highest iodine content is found in brown algae. in most cases, red and green algae have lower iodine content but still a high value compared to terrestrial plants. �e daily adult requirement for iodine is currently 150 µg per day, and this dose can be reached by very small amounts of macroalgae. just one gram of dried brown algae contains 500 to 8,000 µg of iodine, and green algae and red algae provide 100–300 µg in one gram (kandale et al., 2011). it follows that the introduction of a small number of macroalgae into the diet is enough to maintain a healthy thyroid. furthermore, a large part of the world population receives insu�cient iodine because other food sources (plants, animals) contain very low levels of this element. an interesting feature of macroalgae is their richness of proteins and bioactive peptides. �e composition and concentration of proteins depend on the species, place, and time of harvest. �e protein fraction of brown algae is relatively low (1–24% of dry matter) compared to green algae (4–44% of dry matter) and red algae (5–50% of dry matter) (fitzgerald et al., 2011). most macroalgae contain all necessary amino acids and acidic amino acids, e.g. glutamic and aspartic acids, and as is well known, protein is a critical nutrient due to its very important role in the human body. macroalgae are a very good source of nutrients and have a relatively low-fat content (0.3–5% of dry matter) (bedoux et al., 2014). b altic m acroalgae as a potential source for com m ercial applications – review 227 macroalgae also contain large amounts of polysaccharides (which include hydrocolloids), especially in their cell walls. hydrocolloids have gained commercial signi�cance as food additives. �e food industry uses their gelling, water-retaining, emulsifying, and other physical properties. agar is used in food products such as confectionery, meat and poultry products, desserts and drinks, and molded foods. carrageenan is used in salad and sauces, dietary foods, and as a preservative in meat and �sh products, dairy products, and baked goods (kandale et al., 2011). regular consumption of macroalgae has many positive health bene�ts which were describe in the previous chapter. macroalgae are also widely used in the food industry as thickening aqueous solutions, for gel production, and in the production of items such as toothpaste, ice cream, and anti-cancer mayonnaise (milledge et al., 2015). macroalgae have gained great popularity as an interesting addition to dishes or as basic ingredients of dishes. �e best example is sushi, which is a well–known food across the world, for which nori (red algae porphyra umbilicalis) is needed. other examples are kombu (brown algae laminaria japonica areschoug) and wakame (brown algae undaria pinnati�da (harvey) suringar), which are used in various types of salads, soups, and starters (kolb et al., 2004). although the mentioned species do not occur in the baltic sea, other common baltic species of brown algae (e.g. fucus sp., ascophyllum sp.) or red algae (e.g. furcellaria sp., polysiphonia sp.) can be successfully used in the food industry. possible use of baltic macroalgae in the energy industry �e energy industry is of vital importance for the proper functioning of the world’s economies. alternative energy sources are being sought out as non-renewable resources are becoming depleted. carbon, which is the most basic energy source in poland, is not an ideal source of energy due to the negative impact on the environment during its combustion or extraction, similar to oil. �erefore, alternative raw materials are sought, with one being biodiesel obtained from macroalgae. for now, it is quite a  promising material that has great market potential, as its production can be very pro�table with low environmental losses. at present, macroalgae are not widely used as energy sources; however, many promising studies are being conducted in this direction (filipkowska et al., 2008). several solutions have been proposed for the production of biofuels to replace fossil fuels. biodiesel and bioethanol produced by terrestrial plants have attracted the world’s attention as potential alternative solutions. however, due to the other various uses of terrestrial plants, e.g. for consumption, doubts have arisen about converting them to biofuels. �erefore, it is necessary to �nd a new raw material for the production of biofuels that can generate similar by-products as terrestrial plants but can be s yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 228 produced solely for energy purposes. macroalgae are one alternative to conventional terrestrial plant cultivation. �e production of biofuels from macroalgae is widely considered to be one of the most suitable methods. �ese plants are an economically and environmentally sustainable renewable biomass source for biofuel production because they are simple aquatic organisms that carry out photosynthesis and their diversity is much greater than terrestrial plants. almost all of macroalgae biomass can be converted into energy and their waste residues into methane and hydrogen. �ey also contain oils that can be used as a raw material for biodiesel and carbohydrates that can be converted into bioethanol. macroalgae have the advantage of not interfering with food production and have the potential to cover global demand for transport fuels (suganya et al., 2016). anaerobic digestion is a biological process used to produce methane-rich gas from biomass. �is digestion process involves the following steps: i) hydrolysis of organic matter, ii) the synthesis of acetic acid from hydrolysis products, and iii) the production of methane by the relevant bacteria. macroalgae have a similar composition as other sources of organic materials and should be adapted to existing anaerobic digestion installations. however, the content of seaweed-related compounds, such as salts, can inhibit the fermentation process. �erefore, they need to be pre-processed, including at minimum washing, to prevent inhibition of the fermentation process (milledge et al., 2014). anaerobic fermentation of macroalgae allows for the production of biogas consisting of about 60% methane and 40% co 2 (and trace amounts of co, h2s, h2, n2, and o2). �e composition of macroalgae may also vary depending on the season and location, which may a�ect production performance. for example, the methane yield of saccharina latissima (linnaeus) c.e.lane (syn. laminaria saccharina (l.) lamouroux) algae is twice as high from material collected in autumn than material collected in spring (roesijadi et al., 2010). s. latissima occurs in the west area of the baltic sea; however, this species is poorly adapted to the low salinity. it is assumed that at the inner baltic distributional limit of s. latissima in the southern baltic sea proper, its upper limit lies at the water depth of about 20 m (wærn, 1965). fermentation of biomass to ethanol consists of two reactions: i) hydrolysis of organic material to simple sugars (using high temperature, enzymes, or acids) and ii) fermentation of sugars to ethanol. macroalgae use polysaccharides such as sulfated polysaccharides, mannitol, alginates, agar, and carrageenan to produce ethanol. brown algae, especially those of the genus laminaria, are the most desirable macroalgae for alcoholic fermentation because they contain the most polysaccharides. macroalgae biomass has great potential for producing biofuels, although there are challenges in its processing. however, much research is currently underway to identify methods for maximizing its potential for energy generation (milledge et al., 2014). b altic m acroalgae as a potential source for com m ercial applications – review 229 baltic macroalgae as natural fertilisers and products for wastewater treatment macroalgae are becoming increasingly popular as natural fertilisers. �is is due to the presence of nitrogen, magnesium, potassium, and some trace elements in their composition (mohy el din, 2015). �ese plants also have a great ability to absorb nitrogen and phosphorus, which are very important to their growth. in addition to macroelements and microelements, they also contain phytohormones, which positively a�ect growth of macroalgae. liquid macroalgae extracts and suspensions have gained wider application than microalgae meal due to their ease of use. most o�en, fertilisers are based on algae such as ascophyllum sp. and fucus sp. �ese macroalgae are found mainly in the western part of the baltic sea. �e intertidal species f. vesiculosus lives permanently submerged in the kattegat. it grows just below the water surface, with f. spiralis above it and ascophyllum nodosum (linnaeus) le jolis and fucus serratus l. below it (snoeijs-leijonmalm et al., 2017). large amounts of insoluble carbohydrates act as soil improvers (better aeration, structure) and also help retain soil moisture. it has been shown that they not only provide nutrients needed for growth but also increase seed germination, improve tolerance to low temperatures, and are harmful to some insects (milledge et al., 2015). �ey can, therefore, be an alternative to abused chemical fertilisers that have a negative impact on the environment �ere are two main areas where macroalgae have the potential to be used in wastewater treatment. �e �rst is for wastewater and some agricultural waste treatments to reduce total nitrogenand phosphorus-containing compounds before release of these puri�ed waters into rivers or oceans. �e second is for the removal of toxic metals from industrial wastewater. macroalgae can be used to reduce nitrogen and phosphorus content in wastewater a�er treatment. many microalgae take up ammonium as an absorbable form of nitrogen for growth, and this ion is a common form of nitrogen in most domestic and agricultural wastewater. another important feature of many macroalgae is their ability to absorb large amounts of phosphorus (mchugh, 2003). �e usefulness of macroalgae in wastewater treatment is increasingly known but its largescale application has not yet been implemented, although this may change with growing awareness of the need to protect the marine environment. �e accumulation of heavy metals such as copper, nickel, lead, zinc, and cadmium by macroalgae became apparent when they were consumed and their chemical composition studied. �e content of heavy metals, especially in large brown algae, depends on their geographical location as well as their proximity to industrial points. for this reason, the idea of using macroalgae as biological indicators of heavy metal pollution, both from natural sources and from activities such as the extraction or disposal of industrial waste, was proposed (mchugh, 2003; żbikowski et al., 2006, 2007). removal of toxic metals was s yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 230 implemented using brown algae of the genera sargassum, laminaria and ecklonia and the green algae ulva sp. �ey were also used to remove heavy metals in wastewater treatment (mchugh, 2003). among these organisms, sargassum muticum occurs in the baltic sea but only in the more saline areas in the arkona sea, the belt sea, and the öresund (nyberg, 2007). ulva sp. is commonly found in the entire baltic sea. it is possible to use baltic macroalgae to remove heavy metals near factories, processing plants, and sewage treatment plants as one of the stages in this process. macroalgae as a biological factor limiting the occurrence of massive cyanobacterial blooms in the coastal area of the baltic sea �e presence of macroalgae is of great importance for the biological structure, ecosystem functioning, and water quality in shallow water bodies (van donk, van de bund, 2002; hilt, gross, 2008), as well as in the baltic sea. underwater meadows provide shelter for the development of numerous animals and are the basis of the diet for aquatic organisms (rybak, 2016; wells et al., 2017). �ey can also be used to assess the ecological state of the marine environment (saniewski, 2013). macroalgae e�ectively compete with phytoplankton species for available nutrients (rybak, gąbka, 2018). it has been shown that in a  large number of cases, allelopathic compounds produced by macroalgae act on organisms in a  di�erent way than most synthetic herbicides (ohad, hirschberg, 1990; huppatz, 1996). �ese compounds have a broader range of action on target organisms than synthetic inhibitors (duke et al., 2001). �erefore, the importance of the allelopathic e�ects of macroalgae in aquatic ecosystems is gaining interest due to their potential ability to control or limit the occurrence of organisms forming massive blooms, including harmful cyanobacteria (berger, schagerl, 2003; wang et al., 2007; tang, gobler, 2011; pakdel et al., 2013; ghobrial et al., 2015). złoch et al. (2018) tested the allelopathic activity of several representatives of baltic macroalgae from the genus chara on the growth of picocyanobacteria synechococcus sp. it was shown that the addition of aqueous extracts from chara aspera c.l.willdenow, c. baltica (c.j.hartmann) bruzelius, and c. canescens loiseleur signi�cantly a�ected the cell number in synechococcus sp. culture. �e addition of aqueous extracts from c. canescens in a concentration of 5, 25, and 50 μl ml–1 caused the cell number of picocyanobacteria to decrease to 86%, 78%, and 48%, respectively, a�er a week of exposure, compared to control conditions. on the other hand, the addition of 25 and 50 μl ml–1 extracts obtained from c. baltica caused a reduction in the cell number of synechococcus sp. to 55% and 44%, respectively. in turn, the addition of 50 μl ml–1 of an extract from chara aspera inhibited the growth of picocyanobacteria a�er a week of exposure. in this experiment, the number of picocyanobacteria cells decreased to 90% compared to the control treatment. it is believed that the chemical composition b altic m acroalgae as a potential source for com m ercial applications – review 231 and proportions of allelopathic compounds may play a signi�cant role in determining their e�ect on the growth of various phytoplankton species, including cyanobacteria (pakdel et al., 2013). �erefore, weaker inhibition of the growth of picocyanobacteria by c. aspera extract, compared to c. baltica and c. canescens, may be due to the presence of various allelopathic compounds and their concentrations. �is varied sensitivity to allelopathic compounds indicated that species belonging to the genus chara may a�ect the growth and occurrence of synechococcus sp. moreover, the negative effects may be stronger in the natural environment, since these macroalgae occupy large areas of the bottom of the southern baltic sea and allelopathic compounds can be constantly released into environment (granéli, hansen, 2006). studies demonstrating the allelopathic activity of c. aspera, c. baltica, and c. canescens on synechococcus sp. indicated that these macroalgae may potentially control the extent of picocyanobacteria in coastal baltic waters. as mentioned earlier, the lack of recognition of the allelopathic activity of baltic macroalgae from the southern baltic region on the associated species of �lamentous cyanobacteria prompted us to conduct relevant laboratory experiments. representatives of typical the baltic macroalgae ulva intestinalis, as well as �lamentous cyanobacteria nodularia spumigena and nostoc sp., were selected for the study. �e cyanobacteria strains were maintained as a non-axenic batch culture in the culture collection of baltic algae (ccba) at the laboratory of marine plant ecophysiology, university of gdańsk, poland. �e brackish macroalga was selected, based on the allelopathic potential of species from the order of ulvales (e.g. wang et al., 2007). baltic u. intestinalis was collected manually from the coastal zone of the gulf of gdańsk, poland (54°47ʹ13.8ʺn 18°25ʹ33.7ʺe) and carefully washed several times with distilled water in the laboratory to remove attached organisms as described by previous researchers (złoch et al., 2018). �e allelopathic e�ects of u. intestinalis were tested according to a method proposed by wang et al. (2007) with modi�cations. �e monocultures of cyanobacteria nodularia spumigena mertens ex bornet & flahault and nostoc sp. were exposed to three concentrations of both water extracts and �ltrate originating from u. intestinalis. �e �nal concentrations of extract and �ltrate were: 5, 25 and 50 µl ml–1. moreover, to compare the e�ects on the two cyanobacteria of di�erent concentrations of fresh tissue from the u. intestinalis, coexistence assays were performed using a mixed culture system of one macroalga and one cyanobacteria. di�erent initial inoculation concentrations (0, 0.5, 2.5, and 5 g-wet wt l−1) of fresh macroalgal tissues were inoculated into 25 ml erlenmeyer �asks containing 20 ml f/2 culture medium. selection of these concentrations was based on preliminary experiments to determine the most relevant e�ectiveness. it was found that the growth of target cyanobacterium n. spumigena, a�er addition of 5, 25, and 50 µl ml–1 �ltrate and the presence of 0.5, 2.5, and 5 g-wet wt l−1 live tiss yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 232 sue from u. intestinalis, was signi�cantly reduced to approximately 40%, compared to the control treatment (fig. 4). on the other hand, the addition of �ltrate and the presence of live tissue of u. intestinalis had no statistical e�ect on the number of nostoc sp. cells (p > 0.05; fig. 5). slight stimulation of the tested cyanobacteria was noted a�er the addition of aqueous extract, which may be the result of hormesis or the accumulation of nutrients in macroalgae tissue. based on the results, it can be concluded that baltic macroalgae of the genus ulva have the potential to a�ect the occurrence and abundance of some bloom-forming species of cyanobacteria. research on the allelopathic e�ect of macroalgae on the growth of �lamentous and bloom-forming cyanobacteria occurring in the southern baltic sea region is an interesting pursuit that should be recognized as requiring continuation. fig. 4. �e number of nodularia spumigena mertens ex bornet & flahault cells (105 cell ml−1) for controls and experiments with addition of (a) extracts: 5 (a), 25 (b), and 50 (c) (μl ml−1), (b) �ltrate: 5 (a), 25 (b), and 50 (c) (μl ml−1), and (c) tissue: 0.5 (a), 2.5 (b), and 5 (c) (g-wet wt l−1) obtained from ulva intestinalis l. a�er 14 days of exposure. �e values refer to means (n = 3, mean ± sd). asterisks indicate statistically signi�cant di�erence compared to the control obtained with the anova. levels of signi�cance were *p < 0.05; **p < 0.01; ***p < 0.001. b altic m acroalgae as a potential source for com m ercial applications – review 233 conclusions baltic macroalgae are organisms which may be used as industrial resources thanks to their unique properties. due to their content of vitamins, as well as macroand microelements, they are becoming increasingly popular as foods and supplements but also have great potential to gain signi�cance in the pharmaceutical industry. similar trends can also be observed in the cosmetic industry, where macroalgae are an increasingly important ingredient in anti-aging, anti-cellulite, and moisturizing products. �e energy industry has also become interested in macroalgae due to the search for natural solutions for energy production, i.e. biofuel. baltic macroalgae can also be used as natural fertilisers and products for wastewater treatment. finally, based on the research, it can be concluded that some baltic macroalgae may allelopathically a�ect bloom-forming cyanobacteria. �us, the study of the allelopathic activity of macroalfig. 5. �e number of nostoc sp. cells (105 cell ml−1) for controls and experiments with addition of (a) extracts: 5 (a), 25 (b), and 50 (c) (μl ml−1), (b) �ltrate: 5 (a), 25 (b), and 50 (c) (μl ml−1), and (c) tissue: 0.5 (a), 2.5 (b), and 5 (c) (g-wet wt l−1) obtained from ulva intestinalis l. a�er 14 days of exposure. �e values refer to means (n = 3, mean ± sd). asterisks indicate statistically signi�cant di�erence compared to the control obtained with the anova. levels of signi�cance were *p < 0.05; **p < 0.01; ***p < 0.001. s yl w ia ś liw iń sk aw ilc ze w sk a, g ra cj an a b ud za łe k, z uz an na k ow al sk a, m ar ek k lin , a da m l at ał a 234 gae may help determine their possible role as an important biological factor a�ecting cyanobacterial blooms. our review shows that the most economically important macroalgae species are found in the western part of the baltic sea. �is is due to the greatest richness and variety of species. although it is worth noting that cosmopolitan species, such as ulva, occur throughout the baltic sea, so in every region of this sea it is possible to utilize these organisms for commercial purposes. �us, in our opinion, there are su�cient species resources in the waters of the polish part of the baltic sea for commercial use. �e question that arises is whether the degree of pollution of the baltic sea waters allows for the use of registered algae for cosmetic, pharmaceutical, and food purposes. unfortunately, the current literature does not provide unambiguous answers to this question. lack of su�cient data on the amount of accumulated heavy metals and other anthropogenic pollutants in macroalgae tissues and the speci�c limits that must be met leave this question open. �e authors can only speculate that the use of algae that come from open water near the danish straits, where the amount of pollution is less due to greater water exchange, is more favourable. studies on the practical use of baltic macroalgae are still ongoing. �e results of previous studies are quite promising because baltic macroalgae are easy to grow, relatively inexpensive, and less invasive, and these features are highly desirable in order to protect the environment and natural resources. acknowledgements �e authors would like to thank the anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. con�ict of interest �e authors declare no con�ict of interest related to this article. references bedoux, g., hardouin, k., burlot, a.s., bourgougnon, n. 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(2006). distribution and relationships between selected chemical elements in green alga enteromorpha sp. from the southern baltic. environmental pollution, 143(3), 435–448. https://doi.org/10.1016/j.envpol.2005.12.007 b altic m acroalgae as a potential source for com m ercial applications – review 237 żbikowski, r., szefer, p., latała, a. (2007). comparison of green algae cladophora sp. and enteromorpha sp. as potential biomonitors of chemical elements in the southern baltic. science of the total environment, 387(1–3), 320–332. https://doi.org/10.1016/j.scitotenv.2007.07.017 złoch, i., śliwińska-wilczewska, s., kucharska, m., kozłowska, w. (2018). allelopathic e�ects of chara species (c. aspera, c. baltica, and c. canescens) on the bloom-forming picocyanobacterium synechococcus sp. environmental science and pollution research, 25(36), 36403–36411. https://doi. org/10.1007/s11356-018-3579-5 bałtyckie makroglony jako potencjalne źródło zastosowań komercyjnych streszczenie morze bałtyckie jest wyjątkowym ekosystemem wodnym, charakteryzującym się wyraźnymi zmianami w środowisku, szczególnie w odniesieniu do zasolenia i klimatu. jest to także miejsce występowania morskich i słodkowodnych organizmów roślinnych, które od stuleci fascynują naukowców. niewiele jest jednak prac prezentujących bałtyckie glony makroskopowe, jako potencjalne źródło dla zastosowań komercyjnych. celem niniejszego opracowania było przedstawienie bałtyckich makroglonów, jako źródła zasobów przemysłowych. w  przeglądzie uwzględniono, m.in. potencjał wykorzystania tych organizmów w  przemyśle kosmetycznym i medycznym, w tym najważniejsze składniki, które czynią je cennym produktem spożywczym. zwrócono także uwagę na ich rosnącą popularność i potencjalne wykorzystanie w przyszłości, np. jako biopaliwa, nawozy naturalne lub składniki oczyszczalni ścieków. przedstawiono także możliwość wykorzystania makroglonów jako biologicznego czynnika, ograniczającego występowanie masowych zakwitów sinic w morzu bałtyckim. key words: baltic macroalgae, baltic sea, bioactive compounds, biomass, industrial resources received: [2020.02.27] accepted: [2020.06.04] annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x landscape engineering and aesthetics approaches to design a community garden: social activity zones appraisal in the envisaged drwinka river park (kraków, poland) wiktor halecki institute of nature conservation, polish academy of sciences, adama mickiewicza ave. 33, 31-120 kraków, poland; halecki@iop.krakow.pl introduction many countries are placing an increasing emphasis on conservation services as urbanization continues/ progresses (ghelfi, papadopoulos, 2022). the goal is to improve the quality of life in cities, including the economy. this is achieved by building a network of greenery. additionally, agents operate both on and off the street. in new green areas in cities, “natural landscapes” are more and more often drawn (kittiprapas, 2022). in terms of plant species, a wide range of different plants is used, including grasses, forest plants, aquatic plants, etc. to relocate non-urban landscapes into cities, the so-called naturalistic styling is used (brown, 2008). the greenery concept can be used in all kinds of places, including small spaces, benches in small architecture, as well as in water bodies (jelks et al., 2021). landscape architecture designed specifically for drought-prone or water-conserving areas is knows as xeriscaping (çetin et al., 2012). green spaces must be included in the structure of a city (gorham et al., 2009; rodriguez-plesa et al., 2022). each area is distinguished by its size, composition, and plant diversity, as well as the function they serve for sustainable cities (albino et al., 2020). among green spaces, there are city parks, historic parks, city forests, squares, rooftop gardens, home gardens, pocket gardens, allotment gardens, and cemeteries. in addition, one can distinguish street greenery i.e. vegetation growing along roads and pedestrian routes, temporary gardens, and climbing plants, which are called “green walls” (matoga et al., 2015; joshi et al., 2022). large areas of green space with a predominance of biologically active areas act as ecological corridors for wildlife and plants (kondo et al., 2018; halecki et al., 2022). these primarily include urban forests, parks, and vegetation in the valleys of unregulated, natural rivers. some areas have been “adapted” and incorporated into green areas. common examples are remnants of ancient, natural forests that have been converted into urban spaces (wang, 2022). in addition to serving as biodiversity refuges, these forests also contribute to recreational areas in cities. some of them are located within the administrative boundaries of cities and are distinguished by arranged vegetation, planted by man, as well as fragments of spontaneous and natural vegetation, which is a remnant of forests. these areas serve many functions, including aesthetic, cultural, social, tourist, and educational (wolch, 2014, yan et al., 2018). moreover, educational paths, such as those in city forests, are very appealing. as a part of an educational trail, participants may learn more about the genesis of a particular forest, the species composition of plants, animals, and insects as well as issues related to management and protection in that area (kaletová et al., 2022). it was planned to assess the development of green areas of the “na kozłówce” housing estate, located in the xii bieżanów-prokocim district, in the south-eastern part of kraków (50°03′41″n 19°56′18″e, southern poland; fig. 1). fig. 1. the figure shows the protection zones of river parks in kraków (in red); the drwinka river park is marked with yellow ellipse in this study, i present an analysis of the area between the street and the planned drwinka riverfront park area, located south of the neighbourhood. an attempt has also been made to develop a project for the recomposition of this area, and in particular to unify and create a convenient and accessible space for residents. the (main) aim of this project has been designing the river park as an environmentally friendly area that tourists and residents could use all year round. by this design, it will positively affect the aesthetics of the neighbourhood and the quality of life of its residents. characteristics of the project area kraków experiences both hot summers and cold winters, as the area is under the pronounced influence of a continental climate. the city records an average annual air temperature of 8.7 °c. the warmest months are july and august with an average temperature of about 19 °c. while the coldest month is january with an average temperature of 0 °c to -5 °c. the study area is located on a slope to/towards the northwest, separated by two road arteries. the drwinka valley encloses nowosądecka street and wielicka street to the south. since the site is a part of the vistula catchment area, the groundwater is of tertiary origin. a small stream, the drwinka, runs nearby. in drwinka valley, a river park is planned (fig. 1). in the study area, the office of spatial planning of kraków city found the presence of urban soil (urbisol), or anthropogenic soil. urban soils and old parks soil are common in the study area. in these soil profiles, some layers display traces of earlier use (e.g. remnants of pavements and foundations). the area surrounding the settlement is mostly covered with dense buildings (fig. 2–3). industrial and commercial buildings are located to the north and east of the settlement. along the estate, there are located gardens, allotments, urban green areas, and small deciduous forests. residents take care of the greenery and many types of bedding plants, shrubs, and woody plants can be seen here. thus, there is both arranged and unarranged vegetation throughout the analysed area, along with neighbourhood greenery. there are a lot of trees surrounding buildings, green squares, and greenery near sports facilities in areas of multi-family housing. in addition, traffic routes are planted with low hedges, and streetcar tracks are covered with grass and are known as “green tracks”. arterial roads and local roads form a dense network in the part of the city near the drwinka stream, where the river park is being developed. to the northeast lies jerzmanowski park with its old trees. pedunculate oaks have a long history here. at the same time in the drwinka valley itself, situated south of the settlement, there are deciduous trees and shrubs along the river. there is an ash-elm riparian forest in this region, while most of the land directly in the drwinka valley is occupied by alder forests. fig. 2. developing green spaces in the analysed area “na kozłówce” (south-eastern kraków) fig. 3. spatial development in the analysed area “na kozłówce” (south-eastern kraków) the main objectives of the project greenery analysis tall greenery is abundant in the development area, which consists mainly of coniferous and deciduous trees. inter-block greenery enhances the aesthetic qualities/ aesthetic value of the adjacent blocks (fig. 4 – appendix 1). a community garden can be beneficial for residents, including the elderly (fig. 5 – appendix 1). there are several assumptions that community gardens should meet:  integration – it is one of the most important factors affecting the quality of life of residents living in densely populated areas. as a result of integration the sense of belonging to a given place increases leading to a greater sense of security and responsibility for the environment (joshi et al., 2022);  education – nowadays, it is crucial to raise awareness of the value of nature, environmental protection, and ecology. thanks to ecological solutions, such as permeable surfaces, retention reservoirs, apiaries, insect hotels, or birdhouses, residents can be introduced to nature and increase their ecological awareness. the communit y garden allows conducting workshops on various activities both for schoolchildren, students, and pre-schoolers from nearby institutions, as well as for everyone interested in the subject. thanks to the gardener’s house, the activities can be conducted throughout the year, not limited to the season and weather conditions (kou, zhang, 2019);  recreation – leisure is an integral part of everyone’s life. it will certainly be much more pleasant to relax in a semi-private space, enclosed in the beautiful scenery, than within the dense and monotonous block of flats. visitors are attracted to this friendly and peaceful place in increasing numbers, and some may even discover a passion for social gardening, naturism, or just having a good time in a natural environment (asensio, 2022). detailed instructions for shaping the plant cover the richness of dendroflora in the “na kozłówce” housing estate, fulfills its functions both in terms of aesthetics and functionality. apartments need to be exposed to greenery to suppress noise and protect it from excessive snowfall. in this regard, local authorities can create a unified plan of action to coordinate different types of greenery throughout the neighbourhood. most city dwellers live in urban environments that favour the creation of community gardens. however, not everyone can afford or maintain a community garden, which negatively affects the appearance of the home. households cannot maintain gardens. by separating green areas and ameliorating soil quality, community gardens (kirsch et al., 2022) and forested urban parks (millward, 2011) can be improved. a plan for land development composition in the conceptual design for the redevelopment of the “na kozłówce” housing estate, the leading form is the square. the plan mainly includes elements of small architecture, such as kiosks, bicycle playgrounds, flower beds, gazebos, and even garbage cans. the leitmotif is intended to enliven the monotonous landscape and, in a way, unify it in the neighborhood structure. the study area contains public (“na kozłówce” park) and semi-public (semi-private and private) zones. in the central part of the estate, a community garden was placed both for integration as well as educational, recreational, and representative purposes. flowerbeds, greenhouses, gazebos, and outbuildings are arranged in squares throughout the estate. fig. 6. illustration of the conceptual design. honey plants, perennial plants, green roof, hives, and picnic and meeting areas are included in this project a community garden was proposed as a place for learning, relaxation, and contact with neighbours. a detailed plan, selection of plants, and landscaping elements were developed. with the use of insect hotels, birdhouses, beehives, and, in a technical context, retention tanks, the site was also able to cope with the ecological aspect of the project. a detailed analysis of the district and neighbourhood was carried out, which allowed a solid understanding of the study area (fig. 2). many factors were taken into account, including terrain, development, communications, historical and cultural connections, greenery, and the functionality of the neighbourhood. based on the above analyses, design guidelines for the recomposition of the estate were developed. raise beds, greenhouses, gazebos, and farm buildings are all arranged in squares throughout the estate. fig. 7. elements of social garden equipment used during picnics on the grass; the infrastructure of the estate garden consists of: a) bicycle room, b) seats, c) exhibition stand functional zones the green zone combines both passive and active activities. a section of the establishment has been divided into subzones so that the function meets the needs of the residents. green, yellow and blue residential zones have been separated. residents of the blue zone primarily benefit from the community garden located nearby. in the green zone, there are places for relaxation such as gazebos. with ornamental plants more elderly people with mobility problems have the opportunity to take care of their places near their apartments, thus integrating with others (fig. 6). small architecture: a selection of elements concrete buildings mainly constructed in the second half of the 20th century, have monotonous and uninteresting aesthetics, especially if they do not contain greenery. “na kozłówce” estate is a place of both daily recreation and leisure, which has been transformed thanks to a conceptual design based on a number of distinctive and functional elements. these elements were planned to meet residents’ needs. community gardens play a crucial role in the integration of residents, but also in education and recreation. by organizing the space and unifying it, the estate will be renewed and the residents will be more inclined to spend their free time near their apartments, thus improving their quality of life. this concept includes elements of small architecture. an amphitheatre, a bicycle storage, and an exhibition stand are included (fig. 7). all these elements are composed in a square plan. planned plantings plots of land have been set aside in the residential area for ornamental plants, which will be arranged by residents according to their tastes and gardening skills. in turn, the community garden proposes planting fruit trees, melliferous and ornamental plants. a variety of honeyproducing plants should be planted so that the nectar can be collected throughout the summer (tab. 1). table 1. four plant categories recommended for shaping the plant canopy in a community garden no. plant category plant names (common and scientific*) 1. herbaceous honey e.g. alfalfa (medicago sp.), blue tansy (phacelia tanacetifolia benth.), broadleaved thyme (thymus pulegioides l.), catnip proper (nepeta cataria l.), clover (trifolium sp.), columbine (aquilegia sp.), common bugle (ajuga reptans l.), common dandelion (taraxacum officinale coll.), common sage (salvia officinalis l.), european goldenrod (solidago virgaurea l. s.str.), lemon balm (melissa officinalis l.), lungwort (pulmonaria sp.), monkshood (aconitum sp.), primrose (primula sp.), trefoil (lotus corniculatus l.), white melilot (melilotus alba medik.), wild thyme (thymus serpyllum l.) 2. fruit trees e.g. apple tree (malus sp.,), cherry plum (prunus cerasifera ehrh.) 3. fruit bushes and e.g. blackberry (rubus hybridus vill.), blackcurrant (ribes nigrum l.), european shrubs gooseberry (ribes uva-crispa l.), northern highbush blueberry (vaccinium corymbosum l.), quince (cydonia oblonga mill.), redcurrant (ribes rubrum l. s.l.); 4. herbaceous fruit e.g. strawberry (fragaria ×ananassa, duchesne), wild strawberry (fragaria vesca l.) * scientific nomenclature of plants according to: www.atlas-roslin.pl functional program of the community garden the community garden initiative which includes a series of progressive actions designed to increase community engagement and reduce external intervention intensity was formulated by researchers from around the world. for example, a community-university partnership has been developed in shanghai since 2014 as part of a shift in urban regeneration paradigms in china that emphasizes people-centred strategies. social participation was quickly accepted by the public. moreover, community gardeners' attitudes toward other neighbours were positively rated (hencelova et al., 2021). local farms and gardens can provide healthy diets for residents in metropolitan areas. for every £1 invested in a london community garden, the return on investment is calculated at £3. as a contribution to social well-being in cities, community gardens should be included in municipal planning policies (schoen et al., 2020). an innovative landscape design was reported e.g. in the case of the alex wilson community garden in toronto (canada). this kind of design approach reconstructs the natural and cultural history of southern ontario, encompassing lakeshores, agriculture, and woodlands. gardening in the design exclusively uses native plants, which should increase biodiversity. residents monitor and assess the ecological health of the naturalized area as part of their community life. while both of these movements are popular today, they are rarely combined to demonstrate sustainable land use and community planning principles (irvince et al., 1999). the chitora community gardens (zimbabwe’s rural region) were found to improve the lives of many households in rural communities, including wetland revitalization, increased food security, livestock, crop management, and value-added initiatives. knowledge transfer, resilience building, sustainable asset building, and climate change mitigation and adaptation programs benefited rural communities (matsa et al., 2022). additionally, rural community gardens provide locals with opportunities to grow alternative food and engage in activit ies that promote social development and tackle neoliberal inequities (ghose, pettygrove, 2014). consequently, their users are at a lower risk of chronic and non-infectious diseases and stress. growing vegetables change their eating habits. the benefits of gardening include therefore improved health, education, and social connections (janowska et al., 2022). urban horticulture is positively recognised, e.g. by residents of a housing estate in lublin (poland). the study concluded that food security, the primary purpose of urban horticulture, is not essential to its inhabitants. urban gardens are not only functional, but they have secondary functions as well, such as recreation, social integration, aesthetics, and others (sosnowska et al., 2022). the next example is the “szeląg garden”, which was founded in 2018 by the “winogrady” estate council and local activists in poznań (poland). within the garden, there is a wooden pavilion. as the terrain was sloping, the pavilion was mounted on concrete poles, which allows people in wheelchairs to access it easily. apart from the pavilion and the community garden, there is also a catering area, a kids’ zone, a leisure area with wooden terraces, and a marina for riverboats on the plot. on the other hand, the abandoned orchard on the site of the former church was transformed into the “fort bema city garden” (warsaw, poland) in 2015. in the centre of this garden, apple trees are interspersed with a vegetable garden. a relaxation area can be found in the orchard. also in warsaw, in the same year, the “motyka i słońce” community garden was established. throughout the season, it is mainly used for growing herbs and vegetables. sponsors also made available a winter garden, which provides plants with more light in autumn and winter. numerous educational workshops, picnics, and cultural events are held in the garden in addition to relaxing. designing urban community gardens has the potential to generate a new city model, in terms of both the physical and human dimensions (maćkiewicz et al., 2022). the residents of kraków are interested in using community gardens and creating new ones near their estates. the following community gardens are the most popular among users in kraków: (a) kleparski park community garden (b) azory community garden (c) reduta community garden. based on the survey, over 80% of respondents believe that newly emerging housing estates should establish community gardens for social, economic, and revitalization purposes (rajca, kajzar, 2022). a new city plan is included in both the study of conditions and directions of spatial planning and the local plan of spatial development. a local plan of adaptation has been adopted by kraków and 43 other polish cities to mitigate and adapt to climate change. moreover, the city has also adopted a plan to increase afforestation by at least 8% through its “powiat programme to increase afforestation in the city of kraków for the years 20182040”.”development and management of greenery in kraków, 2017-2030” is the most important document and planning initiative of the city. the document discusses land use, cultural heritage, and the social requirements of green spaces as aspects of blue-green infrastructure. it also highlights many real estate management, spatial planning, and cultura l heritage management activities. the design of integration and management of green areas is found in “directions for development and management of green areas in krakow for 20172030”. defining a coherent, planned, and long-term policy for the development of green spaces is its primary goal. as part of the urban greenery and nature-based solutions (nbs) system developed since 1996, a great deal of emphasis is placed on the preservation and development of the kraków river parks system, which also function as community gardens. in urban environments, community gardens are a response to the worldwide trend towards green spaces that are tended by residents (lux 2001; kou et al., 2019). therefore, among others, the cultivation and integration of plants along the drwinka river are planned. aesthetic reasons and the ability to commune with nature lead residents to seek out such places, when they do not have direct access to them. summary and conclusion trees, shrubs, and climbers play a very important role in the design of the estate’s green spaces. the design of the green areas in a well-planned housing estate should take into account several principles of planting trees in such a way that they are useful. the placement of greenery is beneficial in case of the construction of residential blocks since tall buildings tend to shade them. in residential areas, lawns are mainly used for recreation by the elderly and families with young children. this depends largely on the scale of the building and the architectural form to which the surrounding greenery should be adapted. in this type of green space, you can find a variety of social garden spaces. among others, carpet lawns, flower beds or vases with flowers, ornamental shrubs, colourful varieties of trees, etc. are used here. nevertheless, the path system must be very simple and easy to navigate. neighbourhood greening programs should be comprehensive and tailored to the needs of all residents. the shortest distances between stops occur on pedestrian routes. to create more privacy and intimacy, it is beneficial to separate interiors with vegetation that is decorated with plants of different heights. maintaining an effective community garden involves providing favourable growing conditions. 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(2014). urban green space, public health, and environmental justice: the challenge of making cities ‘just green enough’. landscape and urban planning, 125, 234–244. https://doi.org/10.1016/j.landurbplan.2014.01.017 kształtowanie estetyki przestrzeni i krajobrazu dla ogrodu społecznego oraz ocena stref aktywności społecznej w projektowanym parku rzecznym drwinka (kraków, polska) streszczenie w związku z tym, że miasta rozwijają się zgodnie z zasadami zrównoważonego rozwoju, przestrzeń miejska musi być efektywnie wykorzystywana. aby prawidłowo kształtować środowisko miejskie, należy osiągnąć równowagę między terenami inwestycyjnymi, a obszarami cennymi przyrodniczo. w ostatnich latach uwagę rządu przykuwa planowanie przestrzenne, zwłaszcza rozwój zieleni miejskiej. ochrona krajobrazu miejskiego jest istotnym elementem rozwoju gospodarczego, dlatego niezbędne jest odpowiedzialne zarządzanie terenami zielonymi. jednak racjonalne zarządzanie zielenią miejską wydaje się być ciągle dużym wyzwaniem dla władz miasta. w planie zagospodarowania przestrzennego, można wyznaczyć miejsca pod nową roślinność, odnowić zniszczone tereny zielone lub zmienić przeznaczenie terenów poprzemysłowych. celem artykułu jest przedstawienie spójnego projektu odnowienia osiedla, ze szczególnym uwzględnieniem zieleni w planowanym parku rzecznym w krakowie. utworzenie ogrodu społecznego sprzyja spotkaniom towarzyskim, a także podnosi estetykę przestrzeni miejskiej i jakość życia. parki i ogrody społeczne są wymienione, jako strategiczne elementy wyposażenia infrastruktury miasta i komponent przestrzeni publicznej do spotkań i rekreacji lub innych aktywności. key words: blue and green infrastructure, green areas, river park, social life received: [2022.05.01] accepted: [2022.10.16] https://doi.org/10.1016/j.landurbplan.2014.01.017 appendix 1 fig. 4. the analysed area “na kozłówce” (south-eastern kraków) fig. 5. proposal to redesign green spaces and build a community garden 7 annales universitatis paedagogicae cracoviensis studia naturae, 6: 7–24, 2021, issn 2543-8832 doi: 10.24917/25438832.6.1 andriy novikov1*, józef mitka2, mariia sup-novikova3 1state museum of natural history, national academy of sciences of ukraine, teatralna st. 18, 79008 lviv, ukraine;*noviko�av@gmail.com 2institute of botany, jagiellonian university, gronostajowa st. 3, 30-387, kraków, poland 3ukrainian catholic university, stryiska st. 29a, 79000 lviv, ukraine hot-spots of the genus aconitum in the ukrainian carpathian region introduction �e genus aconitum in the world �ora comprises about 300–400 species distributed mainly in the temperate regions of the northern hemisphere (li, kadota, 2001; mitka et al., 2021). in the carpathians is located one of the main centres of the genus diversity in europe (mitka, 2003). �e ukrainian carpathian region (ucr), being part of the eastern carpathians and including surrounding foothills and lowlands, is subdivided into 57 geomorphological mesoregions (novikov, 2021a). in general, in the ucr, there are present 21 unequally distributed aconitum taxa of the speci�c and infraspeci�c level, including ten species of the native �ora (noviko� et al., 2016; onyshchenko et al., 2021). �e preliminary analysis of the genus aconitum distribution in the ukrainian carpathians was published in 2011 (noviko�, mitka, 2011). however, only some of the localities were included in our previous research, and no modern gis-based approaches were applied. �e aim of this research was to reveal main centres of the genus aconitum diversity and distribution (so-called hot-spots) in the ucr. here we represent the results of the genus’ hot-spot analysis basing on the most comprehensive data gathered from herbaria, �eld surveys, and published sources. material and methods in general, 2054 aconitum occurrences from the ucr were included in the analysis and shared as a  gbif dataset (novikov, 2021b). �e data mining was conducted from all available data sources, including published materials, databases, herbarium a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 8 collections, and personal �eld surveys during 2007–2019. in particular, data gathered personally or distantly from the 27 herbaria (see tab. 1 – appendix 1) were involved in the analysis. �e spatial distribution analysis was entirely performed in the qgis 3.10.2 environment (qgis development team, 2020) with an installed hot-spot analysis plug-in (oxoli et al., 2017) and using getis-ord gi* statistics implementation (getis, ord, 1992; ord, getis, 1995). for the purposes of the hot-spot analysis, a 10×10 km hexagonal grid was preliminarily constructed in qgis. hexagons were chosen due to the most reduced edge e�ect biasing the spatial data (birch et al., 2007). �e species richness (sr) was calculated per each of 57 delimited geomorphological mesoregions of the ucr that served as operational geographical units (ogus) throughout all the investigation. however, sr poorly performs for analysis of ogus (albuquerque, beier, 2015). �erefore, an analysis of rarity-weighted richness (rwr) following williams et al. (1996) has been performed too. similarly rwr, which has found its wider application for the calculation of the weighted endemism (crisp et al., 2001), we applied another technique that is also widely used in the analysis of endemism areas. �e principles of parsimony analysis of endemism (pae) allow delimiting the areas of endemism (in our case – areas of aconitum species concentration) and analysing the ogus similarity basing on the matrices with taxa presence/absence serving as character states (morrone, 1994; oliveira et al., 2015; fattorini, 2017; li et al., 2017) were extrapolated to our research. to perform pae, the ogus having zero presence (eight ogus) of the aconitum were excluded, and, at the same time, a  hypothetical ogu having zero presence of investigated taxa was introduced as a  zero-outgroup. �e maximum parsimony analysis was performed in the tnt 1.5 environment (golobo�, catalano, 2016) with tree bisection reconnection (tbr) set as collapsing rule and 1000 replications. �e consensus tree was obtained using the majority rule set to 50% cut-o�. a�er that, the consensus tree was exported to mesquite 3.70 (maddison, maddison, 2021), where it was rooted to zero-outgroup, set up to display the bootstrap support, visualised, and clades were coloured. results and discussion species richness and general distribution pattern most of the registered occurrences were distributed within the outer (76.77%) and inner eastern carpathians (20.16%), considering together the carpathian mountain range in the strict sense, while the adjacent sub-mountainous territories comprised only 3.07% from a total number of occurrences. namely, 54 occurrences (2.63%) were scattered 9 h ot-spots of the genus aconitum in the u krainian c arpathian r egion in the ciscarpathia and only nine (0.44%) – in the pannonian province corresponding to the transcarpathia (fig. 1). �e ogus oc23 (chornohora – 720 occurrences, all ten species registered), oc22 (svydovets – 234 occurrences, all ten species registered), ic10 (maramures – 363 occurrences, nine species registered), oc6 (syvuliansko-stanymyrski gorgany – 60 occurrences, eight species registered), oc14 (torunsko-bertianski gorgany – 77 occurrences, eight species registered), and oc19 (polonyna rivna – 51 occurrences, eight species registered) demonstrated the highest sr (fig. 2a–b – appendix 2). as we can see, the number of occurrences per ogu (in other words – sampling frequency) does not strictly correlate with sr. for example, chornohora demonstrated both the highest sampling frequency and species richness. however, another ogu with a  relatively high sampling frequency (oc12 – waterdivided mountain range) having 69 registered occurrences comprises only �ve di�erent aconitum species. it is important to note that waterdivided mountain range is o�en considered the part of eastern beskyds (= bieszczady wschodnie) following the classi�cation of kondracki (1989, 2002). however, here waterdivided mountain range is considered as independent mesoregional fig. 1. distribution of all registered aconitum occurrences in the ucr (for abbreviations see tab. 2. – appendix 1) a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 10 ogu, following ukrainian approach (slyvka, 2001; hiletskiy, 2012; novikov, 2021a). similarly, oc7 (dovbushanski gorgany) having 60 registered occurrences comprises only �ve di�erent aconitum species too. sampling frequency and sr can be a�ected by several distorting factors, i.e., higher popularity of certain ogus among collectors; repeated collection from the same populations; absence of the information from the hardly accessible places (baltanás, 1992). to correctly analyse the distribution data, it is important to treat all test areas equally, i.e. assuming that all species are detected and that the detectability of the di�erent species is the same (boulinier et al., 1998). we could not be sure that all aconitum species were detected for each certain ogu. however, the high number of registered occurrences relating to a  low number of tested species analysed at the high spatial resolution allowed to minimise the distortion e�ect (graham, hijmans, 2006; gotelli, colwell, 2011; chao, chiu, 2016). considering the limitations of the sr analysis and unequal rarity of the analysed aconitum species (from ten studied species, six are threatened and with limited distribution in the ucr – see novikov (2021b) for details), other advanced data processing techniques were applied. we calculated the rwr index for all ogus and found that oc23, oc22, and ic10 had the highest rwr values, while oc6, oc14, and oc19 also demonstrated a relatively high rwr level (over 0.5). hence, the rwr results were found to be concordant with sr outcomes. additionally, oc21 (polonyna krasna) showed the rwr value very close to 0.5 points (0.48), while other analysed ogus di�ered much distinctly and did not reach 0.3 rwr value (fig. 2c – appendix 2). hot-spot analysis getis-ord gi* statistics allowed to primarily reveal four hot-spot centres in the ucr (fig. 3a – appendix 2): 1 – located mainly in the ogus oc23, oc22, and ic10 (i.e., chornohora, svydovets, and maramures); 2 – located mainly in oc12 and oc19 (i.e., waterdivided mountain range and polonyna rivna); 3 – located mainly in the oc6 (i.e., syvuliansko-stanymyrski gorgany); 4 – located mainly in the oc14 (torunsko-bertianski gorgany). normalisation of the hot-spot analysis eliminated the last two hot-spots (fig. 3b – appendix 2). hence, also taking into account the data on the species richness and sampling frequency, we can conclude that there are two centres of the genus aconitum diversity and distribution within the ucr. �e main centre is located in the south-eastern part of the ucr and covers svydovets, chornohora, and maramures. at the same time, the second small isolated centre exists in the polonyna rivna and waterdivided mountain range. a similar result was obtained while studying the centres of phenetic coherence of the aconitum sect. aconitum (= napellus) in the eastern carpathians, based on mahanalobis distances among regions’ centroids in discriminant function analysis (mitka, 2002). here, the svydovets-negrovets region and chornohora formed 11 closely related regions in terms of the taxon morphological variability. another centre (partially outside the ukrainian carpathians) is in maramures and bistrica mts-čeahlău region (mitka, 2002). �e high aconitum diversity and distribution in the svydovets, chornohora, and maramures are not surprising due to the highest altitudes in the region. �ese mountain ranges reach the alpine belt and generally have the highest �oristic diversity (malynovskiy, 1991; tasenkevich, 2003; cherepanyn, 2017). moreover, in the maramures is located one of the main areas of plant endemism in the carpathians, partly spread to the north-west and separated by a strong barrier from surrounding mountain ranges (pawłowski, 1970; tasenkevich, 2014; hurdu et al., 2016). much engaging is the presence of a small hot-spot of the genus aconitum in the waterdivided mountain range and polonyna rivna. in the ukrainian carpathians, waterdivided mountain range is not as high as other surrounding ranges and is considered a relatively older geomorphological construction hosting the relict �oristic complexes and unique plant taxa (popov, 1949; stoyko et al., 1997). moreover, mt. pikui, located in this mountain range, has the beech dwarf forest delimiting a short subalpine belt with rocky outcrops, making unique isolated habitats (stoyko et al., 1997). in particular, here exists isolated and the most abundant in the ucr population of the aconitum bucovinense zapał. (noviko�, mitka, 2011). �ere are no reports about such unique �oristic conditions on the polonyna rivna, and most analysed aconitum occurrences related to runa-plai mt. and its slopes. cladistics analysis maximum parsimony analysis following the principles of pae has been performed to reveal the similarity of the analysed ogus basing on their species composition. li et al. (2017) reported that pae analysis at the multiple taxa level is not well-performing. �erefore we cut o� all taxa at the same (i.e., species) level. most of the analysed ogus did not demonstrate a statistically signi�cant di�erence in pae (perhaps, due to a low number of studied species). it showed eight well-distinguishing clades of aconitum diversity in the ucr (fig. 4a – appendix 2, marked on by color). �e ogus oc23 (chornohora) and oc22 (svydovets) were found to be the most outstanding (bootstrap support = 1) and sharing the common node with clades ic10 (maramures) and oc14 (torunsko-bertianski gorgany). interestingly, that �oristically poor regions oc19 (polonyna rivna) and oc21 (polonyna krasna) appeared to be distinctly separated from oc20 (polonyna borzhava) that belongs to the same mountainous group. moreover, pae did not allow for the distinguishing oc12 (waterdivided mountain range) that was found to be distinct in the previous analyses. despite this, pae ampli�ed the clades of ogus oc5, oc6, and oc14 belonging to di�erent mesoregions of the gorgany mts. extrapolation of the pae results on h ot-spots of the genus aconitum in the u krainian c arpathian r egion a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 12 the map (fig. 4b – appendix 2) con�rmed that there exists the main centre of the genus aconitum diversity in the ucr with its core in the chornohora and svydovets that is spread to adjacent mountain ranges. �e second isolated centre of aconitum diversity was revealed in the polonyna rivna, which is also partly concordant with mentioned above �ndings. perhaps, a  continuous distribution pattern of the genus aconitum from the se toward nw of the ucr existed before. �e gap in the oc20 (polonyna borzhava) could arise as a  result of anthropogenic pressure. in the polonyna borzhava, just like in polonyna rivna and polonyna krasna, the forest level was arti�cially lowered, and open spaces were overgrown by vaccinium myrtillus l. (felbaba-klushina, bizilya, 2015). conclusions di�erent methods of spatial distribution analysis were tested for the genus aconitum in the ucr. it was found that chornohora and svydovets mts., with a  signi�cant contribution of maramures, form the core of the genus diversity and distribution in this region. basing on maximum parsimony analysis (adopted pae), it was found that chornohora and svydovets mts. form the most distinct clades with high bootstrap support. �e polonyna rivna was found to have surprisingly high rwr value and clustering together with polonyna krasna, while polonyna borzhava disrupts these two ogus from the main hot-spot of the genus in the se part of the ucr. perhaps, a  continuous gradient of aconitum diversity and/or distribution from se toward nw of the ucr existed before, and the gap in borzhava secondarily resulted from anthropogenic transformation. acknowledgment �e authors thank all supporting agencies that allowed performing this investigation. in particular, this investigation was partly supported by the grant of the queen jadwiga fund (jagiellonian university, krakow, poland) given to an and supervised by jm in 2009, 2011, 2013, and 2016. �e �eld surveys were performed by an and msn in frames of the grants of the ru�ord foundation (rsg-16667-1 and rsg-21313-2) in 2015 and 2017. �e distribution data also were partly obtained during the visit of an to slovakia in frames of the national scholarship program of the slovak republic in 2012 and 2018. con�ict of interest �e authors declare no con�ict of interest related to this article. references albuquerque, f., beier, p. 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te r l ow la nd o ve rs ia n ba si n 1 1 cc 4 st ry vi go rbo lo zi vk a h ig h la nd st ry vi go r h ig hl an d 0 cc 5 u pp er -d ni st er d ep re ss io n d ni st er -s vi ch a lo w la nd 1 4 cc 6 d ro ho by ch h ig hl an d 3 2 3 1 cc 7 m or sh yn h ig hl an d bo le kh iv -z hu ra vn e re gi on 2 5 1 cc 8 z al is si a h ig hl an d d ol yn obo lo kh iv sk yi re gi on 1 2 cc 9 r oz hn ia tiv -k al us h d e pr es si on li m ny ts ia re gi on 1 1 cc 10 pr yl uk va h ig hl an d lu kv a h ig hl an d 2 3 3 cc 11 by st ry ts ia d ep re ss io n 0 cc 12 in te rb ys tr yt si a h ig hl an d m iz hb ys tr its ke h or bo gi r ri a, g vi zd ts ke h or bo gi rr ia 1 2 17 cc 13 d el ia ty nn ad vi rn a h ig hla nd pr ut -b ys tr its ia h ig hl an d 0 cc 14 pr ut -l yu ch ka h ig hl an d 1 1 cc 15 t lu m ac ho be rt yn h ig h la nd 2 2 1 cc 16 k ol om yi ac he rn iv ts i p la in 4 3 2 3 6 cc 17 po ku tti a h ig hl an d 0 cc 18 se re tc he re m os h h ig hl an d 1 1 cc 19 se re tpr ut h ig hl an d 1 1 cc 20 bu ko vi na h ig hl an d 1 1 ic 1 u pp er d ni st er b es ky ds ve rk hn io dn is te rs ki b es ky dy 1 20 1 1 ic 2 sk ol e be sk yd s 4 4 1 ic 3 sv its ko -m iz un sk i g or ga ny 1 ic 4 a rs hy ts ia -i le m sk i g or ga ny 5 1 ic 5 ve rk hn io lim ny ts ki g or ga ny 5 5 ic 6 sy vu lia ns ko -s ta ny m yr sk i g or ga ny 8 6 ic 7 d ov bu sh an sk i g or ga ny 5 1 ic 8 z ap ru ts ki g or ga ny 3 2 1 1 1 2 ic 9 po ku tti a m ts . 1 ic 10 bu ko vy na m ts . 3 12 6 17 75 81 29 1 15 8 10 oc 1 st ri ysi an v er kh ov yn a 4 3 oc 2 w at er di vi de d m ou nt ai n r an ge 5 17 10 5 2 oc 3 vo lo ve ts -m iz hg ir iy a ve r kh ov yn a 3 7 oc 4 to ru ns ko -b er tia ns ki g or ga ny 8 8 2 9 1 1 h ot-spots of the genus aconitum in the u krainian c arpathian r egion a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 18 oc 5 br at ki vs ki g or ga ny 2 15 1 3 1 1 oc 6 ya si ni a d ep re ss io n 4 14 2 21 2 3 8 4 6 oc 7 vo ro kh ta -p ut yl a va lle y 4 33 4 17 3 3 oc 8 h ig h bi es zc za dy bi es zc za dy w ys ok ie 4 4 1 1 oc 9 bu ko vs ke v rc hy bu ko vs ké v rc hy 8 1 oc 10 po lo ny na r iv na po lo ny na r un a 4 11 1 1 oc 11 po lo ny na b or zh av a 6 8 1 7 2 oc 12 po lo ny na k ra sn a 10 26 5 1 33 3 oc 13 sv yd ov et s 10 3 2 1 oc 14 c ho rn oh or a 4 12 1 15 1 7 4 17 19 oc 15 g ry ni av a 5 3 7 oc 16 ts yr ok hbo rz av a va lle y be re zn eli ps ha ns k va lle y 3 34 2 14 1 oc 17 v yg or la t 2 10 2 10 1 oc 18 m ak ov yt si a 0 7 12 5 1 oc 19 sy ni ak 1 17 8 4 6 1 6 2 5 oc 20 ve ly ki y d il 1 17 1 5 1 oc 21 tu pi y 1 7 4 2 2 15 1 oc 22 o as g ut yn 1 70 2 8 37 7 3 2 16 55 34 oc 23 te re bl ia m as si f 5 17 8 7 24 13 7 32 1 6 17 8 63 83 oc 24 a ps hy ts ia m as si f 0 13 1 10 3 oc 25 m ar am ur es m ts . m ar m ar os h m ts . 9 25 3 1 8 1 tc 1 tr an sc ar pa th ia n lo w la nd ty sa l ow la nd , p ry ty se ns ka d ol in a, t ys en sk a d ol in a 3 6 2 1 19 h ot-spots of the genus aconitum in the u krainian c arpathian r egion appendix 1 tab. 1. list of applied herbaria’s acronym following �iers (2021) no. acronym herbarium location 1. b botanischer garten und botanisches museum berlin, zentraleinrichtung der freien universität berlin germany, berlin 2. bm �e natural history museum u.k. england, london 3. bp hungarian natural history museum hungary, budapest 4. brnu masaryk university czech republic, brno-bohunice 5. cher yu. fedcovich chernivtsi national university ukraine, chernivtsi 6. cl babes-bolyai university romania, cluj-napoca 7. gjo universalmuseum joanneum austria, graz 8. hbg university of hamburg germany, hamburg 9. je friedrich schiller university jena germany, jena 10. kra jagiellonian university poland, kraków 11. kram w. szafer institute of botany, polish academy of sciences poland, kraków 12. kw m.g. kholodny institute of botany, na-tional academy of sciences of ukraine ukraine, kyiv 13. kwha national academy of sciences of ukraine ukraine, kyiv 14. kwhu o. v. fomin botanical garden of the taras shevchenko national university of kyiv ukraine, kyiv 15. li upper austrian state museum austria, linz 16. lw ivan franko national university of lviv ukraine, lviv 17. lwks institute of ecology of the carpathians ukraine, lviv 18. lws state museum of natural history ukraine, lviv 19. o university of oslo norway, oslo 20. ohn biological museum, oskarshamn sweden, oskarshamn 21. pr national museum in prague czech republic, praha 22. prc charles university, prague czech republic, praha 23. sav slovak academy of sciences slovakia, bratislava 24. sib natural history museum romania, sibiu 25. uu uzhhorod national university ukraine, uzhhorod 26. wu universität wien austria, wien 27. zt eidgenössische technische hochschule zürich switzerland, zürich a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 20 description fig. 2ab on the next page appendix 2 21 h ot-spots of the genus aconitum in the u krainian c arpathian r egion fig. 2. �e number of registered occurrences (a), species richness (b), and rarity weighted richness (c) of the genus aconitum in the ukrainian carpathian region a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 22 fig. 3. standard (a) and normalised (b) getis-ord hot-spot analysis of the aconitum distribution in the ucr. h ot-spots of the genus a conitum in the u krainian c arpathian r egion 23 fig. 4. cladogram of the adopted pae analysis of the aconitum diversity in the ucr (a) extrapolated to the map (b) a nd riy n ov ik ov , j óz ef m itk a, m ar iia s up -n ov ik ov a 24 abstract basing on 2054 georeferenced occurrences of ten aconitum species, the initial map of the species richness per each of 57 operational geographic units (ogu) of the ukrainian carpathian region (ucr) has been developed. next, to avoid unequal data distribution, we counted absolute species presence per ogu, based on which we calculated ‘rarity-weighted richness’ (rwr) and performed parsimony analysis. as a result, we found that chornohora and svydovets made a strongly supported cluster with the highest aconitum species diversity and demonstrated the highest rwr values, hence being the main hot-spot of the genus distribution in the ucr. �is main hot-spot also spreads to adjacent mountain ranges, including maramures and gorgany. probably, a continuous gradient of aconitum taxonomic richness from southeast toward northwest of the ucr existed before. �e gap in polonyna borzhava, located between polonyna rivna and polonyna rivna, can be secondarily resulted by the high level of anthropogenic transformation of this region. contrary, the high rwr value and distinct clustering of the polonyna rivna supported the presence of a local isolated aconitum hot-spot related to two prominent mountain peaks located here – runa-plai mt. (in polonyna rivna) and pikui mt. (in adjacent waterdivided mountain range). key words: aconitum, biogeography, hot-spot analysis, ranunculaceae, ukrainian carpathians received: [2021.09.21] accepted: [2021.10.18] hot-spoty rodzaju aconitum w ukraińskim karpackim regionie streszczenie na podstawie analizy 2054 georeferencyjnych wystąpień dziesięciu gatunków rodzaju aconitum opracowano wstępną mapę bogactwa gatunkowego w każdym z 57 operacyjnych jednostek geogra�cznych (ogu) ukraińskiego karpackiego regionu (ukr). następnie, aby uniknąć nierównego rozkładu danych, policzyliśmy bezwzględną obecność gatunków w każdym ogu, obliczyliśmy bogactwo ważone rzadkością (rwr) i wykonaliśmy analizę parsymonii. w rezultacie stwierdziliśmy, że czarnohora i svydovets utworzyły silnie poparte ugrupowanie o najwyższym bogactwie gatunków aconitum i wykazywały najwyższe wartości rwr, będąc tym samym głównym hot-spotem rozmieszczenia rodzaju w ucr. ten główny hots-pot rozciąga się również na sąsiednie pasma górskie, w tym maramures i gorgany. prawdopodobnie w ukr istnieje gradient bogactwa taksonomicznego aconitum z południowego wschodu na północny zachód. luka w borżawie, położonej między równą a krasą, może być wtórnie spowodowana wysokim stopniem antropogenicznej transformacji tego regionu. natomiast wysoka wartość rwr i wyraźna odrębność połoniny rivna wskazuje na obecność lokalnego, izolowanego hot-spotu rodzaju aconitum, związanego z dwoma jej głównymi szczytami górskimi – runa-plai (na połoninie rivna) i pikui (na werchowynśkim wododilnim chrebetie). słowa kluczowe: aconitum, biogeogra�a, analiza hot-spotów, ranunculaceae, karpaty ukraińskie information on the authors andriy novikov https://orcid.org/0000-0002-0112-5070 ukrainian scientist specialising in plant morphology and phytogeography in the carpathians. his interests are framed by but not limited to the taxonomy, conservation, and distribution of genus aconitum in this region. józef mitka https://orcid.org/0000-0001-8472-1742 he is a specialist in the taxonomy, morphology and genetics of the genus aconitum. specialised in biodiversity, conservation, and ecology of plants. mariia sup-novikova https://orcid.org/0000-0002-8542-3605 professor of the ukrainian catholic university, assisting in the data mining, transcription, and processing, as well as during the botanical �eld surveys. she has no special scienti�c interests, contributes sporadically but signi�cantly to di�erent research activities of andriy novikov. 127 annales universitatis paedagogicae cracoviensis studia naturae, 6: 127–145, 2021, issn 2543-8832 doi: 10.24917/25438832.6.8 anna sołtys-lelek1*, zbigniew caputa2 1ojców national park, ojców 9, 32-045 sułoszowa, poland; *ana_soltys@wp.pl 2a. ficka st. 4/1, 40-421 katowice, poland the influence of solar radiation on selected physiological processes of mosses in karst conditions of the spring niches of the ojców national park (southern poland) solar radiation has various and multidirectional e�ects on the growth and development of plants. it a�ects, incl. on the anatomical and morphological structure of individual organism structures, photosynthetic apparatus, quantity and quality of plant pigments (öpik, rolfe, 2005; pilarski et al., 2012; możdżeń, 2019). depending on the season and day, 15 to 30% of solar radiation reaches the forest layer. however, a signi�cant part of this solar radiation is scattered or re�ected at di�erent angles by plants from di�erent layers of the forest (swanson, flanagan, 2001). mosses, as poikilohydric organisms, may remain metabolically inactive when the solar radiation intensity is high or the water level is too low. due to their small size and tolerance to low light, mosses o�en colonise habitats that are characterised by low solar radiation intensity (niinemets, niinemets, 2009). �e photosynthesis process is highly sensitive to abiotic stress factors, including light (rzepka, 2008; możdżeń, 2019). �e intensity of photosynthetically active solar radiation is a factor that directly or indirectly in�uences the photochemistry of photosynthesis (marschall, proctor, 2004). �e �rst symptom of high intensity of solar radiation is the degradation of chlorophylls and the reduction of photosynthesis (tallis, 1959; rastorfer, 1970). sensitivity to light is the result of phyloand ontogenetic adaptation. for plants growing in extreme shaded conditions, light with an intensity of several thousand lux can inhibit photosynthesis, while in other plants this e�ect occurs only at intensities exceeding 100,000 lux. plants adapt to light through structural adaptations and plant pigments. bryophytes, unlike tracheophytes, do not have a protective epidermis layer, under which there is a layer of palisade crumb, which additionally absorbs light before it reaches the phoa nn a s oł ty sle le k, z bi gn ie w c ap ut a 128 tosynthetic tissue of the spongy crumb. duckett and renzaglia (1988) and nasrulhaq-boyce and duckett (1991) described a large variation in the number of chloroplasts per cell between mosses exposed to direct and di�use radiation. bryophytes need to “invest” more in protection at the cellular level to mitigate the harmful e�ects of high levels of light (robinson, waterman, 2014). mosses are found in many habitats around the world, but knowledge of their responses to various environmental factors is still largely unexplored. �e aim of this study was to investigate the in�uence of solar radiation reaching the karst spring niches, on selected physiological parameters of the spring mosses: cratoneuron �licinum (hedw.) spruce (obligatory krenophyte) and brachythecium rivulare schimp. (facultative krenophyte). �e impact of habitat conditions on their photosynthetic activity was investigated by: (1) imaging the parameters of chlorophyll a  �uorescence, (2) determining biomass and water content in gametophores and (3) analysing the degree of destabilisation of cell membranes. material and methods study area in the ojców national park (onp) (southern poland – 50°12′24″n 19°49′45″e ) the geological substrate consists of limestones and upper jurassic marls, with a thickness of over 250 m. �e network of vertical and horizontal fractures in them enables easy migration of water (aleksandrowicz, wilk, 1962). �e prądnik and sąspowska valleys have the character of a deep ravine with asymmetrical slopes. �e eastern slopes are steeper and the rock formations on them are more numerous. only two permanent watercourses �ow along the valley bottoms – prądnik and sąspówka. all side valleys, connecting with the prądnik and sąspówka valleys, are dry, without any permanent watercourses. �ere are numerous springs in this area – karst spring (gradziński et al., 2008). most of the springs are located in the sąspówka and prądnik alluvia, and the water �ows directly from the rock-mantle jurassic limestones (różkowski, pawlik, 2001). �e morphological diversity of the park’s terrain in�uences di�erent solar and microclimatic conditions (caputa, 2009; caputa, wojkowski, 2015; wojkowski, caputa, 2015). �e region is characterised by a warm plateau region, a region of thermally diverse slopes and a region of cold valley bottoms (brzeźniak, 1974; kliein, 1972). �e study area located at the valley �oors is characterised by frequent thermal inversions, fogs and cold air (brzeźnik, partyka, 2008). �e �ve karst spring niches (4 in the prądnik valley, 1 in the sąspowska valley) were selected for the plant material collection, in which two of the moss species tested occurred simultaneously (fig. 1–2). the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 129 spring of “orczyk” in sułoszowa a �ssure spring, located on the right side of the prądnik stream, 390 m a.s.l. exposition ne. it is made up of two rivers �owing from under a limestone slope. one of them has a concrete housing that distorts its natural character. it is a high capacity spring of 14.5 l/s (main �ow), 5.6 l/s (secondary �ow). its waters are bicarbonate-calcium, sweet, slightly alkaline, hard (pawlik, 1998). �e spring is surrounded by a forest, a meadow and a roadside. fig. 1. distribution of study stands: a  – rivers, b – prądnik river, c – study stands, d – second-order catchment, e – park onp border, f – boundary of the park’s bu�er zone; 1–4 spring niches in prądnik valley: 1 – spring of “orczyk” in sułoszowa, 2 – spring of a car park in pieskowa skała, 3 – spring near the chapel “na wodzie” in ojców, 4 – spring opposite the wapiennik rock in ojców, 5 – spring in the sąspowska valley a nn a s oł ty sle le k, z bi gn ie w c ap ut a 130 spring of a car park in pieskowa skała a �ssure-karst spring, near the riverbed, located on the le� side of the prądnik stream, 390 m a.s.l., with an ssw exposure. it is made up of three out�ows. �e largest of them, being a spring niche, has a concrete lining; the other two �ow naturally, almost in the very channel of the stream. �is spring has a  low capacity of 0.5 l/s. its waters are bicarbonate-calcium, sweet, slightly alkaline, medium hard (dynowska, 1983; pawlik, 1998). �e spring is surrounded by a forest and a parking lot. spring near the chapel “na wodzie” in ojców a �ssure-karst spring, terraced, by the riverbed, located on the right side of the prądnik stream, 325 m a.s.l., with ene exposure. it is enclosed in an unworked limestone casing. �e spring niche is an arti�cially directed out�ow of water, which �ows into prądnik along a  natural slope of limestone rubble. �is spring has a  low capacity of 1.2–0.5 l/s. its waters are bicarbonate-calcium, sweet, slightly alkaline, medium hard (aleksandrowicz, wilk, 1962; sadowski, różkowski, 1989; pawlik, 1998). �e spring is surrounded by meadows and a small tourist infrastructure. fig. 2. selected study stands: a – spring the sąspowska valley, b – spring of “orczyk”, c – spring opposite the wapiennik rock, d – spring near the chapel “na wodzie” (photo. a. sołtys-lelek) the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 131 spring opposite the wapiennik rock in ojców fissure-karst spring, near-channel, located on the right side of the prądnik stream, 310 m a.s.l., with ene exposure. it is enclosed in a concrete housing, and part of its water supplies the farm. part of the spring also �ows down to prądnik in a small stream. �is spring has a low capacity of 1.4 l/s. its waters are bicarbonate-calcium, sweet, slightly alkaline, medium hard (aleksandrowicz, wilk, 1962; pawlik, 1998). �e spring is surrounded by meadows. spring in the sąspowska valley a �ssure-karst spring, near-channel, located on the right side of the sąspówka stream, 350 m a.s.l., with ne exposure. �ey are two out�ows of a natural character that merge and �ow into the sąspówka stream. �is spring has a low capacity of 3.9 l/s. its waters are bicarbonate-calcium, sweet, slightly alkaline, medium hard (pawlik, 1998). �e spring is surrounded by a forest. �is spring is the only one of the surveyed, characterised by a completely natural spring niche, devoid of anthropogenic factors. measurements of solar radiation and temperature global solar radiation (kc) and re�ected solar radiation were measured with the sp lite pyranometer (weiz, austria). nr lite kipp & zonen radiometer was used to measure net all-wave radiation (nr) for individual springs. based on the quotient of re�ected to total radiation (kodb/kcal), the re�ectance coe�cient – albedo was determined, which indirectly determines the ability of the surface to re�ect radiation. air temperature (2 m above the surface) – tp, water – tw, on the spring surface – tpo and inside moss turf – tw was measured by thermometer lb401 with a pt100 sensor with an accuracy of 0.1°c (lab-el, krakow, poland). solar radiation and temperature measurements were made on a sunny day from 11:00 to 16:00. plant material – short botanical characteristic brachythecium rivulare schimp. in bruch, schimp. & w. gümbel – (canal moss) is green or yellow-green plagiotropic moss. it occurs all over poland, in the mountains up to 1,800 m a.s.l. �e upward stems are usually pinnate, 3 to 6 cm long. �e leaves are ovoid-triangular, suddent at the top, sharpened brie�y, narrowed at the base, concave, irregularly longitudinally folded, and �nely, sharply serrated on the edge. �e leaf rib is single, tapering towards the top. �e leaf cells are elongated, prozenchymatic, with a small amount of chloroplasts, with thickened cell walls, orange in colour (jusik, 2012). cratoneuron �licinum (hedw.) spruce – (triangle moss) it is a plagiotropic moss that forms a pale green turf. �is species is common throughout poland. it occurs mainly in the carpathians, sudetes, lakeland belt, jura krakowsko-częstochowska and roztocze. its gametophores are creeping at the bottom, singly branched at the top, reaching 2 to 5 a nn a s oł ty sle le k, z bi gn ie w c ap ut a 132 cm in length. �e leaves are wide, heart-triangular and ovate-lanceolate. �e rib is single, massive, biconvex, made of undi�erentiated cells. �e leaf cells are short-rectangular or 6-sided, and a few elongated, thick-walled, more elongated at the tip (jusik, 2012). chlorophyll a �uorescence chlorophyll a �uorescence measurements were performed on 5 di�erent gametophores of mosses. during the study, the gametophores of mosses were put on �lter paper wet of distilled water and adopted for darkness for 20 min in a closed chamber fluorcam fc 800c (photon systems instruments, czech republic) according to the method used by możdżeń (2019). among the results obtained, the following parameters were analysed: the zero �uorescence (f0), the maximum �uorescence (fm), the maximum e�ciency of the photochemical psii (fv/fm), the non-photochemical quenching (npq) and the vitality of psii (rfd). fresh and dry mass, water content fresh and dry masses (fm and dm, respectively) were determined on a  laboratory balance (ohaus adventurer pro, usa) with an accuracy of 0.0001 g. single moss gametophytes were dried for 48 h at 105°c temperature (dryer – wamed sup-100, poland). on the basis of the masses obtained, the ratio of dry mass to fresh mass and the percentage of water content were determined according to the following formula (1): wc [%] = 100 – [(dm × 100) / fm] (1) where: wc – water content, dm – dry mass, fm – fresh mass electrolyte leakage separately gametophytes of two mosses were placed in vials containing 10 ml of deionised water with a  speci�c conductivity of 0.05 µs. plant material was incubated on a shaker (labnet, rocker, usa) for 3 h at 25°c to determine the electrolytes leakage (e1). �en vials with mosses were frozen for 24 h at −80°c (platilab 500next, angelantoni industrie, italy). next day, samples were thawed and subjected to the shaking procedure described above. a�er this time, the �ow of electrolytes (e2) were measured. �e percentage of electrolyte leakage (el) was calculated according to the formula: el = (e1 / e2) 100%. �e electrolyte leakage measurements were made using a cx701 conductometer (elmetron, poland) with an electrode with a  constant k = 1.02 (elmetron, poland). data analysis �e mean results from 5 replicates were analysed in microso� excel and statso�, inc. the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 133 2018. statistica (data analysis so�ware system), version 13.1. �e signi�cance of di�erences between means (± sd) were analysed by duncan’s test at p ≤ 0.05. results �e in�ow of solar radiation and temperature total shortwave (kc) and re�ected (ko) radiation were the highest on stand 3, and the lowest on stands 1 and 5 (tab. 1). albedo achieved the highest values on stand 2, and the lowest on stands 3, 4, 5. air temperature (tp) was the highest on stand 4, and the lowest on stand 1. tab. 1. characteristics of environmental factors on the researched springs in ojców national park (wojkowski, caputa, 2009) no. of spring radiation coe�cients spring temperature [°c] annual sum of total radiation �e annual amount of potential insolation measurement data [mv] radiation [w × m–2] kc ko nr kc ko nr albedo tp tpo tm tw [mj × m–2] [h] 1 65 9 56 77 155 36 0.15 14.35 11.65 8.15 8.65 3484 3235 2 103 12 91 102 122 51 0.19 15.85 16.85 16.1 8.35 2871 2570 3 603 80 523 68 150 354 0.13 17.2 22.93 13.7 8.75 3566 3156 4 392 50 342 86 117 292 0.13 18.1 15.7 10.4 9.30 3302 2746 5 71 9 62 82 126 46 0.13 17.5 16.0 11.4 8.50 3200 < 2700 1 – spring of “orczyk” in sułoszowa, 2 – spring of a car park in pieskowa skała, 3 – spring near the chapel “na wodzie” in ojców, 4 – spring opposite the wapiennik rock in ojców, 5 – spring in the sąspowska valley; kc – total short-wave radiation, ko – re�ected short-wave radiation, nr – balance of shortand long-wave radiation, albedo – determines the ability of a surface to re�ect short-wave radiation: albedo = kodb/kcal, tp – air temperature, tpo – surface temperature above the spring, tm – temperature in the moss, tw – water temperature �e surface temperature above the spring was the highest at stand 3, and the lowest at stand 1. �e temperature in the moss turf (tm) was the highest on stand 2, and the lowest on stand 1. �e water temperature (tw) was the highest at stand 4, and the lowest at stand 2. �e annual sum of total radiation reached the highest values at stand 3, and the lowest at stand 2. �e annual sum of potential insolation showed the highest values at stand 1, and the lowest at stand 2. chlorophyll a �uorescence �e zero �uorescence (f0) brachythecium rivulare was the highest in plants growing on stand 3 (the most yellow and red colour), compared to the gametophores from stands 1 and 5 (fig. 3). intermediate values of f0 were reached by mosses from stands 2 and a nn a s oł ty sle le k, z bi gn ie w c ap ut a 134 4. �e maximum �uorescence (fm) was similar between specimens of b. rivulare from stands 2 and 3 (the most yellow and red). slightly lower values of this parameter were found for mosses collected from stands 1 and 4. �e lowest values of fm were found for plants from stand 5 (green colour is dominant). �e maximum e�ciency of the photochemical psii (fv/fm) was clearly the highest for the gametophores collected from stands 2 and 4 (intense red colour dominated). intermediate values of this parameter were observed in plants from the remaining 3 stands (1, 3 and 5 – half yellow and red, respectively). �e non-photochemical quenching (npq) reached the highest values in b. rivulare taken from stand 5 (yellow and red predominance). �e lowest npq values were observed for gametophores from stand 3 (green and yellow colours dominate). intermediate values of this parameter were found for plants from stands 1, 2 and 4 (mostly yellow and green, a small proportion of red). �e vitality of psii (rfd) reached the lowest values in b. rivulare from stand 1 (green was the dominant colour). �e rfd values were similar in plants from the remaining 4 stands (fig. 3). �e zero �uorescence (f0) cratoneuron �licinum was similar in the 5 tested stands (red and green colours). maximum �uorescence values were similar in plants collected from stands 1, 3 and 5. clearly lower values were observed for plants from stands 2 and 4 (a small proportion fig. 3. imaging of �uorescence parameters of brachythecium rivulare schimp. from the �ve karst spring niches (1–4 in the prądnik valley, 5 in the sąspowska valley); f0 – the zero �uorescence, fm – the maximum �uorescence, fv/fm – the maximum e�ciency of the photochemical psii, npq – the non-photochemical quenching, rfd – the vitality of psii the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 135 fig. 4. imaging of �uorescence parameters of cratoneuron �licinum (hedw.) spruce from the �ve karst spring niches (1–4 in the prądnik valley, 5 in the sąspowska valley); f0 – the zero �uorescence, fm – the maximum �uorescence, fv/fm – the maximum e�ciency of the photochemical psii, npq – the non-photochemical quenching, rfd – the vitality of psii of red, the dominant colour is green). �e maximum e�ciency of the photochemical psii for gametophores from stands 2, 3 and 4 were similar (red and yellow colours were dominant). �e lowest fv/fm values were recorded for mosses from the stand 5 (a large share of red colour; however, a green colour also appeared). �e non-photochemical quenching values were similar in plants from standns 1, 2 and 5. c. �licinum gametophores from stands 3 and 4 achieved similar npq values (small proportion of red colour; green was the dominant colour). �e vitality of psii (rfd) was similar in plants harvested from all 5 stands (fig. 4). fresh and dry mass, water content �e fresh mass of brachythecium rivulare gametophytes was signi�cantly the highest in plants growing on stand 5, compared to the other 4 stands. intermediate values were observed for plants from stand 4, in relation to the other results. �e smallest increase in fresh mass was recorded in mosses collected from stands 1, 2 and 3. in the case of cratoneuron �licinum gametophytes, fresh mass was the largest for plants harvested from stands 3 and 5. on stand 4, the fresh mass was smaller than the mass of gametophytes from stands 3 and 5, but larger than that of plants growing on stands 1 and 2. a nn a s oł ty sle le k, z bi gn ie w c ap ut a 136 mosses from stands 1 and 2 showed the lowest values of this parameter in relation to the remaining stands (fig. 5a). �e dry mass of b. rivulare gametophytes was similar and was the largest in plants from stands 1, 2 and 5. on the other two stands (3 and 4), the dry mass values were signi�cantly lower. for c. �licinum, the signi�cantly largest dry mass increase was obtained for plants collected from stands 3, 4 and 5, compared to other stands (fig. 5b). �e percentage of water content in b. rivulare cells was signi�cantly the highest in plants from stand 4, compared to the other stands. �e lowest values of this parameter were found for mosses from stand 2. for c. �licinum, the highest water content was found in plants from stand 5. intermediate values are shown in stand 3. �e lowest water concentration in the tissues of this moss was observed in plants growing on stand 2 (fig. 5c). �e ratio of dry mass to fresh mass in b. rivulare was the highest for plants collected from stands 1 and 2, compared to the other 3 stands. intermediate values of this coef�cient were shown for mosses from stands 3 and 5. �e smallest ratio was calculated for the gametophytes from stand 4. for c. �licinum, the value of this parameter was clearly the highest in the plants from stand 4, compared to the other stands (fig. 5d). �e percentage of electrolyte leakage from b. rivulare gametophores was the highest in plants from stand 2. intermediate values were observed for the gametophores from fig. 5. fresh and dry mass, and water content in gametophytes of brachythecium rivulare schimp. – a and cratoneuron �licinum (hedw.) spruce – b. �e �ve karst spring niches (1–4 in the prądnik valley, 5 in the sąspowska valley); mean values (n = 5, ± sd) marked with di�erent letters di�er signi�cantly according to duncan’s test at p ≤ 0.05 the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 137 stands 4 and 5. signi�cantly, the lowest destabilisation of cell membranes was demonstrated for plants from stands 1 and 3. for c. �licinum, the signi�cantly highest electrolyte leakage was found for plants from stand 1, in relation to the remaining 4 stands. �e lowest degree of destabilisation of cell membranes was observed for gametophores from stands 4 and 5 (fig. 6). discussion �e analysed stands in the ojców national park are located in the bottoms of deep karst valleys (fig. 1–2). �e concave terrain forms such as the bottoms of valleys and canyons are characterised by much lower incoming solar radiation values compared to plateaus (caputa, wojkowski, 2009, 2013, 2015). according to bárány-kevei (2011), microclimatic di�erences are responsible for the diversity of vegetation in the karst ecosystem. in such ecological conditions, pioneering plants, such as bryophytes, �nd their refuge. �ese are the conditions of the variable ratio of short-wave and long-wave radiation and direct radiation that appear systematically in the same place during the day such as “sun �akes” (sołtys-lelek, 2009; pilarski et al., 2012). in the conducted research, the solar radiation �ux at individual measurement stands was strongly di�erentiated due to the topography and vegetation growing there (fig. 1–2; tab. 1). high values of solar radiation above the sunlit surface proved that mosses strongly absorbed light. lower values obtained in the immediate vicinity of shaded trees indicated limitations in absorption of di�use radiation (caputa, wojkowski, 2015). �ese types of changing conditions a�ect the formation of speci�c morphological and physiological features of mosses and allow them to occur in a variety of ecosystems. fig. 6. electrolyte leakage of brachythecium rivulare schimp. and cratoneuron �licinum (hedw.) spruce. �e �ve karst spring niches (1–4 in the prądnik valley, 5 in the sąspowska valley); mean values (n = 5, ± sd) marked with di�erent letters di�er signi�cantly according to duncan’s test at p ≤ 0.05 a nn a s oł ty sle le k, z bi gn ie w c ap ut a 138 bryophytes have developed �lters that help protect them from high levels of solar radiation. for example, some species of the genus polytrichum have lamellae, surrounded by a curled leaf blade, so that their structure does not di�er much from that of a tree leaf. others have leaves with �laments – crossidium, hyaline tips (hedwigia ciliata, bryum argenteum), and awns (tortula sp., syntrichia sp.), that overlap the next leaf and help to de�ect light before it reaches the cell interior. other mosses have warts that become more transparent when wet, doubling their ability to absorb solar radiation (glime, 2017). �e anatomical and morphological di�erences of mosses are treated as an expression of the adaptation arising in the phylogenetic development, allowing them to operate the photosynthetic apparatus in the most favourable conditions in the given habitat (krupa, 1974). in many studies, the response of bryophytes – e.g. bryum argenteum (rastorfer, 1970), racomitrium lanuginosum (kallio, heinonen, 1975), grimmia pulvinata and tortula ruralis (alpert, oechel, 1987) – to solar radiation showed saturation of photosynthesis even at low, 20% light intensity. murray et al. (1993) showed that mosses from shady areas, exposed to hight intensity of light, lost their ability to photosynthesize. on the other hand, mosses that had been transferred from full sun to shade grew at a rate of 2–3 times higher. �e photosynthesis in bryophytes is limited at very low light levels, yet these plants can survive in low light conditions, as previously mentioned. according to smith et al. (2009) and niinemets and niinemets, (2014) the total leaf area can absorb much more light and increase photosynthetic activity. �e leaf structure of bryophytes, with lower plant density, seems to provide the plant with an increased e�ciency of light uptake (niinmets, niinmets, 2009; rice et al., 2008). �e phenomena accompanying the disintegration of the photosynthetic apparatus are re�ected in changes in the values of chlorophyll a  �uorescence parameters (lichtenthaler et al., 1986; bolhàr-nordenkampf, öquist, 1993; �ach et al., 2007; demmig-adams et al., 2014; ruban, 2016). in this study, the variability of chlorophyll �uorescence may suggest that it was the result of photoinhibition (murray et al., 1993). �e photoinhibition was indicated by lower fv/fm values and activated photoprotection mechanisms, as evidenced by higher npq values, depending on the habitat conditions in the analysed stands (marschall, proctor, 2004). according to proctor et al. (2007) the increase in npq can be explained by the degree of cell hydration, which decreased in brachythecium rivulare and cratoneuron �licinum specimens from stands 1 and 2 (fig. 3–5). �e ability of the protective mechanisms that make up npq is speci�c and characteristic of a given moss species. most o�en it is related to the psii antenna system, i.e. performance of two essential light-harvesting complex (lhc)-like proteins, photosystem ii subunit s (psbs) in plants and light-harvesting complex stress-related (lhcsr) (rintamaki et al., 1994; pinnola et al., 2015; dikaios et al., 2019). mosses, depending on the species, are able to quickly regenerate physiological activity disturbed the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 139 by a stress factor (proctor, tuba, 2002; ciu et al., 2008; możdżeń, 2019). �ey share many photoprotective mechanisms with vascular plants. however, there are some key di�erences in the photoprotection available (robinson, waterman, 2014). �e state of hydration of the cells a�ects the moss ability to absorb or re�ect light (lappalainen et al., 2008). �is suggests that the mosses have developed mechanisms that allow them to scatter light internally and/or externally. in dried mosses, increased re�ection (albedo) and reduced solar radiation absorption (low nr values) should provide them with some protection against the harmful e�ects of light. in fully hydrated mosses (low albedo and high nr), the surface and interior are more homogeneous and absorb solar radiation more e�ectively (lovelock, robinson, 2002). in the analyses carried out here, the di�erences in the fresh and dry mass in�uenced the water content in the gametophores of the studied species (fig. 5). �e water content was signi�cantly the lowest in both mosses collected from stands 1 and 2, compared to the 3 remaining stands. �is was most likely due to the fact that mosses are highly �exible in relation to environmental conditions. at these two stands, the measurements show a high temperature and strong absorption of solar radiation by the surface of the mosses (tab. 1). �is is probably why the tested mosses were characterised by the lowest masses and water content values (fig. 5). �is result could also depend on other environmental factors (tab. 1). it can be assumed that the substrate, species composition of other plants, temperature and humidity of the environment played an important role here. at stands 1 and 2, the spring niche is created by a semi-circular concrete lining, which makes the substrate highly exposed to drying out. in the remaining stands, mosses grew on soil or calcareous rock-mantle mixed with soil (fig. 1–2). mosses are characterised by di�erent hydration depending on, among others light, the availability of surface water, the action of capillary forces, supported by the setting of gametophore leaves (proctor et al., 2007; romańska, 2020). �is directly a�ects the rate of their elongation growth, masses gain and changes in the chemical composition of plant pigments and other metabolites. such interactions are also perceived at the cellular and molecular level (ueneka, 2005). �e e�ects of stress caused by di�erent spectral composition and light intensity destabilise the proper functioning of mosses, causing changes in the permeability of cell membranes (możdżeń, 2019). cell membranes react the fastest to the in�uence of a stress factor. �ey are highly selective permeation barriers but do not completely isolate because they contain speci�c channels, conveyors and pumps. undamaged cell membrane allows water molecules to enter the cell interior and is a barrier for molecules of substances dissolved in the cell. �e higher the degree of damage to membranes by stress factors, the higher part of the cell contents �ows out (kocheva et al., 2014). �e stress-induced electrolyte leakage is accompanied by changes in the structure of proteins and lipids, the concentration of ions in the vacuole and cytoplasm, and the generation and accumulation of reactive oxygen species, which in extreme cases leads to cell death a nn a s oł ty sle le k, z bi gn ie w c ap ut a 140 (demidchik et al., 2014; scotti-campos, pham-�i, 2016). �e studies of electrolyte leakage carried out here in b. rivulare and c. �licinum showed the highest membrane destabilisation in plants from stands 1 and 2, respectively, compared to other stands (tab. 1; fig. 6). �e obtained results prove the most unfavourable environmental conditions for the existence of mosses in these stands. within them, mosses do not occur on natural ground, because the spring niches have been quite strongly transformed by human – the surrounding of the out�ow with a concrete lining (fig. 2). comparing the photosynthetic activity of moss gametophores is not easy, because the response of plants is o�en the result of many di�erent environmental factors. from an ecological point of view, an important role is played by the leaf area to volume ratio and the percentage of assimilation tissues in the total leaf mass (miyata et al., 2015). �e comparison of photosynthesis e�ciency to mass is relative and it seems that there is no adequate relationship in terms of the total photosynthetic production (krupa, 1974). �e analysis of selected physiological parameters did not clearly indicate which of the studied stands is the most optimal for mosses. �erefore, further research in this area is necessary in order to distinguish both the most convenient stands of spring mosses in the onp area and to verify which environmental factors play a key role in their physiology. conclusion (1) �e photosynthetic activity of mosses was speci�c and depended on the stand and species. such reactions most likely resulted from the structure of mosses and their di�erent adaptations to environmental conditions. (2) �e fresh mass of gametophytes was signi�cantly the largest in plants growing on stand 5, where there is the only completely natural spring niche among the studied. mosses from stands 1 and 2 showed the lowest values of this parameter. �e dry mass of mosses varied and also depended on the species and stand. �e percentage of water content of both tested mosses was the lowest for plants growing in stand 2 with the most transformed spring niche; bryophytes appeared here only on the arti�cial substrate of concrete lining. (3) �e degree of destabilisation of the cell membranes of brachythecium rivulare and cratoneuron �licinum gametophores was speci�c and, as in the previous parameters, depended on the species and location. in general, the highest percentage of electrolytes leakage was found in plants harvested from stands 1 and 2, which proves the highest environmental stress in these two examined positions. the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 141 acknowledgements �e authors wish to express their gratitude to phd katarzyna możdżeń for her help in plant physiological analyses. we are very grateful to the reviewers for their valuable comments to improve the manuscript. con�ict of interest �e authors declare no con�ict of interest related to this article. references aleksandrowisz, s.w., wilk, z. 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(2016). �e impact of karst relief on the diversity of insolation conditions and mesoclimate variation: case study of the ojców national park, poland. international journal of geoheritage, 4(1), 33–43. abstract �e availability of light is one of the most important environmental factors in�uencing the �oristic diversity of spring niches, especially in the speci�c conditions of deep, karst valleys occurring in the ojców national park (southern poland). �e aim of this study was to investigate the in�uence of solar radiation reaching the karst spring niches, on selected physiological parameters of the spring mosses: cratoneuron �licinum (hedw.) spruce (obligatory krenophyte) and brachythecium rivulare schimp. (facultative krenophyte). �e �ve karst spring niches (4 in the prądnik valley, 1 in the sąspowska valley) were selected for the plant material collection, in which two of the moss species tested occurred simultaneously. on sunny days, measurements of total and re�ected radiation, as well as the radiation balance in the full spectrum range over the vegetation were made. �e temperature was measured for air, water, and on the surface and inside the plants. �e collected biological material was subjected to laboratory analysis. fresh mass of moss gametophytes was signi�cantly the highest from plants growing on stand 5 (intermediate values of light and temperature parameters), and the lowest from mosses on stands 1 and 2 (including lowest air temperatures). dry mass varied depending on the species and stand. �e percentage of water in b. rivulare was highest in plants from stand 4 (highest air and water temperature), and in c. �licinum from stand 5. signi�cantly the lowest values of this parameter were found for plants growing in stand 2 (lowest temperature of water). �e electrolytes leakage from moss cells was speci�c and depended on the species. �e greatest destabilisation of cell membranes was demonstrated in plants harvested from stands 1 and 2, where it was the coldest. �e �uorescence of chlorophyll a varied depending on the moss species and the habitat of spring niches. �is paper, presenting of preliminary results, is a kind of introduction to wider research in this topic. the influence of solar radiation on selected physiological processes of m osses in karst conditions of the spring niches of the o jców n ational p ark (s outhern p oland) 145 key words: albedo, biomass, brachythecium rivulare, cratoneuron �licinum, chlorophyll a �uorescence, electrolyte leakage, net all-wave radiation received: [2021.02.22] accepted: [2021.05.15] wpływ promieniowania słonecznego na wybrane procesy fizjologiczne mchów w warunkach krasowych nisz źródliskowych ojcowskiego parku narodowego (południowa polska) streszczenie dostępność światła jest jednym z najważniejszych czynników środowiskowych wpływających na różnorodność �orystyczną nisz źródliskowych, zwłaszcza w specy�cznych warunkach głębokich, krasowych dolin, występujących w ojcowskim parku narodowym (południowa polska). celem niniejszej pracy było zbadanie wpływu promieniowania słonecznego, docierającego do nisz źródliskowych, na wybrane parametry �zjologiczne mchów źródliskowych: cratoneuron �licinum (hedw.) spruce (kreno�t obligatoryjny) i brachythecium rivulare schimp. (kreno�t fakultatywny). do zbioru materiału roślinnego wytypowano 5 krasowych nisz źródliskowych (4 w dolinie prądnika, 1 w dolinie sąspowskiej), w których występowały jednocześnie obydwa gatunki mchów. w słoneczne dni dokonano pomiarów promieniowania całkowitego, odbitego oraz salda promieniowania w pełnym zakresie widma nad roślinnością. pomierzono temperaturę powietrza, wody oraz na powierzchni i wewnątrz roślin. zebrany materiał biologiczny poddano analizom laboratoryjnym. świeża masa gameto�tów była istotnie największa u roślin rosnących na stanowisku 5 (pośrednie wartości parametrów światła i temperatury), a najmniejsza u mchów ze stanowisk 1, 2 (m.in. najniższe temperatury powietrza). sucha masa zmieniała się w zależności od gatunku i siedliska. procentowa zawartość wody u b. rivulare była największa dla okazów ze stanowiska 4 (najwyższa temperatura powietrza i wody), a u c. �licinum ze stanowiska 5. istotnie najmniejsze wartości tego parametru stwierdzono dla roślin rosnących na stanowisku 2 (najniższa temperatura wody). wypływ elektrolitów z komórek mchów był specy�czny i zależał od gatunku. największą destabilizację błon komórkowych wykazano u roślin zebranych ze stanowisk 1 i 2, gdzie było najchłodniej. fluorescencja chloro�lu a zmieniała się w zależności od gatunku mchu i źródliska. niniejsza praca prezentująca pilotażowe wyniki, stanowi niejako wstęp do szerzej zakrojonych badań w tym zakresie. slowa kluczowe: albedo, biomasa, brachythecium rivulare, cratoneuron �licinum, �uorescencja chloro�lu a, wypływ elektrolitów, saldo promieniowania krótko i długofalowego information on the authors anna sołtys-lelek https://orcid.org/0000-0002-9595-3167 author of numerous scienti�c and popular science studies in the �eld of botany and environmental protection. her research interests relate particularly to the critical type of rose (rosa) and hawthorn (crataegus). member of the polish and slovak botanical society. zbigniew caputa he is a climatologist, polar explorer, former employee of the silesia university. his research focused in particular on the long-term variability of the climate and modelling the in�ow of solar radiation. 63 annales universitatis paedagogicae cracoviensis studia naturae, 6: 63–80, 2021, issn 2543-8832 doi: 10.24917/25438832.6.4 elke bloem1*, jowita ciaranek2 1institute for crop and soil science, federal research centre for cultivated plants (jki), bundesallee 69, 38116 braunschweig, *elke.bloem@julius-kuehn.de 2institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland the influence of lead compounds on selected morphological features and the physiological processes of zea mays l. introduction soil is a valuable, non-renewable environmental resource, essential for the development and continuity of plant growth and life. �e soil stores, �lters and transforms various substances, including nutrients and carbon. it contains mineral salts and water – substances necessary for the seed germination process, for the growth and development of most plants. it is a heterogeneous mixture of organic and inorganic compounds with various particle sizes (kabata-pendias, pendias, 2001). soils should be protected because of their importance, both for the natural environment and in the economic and social context. however, in the modern world, soil degradation is a serious problem. �e reason for this degradation is anthropopression manifested by: inadequately conducted agricultural and forestry works, industrial activity, tourism, uncontrolled development of cities and industrial regions, and improper land development (marcinek et al., 1995; ettler, 2016; możdżeń et al., 2017; huo et al., 2020). as side e�ect of the process of industrialisation, urbanisation and technology advances, one of the many problems around the world is the release of heavy metals into the soil. heavy metal pollution causes adverse changes in soil properties, has a direct impact on water quality, and negatively a�ects biodiversity and climate change (rodriguesa et al., 2017). such di�use pollution also threatens the production of organic food (konieczna et al., 2018a, b). among heavy metals, lead (pb) is an element that accumulates easily in soils and sediments (kabata-pendias, pendias, 2001). �e main sources of its pollution are emissions from copper, iron and steel smelters, smelted zinc ores and cement production (pattee, pain, 2002; carocci et al., 2016). in many countries around the world, the release of lead has been reduced, but unfortunately it is still used in the automotive industry, el ke b lo em , j ow ita c ia ra ne k 64 in the production and recycling of batteries, boat building, pottery and book printing, ammunition for hunting. due to its non-biodegradable nature, its concentration accumulates in the environment, which contributes to an increase in the threat to health and life of organisms (jiao et al., 2012; li et al., 2012; arnemo et al., 2016; zhou et al., 2018). although lead is not an essential element for plants, it is easily absorbed and accumulates in di�erent plant parts. �e highest amounts of lead were found in the roots of plants, with the simultaneous limitation of its translocation to the above-ground parts (seregin, ivanov, 2001). lead uptake by plants is regulated by the ph, particle size and cation exchange capacity of the soil. �e excess of lead causes blackening of the root system of plants, growth inhibition and chlorosis. moreover, photosynthesis is disturbed, mineral nutrition and water balance are disrupted, the hormonal status is changed and the structure and permeability of cell membranes is a�ected (sharma, dubey, 2005; puła et al., 2019). �e toxic e�ect of lead on living organisms is mainly related to the reaction of lead ions with sul¶ydryl groups of enzymes and proteins (kabata-pendias, pendias, 2001; kabata-pendias, mukherjee, 2007). �e extent to which this metal a�ects plants varies with its concentration as well as with plant species. some plants can tolerate levels of heavy metals that are already toxic to other plants (kranner, colville, 2011). although studies of the lead-plant interactions are quite frequent, few experiments have been performed simultaneously at the stage of seed germination and plant growth. �erefore, the aim of the current study was to determine the germination of grains and the growth of maize (zea mays l.) plants that are equally exposed to lead in the form of nitrate compounds. it was hypothesised that the percentages of the lead solution used had a  di�erent e�ect on the physiological parameters of maize. �erefore, analyses were carried out on (1) grains germination capacity, (2) plant growth, (3) fresh and dry mass of maize organs and (4) photosynthetic activity by measuring photosynthesis and transpiration. material and methods plant material grain of maize (zea mays l.) was purchased in the polan garden store. �ese grains were used in petri dishes germination biotests and for cultivation in the growth chamber. solution used in experiment water solutions of pb(no3)2 × 4h2o were prepared according to the procedure for 0.1%, 1% and 3% solutions. a solution containing 1% was prepared from 1 g of a chemical compound dissolved in 99 ml of distilled water. �e other solutions were prepared accordingly and stored in the refrigerator for the duration of the experiment. 65 germination condition maize grains (zea mays l.), rinsed under running distilled water by 30 min., were placed in sterile petri dishes with a diameter of 15 cm with three layers of �lter paper (100 grains per petri dish). each petri dish was soaked with 10 ml of the appropriate solution and 3 ml was watered every other day. �e control consisted of petri dishes with grains watered with distilled water in the same amounts as the petri dishes with lead solutions. �e germination test in petri dishes was stored in the dark at a temperature of 25°c (day / night) and under a relative air humidity of 60–70%. a�er 24 hours, the germinated grains were counted. �e operation was repeated a�er 48 h, 72 h and 96 h. plant cultivation maize grains germinated in petri dishes were planted in 0.5 litre pots with sand and were transferred into a growth chamber with a light intensity of 200 μmol × m–2 × s–1 (li-cor, usa), during the photoperiod of 12 h and with a temperature of 25°c and a temperature of 20°c. �e relative humidity (rh) was 70–80%. two di�erent experimental setup were established. in one group of pots plants were grown from grains which germinated already on substrates containing lead(ii) nitrate solution and which were watered during their further growth with distilled water. �e second group of pots included plants grown from grains germinated in distilled water but which were watered during their growth with lead(ii) nitrate solution. �e control group consisted of plants, which were watered with distilled water, both during germination and further growth. once a week, all plants were watered with steiner medium (steiner, 1961). biometric analysis �e e�ect of di�erent concentrations of pb(no3)2 solutions on the growth of maize plants was measured with a calliper, with an accuracy of ±0.1 cm. �e length of the roots, blades, leaves (�rst to ��h leaf ) and the remaining part of the shoot was measured. based on the formula of mominul islam et al. (2012), the length of the organs expressed as a percentage of control organ was determined (ip): ip [%] = [1 – (ls / lc)] × 100 ip – inhibition percentage [%], ls – organ length [cm] treated with the water solution type, lc – organ length [cm] treated with the distilled water (control group) plant biomass �e fresh mass (fm) of the maize organs was determined on a  laboratory balance (ohaus adventurer pro, usa) with an accuracy of 0.0001 g. in order to determine the the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 66 dry mass, plant organs were dried in a laboratory dryer (wamed sup 100, poland) for 48 h at a temperature of 105°c. based on the masses obtained, the percentage of water content was determined according to the formula: wc = [(fm – dm) / fm] × 100 and percentage of dry mass: dm = (dm / fm) × 100; where wc – water content, fm – fresh mass, dm – dry mass. photosynthesis and transpiration �e intensity of photosynthesis (pn) and transpiration (e) was determined for the third maize leaf. for this purpose, the ciras-2 infrared gas analyser was used. a plc 4 board chamber with an area of 2.5 cm2 was used for measurements. �e intensity of light reaching the leaf during photosynthesis measurements was 200 μmol × m–2 × s–1, 60–70% humidity, and temperature was about 25°c. statistical analysis �e signi�cance of di�erences between the subjects was tested by duncan’s test for homogeneous groups, at the level of p ≤ 0.05; mean values (n = 5, ± sd) marked with di�erent letters in rows or columns di�er signi�cantly. �e calculations were made in excel and statistica for windows 13.1. statso�, inc. (2021). results germination of maize was a�ected by the higher concentrated lead solutions (tab. 1). a�er 48 h of germination of zea mays l. grains it was observed that grains germinated to a  higher extent in the lead salt solution containing 0.1% pb(no3)2, compared to the control and the higher concentrations of lead solutions. �e lowest percentage of germinated grains was found on the substrates containing 3% pb(no3)2. on the third day of germination, the number of germinated grains was lower in each concentration of lead solutions, compared to the germination rate in the control petri dishes. �e signi�cantly, lowest number of seeds germinated in the petri dishes containing the 3% lead solution. in the next two days of germination, the percentage of germinated grains was the highest in the control, in relation to the lead salt solution used. along with the increase in the concentration of the solution, a signi�cant inhibition of the germination of maize grains was demonstrated (tab. 1). maize plants developed very di�erently in relation to the applied lead treatment when grown in pots (tab. 2). plants grown from grains germinated already on lead water solutions developed signi�cantly shorter roots compared to plants that germinated without lead but were watered with metal solutions during their growth a�er germination (fig. 1; tab. 2). 67 tab. 1. percentage of germinated maize grains (zea mays l.) treated with solutions of pb(no3)2 at concentrations of 0.1%, 1% and 3% in the germination test in petri dishes time [h] control 0.1% 1% 3% 48 56 b 66 a 22 e 2 f 72 92 a 82 b 36 d 12 f 96 94 a 90 b 66 d 20 e 120 99 a 90 b 67 d 21 f mean values (n = 5) marked with di�erent letters (within the row) di�er signi�cantly according to duncan’s test at p ≤ 0.05 tab. 2. development of di�erent plant organs of zea mays l. in length [cm] in relation to lead application at di�erent concentrations during germination in comparison to treatment a�er germination (control – seeds and plants treated with distilled water; lead application: pb(no3)2 water solutions at 0.1%, 1% and 3% concentrations during the germination stage or as irrigation during growth) organ control lead application during germination lead application during growth 0.1% 1% 3% 0.1% 1% 3% root 20.80 c 19.82 d 18.32 de 19.70 d 23.80 a 21.40 bc 14.60 g blade 6.40 a 3.54 e 3.14 g 4.80 d 5.60 bc 5.70 b 3.30 f i leaf 5.60 a 4.52 c 3.42 e 4.80 b 5.30 ab 5.50 ab 4.80 b ii leaf 15.80 a 10.06 de 9.04 e 12.50 cd 15.20 ab 15.30 ab 13.10 c iii leaf 23.30 d 18.10 f 17.36 g 21.60 d 26.50 b 28.20 a 18.40 e iv leaf 13.60 cd 19.56 b 19.05 b 14.50 c 19.70 b 21.70 a 11.40 e v leaf 9.00 a 7.14 b 6.28 c 0.00 h 2.80 f 6.00 c 0.00 h rps 5.26 a 5.02 ab 5.24 a 3.00 c 3.60 c 4.10 b 3.40 c i leaf, ii leaf, iii leaf, iv leaf, v leaf – leaf number, rps – remaining part of the shoot; mean values (n = 5) marked with di�erent letters (within the row) di�er signi�cantly according to the duncan test at p ≤ 0.05 compared to the control, the roots of z. mays were the longest in plants grown from grains germinated in distilled water and watered with 0.1% pb(no3)2 during growth, and the shortest in plants treated with 3% pb(no3)2. biometric analyses of blades and leaf i and ii showed that each of the solutions inhibited the growth of these organs, regardless of the stage in which lead was applied. in case of the third leaf shorter leaves in comparison to the control were developed in plants which germinated in lead solutions, but longer ones when lead application occurred during growth. �e iv leaf was longer in lead-treated plants regardless of the application phase. compared to the control shorter leaves were observed only in the plants that received 3% pb(no3)2 during growth. �e length of ��h maize leaf was the longest for the control plants. plants germinated in lead solution or watered with 3% pb(no3)2 solution did not develop a ��h leaf. �e remaining part of the shoot was signi�cantly the longest in the control plants, compared to the plants receiving lead at germination or during growth (tab. 2). the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 68 �e percentage of growth inhibition (ip value) for the di�erent plant organs in relation to lead application is shown in �gure 1. �e ip clearly indicate that the strongest negative e�ect was observed for the length of the ��h leaf. biomass development of maize in relation to lead application is shown in table 3. �e fresh mass of root of the maize plants varied depending on the concentration of the solution. fig. 1. length, expressed as the ip (inhibition percentage) index in percentage of the control group, of the individual plant organs of zea mays l. (treatments: control – watered with distilled water in comparison to pb(no3)2 solutions at concentrations 0.1%, 1% and 3% applied at (a) – germination or (b) – during growth; mean values (n = 5), positive values indicate a negative e�ect, negative values indicate a positive e�ect; 1 (blue): 0.1% pb(no3)2, 2 (yellow): 1% pb(no3)2, 3 (green): 3% pb(no3)2; rps – remaining part of the shoot 69 tab. 3. development of fresh biomass [g] of individual plant organs of zea mays l. in relation to lead application at di�erent concentrations during germination in comparison to treatment a�er germination (control – seeds and plants treated with distilled water; lead application: pb(no3)2 water solutions at 0.1%, 1% and 3% concentrations during the germination stage or as irrigation during growth) organ control lead application during germination lead application during growth 0.1% 1% 3% 0.1% 1% 3% root 2.0122 ab 2.1154 a 1.9886 b 1.7870 c 2.1014 a 1.7925 c 0.7218 f blade 0.4916 d 0.4830 d 0.4022 e 0.4906 d 0.7736 b 0.8453 a 0.2836 f i leaf 0.0440 f 0.0710 d 0.0640 e 0.0886 cd 0.1210 a 0.1128 b 0.0190 g ii leaf 0.1518 e 0.1550 e 0.1536 e 0.2216 d 0.3054 a 0.2948 ab 0.1116 g iii leaf 0.3014 ef 0.3111 e 0.3077 f 0.3700 d 0.5555 b 0.6220 a 0.2082 h iv leaf 0.3646 b 0.3706 b 0.3506 b 0.1950 d 0.3934 a 0.4320 a 0.1108 f v leaf 0.0924 a 0.0754 b 0.0600 c 0 h 0.0334 f 0.0428 e 0 h rps 0.4004 b 0.4298 ab 0.4320 ab 0.2030 d 0.3558 c 0.4123 ab 0.1452 f i leaf, ii leaf, iii leaf, iv leaf, v leaf – leaf number, rps – remaining part of the shoot; mean values (n = 5) marked with di�erent letters (within the row) di�er signi�cantly according to the duncan test at p ≤ 0.05 �e lowest biomass development was recorded for all investigated plant parts in plants treated with solutions containing 3% of lead, compared to the control, regardless of the application stage. �e values of the fresh mass of the blades were clearly higher for plants treated with lead solutions during growth in comparison to application at germination stage. compared to the control, the fresh mass of the �rst three leaves was higher in plants watered with lead solutions during growth while that of the plants that received lead during the germination stage was comparable. �e fourth leaf achieved the highest fresh mass in maize plants watered during growth with 0.1% or 1% pb(no3)2, relative to the control. �ere were no di�erences in the values of this parameter between the control plants and the plants germinating in 0.1% and 1% of pb(no3)2. biomass of the ��h leaf was lower in all treatments in comparison to the control. �e fresh mass of the remaining part of the maize shoot was greater in plants grown from grains watered with lead solutions during the germination stage. generally, in the remaining cases, these values were lower than the control. �e lowest fresh mass values were recorded in plants watered with 3% pb(no3)2 during growth. �e percentage of dry mass in the roots of z. mays grown from grains watered with lead solutions was the highest in the control, in relation to all solutions used (fig. 2). in the case of plants treated with lead solutions during growth, the highest percentage of root dry mass was found in those watered with 3% pb(no3)2. �e percentage of dry mass of the blade was lower in each of the lead solutions, in relation to the control, regardless of the watering stage. the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 70 exceptions were noted for plants from the germination stage treated with 0.1% pb(no3)2 and in the growth stage treated with 3% pb(no3)2. for the �rst maize leaf, lower values of this parameter were found, compared to the values from the control plants. �e percentage of dry mass for the 2nd and 3rd leaves di�ered from the control and depended on the concentration and time of application. in the case of the ��h leaf, lower values of this parameter were observed compared to the control sample. dry mass for the remaining part of the shoot showed di�erent values compared to the control. �e dry mass of z. mays roots was greatest in the control plants (tab. 4). regardless of the treatment stage, a decrease in the mass gain of this organ was observed once the concentration of lead solutions increased. �e dry mass values for the blade varied fig. 2. development of dry biomass [g] of individual plant organs of zea mays l. in relation to lead application at di�erent concentrations during germination (a) in comparison to treatment a�er germination during growth (b) (mean values (n = 5); 1 (blue) – control (distilled water), 2 (red) – 0.1% pb(no3)2, 3 (green) – 1% pb(no3)2, 4 (yellow) – 3% pb(no3)2, rps – remaining part of the shoot) 71 depending on time of lead application and concentration of the solution. in most cases, the dry mass of the �rst maize leaf was higher in plants watered with lead solutions in comparison to the control. for the �rst leaf no di�erences were found between the control plants and plants that received 3% pb(no3)2 solution at germination stage or during growth. for the second leaf, the dry mass value was higher in the plants treated with lead solutions compared to the control. �e lowest value was observed for plants watered with 3% pb(no3)2 during growth. dry mass of the third leaf was not a�ected by lead application during germination while that of the fourth leaf was clearly lower in plants watered with lead solutions and at the highest lead concentration during germination stage. �e dry mass of the ��h leaf was signi�cantly lower in each lead treatment what was already obvious in the results presented before. compared to the control, the dry mass of the remaining part of shoot was lower in all plants watered with lead solutions during growth. during the germination stage, the dry mass of this organ was higher in the treatment with 1% pb(no3)2. tab. 4. development of dry biomass [g] of individual plant organs of zea mays l. in relation to lead application at di�erent concentrations during germination in comparison to treatment a�er germination (control – seeds and plants treated with distilled water; lead application: pb(no3)2 water solutions at 0.1%, 1% and 3% concentrations during the germination stage or as irrigation during growth) organ control lead application during germination lead application during growth 0.1% 1% 3% 0.1% 1% 3% root 0.2976 a 0.2618 c 0.2666 c 0.2274 d 0.2290 d 0.2300 d 0.1402 g blade 0.0548 b 0.0628 a 0.0464 c 0.0476 c 0.0550 b 0.0643 a 0.0332 d i leaf 0.0078 c 0.0110 a 0.0092 b 0.0076 c 0.0106 a 0.0110 a 0.0070 c ii leaf 0.0244 c 0.0256 bc 0.0312 a 0.0254 bc 0.0302 a 0.0308 a 0.0202 d iii leaf 0.0523 b 0.0560 b 0.0554 b 0.0500 bc 0.0586 ab 0.0686 a 0.0350 d iv leaf 0.0660 ab 0.0700 a 0.0626 ab 0.0300 d 0.0434 c 0.0523 b 0.0174 e v leaf 0.0146 a 0.0080 c 0.0088 bc 0 g 0.0028 e 0.0053 d 0 g rps 0.0332 b 0.0150 d 0.0438 a 0.0208 c 0.0242 c 0.0300 b 0.0198 d i leaf, ii leaf, iii leaf, iv leaf, v leaf – leaf number, rps – remaining part of the shoot; mean values (n = 5) marked with di�erent letters (within the row) di�er signi�cantly according to the duncan test at p ≤ 0.05 water content in the roots and blades of maize was highest in plants watered during growth with lead solutions of 0.1% and 1%, in relation to the remaining concentrations and the control (tab. 5). in the case of the blade and the examined leaves, the lead solutions partly caused an increase in the percentage of water content. statistically signi�cant di�erences were shown especially in maize plants watered with lead solutions during growth. in the case of the remaining part of the shoot, there were no di�erences in the water content in relation to lead application. the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 72 �e intensity of photosynthesis of maize leaves changed in relation to the concentration of lead solutions. with an increase in lead concentration, a  decrease in the photosynthetic e�ciency of plants was observed (fig. 3). tab. 5. total water content [%] of individual plant organs of zea mays l. in relation to lead application at di�erent concentrations during germination in comparison to treatment a�er germination (control – seeds and plants treated with distilled water; lead application: pb(no3)2 water solutions at 0.1%, 1% and 3% concentrations during the germination stage or as irrigation during growth) organ control lead application during germination lead application during growth 0.1% 1% 3% 0.1% 1% 3% root 85.21 bc 87.62 b 86.59 bc 87.27 b 89.10 a 87.17 a 80.58 c blade 88.85 c 87.00 c 88.46 bc 90.30 b 92.89 a 92.39 a 88.29 b i leaf 82.27 b 84.51 b 85.63 b 91.42 a 91.24 a 90.25 a 63.16 c ii leaf 83.93 b 83.48 b 79.69 bc 88.54 a 90.11 a 89.55 a 81.90 a iii leaf 82.65 ab 82.00 ab 82.00 ab 86.49 a 89.45 a 88.97 a 83.19 ab iv leaf 81.90 b 81.11 b 82.14 ab 84.62 ab 88.97 a 87.89 a 84.30 a v leaf 84.20 c 89.39 b 85.33 c n.d. d 91.62 a 87.62 bc n.d. d rps 91.71 a 96.51 a 89.86 ab 89.75 ab 93.20 a 92.72 a 86.36 ab i leaf, ii leaf, iii leaf, iv leaf, v leaf – leaf number, rps – remaining part of the shoot, n.d. – not developed; mean values (n = 5) marked with di�erent letters (within the row) di�er signi�cantly according to the duncan test at p ≤ 0.05 fig. 3. �e intensity of photosynthesis (pn) and transpiration (e) of the third leaf of zea mays l. grown from grains treated with lead solutions during germination stage and distilled water during growth (a, c), and distilled water during germination and lead solutions during growth (b, d); pb(no3)2 solutions at concentrations 0.1%, 1% and 3%; mean values (n = 5, ± sd) marked with di�erent letters di�er signi�cantly according to the duncan test, with p ≤ 0.05 73 in plants grown from grains treated with pb(no3)2 solutions during germination, signi�cant di�erences were found between 3% solutions and the control sample (fig. 3a). in plants grown from grains treated with distilled water, and with lead solutions during growth, a negative e�ect of lead was demonstrated at concentrations of 1% and 3% (fig. 3b). �e intensity of transpiration of z. mays leaves was signi�cantly lower in each of the treatments with lead solutions at germination as well as during growth (fig. 3.c–d) with the only exception of plants that received 0.1% pb(no3)2 during germination (fig. 3c). discussion contamination of soils with heavy metals causes the accumulation of these metals in plant organs, which reduce crop yield (saifullah et al., 2015). �is is con�rmed by scienti�c reports from various regions of the world (adekunle et al., 2009; singh et al., 2010; bigdeli, seilsepour, 2010; zhuang et al., 2009; kachenko, singh, 2006; schreck et al., 2013). one of the �rst stages of plant contact with heavy metals is seeds sown into contaminated soil. during germination, the seed coat becomes permeable not only to water, but also to other environmental factors. it is the �rst protective barrier that metals pass to get inside the embryo (kranner, colville, 2011; możdżeń, rzepka, 2016). �is study showed the inhibitory e�ect of lead compounds on the germination of maize (zea mays l.). �e negative e�ect of lead, in the form of nitrate compounds, increased in proportion to the concentration of their solutions (tab. 1). �is kind of response could be related, i.e. with limited water uptake and transport in the presence of lead compounds (abraham et al., 2013). it could also result from the interference of lead with the enzymes of proteases and amylases (sengar et al., 2008). heavy metals in high concentrations contribute to the destabilisation of metabolic processes and the production of excess compounds, harmful to further growth and development (brewer, 2010; możdżeń et al., 2017). lead, regardless of the form in which it occurs, not only limits seed germination, but also adversely a�ects plant growth. it inhibits root growth to a greater extent than growth of aboveground plant parts (greger, 2004; liu et al., 2009; shah et al., 2010; yang et al., 2010; seregin, ivanov, 2011). lead absorption by the roots takes place through the apoplastic route or through calcium ion-permeable channels. a�er absorption, lead accumulates primarily in the root cells due to the blockage of the “casparian strips” in the endoderm. additionally, lead can be picked up by the negative charges on the root cell walls. as the concentration of lead in cells increases, there are also a number of changes at their ultrastructural level (piechalak et al., 2002). in the studies carried out here, the negative e�ect of lead solutions on the growth of maize organs was demonstrated (fig. 1, 4; tab. 2). �e highest negative e�ect of lead the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 74 was found in plants grown from grains germinated on substrates with the analysed solutions, compared to plants treated with lead solutions only during growth. for example, shah et al. (2010) found that the seedling stage is more susceptible to heavy metal stress than the later vegetative stages. �e positive values of the ip index indicated the percentage of inhibition of organ growth relative to the control (fig. 1). �ese di�erences, with respect to control, were highest for the ��h leaf and the remaining part of the shoot, and not for the roots. �e observed stimulating e�ect of lead on root growth was most likely a secondary e�ect, caused by damage to the selective function of cell membranes (jasiewicz, 1996). lead uptake by the root system is a passive process and is proportional to the presence of soluble forms in the substrate. �e factors that signi�cantly increase its phyto-bioavailability are the acidic ph of the soil and high ambient temperature. atmospheric lead is an important source of contamination of plants, especially aboveground parts and soils. it is estimated that approximately 73–95% of the total lead content in plants comes from this source (kabata-pendias, pendias, 2001). fig. 4. maize (zea mays l.) plants grown from grains treated with solutions of pb(no3)2 at concentrations of 0.1%, 1% and 3% at (a) germination and (b) during growth (photo. e. bloem) 75 ahmad et al. (2011) found the phytotoxic e�ect of lead on germinating, growth, and the value of fresh and dry mass of maize seedlings. in this study, a decreasing fresh and dry mass and changes in the water content in maize organs was observed (fig. 2, tab. 2–5) in relation to lead concentration. �e di�erences in the values of these parameters were proportional to the concentrations of the lead solutions and slightly di�ered between the chemical compounds from which they were prepared (ahmad et al., 2011; malar et al., 2014; iqbal et al., 2017). �e obtained results clearly indicate that maize belongs to plants with poorly developed adaptive mechanisms in relation to high concentrations of lead. �e mechanism of tolerance consists primarily in changes in the properties of cell membranes, which, due to the secretion of more pectin’s, increase their sorption capacity in relation to lead. lead can also be excluded from metabolism by retaining it in endoderm cells. one of the processes of lead immobilisation in plants is its precipitation in the form of orthoand pyrophosphates on the cell membranes of the roots or stems (jasiewicz, 1996; sharma, dubey, 2005). lead has a strong in�uence not only on the morphology but also on the physiology of plants. it causes changes in the structure and functioning of chloroplasts, blocks the electron transport chain, the enzymes of the calvin cycle, impairs the uptake of essential elements – mg and fe, and causes co2 de�ciency due to the closure of the stomata. under environmental stress, stomata as well as non-stomata factors can reduce the photosynthetic abilities of plants (kodera et al., 2008; oliwa et al., 2016). in the present experiment it was observed that the rate of transpiration (e) in�uences the photosynthetic ability (fig. 3). with the increase in the concentration of lead solutions, a  decrease in the photosynthetic e�ciency of plants was observed. lower values of parameters were demonstrated in maize plants treated with lead solutions during growth, compared to the control (fig. 3b). �is type of reaction was likely due to excessive evaporation, which can transport the lead ions up the plant and binds them in the surrounding vascular tissue. perhaps it was also due to the accumulation of lead in the cuticular layer of the leaves (verbruggen et al., 2009; puła et al., 2019). �e chemical forms of lead have di�erent e�ects on plants, changing their biological properties. although they are similarly taken up from the soil, transposed and stored in plant organs, they a�ect the metabolism to a di�erent extent, in extreme cases leading to death (iqbal et al., 2017). plants have developed strategies to combat heavy metal stress (hong-bo et al., 2010). �is include, e.g., reduced uptake of metals into the cell, binding of lead to phyto-chelatins, glutathione and amino acids in the vacuole by forming complexes. to achieve nutrient homeostasis, plants have developed various strategies for mobilization and uptake, chelation, storage, and transport between cells and organs. plants regulate the uptake of nutrients, while responding to changes in the soil and inside (williams et al., 2000). as a secondary defence system, they have developed antioxidant activation the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 76 abilities to combat the increased production of reactive oxygen species (reddy et al., 2005; pourrut et al., 2011). conclusion in response to toxic soil conditions, plants struggle to survive by reduced absorption of heavy metals, activating mechanisms that exclude their absorption in metabolically inactive cells. despite this, they still cannot cope with this type of stress, which is manifested by disturbances in their life processes. in this experiment, it was observed that lead contamination inhibited the germination of maize grains (zea mays l.) 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(2009). health risk from heavy metals via consumption of food crops in the vicinity of dabaoshan mine. south china science total environment, 407, 1551–1561. https://doi.org/10.1016/j.scitotenv.2008.10.061 abstract soil contamination with heavy metals leads to the accumulation of signi�cant amounts of these elements in plants and disrupts their growth and development. �e current experiment investigated the e�ect of lead in the form of pb(no3)2 in water solutions of various percentages (0.1%, 1%, 3%) on the germination of maize grains (zea mays l.), plant growth (fresh and dry mass) and their photosynthetic activity. �e experiment was performed on plants grown from grains germinated on lead solutions and on plants germinated in distilled water, and watered with lead solutions during growth. �e negative in�uence of lead solutions on the germination capacity of grains was demonstrated. regardless of the timing of lead application, maize elongation growth was clearly inhibited. similar results were obtained for the masses of the examined plant organs. �e rate of transpiration in�uenced the photosynthesis intensity and depended on the concentration of the lead solution. along with the increase in lead concentrations a negative e�ect of lead on all the parameters tested was observed. in general, it can be concluded that only proper management of arable soils can limit the uptake of heavy metals by plants and thus improve their growth and development. key words: elongation growth, germination, masses, photosynthesis, transpiration the influence of lead com pounds on selected m orphological features and the physiological processes of zea m ays l. el ke b lo em , j ow ita c ia ra ne k 80 received: [2021.08.01] accepted: [2021.10.06] wpływ związków ołowiu na wybrane cechy morfologiczne i procesy fizjologiczne zea mays l. streszczenie skażenie gleb metalami ciężkimi prowadzi do kumulowania znacznych ilości tych pierwiastków w roślinach oraz zaburzeń ich wzrostu i rozwoju. w doświadczeniu zbadano wpływ ołowiu, w postaci wodnych roztworów pb(no3)2 o różnych stężeniach procentowych (0.1%, 1%, 3%), na kiełkowanie ziarniaków kukurydzy (zea mays l.), wzrost roślin (świeżą i suchą masę) oraz ich aktywność fotosyntetyczną. eksperyment wykonano na roślinach wykiełkowanych na roztworach z ołowiem i na roślinach wykiełkowanych na wodzie destylowanej, a podlewanych roztworami ołowiu w czasie wzrostu. wykazano negatywny wpływ roztworów ołowiu na zdolność kiełkowania ziarniaków. niezależnie od etapu podlewania, wzrost elongacyjny kukurydzy był wyraźnie hamowany. podobne rezultaty uzyskano dla mas badanych organów roślin. szybkość transpiracji wpływała na intensywność fotosyntezy i zależała od stężenia roztworu ołowiu. wraz ze wzrostem stężeń ołowiu, obserwowano jego negatywny wpływ na wszystkie badane parametry. generalnie można stwierdzić, że tylko właściwe gospodarowanie glebami uprawnymi może ograniczyć pobieranie metali ciężkich przez rośliny i tym samym poprawić ich wzrost oraz rozwój. słowa kluczowe: kiełkowanie, wzrost elongacyjny, masa, fotosynteza, transpiracja information on the authors elke bloem https://orcid.org/0000-0002-5597-1969 her research focuses on the metabolism of sulfur in plants and soils. �e investigation of secondary sulfur-containing compounds and valuable plant ingredients in relation to biotic (fungal pathogens) and abiotic stress (salt and drought) are topics of collaboration. it is the main target to understand which factors (biotic and abiotic) are important in secondary plant metabolism and how this knowledge can be transferred to agriculture to deliver high-value medical crops and healthy plants. jowita ciaranek she was am msc degree student of biology of the pedagogical university in kraków (poland). she is interested in the impact of heavy metals on plant physiology.w 81 annales universitatis paedagogicae cracoviensis studia naturae, 6: 81–94, 2021, issn 2543-8832 doi: 10.24917/25438832.6.5 gracjana budzałek, sylwia śliwińska-wilczewska* institute of oceanography, university of gdansk, gdynia, poland, *ocessl@ug.edu.pl allelopathic effect of macroalgae fucus vesiculosus (ochrophyta) and coccotylus brodiei (rhodophyta) on the growth and photosynthesis performance of baltic cyanobacteria introduction �e term “allelopathy” was introduced to science by hans molisch (molisch, 1937), who used this appellation to describe the chemical interaction between plants. with the development of allelopathic research, this de�nition includes negative and positive e�ects of compounds secreted by various plant and animal organisms (rice, 1984). inderjit and dakshini (1994) have also shown allelopathic activity in aquatic environments between cyanobacteria (aphanizomenon gracile (lemmerm.) lemmerm.) and microalgae (pediastrum boryanum (turpin) menegh., cosmarium lundellii delp., micrasterias sp.) (guiry, guiry, 2021). with the end of the 20th century, the de�nition of allelopathy was standardised by the international allelopathy society (ias, 1996), describing any process in which bioactive metabolites (so-called allelochemicals or allelopathic compounds) secreted by organisms a�ect the development of other plant and animal species (legrand et al., 2003). currently, allelopathy is considered to be a unique strategy to deter or eliminate competitors and predators coexisting in the same ecosystem (granéli et al., 2008; gomes et al., 2017; śliwińska-wilczewska et al., 2021). in aquatic ecosystems, the allelopathy of donor organisms depends on the production and secretion of active allelopathic compounds and their e�cient dispersal to target organisms a�ected by these compounds (śliwińska-wilczewska et al., 2021). for photoautotrophic organisms, the production of active allelopathic compounds becomes a crucial adaptation in achieving a competitive advantage over other primary producers (legrand et al., 2003). red and brown macroalgae are the important source of many biologically active metabolites (el gamal, 2010). for instance, the tichocarpols a and b isolated from the red alga tichocarpus crinitus (gmel.) rupr. exhibit antifeedant activity against the sea urchin (ishii et al., 2004). on the other hand, dictyterepenoids a and g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a 82 b isolated from the brown algae dilophus okamurae e.y.dawson display antifeedant activity against the abalone (suzuki et al., 2002). in benthic communities, organisms are o�en located at shorter distances from each other. �us, direct contact between donor and target cells may occur. �is also leads to constant competition for space, nutrients, and light. competition for resources may then be crucial in the occurrence of allelopathic interactions (gross, 2003). it has been found that target organisms can be inhibited, resisted, or even stimulated by allelopathic compounds present in water (suikkanen et al., 2004; śliwińska-wilczewska et al., 2021). consequently, allelopathic interactions may contribute to changes in phytoplankton and phytobenthic structure in aquatic ecosystems. compared to the intensive and extensive research on allelopathic interactions among terrestrial plants, knowledge of allelopathy in aquatic plant communities is still not fully demonstrated. �is is partly because it is di�cult to provide direct evidence for allelopathic interactions in water ecosystems under natural conditions. it is challenging to study allelopathic e�ects among aquatic organisms under natural conditions because factors such as competition for nutrients and light, temperature, and ph change can mask allelopathic e�ects (legrand et al., 2003). �erefore, it is important that attempts to identify allelopathic interactions among aquatic organisms be conducted in a controlled system, such as by conducting a series of laboratory experiments. �erefore, in the present study, the allelopathic e�ect of aqueous extracts from the baltic red alga coccotylus brodiei (turner) kütz. and the brown alga fucus vesiculosus l. (guiry, guiry, 2021) on the growth and chlorophyll �uorescence of two bloom-forming cyanobacteria from the genus aphanizomenon and nostoc were investigated. material and methods material and place of sampling �e material used in the experiments consisted of strains of baltic cyanobacteria aphanizomenon sp. (ccba-69) and nostoc sp. (ccba-81) (fig. 1). strains of cyanobacteria were isolated from the natural phytoplankton communities of the coastal waters of the gulf of gdansk (southern baltic sea: 54°30’53.7”n 18°54’23.5”e). currently, these strains are maintained as monocultures in the culture collection of baltic algae (ccba) at the laboratory of marine plant ecophysiology at the university of gdańsk (latała et al., 2006). �e macroalgae used in the study were collected from the coastal zones of the gulf of gdańsk (54°30’08.7”n 18°33’32.3”e). determination of macroalgae based on the examination of morphological features. �e herbarium sheets were deposited at the institute of oceanography, university of gdansk (poland) and are available on the website (https://zielnik.ug.edu.pl/en/home/). 83 fig. 1. photographs of the cyanobacterial culture in 25-ml erlenmeyer �asks (a) of strain: ccba-69 aphanizomenon sp. (aa) and ccba-81 nostoc sp. (ba); light microscope photographs (b) of cyanobacterial strain; scale = 10 μm, (photo. s. śliwińska-wilczewska) cyanobacteria cultivation �e studied cyanobacteria were cultured on sterile mineral medium f/2 (guillard, 1975) prepared with baltic sea water �ltered through glass �ber �lters (whatman gf/c) and autoclaved. �e salinity was 8 psu as measured with a salinometer (inolab cond level 1, weilheim in oberbayern, germany). �e cyanobacterial strains used in the experiments was maintained in 25ml glass erlenmeyer �asks. cyanobacteria were cultured at a par intensity of 10 μmol photons m–2s–1 (16:8 h light: dark cycle) and a temperature of 18°c. photosynthetically active irradiance (par) was measured using a  quantum meter (li-cor, nebraska, usa). �e light sources used in the experiment were lamps (cool white 40w, sylvania, usa). �e culture was acclimated to these conditions for 7 days, and these growth conditions were used for the experiments. a llelopathic effect of m acroalgae fucus vesiculosus (o chrophyta) and coccotylus brodiei (r hodophyta) on the grow th and photosynthesis perform ance of b altic cyanobacteria g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a 84 determination of the allelopathic e�ect of an aqueous extract obtained from macroalgae �e allelopathic e�ect of baltic macroalgae was investigated by adding di�erent concentrations of aqueous extract to the target cyanobacteria monocultures. �e allelopathic e�ect was examined according to the method proposed by złoch et al. (2018) with modi�cations. first, dried macroalgae material was mechanically ground to powder in a mortar. in the next step, 50 g of dry matter was weighed and �ltered in 50 ml of culture medium. finally, the extract was �ltered with a glass �bre �lter (whatman gf/c) using a vacuum pump (400 mbar) to remove suspended particles. �e concentration of major nutrients in the control and experimental samples were adjusted to the same level as in the f/2 standard. �erefore, the in�uence of nutrients, micronutrients and vitamins on the experimental result can be excluded. target cyanobacterial strains were maintained in 25-ml erlenmeyer �asks. in all experiments, the starting concentration of chlorophyll a in cyanobacterial cultures was 0.4 μg chl a ml–1. �e experiment was conducted by adding 100, 500, and 1000 µl of macroalgae extract to erlenmeyer �asks containing 20 ml of the target cyanobacteria. �us, the �nal extract concentrations were: 5, 25, and 50 µl ml–1. control cultures were prepared analogously, but f/2 medium was added instead of extract at 100, 500, and 1000 µl. a�er 7 days of the exposure, the cells concentration as well as chlorophyll a �uorescence parameters were determined. each experiment was performed in triplicate. determination of cyanobacterial number of cells �e number of cells (n) in aphanizomenon sp. and nostoc sp. cultures was estimated with previously determined linear correlations between cell abundance (n ml–1) and optical density (od). n was counted using a bürker chamber (48 squares per count) and light microscope following a procedure according to guillard and sieracki (2005), and the od was measured spectrophotometrically at 750 nm with a multiskan go uvvis spectrophotometer (�ermo scienti�c, massachusetts, usa). �e linear correlation between n and od for aphanizomenon sp. was described by śliwińska-wilczewska et al. (2017) whereas for nostoc sp. was examined by budzałek et al. (2018). od measurements were performed on the 7th days of the experiment and control and converted to cyanobacteria cells. determination of the chlorophyll a �uorescence in the conducted experiments, the pulsed amplitude modulation (pam) method was used to measure the chlorophyll a  �uorescence (fms1, hansatech). �is method is widely used to measure chlorophyll a �uorescence in cyanobacteria, both in the laboratory and in the natural environment (schreiber et al., 1995; campbell et al., 1998). samples were taken for chlorophyll �uorescence analysis a�er the 7th days of the ex85 periment. about 5 ml of target cyanobacteria were �ltered through 13-mm glass �ber �lters (whatman gf/c). in the next step, the �lters were placed in holders. �e samples were kept in the dark for 5 min before measurement. in this study, the maximum psii quantum e�ciency (fv/fm) and the e�ective quantum yield of psii photochemistry (φpsii) were determined (campbell et al., 1998). statistical analysis to con�rm the e�ect of the extracts obtained from macroalgae on the number of cells and chlorophyll a  �uorescence parameters of target cyanobacteria, a  t-test was performed at three levels of signi�cance: *p < 0.05, **p < 0.01, ***p < 0.001. data are reported as the means ± standard deviations (sd). �e statistical analyses were performed using statistica® 13.1 so�ware. results fig. 2. �e number of cells (n 105 ml–1) of aphanizomenon sp. (a) and nostoc sp. (b) in controls and cultures to which were added: 5 (a), 25 (b), and 50 (c) (µl ml–1) of aqueous extract obtained from fucus vesiculosus l. a�er 7 days of the experiment. �e values shown are mean (n = 3, mean ± sd). * indicates statistically signi�cant di�erences compared to control based on t-test at: * p < 0.05; ** p < 0.01; *** p < 0.001 allelopathic e�ect of aqueous extract on cyanobacterial abundances in this work, the number of cells (n 105 ml–1) of aphanizomenon sp. and nostoc sp. in controls and cultures to which were added: 5, 25, and 50 (µl ml–1) of aqueous extract obtained from fucus vesiculosus and coccotylus brodiei a�er 7 days of the experiment were determined. it was found that aqueous extracts obtained from f. vesiculosus had no statistically signi�cant e�ect on the number of cells of the cyanobacteria aphanizomenon sp. and nostoc sp. (fig. 2a–b). on the other hand, it was examined a stimulating e�ect of 5, 25, and 50 µl ml–1 of the aqueous extract obtained from c. brodiei on the number of a llelopathic effect of m acroalgae fucus vesiculosus (o chrophyta) and coccotylus brodiei (r hodophyta) on the grow th and photosynthesis perform ance of b altic cyanobacteria g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a 86 nostoc sp. cells which constituted 108% (p < 0.01), 140% (p < 0.01), and 147% (p < 0.001), respectively, relative to the control treatment (fig. 3 ba-bc). moreover, the c. brodiei extracts had no signi�cant e�ect on the growth of aphanizomenon sp. (fig. 3aa–ac). fig. 3. �e number of cells (n 105 ml–1) of aphanizomenon sp. (a) and nostoc sp. (b) in controls and cultures to which were added: 5 (a), 25 (b), and 50 (c) (µl ml–1) of aqueous extract obtained from coccotylus brodiei (turner) kütz. a�er 7 days of the experiment. �e values shown are mean (n = 3, mean ± sd). * indicates statistically signi�cant di�erences compared to control based on t-test at: * p < 0.05; ** p < 0.01; *** p < 0.001 allelopathic e�ect of an aqueous extract on �uorescence parameters �e values of the �uorescence parameter fv/fm (the maximum psii quantum e�ciency) and φpsii (the e�ective quantum yield of psii photochemistry) for aphanizomenon sp. and nostoc sp. in control and cultures to which were added a di�erent concentrations (5, 25, and 50 µl ml–1) of aqueous extract obtained from f. vesiculosus and c. brodiei a�er 7 days of experiment were examined. a stimulating e�ect of concentrations of 5 and 25 µl ml–1 of f. vesiculosus extract on the values of �uorescence parameters of cyanobacteria aphanizomenon sp. were observed and in this conditions these parameters constituted 119% (t-test, p < 0.01) and 113% (t-test, p < 0.05), respectively, for fv/fm and 122% (t-test, p < 0.001) and 118% (p < 0.001), respectively, for φpsii, relative to the control culture (fig. 4aa–ab). moreover, a stimulatory e�ect of 25 and 50 µl ml–1 of f. vesiculosus extract on �uorescence parameters fv/fm and φpsii of tested cyanobacterium nostoc sp. was observed. it was found that the values of fv/fm was 109% (t-test, p < 0.01) at 25 µl ml –1 and 109% (t-test, p < 0.05) at 50 µl ml–1, relative to the control culture (fig. 4bb–bc). furthermore, in the same experiment conditions, the values of φpsii were 111% (t-test, p < 0.01) and 87 116% (t-test, p < 0.001), respectively, compared to control (fig. 4bb–bc). on the other hand, an inhibitory e�ect of 50 µl ml–1 of f. vesiculosus extract on fv/fm was noted for cyanobacteria aphanizomenon sp. which was 92% (t-test, p < 0.01) relative to the control (fig. 4ac). similarly, an inhibitory e�ect of this brown alga extract at a concentration of 5 µl ml–1 on nostoc sp. was observed. in this condition, the fv/fm constituted 92% (t-test, p < 0.01) relative to the control (fig. 4ba). fig. 4. �e values of the �uorescence parameter fv/fm and φpsii for aphanizomenon sp. (a) and nostoc sp. (b) in control and cultures to which were added: 5 (a), 25 (b), and 50 (c) (µl ml–1) of aqueous extract obtained from fucus vesiculosus l. a�er 7 days of the experiment. �e values shown are mean (n = 3, mean ± sd). * indicates statistically signi�cant di�erences compared to control based on t-test at: * p < 0.05; ** p < 0.01; *** p < 0.001 it was found a stimulating e�ect of concentrations of 5 µl ml–1 of c. brodiei extract on the fv/fm and φpsii parameters of cyanobacteria aphanizomenon sp. in these conditions these parameters constituted 133% (t-test, p < 0.001) and 130% (t-test, p < 0.001), respectively, relative to the control culture (fig. 5aa). a stimulatory e�ect of 5 and 25 µl ml–1 of c. brodiei extract on �uorescence parameter fv/fm of nostoc sp. was also examined. it was found that the values of fv/fm was 147% (t-test, p < 0.001) and 106% (t-test, p < 0.01), respectively (fig. 5ba–bb). furthermore, a stimulating e�ect of concentrations of 5, 25, and 50 µl ml–1 of c. brodiei extract on the φpsii values of cyanobacteria nostoc sp. was observed and in these conditions these parameters constituted 164% (t-test, p < 0.001), 121% (t-test, p < 0.001), and 107% (t-test, p < 0.001), respectively, relative to the control culture (fig. 5ba–bc). on the other hand, an inhibitory e�ect of 25 and 50 µl ml–1 of c. brodiei extract on fv/fm was demonstrated for aphanizomenon sp. which was 72% (t-test, p < 0.001) and 93% (t-test, p < 0.01), respectively, of the control (fig. 5ab–ac). furthermore, an inhibitory e�ect of the highest concentration of the c. brodiei extract (50 µl ml–1) on the φpsii values of the cyanobacterium aphanizomenon sp. was noted and constituted 88% (p < 0.001) of control (fig. 5ac). a llelopathic effect of m acroalgae fucus vesiculosus (o chrophyta) and coccotylus brodiei (r hodophyta) on the grow th and photosynthesis perform ance of b altic cyanobacteria g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a 88 fig. 5. �e values of the �uorescence parameter fv/fm and φpsii for aphanizomenon sp. (a) and nostoc sp. (b) in control and cultures to which were added: 5 (a), 25 (b), and 50 (c) (µl ml–1) of aqueous extract obtained from coccotylus brodiei (turner) kütz. a�er 7 days of the experiment. �e values shown are mean (n = 3, mean ± sd). * indicates statistically signi�cant di�erences compared to control based on t-test at: * p < 0.05; ** p < 0.01; *** p < 0.001 discussion allelopathic activity of red and brown algae on cyanobacteria growth in the present study, the allelopathic e�ect of baltic red alga coccotylus brodiei and brown alga fucus vesiculosus extract on growth and chlorophyll �uorescence of two cyanobacteria aphanizomenon sp. and nostoc sp. was investigated. it was noted that aqueous extracts obtained from f. vesiculosus had no e�ect on the change in cell concentrations of the cyanobacteria aphanizomenon sp. and nostoc sp. additionally, there was no signi�cant e�ect of extract from c. brodiei on the cell abundance of cyanobacteria aphanizomenon sp. only a stimulating e�ect of each of the three tested concentrations (5, 25, and 50 µl ml–1) of the aqueous extract of c. brodiei on the cell abundance of nostoc sp. was observed. it is worth noting here that the slightest e�ect was observed when the allelopathic e�ect was tested at the lowest extract concentration (5 µl ml–1), while the ,most substantial e�ect was observed at the highest tested concentration of 50 µl ml–1. it may be due to the fact that the extract obtained from the dry thallus of tested macroalgae contained certain mineral compounds that stimulated the growth of baltic cyanobacteria. it should be emphasized here that there are very few studies on the allelopathic activity of brown and red algae on associated cyanobacteria and microalgae species. kakisawa et al. (1988) showed inhibition or no signi�cant e�ect of the brown alga cladosiphon okamuranus tokida which depended on the target species (cyanobacteria: microcyctis wesenbergii (komárek) komárek ex komárek, oscillatoria raciborskii woloszynska; rhodophyta: cyanidium caldarium (tilden) geitler; bacillariophyta: aulacosira ambiqua (grunow) sim., chaetoceros debilis p.t. cleve, coscinodiscus granii 89 l.f. gough, skeletonema costatum (greville) p.t. cleve, tabellaria �occulosa (roth) kütz.; miozoa: gymnodinium nagasakiense h. takayama & m. adachi, g. sanguineum k. hirasaka, heterocapsa triquetra (ehrenb.) f. stein, prorocentrum micans ehrenb.; ochrophyta: chattonella antiqua (y. hada) c. ono, c. marina (subrahmanyan) y. hara & m. chihara, olisthodiscus luteus n. carter, heterosigma akashiwo (y. hada) y. hada ex y. hara & m. chihara; haptophyta: cricosphaera rosco�ensis (p.a. dangeard) gayral & fresnel; cryptophyta: cryptomonas sp.; chlorophyta: hafniomonas reticulata (korshikov) ettl & moestrup, nephroselmis sp., pterosperma cristatum schiller, plymnesium parvum n. carter, pyramimonas sp., tetraselmis cordiformis (h.j. carter) f. stein, tetraselmis chui butcher, chlamydomonas augustae skuja, chlamydomonas sp., chlorella pyrenoidosa h. chick, chlorosarcinopsis sp., chlorosarcinopsis delicata shin watanabe, haematococcus lacustris (girod-chantrans) rosta�nski, oltmannsiella sp., scenedesmus quadricauda (turpin) bréb. in bréb. & godey, volvox aureus ehrenb., closterium acerosum ehrenb. ex ralfs, and also euglenozoa: euglena gracilis g.a. klebs, eutreptia sp.) (guiry, guiry, 2021). on the other hand, nagayama et al. (2003) investigated the e�ect of the extract of brown alga ecklonia kurome okamura on bloom-forming dino�agellates karenia mikimotoi (miyake & kominami ex oda) gert hansen & moestrup, cochlodinium polykrikoides margalef, and chattonella antiqua. �e authors showed a very high inhibition of dino�agellate growth, morphological changes, and even complete cell death. in turn, suzuki (1998) studied the inhibitory e�ect of extract obtained from red alga lithophyllum sp. on dino�agellate heterosigma akashiwo. wang et al. (2007) investigated the inhibitory e�ects of extract, �ltrate, and live thallus of the brown alga sargassum thunbergii (mertens ex roth) kuntze and the red alga corallina pilulifera postels & ruprecht on the growth of two dino�agellates heterosigma akashiwo and alexandrium tamarense (lebour) balech. live material from both macroalgae caused lysis of h. akashiwo cells. growth of a. tamarense was inhibited, but cells were not lysed. �e �ltrate caused an overall decrease in both macroalgae, but only the �ltrate from corallina caused lysis of h. akashiwo cells. �e extract caused the least e�ect on the cell concentration of the tested dino�agellates. an exception was an experiment where the in�uence of s. thunbergii �ltrate on h. akashiwo was tested where cells were lysed a�er the �rst day of culture. later, lu et al. (2011) demonstrated both inhibitory and stimulatory e�ects of the extract obtained from red alga gracilaria lemaneiformis (bory) greville on the diatom skeletonema costatum. to the best of the author’s knowledge, there are no more papers con�rming the allelopathic activity of brown and red algae on phytoplankton representatives. �e results obtained in this work constitute an important contribution to the knowledge on the allelopathic activity of baltic red and brown algae on certain bloom-forming species of �lamentous cyanobacteria. a llelopathic effect of m acroalgae fucus vesiculosus (o chrophyta) and coccotylus brodiei (r hodophyta) on the grow th and photosynthesis perform ance of b altic cyanobacteria g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a 90 e�ects of allelopathic compounds produced by baltic macroalgae on photosynthesis performance analysis of chlorophyll a �uorescence parameters allows for the assessment of photosynthetic activity (machado et al., 2015; song et al., 2017). fluorescence measurements are a useful tool in physiological studies and are a highly sensitive method of studying photosynthetic reactions in cyanobacteria (campbell et al., 1998). in this study, we investigated the e�ect of the allelopathic e�ect of macroalgal extracts on the maximum psii quantum e�ciency (fv/fm) and the e�ective quantum yield of psii photochemistry (φpsii). it was found that fucus vesiculosus and coccotylus brodiei had the allelopathic e�ect on �uorescence parameters of tested cyanobacteria. �e low and medium concentrations of aqueous extract (5 and 25 µl ml–1) from f. vesiculosus stimulated the values of fv/fm and φpsii of cyanobacterium aphanizomenon sp. compared to the control. �e inhibitory e�ect of the highest concentration of aqueous extract (50 µl ml–1) on the fv/fm parameter of aphanizomenon sp. was demonstrated. moreover, a stimulating e�ect of medium and high extracts concentrations of f. vesiculosus on cyanobacterium nostoc sp. on the �uorescence parameters fv/fm and φpsii was observed. in addition, the inhibitory e�ect of this brown alga at the lowest extract concentration on fv/fm of nostoc sp. was demonstrated. on the other hand, the stimulatory e�ect of the lowest extract concentration (5 µl ml–1) obtained from c. brodiei on the �uorescence parameter fv/fm and φpsii of cyanobacterium aphanizomenon sp. was noted. in addition, the inhibitory e�ect of 25 and 50 µl ml–1 extracts on �uorescence parameters of this cyanobacterium was demonstrated. it was also shown that all tested extract concentrations obtained from c. brodiei stimulated the φpsii parameter of nostoc sp. in our study it has been demonstrated that brown alga f. vesiculosus and red alga c. brodiei can release some allelopathic substances which a�ect the �uorescence parameters of cyanobacteria nostoc sp. and aphanizomenon sp. similarly, as in the case of cyanobacterial growth, the stimulation of the �uorescence parameters may be caused by additional mineral compounds contained in the macroalgal extracts. moreover, the high values of the �uorescence parameters indicate a relatively high potential e�ciency of photosystem ii in the studied cyanobacteria. �e low level of these parameters indicates certain disturbances in the photosynthesis process. �ree years earlier, budzałek et al. (2018) demonstrated that the dry powder of ulva intestinalis l. contains water-soluble allelochemical(s) is capable of restricting the �uorescence parameter fv/fm of cyanobacterium nostoc sp. �e authors noted that the highest decrease in fv/fm for nostoc sp. was observed a�er the �rst, third and seventh day of the experiment, a�er the addition of 100 µl ml–1 extracts obtained from u. inestinalis with a magnitude of 69%, 59%, and 49% respectively, compared to the control treatment. to the best of the author’s knowledge, no more works con�rm the allelopathic e�ects of macroalgal extracts on the �uorescence parameter fv/fm of cyanobacteria. moreover, 91 this work is the �rst to indicate the allelopathic e�ect of macroalgal aqueous extracts on the e�ective quantum yield of psii photochemistry (φpsii) of baltic cyanobacteria. �erefore, these studies indicate the need to study the photosynthesis performance in target organisms that are exposed to contact with baltic macroalgae in more detail. �e characterisation of the allelopathic compounds is a time-consuming task. some studies have described novel secondary metabolites, produced by marine red (rhodophyta) and brown (ochrophyta) macroalgae, which have signi�cant biological activity on target organisms (el gamal, 2010) however, to the best of the authors knowledge, there is no information about the allelopathic compounds produced by f. vesiculosus and c. brodiei. only kristinsson and jónsdóttir (2015) described that f. vesiculosus showed antioxidant activity however, the chemical structure of this compound has not been studied. �us, demonstrating the composition of the these macroalgal allelochemicals should become a priority for future work. acknowledgements �is work has been funded by ugrants–start, poland, no. 533-o000-gs12-21. con�ict of interest �e authors declare no con�ict of interest related to this article. references budzałek, g., śliwińska-wilczewska, s., latała, a. 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(2017). allelopathic interactions of linoleic acid and nitric oxide increase the competitive ability of microcystis aeruginosa. �e isme journal, 11, 1865–1876. https://doi.org/10.1038/ismej.2017.45 suikkanen, s., fistarol, g.o., granéli, e. (2004). allelopathic e�ects of the baltic cyanobacteria nodularia spumigena, aphanizomenon �os-aquae and anabaena lemmermannii on algal monocultures. journal of experimental marine biology and ecology, 308, 85–101. suzuki, m., yamada, h., kurata, k. (2002). dictyterpenoids a and b two novel diterpenoids with feeding-deterrent activity from the brown alga dilophus okamurae. journal of natural products, 65, 121–125. https://doi.org/10.1021/np010234b 93 suzuki, y., takabayashi, t., kawaguchi, t., matsunaga, k. (1998). isolation of an allelopathic substance from the crustose coralline algae, lithophyllum spp., and its e�ect on the brown alga, laminaria religiosa miyabe (phaeophyta). journal of experimental marine biology and ecology, 225, 69–77. https://doi. org/10.1016/s0022-0981(97)00208-6 śliwińska-wilczewska, s., maculewicz, j., barreiro felpeto, a., vasconcelos, v., latała, a. (2017). allelopathic activity of the picocyanobacterium synechococcus sp. on �lamentous cyanobacteria. journal of experimental marine biology and ecology 496, 16–21. http://doi.org/10.1016/j.jembe.2017.07.008. śliwińska-wilczewska, s., wiśniewska, k., konarzewska, z., cieszyńska, a., barreiro felpeto, a., lewandowska, a.u., latała, a. (2021). �e current state of knowledge on taxonomy, modulating factors, ecological roles, and mode of action of phytoplankton allelochemicals. science of the total environment, 773, 145681. https://doi.org/10.1016/j.scitotenv.2021.145681 wang, r., xiao, h., zhang, p., qu, l., cai, h., tang, x. (2007). allelopathic e�ects of ulva pertusa, corallina pilulifera and sargassum thunbergii on the growth of the dino�agellates heterosigma akashiwo and alexandrium tamarense. journal of applied phycology, 19, 109–121. https://doi.org/10.1007/ s10811-006-9117-8 złoch, i., śliwińska-wilczewska, s., kucharska, m., kozłowska, w. (2018). allelopathic e�ects of chara species (c. aspera, c. baltica, and c. canescens) on the bloom-forming picocyanobacterium synechococcus sp. environmental science and pollution research, 25, 36403–36411. https://doi.org/10.1007/ s11356-018-3579-5 abstract in aquatic ecosystems, allelopathic activity depends on the production and secretion of allelopathic compounds and their e�ective dispersal in the environment. in addition, macroalgae have been found to produce active metabolites that a�ect other organisms that compete with them for nutrients. however, the allelopathic activity of baltic red and brown macroalgae on �lamentous cyanobacteria is still insu�ciently understood. �erefore, the main objective of this study was to demonstrate and compare the allelopathic e�ects of macroalgae fucus vesiculosus l. and coccotylus brodiei (turner) kütz. on the growth and photosynthetic activity of two baltic cyanobacteria aphanizomenon sp. and nostoc sp. it was found a stimulating e�ect of di�erent concentrations (5, 25, and 50 µl ml–1) of the aqueous extract obtained from c. brodiei on the number of cells of nostoc sp. which constituted 108%, 140%, and 147%, respectively, relative to the control treatment. on the other hand, extracts obtained from f. vesiculosus had no statistically signi�cant e�ect on the number of cells of the cyanobacteria aphanizomenon sp. and nostoc sp. moreover, the c. brodiei extracts had no signi�cant impact on the growth of aphanizomenon sp. furthermore, baltic macroalgae f. vesiculosus and c. brodiei was able to exert allelopathic e�ects on photosynthesis performance of nostoc sp. and aphanizomenon sp. and compounds produced by them had inhibitory, stimulatory, or no signi�cant e�ect on the maximum psii quantum e�ciency (fv/fm) and the e�ective quantum yield of psii photochemistry (φpsii). �e results obtained in this work constitute an important contribution to the knowledge on the allelopathic activity of baltic red and brown algae on certain bloom-forming species of �lamentous cyanobacteria. key words: allelopathy, aqueous extract, brown algae, cyanobacteria, �uorescence, growth, macroalgae, red algae received: [2021.04.29] accepted: [2021.07.06] a llelopathic effect of m acroalgae fucus vesiculosus (o chrophyta) and coccotylus brodiei (r hodophyta) on the grow th and photosynthesis perform ance of b altic cyanobacteria g ra cj an a b ud za łe k, s yl w ia ś liw iń sk aw ilc ze w sk a 94 oddziaływanie allelopatyczne fucus vesiculosus (brunatnica) i coccotylus brodiei (krasnorost) na wzrost oraz aktywność fotosyntetyczną bałtyckich sinic streszczenie aktywność allelopatyczna w ekosystemach wodnych zależy od produkcji i uwalniania związków allelopatycznych oraz ich skutecznego rozprzestrzeniania się w środowisku. stwierdzono, że makroglony wytwarzają aktywne metabolity, które wpływają na inne organizmy, konkurując z nimi o światło i składniki odżywcze. jednak aktywność allelopatyczna bałtyckich krasnorostów i brunatnic na nitkowate sinice jest nadal niedostatecznie poznana. dlatego głównym celem niniejszej pracy było wykazanie i porównanie aktywności allelopatycznej makroglonów fucus vesiculosus l. (brunatnica) i coccotylus brodiei (turner) kütz. (krasnorost) na wzrost i aktywność fotosyntetyczną dwóch bałtyckich sinic aphanizomenon sp. i nostoc sp. w pracy stwierdzono stymulujący wpływ różnych stężeń (5, 25 i 50 µl ml–1) wodnego ekstraktu otrzymanego z c. brodiei na liczebność komórek nostoc sp., które wynosiły odpowiednio: 108%, 140% i 147%, w stosunku do grupy kontrolnej. z drugiej strony ekstrakty uzyskane z  f. vesiculosus nie miały istotnego statystycznie wpływu na liczebność komórek sinic aphanizomenon sp. i nostoc sp. wykazano także, że ekstrakty z c. brodiei nie miały istotnego wpływu na wzrost aphanizomenon sp. ponadto bałtyckie makroglony f. vesiculosus i c. brodiei wpływały allelopatycznie na aktywność fotosyntetyczną u nostoc sp. i aphanizomenon sp., a wydzielane przez nie związki wykazywały hamujący, stymulujący lub brak wpływu na maksymalną wydajność kwantową drugiego fotosystemu (psii) w ciemności (fv/fm) oraz na rzeczywistą wydajność kwantową psii w świetle (φpsii). wyniki uzyskane w niniejszej pracy stanowią ważny wkład w stan wiedzy na temat aktywności allelopatycznej bałtyckich krasnorostów i brunatnic na wybrane gatunki nitkowatych sinic, zdolnych do tworzenia masowych zakwitów. słowa kluczowe: allelopatia, brunatnice, ekstrakt wodny, �uorescencja, krasnorosty, makroglony, sinice, wzrost information on the authors gracjana budzałek �e �eld of her interest is allelopathic interactions between macroalgae and cyanobacteria. in her research she is focusing mostly on baltic species. she is investigating the in�uence of allelopathic compounds produced by macroalgae on bloom-forming cyanobacteria. sylwia śliwińska-wilczewska https://orcid.org/0000-0002-3147-6605 she is interested in allelopathy of cyanobacteria and microalgae; in particular of picocyanobacteria synechococcus sp. allelopathy plays an important role in interspeci�c competition and contributes to cyanobacterial bloom maintenance. in her study, the in�uence of allelochemicals on the growth, chlorophyll �uorescence and photosynthesis irradiance curves of di�erent phytoplankton species was investigated. she is also investigating the in�uences of environmental factors on produced allelopathic compounds on algae and cyanobacteria. anna sołtys-lelek1*, wojciech gruszka2 annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832 doi: 10.24917/25438832.7.x anna sołtys-lelek 1* , wojciech gruszka 2 1 ojców national park, 32-045 sułoszowa, ojców 9, poland; *ana_soltys@wp.pl 2 department of biological sciences, faculty of physical culture in gorzów wlkp., poznań university school of physical education, estkowskiego 13, 66-400 gorzów wlkp., poland occurrence of the red-leaved rose, rosa glauca pourr. (rosaceae), in poland introduction the red-leaved rose, rosa glauca pourr., is a mountain species endemic to europe, found in its southern, western and central areas. its original range included the pyrenees, the alps, the vosges, the swabian jura, the carpathians, the apennines and the mountains of the balkan peninsula (zieliński, 1987; popek, 2007; khapugin et al., 2021). in addition, it is cultivated and feral in poland, germany, the netherlands, great britain, lithuania, latvia, estonia, belarus, ukraine, russia, moldova, scandinavian countries, the united states and canada (tutin et al., 1968; zieliński, 1987; popek, 2007; khapugin et al., 2021; global biodiversity information facility 2022). r. glauca was introduced to poland in 1817. from cultivation, it began to mainly penetrate anthropogenic habitats and communities. it has now been granted the status of a non-invasive kenophyte, permanently established in the national flora (tokarska-guzik et al., 2012). there is practically no information on the distribution of this species in the national botanical literature. this is most likely due to the fact, that its localities, apart from cultivation in botanical gardens or arboretums, are not very numerous. zieliński (1987) reported it as a frequently cultivated species in many places, especially in the west of poland where it may have appeared as an escapee from cultivation. its closest natural sites are in the mountains near bielsko in slovakia (zieliński, 1987). hybrids of the native species rosa pendulina l. with r. dumalis becht or with r. canina l. may be mistaken with r. glauca (zieliński, 1987). characteristic features of this rosa are glaucous leaves with a purplish bloom, red-brown bark, and curved or declined prickles, subulate. leaflets 5–7(–9) have blades narrowly elliptic to ovate, leathery, margins 1-serrate, eglandular or few gland-tipped, glaucous, dull, glabrous. inflorescences are 1–5flowered. pedicels are glabrous, stipitate-glandular. flowers have 2–3 cm diameter (fig. 1). fig. 1. flower of red-leaved rose rosa glauca pourr. (photo. w. gruszka, motylewo village, 2022) petals are single, with color from deep pink to crimson-red, sometimes white basally, styles exsert 1–2 mm beyond stylar orifice (1.5–2 mm diameter) of hypanthial disc (3 mm diameter). hypanthium is narrowly ovoid, glabrous, glandular or eglandular, neck purplish, sepals spreading, lanceolate, margins entire, and sometimes pinnatifid. hips are dark brownish red to crimson red, globose, ovoid, or obovoid. sepals deciduous as hips mature, erect to spreading (popek, 2007; lewis et al., 2014, fig. 2). it is a tall shrub reaching 2–3 m in height, with erect purple-colored shoots, without or only with very short stolons (seneta, dolatowski, 2004, fig. 3). in 2021, during field research carried out in the vicinity of gorzów wielkopolski, the authors came across a site (ac6672 square) where r. glauca was growing in a cluster of approximately 30 m 2 . this finding provided the impetus to undertake this research study, which was aimed at presenting all the currently identified r. glauca sites in poland and determining its ability to establish and spread. fig. 2. red-leaved rose rosa glauca pourr. (photo. 2022. a. sołtys-lelek, specimen from poland, motylewo, 2022, w. gruszka); a: part of a fruiting short shoot; b: part of a long shoot; c: leaf; d: stipule; e: fruit with glandular sepals; f: glandular petiole; h, i: the top part of the hypanthium; j, k: leaflet; l: leaf margin. solid bar = 1 cm. double bar = 0.5 cm fig. 3. red-leaved rose rosa glauca pourr. (photo. w. gruszka, motylewo village, 2022) materials and methods in order to explore discover the distribution of spontaneous localities of rosa glauca in poland, the authors, in addition to their own observations, verified collections gathered in the herbaria of national universities. specimens were found in: university of silesia in katowice (ktu), maria curie-sklodowska university (lbl), institute of biology, faculty of exact and natural sciences, wroclaw university (wrsl), university of warsaw (wa), jagiellonian university (kra), w. szafer institute of botany, polish academy of sciences (kram), adam mickiewicz university (poz), forest research institute (bil) institute of dendrology (kor). data published in the available literature were also taken into account. all the collected data are included on the map (fig. 4). in the list of sites and on the distribution map, only sites that arose spontaneously or those that are remnants of former cultivation (feral) are included. sites in private collections, botanical gardens, arboretums, etc. were not included. these sites were located within 10 km of atpol square (zając, zając, 2001) and the nearest village. fig. 4. current distribution of rosa glauca pourr. in poland on the atpol grid square system. explanation of symbols: ● – squares in which single stands of the species were reported; ● – new stand found by the authors; ▲ – squares in which two stands of the species were reported glossary of geographical names in the study this list includes the geographical coordinates for the villages in the article: bakałarzewo 54°05ʹ30ʹn; 22°39ʹ17ʹʹe (fb16) białka 49°41ʹ36ʹn; 19°40ʹ21ʹʹe (dg07) białowieża 52°42ʹ02ʹn; 23°52ʹ00ʹʹe (gc65) bojszowy 50°03ʹ25ʺn; 19°06ʹ03ʺe (df63) borawskie 54°05ʹ58ʹn; 22°37ʹ48ʹʹe (fb16) dębówka 52°11ʹ14ʹn; 20°20ʹ51ʹʹe (ed22) gołuchów 51°50ʹ55ʹn; 17°55ʹ50ʹʹe (cd65) huwniki 49°39ʹ20ʺn; 22°42ʹ14ʺe (fg09) kamienna góra 50°46ʹ59ʹn; 16°01ʹ59ʹʹe (be82) kępno 51°16ʹ42ʹn; 17°59ʹ16ʹʹe (ce25) klęka 52°04ʹ32ʹn; 17°25ʹ18ʹʹe (cd42) kowary 50°47ʹ27ʹn; 15°50ʹ14ʹʹe (be80) lubawka 50°42ʹ09ʹn; 15°59ʹ57ʹʹe (be91) lublin 51°14ʹ42ʹn; 22°34ʹ00ʹʹe (fe27) luboń 52°20ʹ38ʹn; 16°52ʹ35ʹʹe (bd08) motylewo 52°40ʹ56ʺn; 15°03ʹ06ʺe (ac66) nałęczów 51°17ʹ19ʹn; 22°12ʹ49ʹʹe (fe25) orzeszków 51°24ʹ02ʹn; 16°30ʹ25ʹʹe (be15) ożarów mazowiecki 52°12ʹ37ʹn; 20°47ʹ56ʹʹe (ed25) pacław 49°37ʹ31ʺn; 22°42ʹ24ʺe (fg09) parkowo 52°42ʹ13ʺn; 16°54ʹ59ʺe (bc68) pawłów 50°06ʹ39ʹn; 18°07ʹ43ʹʹe (cf66) piaseczno 53°03'40"n 14°41'59"e (ac24) piastów 52°11ʹ02ʹn; 20°50ʹ24ʹʹe (ed25) piła 53°08'31"n 16°45'44"e (bc28) pogroszew 52°13ʹ48ʹn; 20°44ʹ34ʹʹe (ed24) poznań 52°24ʹ52ʹʹn; 16°55ʹ16ʹʹe (bd08) puszczykowo 52°16ʹ58ʹn; 16°50ʹ54ʹʹe (bd18) radonice 52°10ʹ15ʹn; 20°36ʹ41ʹʹe (ed24) rzeki wielkie 50°52ʹ36ʹʹn; 19°23ʹ19ʹʹe (de75) rzepin 52°20ʹ44ʺn; 14°49ʹ48ʺe (ad04) sompolno 52°23ʹ17ʺn; 18°30ʹ09ʺe (cd09) szczyglice 51°37ʹ47ʹn; 16°07ʹ32ʹʹe (bd83) sieraków 52°18ʹ44ʹn; 20°48ʹ47ʹʹe (ed15) stańczyki 54°17ʹ31ʹn; 22°39ʹ12ʹʹe (fa86) staropole 52°20ʹ44ʺn; 15°26ʹ40ʺe (ad08) stojków 50°19ʹ13ʹn; 16°53ʹ14ʹʹe (bf37) szydłowo 53°04ʹ47ʹn; 20°26ʹ54ʹʹe ec22 turew 52°03ʹ32ʺn; 16°49ʹ38ʺe (bd48) tychy 50°07ʹ34ʺn; 19°00ʹ30ʺe (df53) warszawa 52°14ʹ32ʹn; 21°01ʹ02ʹʹe (ed26) wyry 50°07ʹ33ʺn; 18°53ʹ27ʺe (df52) zachełmie 50°49ʹ32ʹn; 15°39ʹ39ʹʹe (ae79) zawoja 49°40ʹ04ʹn; 19°34ʹ55ʹʹe (dg17) żernica 50°14ʹ47ʺn; 18°36ʹ56ʺe (df40) results unpublished sites in the atpol grid squares ac6672: village of motylewo near bogdaniec, roadside of voivodeship road 132, 2021, leg. w. gruszka, ae79: karkonosze, zachełmie, on the northern slope of kopa, 1982, a. boratyński (kor029803, kor029804, kor029805) bd08: luboń, wielkopolska national park, edge of willow scrub, 1977, m. ratajczak (poz-v-0089053) bd18: puszczykowo, on the warta river wielkopolska national park, 2004, a purcel (kor027974, kor027975) bd83: szczyglice, głogów municipality, in roadside ditch 1973, e. kozioł (wrsl 32303) bd83: krzepów, city of głogów, in scrubland. 1973, e. kozioł (kram 264848, poz-v-0089469) be15: south side of road from małoiwce to orzeszków, near ścinawa, roadside ditch. 1961 k. borowicz (kor027976, kor027977). be80: karkonosze, kowarska pass, 1982, a. boratyński (kor027983, kor027984, kor027985) be82: kamienne mountains, sadowa góra, 1982, a. boratyński (kor027991, kor027992 kor027993, kor027696, kor027970, kor027971, kor027972) be91: krucze mountains near lubawka, on rocks, 1982, a. boratyński (kor027988, kor027989, kor027990) bf37: lądek, stojków, złote mountains, biała lądecka valley, roadside, scrub, near railroad tracks. p. kosiński (kor027973) cd42: klęka, nowe miasto municipality, by the road to książ, 1995, a. czarna (poz-v-0089052). cd65: gołuchów, commune. gołuchów, on the outskirts of the palace in palace park, 2000, a. czarna (poz-v0089050) ce25: kepno, by road south of the village, 1934, f. krawiec (poz-v-0089055) cf66: pawłów district of racibórz, palace park, 1963 (kra 0138864) de75: rzeki wielkie, remnant of a park, 1978, t. muras, (ktu 022280), dg07: białka near maków podhalański, on the road to zawoja (and zubrzyca), 2004, w. bartoszek (kra 0260719, kra 0260718, kra 0260714, kra 0260720, kra 0260721) dg17: zawoja, roadside, 1984, a. mach, (ktu 144448); ec22: between szydłowo a kluszewo, east of mława, scrubby roadside 1980, j. zieliński (kor027965, kor027966). ed15: sieraków, roadside, 1970, k. nowak (wa 0000092957) ed16: warsaw młociny, edge of the forest, 1978, k. wichowicz (wa 0000092956) ed22: dębówka, ditch, 1968, k. nowak (wa 0000092960, wa 0000092961, wa 0000092963, wa 0000092967) ed24: pogroszew, ditch, 1967, k. nowak (wa 0000092965) ed24: radonice, buttress, 1968, k. nowak (wa 0000092968) ed25: ożarówek, roadside fence, 1968, k. nowak (wa 0000092962, wa 0000092966) ed25: piastów, buttress, 1968, k. nowak (wa 0000092964) fa86: village of stańczyki, municipality of gubieniki, scrub on a roadside escarpment near the ruins of an old farmhouse, 2010, a pliszko (kra 0417638, kra 0417637) fb16: bakałarzewo village, edge of pine forest, 2011, a. pliszko (kra 0419605, kra 0419606, kra 0419607) fb16: village of borawskie, municipality of olecko, edge of the forest near the ruins of an old farmhouse, 2009, a. pliszko (kra 0417665, kra 0417664, kra 0417663) fe25: nałęczów, 1959, m. strasburger (wa 0000092954, wa 0000092955) fe27: lublin, by the street in a hawthorn hedge, 1978, w. kraczek (lbl, no specimen number) gc65: białowieża, polana białowieska near pttk building, 1973, a. sokolowski (bil 57199), 1971 (bil 46622, 46623) published sites in the atpol grid squares ac24: vicinity of piaseczno village (startek b., et al. 2020). ad04: beaches between rzepin and boczow (zieliński 1976, kram 408818, kor027964) ad08: between staropol and wysoka, feral in the area of the old park (zieliński 1976, kram 408819, kor 077986, kor 077987) bc28: piła; a cultivated specimen running wild (sołtys-lelek, gruszka 2016) bc68: parkowo near oborniki (stefanek 1984, kor027978; kor027979; kor027980, kor027981; kor027982) bd08: poznań, rusałka lake area (dyderski et al. 2016) bd48: turew, kościan municipality, in the manor park (czarna 2009, poz-v-0089051) cd09: sompolno, old cementary (czarna 2016) df40: north of żernica, scrub on roadside (urbisz 2021) (ktu 0124309) df52: wyry, near the former state farm (urbisz 2021) (ktu 0124308) df53: tychy (urbisz 2021) df63: bojszowy (urbisz 2021) fg09: chyb mountain in huwniki, ubocz wilderness in pacław (piórecki 2013) discussion rosa glauca has the status of an established, non-invasive kenophyte species in poland (tokarska-guzik et al., 2012). in the 1970s, it was reported to be commonly cultivated (kościelny, sękowski, 1971). since the second half of the 20th century, there have been reports in the literature of its spontaneous, sporadic spread from cultivated areas, especially in the west of the country (zieliński, 1987). r. glauca, as an escapee from cultivation, is a rare species in poland. a total of 45 sites have been identified, where r. glauca grows; all of which were either spontaneously established or are remnants of former cultivation. the sites are located in 40 quadrants of the atpol grid (fig. 4). its current range covers the entire country, although it seems to be more frequent in the western portion of the country. this confirms the literature data, which stated that the species was cultivated and wild mainly in the western part of poland (zieliński, 1987). however, it can be assumed that the distribution image of this species in poland is probably affected by the insufficient state of field research. therefore, the list of sites presented in the publication can be expected to be supplemented in the future. the spread of this species in the country may be favored by its wide tolerance, both with regard to climatic and habitat conditions. it can tolerate frost down to -42.8°c (usda zone 2a). it has no special soil requirements. but, it does require a plenty of sunshine although it can tolerate sites in partial shade. r. glauca is a species penetrating plant communities and developing on anthropogenic habitats, i.e. habitats created by humans, semi-natural communities, or communities/habitats partly transformed. its spread may be aided by birds, such as jays (lewis et al., 2014). this rose has been listed as an alien species in poland. although its possible negative impacts on native species of flora or on other elements of the habitats in which it grows has not yet been thoroughly studied. however, such an impact cannot be ruled out in the future, because in the case of many species of foreign origin their invasive behavior becomes delayed (lag phase) (hobbs, humpheries, 1995; richardson, pyšek, 2006). therefore, it is important to know the current distribution of spontaneously established populations of this species and, in the long run, undertake research aimed at determining its impact on native floral elements. in addition, it has been proven that the species tends to hybridise with other rose species spontaneously, including native species. it forms spontaneous hybrids with rosa dumalis, r. pendulina, r. spinosissima and r. gallica (szafer, 1935; szafer et al., 1986). thus, it may pose a potential threat to native floral elements by being able to hybridise with domestic rose taxa. acknowledgments the authors of this paper would like to thank ms. joanna ostapiuk-karolczuk for her help in field research. conflict of interest the authors declare no conflict of interest related to this article. references gatunki obce w polsce (alien species in poland). http://www.iop.krakow.pl/ias (access: 2022) [in polish]. global biodiversity information facility. https://www.gbif.org/species/3003709 (access: 2022). czarna, a. 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[in polish] abstract the red-leaved rose, rosa glauca, has the status of an established, non-invasive kenophyte species in poland. in the 1970s, it was reported to be commonly cultivated, and since the second half of the 20 th century, information about its spontaneous spread, mainly in the western part of the country, has appeared in the literature. the aim of this study was to present the current distribution of rosa glauca sites in poland. the possible invasive potential of this species was also studied. key words: rosaceae, distribution of stands, invasive potential, kenophyte received: [2022.10.11] accepted: [2022.11.15] występowanie róży francuskiej rosa glauca pourr. (rosaceae), w polsce streszczenie róża francuska rosa glauca pourr. jest gatunkiem górskim, endemicznym dla europy. jej pierwotny zasięg obejmował pireneje, alpy, wogezy, jurę szwabską, karpaty, apeniny oraz góry półwyspu bałkańskiego (zieliński 1987; popek 2007; khapugin i in. 2021). r. glauca została sprowadzona do polski w 1817 roku. z uprawy zaczęła przenikać głównie w siedliska i zbiorowiska antropogeniczne. obecnie uzyskała status nieinwazyjnego kenofita, trwale zadomowionego we rodzimej florze (tokarska-guzik i in., 2012). w bibliografii krajowej nie ma wiele informacji na temat rozmieszczenia tego gatunku. dlatego autorzy niniejszej pracy w celu określenia rozmieszczenia spontanicznych stanowisk tego gatunku w polsce zebrali dane z krajowego zielnika i dostępnej bibliografii. r. glauca, jako uciekinier z hodowli, jest w polsce gatunkiem rzadkim. sumarycznie zidentyfikowano 45 jej stanowisk, powstałych samoistnie lub będących pozostałościami po dawnej uprawie (zlokalizowanych w 40 kwadrantach sieci atpol). rozprzestrzenianiu się tego gatunku może sprzyjać jego szeroka tolerancja, zarówno w odniesieniu do warunków klimatycznych, jak i siedliskowych. w związku z tym można się spodziewać, że w przyszłości lista stanowisk będzie wzrastać. słowa kluczowe: rosaceae, rozmieszczenie stanowisk, potencjał inwazyjny, kenophyte information on the authors anna soltys-lelek https://orcid.org/0000-0002-9595-3167 author of numerous scientific and popular science studies in the field of botany and environmental protection. her research interests relate particularly to critical types of roses (rosa) and hawthorns (crataegus). member of the polish and slovak botanical society. wojciech gruszka http://orcid.org/0000-0002-6229-8397 author and co-author of scientific and popular science studies in lichenology and botany. his main research interests relate to the ecology and protection of lichens. in addition, he participates in research on the distribution of representatives of rose (rosa) and hawthorn (crataegus) species in poland. annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x revision of the genus daldinia in the kram herbarium collection: daldinia childiae, d. loculata and d. loculatoides, three new species for poland andrzej chlebicki w. szafer institute of botany of the polish academy of sciences, lubicz 46, 31-512 kraków, poland; email: a.chlebicki@botany.pl introduction the fungi from the genus daldinia were neglected for a long time. the genus was erected by two mycologists (cesati, de notaris, 1863) and devoted to the monk, agostino daldini. the first important monograph was prepared in 1932 (child, 1932) and after that, important elaborations appeared (you et al., 1997; stadler et al., 2014) that enabled progress in the taxonomy of this genus. the species of the genus daldinia, according to cesati and de notaris (1863), are conspicuous because of their relatively large stromata. most of them possess internal alternating zones. the genus comprises ca. 50 species that occur on angiosperm hosts. only d. lloydii, d. eschscholtzii and d. childiae occasionally were noted on gymnosperm hosts (pinus and cryptomeria). daldinia hawksworthii is known only in its anamorph state (stadler et al., 2014). however, anamorph states of other species can occur in a wide range of angiosperms as endophytes. the taxonomy of this genus is based on morphological characters, colour of stromatal pigments, secondary metabolites, anamorph structures and molecular methods (stadler et al., 2001, 2014; guidot et al., 2003; stadler, 2011; helay et al., 2018). the anamorphic structures are most often referred to as the genus nodulisporium preuss that produces conidia from percurrent proliferating conidiogenous cells (petrini, müller, 1986; you et al., 1997). conidia are formed on young stromata or on invaded wood. stromata attached to the tree branches can actively discharge ascospores for more than 100 days (ingold, 1965) and finish their discharge in late autumn. fungi from the daldinia concentrica complex produce volatile antibiotics that are active against plant pathogenic nematodes (liarzi et al., 2016a, b) and d. hawksworthii is an insect-associated endophytic species that can produce volatile, small polyketide dalsymbiopyrone (pažoutová et al. 2013). mycelia of daldinia have been characterised as “early colonisers”. they are present in the tissue of the host plants for a very long time without causing any symptoms of parasitism (stadler et al., 2014). for a long time, d. concentrica was the only species reported from poland (mułenko et al., 2008). however, in light of recent taxonomic revisions of the genus daldinia it appears that this species does not occur in poland! almost all polish collections cited by mułenko et al. (2008) were possibly wrongly determined and it is necessary to check and revise all polish collections of this species. apart from the doubtful collections of d. concentrica, only five species have been reported from poland: d. decipiens (karasiński, 2009; ruszkiewiczmichalska et al., 2015; gierczyk et al., 2017), d. petriniae (wojewoda et al., 2013; stadler et al., 2014), d. pyrenaica (karasiński, 2009), d. vernicosa (stadler et al., 2014; gierczyk et al., 2017) and d. lloydi (stadler et al., 2014). the aim of the present study was the revision of the part of the richest polish collection of daldinia species, preserved in the fungal collection of the herbarium of the w. szafer institute of botany, polish academy of sciences, in kraków (kram f). material and methods morphological characters of stromata were examined under a stereo microscope nikon smz 1500. ascospores were examined in water, glycerol and lactophenol under a light microscope nikon eclipse 80i with an oil immersion lens. twenty spores of each specimen were measured. also, the colour of stromatal pigments in 10% potassium hydroxide (koh) was used to determine the fungal material. fungi presented in this article are deposited in kram f. information about the localities and habitats follow those on the original labels. results daldinia childiae j. d. rogers & y. -m. ju, mycotaxon 72: 512-513, 1999. stromata subglobose and turbinate, measuring up to 2 cm in diameter. the koh-extractable stromatal pigments yellowish or yellow-brown in colour (fig. 1a). ascospores with narrowlyrounded ends, 13–14 × 6.3–7 µm (fig. 2c). the transverse striation of episporium mostly loosely distributed (fig. 3). fig. 1. koh-extractable stromatal pigments: a – daldinia childiae, b – d. concentrica, c – d. eschscholtzii, d – d. lloydii, e – d. loculata, f – d. loculatoides, g – d. decipiens, h – d. vernicosa, i – d. petriniae (photo. a. chlebicki) specimens examined: 1. poland: the sudetes, the bystrzyckie mts., the topieliska nature reserve near zieleniec, on a branch of betula sp., 29 august 1984, leg. a. chlebicki, kram f 40392 (as d. concentrica). 2. poland: western carpathians, orawa-nowy targ basin, near the road from rogoźnik to stare bystre, ca. eight km southwest of nowy targ, ca. 620 m a.s.l., on a trunk of salix sp., 7 june 1970, leg. z. heinrich, kram f 27202 (as d. concentrica). 3. poland: western carpathians, the pieniny mts., the skalice range, in the forest near łapsze niżne, 18 km southeast from nowy targ, ca. 600 m a.s.l., on dead wood (possibly salix sp.), 1 june 1979, collector unknown, kram f 31870 (as d. concentrica). 4. poland: western carpathians, the bieszczady zachodnie mts., nasiczne, on a dead trunk of alnus incana (l.) moench, 26 december 2011, leg. a. wilczek, kram f without number. fig. 2. ascospores: a – daldinia loculata, b – d. decipiens c – d. childiae, d – d. vernicosa (photo. a. chlebicki) comments: the perfect state of d. childiae is host-limited to the angiosperm trees but its anamorph can inhabit a wide range of angiosperms as an endophyte (stadler et al., 2014). it also occasionally occurs on gymnosperm hosts (i.e., cryptomeria). the transverse striation of the spores of d. childiae is conspicuous and loosely distributed as compared to d. concentrica and d. eschscholtzii (fig. 3). in the autumn of 2011, an insect larva inside of d. childiae stromata was noted (eastern carpathians, bieszczady zachodnie mts, nasiczne). in the spring of 2012, the imago of a fly was found inside a box with daldinia (fig. 4). this fly possibly belongs to the old branch of wood-inhabiting agromyzidae (t. zatwarnicki and a. palaczyk, pers. comm.). fig. 3. ascospores of daldinia childiae (photo. a. chlebicki) fig. 4. larva and imago of fly possibly from the family agromyzidae, noted on old stromata of daldinia childiae (photo. a. chlebicki) daldinia concentrica (bolton: fr.) ces. & de not., comm. soc. crittog. ital. 1 (no. 4): 197. 1863. stromata semi-spherical, sessile, widely attached to the substrate, smooth, up to 4.4 cm in diameter. the koh-extractable stromatal pigments dark purple (fig. 1b). ascospores brown, ellipsoid, with narrowly rounded ends, 13.4–16.3 × 6.8–9 µm (fig. 5). fig. 5. ascospores of daldinia concentrica from bulgaria, kram f 47531 (photo. a. chlebicki) specimens examined: 1. bulgaria: stranja mts., the ropotamo national park, 5 km from an estuary, on acer campestre l., 18 may 1981, leg. a. chlebicki, kram f 47531 (as d. concentrica). comments: it is a the only specimen of d. concentrica in the kram f collection. stadler et al. (2014) reported d. concentrica from the same locality in the ropotamo national park. daldinia decipiens h. wollweber & m. stadler, mycotaxon 80: 168. 2001. stromata semiglobose and often subsessile. the koh-extractable stromatal pigment vinaceous purple (fig. 1g). ascospores brown and amygdaliform with hyaline appendages at both ends, (16)17.5–20 × 7.5–9.6 µm (fig. 2b). specimens examined: 1. poland: the ochojec nature reserve, on a betula sp. branch, 12 july 2008, leg. d. karasiński, kram f 5662 (as d. decipiens). 2. poland: wysoczyzny podlasko-białoruskie, in the białowieża national park, section 256, square g4, in a peucedano-pinetum w.mat. (1962) w.mat. et j.mat. 1973 forest, 27 october 1988, leg. a. chlebicki, kram f 41088 (as d. concentrica). 3. poland: pojezierza wschodniobałtyckie, in puszcza augustowska primeval forest, in a birch forest near płaska lake, on twigs of betula pendula roth, 2 may 2007, leg. a. chlebicki, kram f 56344 (as d. concentrica). 4. poland: górny śląsk, wyżyna śląsko-krakowska, jastrzębie zdrój district, wodzisław śląski, in the forest on the branches of betula sp., 3 march 1972, leg. k. filipowska, kram f 12634 (as d. concentrica). comments: the species belongs to the d. petriniae group. it is characterised by the long spores (up to 20 μm) and purple stromatal pigment. it occurs mostly on burnt substrates. the species was already noted by karasiński (2009), ruszkiewicz-michalska et al. (2015) and gierczyk et al. (2017). daldinia eschscholtzii (ehrenb.: fr.) rehm, annals mycol. 2(2): 175. 1904 stromata turbinate and sessile. surface without any conspicuous perithecial outlines and with slightly papillate ostioles. the koh-extractable stromatal pigment vivid vinaceous purple (fig. 1c). ascospores brown, inequilateral with narrowly rounded ends, 11.5–13.4 × 5–6 µm. specimen examined: 1. cameroon: in east province, department lom et djérem, between koumé and koundi, ca. 15 km northwest from bertoua, ca. 710 m a.s.l., in the guineo-congolian tropical rainforest, on a fallen trunk of a deciduous tree, 15 december 2007, leg. j. piątek and m. piątek, kram f 59661 (as daldinia sp.). comments: it is a common species in warmer climates, that occurs frequently in africa and was also noted in cameroon (stadler et al., 2014). daldinia lloydii y.m. ju, j.d. rogers & f. san martín, mycotaxon 61: 273. 1997. stromata fulvous with a surface cracked into polygonal scales. the koh-extractable stromatal pigments olivaceous green (fig. 1d). specimens examined: 1. ukraine: zhulkev region, dublany, the botanical garden, on handrail made of a birch wood (betula sp.), august 1928, leg. k. rouppert, kram f 1588 (as d. concentrica). comments: the species was reported from poland by stadler et al. (2014) from myszyniec. daldinia loculata (lév.) sacc. syll. fung. i: 394 (1882). stromata hemispherical and sessile, from 2.5 up to 4 cm in diameter. the koh reaction of stromatal materials mostly yields a dense purple pigment (fig. 1e). ascospores dark brown, with broadly rounded ends, (13)14–15(16) × 7–8.3 µm (fig. 2a). specimens examined: 1. poland: niziny sasko-łużyckie, zielona góra voivodeship, zasieki and lubsko, near the polish-german border, on burned trunks of betula sp. april 1984, leg. b. ginko, kram f 39306 (as d. concentrica). 2. poland: wyżyna środkowomałopolska, małopolska voivodeship, the ojców national park, sępówka valley, on a burned trunk of fagus sylvatica l., 7 november 1964, leg. w. wojewoda, kram f 13033 (as d. concentrica). 3. poland: pojezierze wschodniobałtyckie, in puszcza augustowska forest, sosnowo island in serwy lake, on a dead trunk of betula pendula, 10 july 2003, leg. a. chlebicki, kramf 55322, (as d. concentrica). comments: this species is associated with betula sp. but was also noted on salicaceae mirb. (stadler et al., 2014). it differs from d. vernicosa by its sessile stromata and the granules in its stromatal morphology. daldinia loculatoides wollw. & m. stadler, mycol. res. 108(9): 1030. 2004. stromata semiglobose, sessile, black, up to 4 cm in diameter. the koh-extractable stromatal pigment vinaceous purple (fig. 1f). ascospores dark brown, shaped like a rugby ball, with broadly rounded ends, 15–17(20) × 7–8 µm (fig. 6). specimens examined: 1. poland: wyżyna środkowomałopolska, małopolska region, kraków, rakowicki cemetery, on a burned trunk of acer platanoides l., 27 october 1993, leg. w. wojewoda, kram f 34899 (as d. concentrica). comments: this is a rare species that differs from d. loculata by its larger ascospores and semiglobose stromata (fig. 6). fig. 6. ascospores of daldinia loculatoides (kram f 34 899) (photo. a. chlebicki) daldinia petriniae y.m. ju, j.d. rogers & f. san martín, mycotaxon 61: 275 (1997) stromata semiglobose, aggregated and sessile. the koh-extractable stromatal pigments vinaceous to vivid purple (fig. 1i). ascospores inequilateral, dark brown, 10–14 × 6–7 µm. specimens examined: 1. poland: western carpathians, the bieszczady zachodnie mts., duszatyn, the przełom osławy reserve, on a standing dead trunk (ca. 6 cm in diameter) of alnus incana, 6 april 2012, leg. a. chlebicki, kram f without number 2. poland: western carpathians, the bieszczady zachodnie mts., prełuki near komańcza and osławica river, on a dead, small trunk (7 cm in diameter) of alnus incana, 9 april 2012, leg. a. chlebicki, kram f without number 3. poland: western carpathians, the bieszczady zachodnie mts., nasiczne, on a dead trunk of alnus incana, 26 december 2011, leg. a. wilczek, kram f without number comments: it was collected and determined by p. drzewiecki in mohle, kujawsko-pomorskie region, on the trunk of a young alder (snowarski, 2014). karasiński (wojewoda et al., 2016) also noted this species in the gorce mts. and domański et al. (1967) reported it from branches of alnus incana (as d. concentrica) in the bieszczady zachodnie mts. the fructification of the species begins in autumn. it is the most common species of daldinia in kram f. this species differs from d. concentrica by its vivid purple pigments and smaller ascospores. daldinia petriniae is similar to d. eschscholtzii but the latter species occurs in tropical zones. daldinia vernicosa ces. & de not., comment. soc. crittog. ital. 1: 198. 1863 stromata subglobose, sessile, shortly stipitate to distinctly stipitate. the koh-extractable stromatal pigment purple, vivid violet to vivid brown, or vinaceous purple (fig. 1h). ascospores dark brown with broadly to narrowly rounded ends: (10,5)12–14 × 6.2–7.3 µm (fig. 2d). specimens examined: 1. poland: eastern podkarpacie, sandomierz dell, przemyśl, near the railway station in przemyśl-pikulice towards sielec, near wiar river, on dead wood, 14 september 1979, leg. w. wojewoda, kram f 33838 (as d. concentrica). 2. poland: niziny sasko-łużyckie, in zielona góra voivodeship, close to zasieki and lubsko, near the polish-german border, on burned trunks of betula sp., april 1984, leg. b. ginko, kram f 39306 (as d. concentrica). 3. poland: lakelands wschodniobałtyckie, in augustowska forest, the perkuć nature reserve, vaccinio myrtylli-pinetum juraszek 1928 forest, on a lying betula trunk, 16 september 1974, leg. w. wojewoda kramf 32456 (as d. concentrica). 4. poland: lower silesia, sudety mts., przedgórze sudeckie, strzelińskie hills, 1 km northeast of biały kościół, on a burned trunk of prunus spinosa l., 26 march 1985, leg & det. a. chlebicki, kram f 55346 (as d. fissa c. g. lloyd). 5. poland: in biebrza valley, the biebrza national park, ciszewo, brzeziny ciszewskie forest, on a frangula alnus mill. host, 17 august 1991, leg. & det. a. chlebicki, kram f 43103 (as d. fissa). 6. poland: małopolska region, tarnów, near lipie forest, on a dead trunk of betula pendula, 31 june 1997, leg. m. piątek, det. a. chlebicki, kram f 56616, (as d. fissa). comments: the species was reported from poland by stadler et al. (2014) and gierczyk et al. (2017). chlebicki (2008) noted this species on prunus spinosa, frangula alnus and betula pendula (as daldinia fissa). stromata of this species are reported also on fire-damaged trees (stadler et al., 2014; chlebicki, 2008). stadler et al. (2014) recognized d. fissa as a synonym of d. vernicosa and suggested that d. fissa is an aberrant form with compressed stromata, damaged by insect larvae. discussion the use of chemotaxonomic evidence, molecular phylogeny and anamorphic characters enable to divide the members of the genus daldinia into five major groups (stadler et al., 2014). the colour of stromatal pigments appears to be very useful in identification of the specimens (stadler et al., 2014) and it was successfully used to determine polish collections from kram f. after the revision, the exsiccata of the following daldinia species are present in the kram f: d. childiae (the d. childiae group), d. eschscholtzii (the d. eschscholtzii group), d. loculata, d. loculatoides and d. vernicosa (the d. vernicosa-d. loculata group), d. decipiens, d. lloidii and d. petriniae (the d. petriniae group). from these species, three are reported here from poland for the first time: d. childiae, d. loculata and d. loculatoides. three species, which specimens are present in kram f were gathered outside of poland: d. lloydii from ukraine, d. concentrica from bulgaria and d. eschscholtzii from cameroon. the most common species of the genus preserved in kram f appears to be d. petriniae noted on the alnus incana. however, in the world d. childiae is the most commonly encountered species (stadler et al. 2014). in the case of one species from kram f, namely d childiae, a larva and an imago of a fly from the family agromyzidae was found inside stromata. insect-fungus interactions are known very well. they include dispersal, protection, nutrition and all kinds of symbiosis. information about insects living on stromata of daldinia is very scary. alex hyde (2020) noted platyrhinus resinosus eating stromata of d. concentrica in peak district national park, derbyshire, uk. some insects, such as xiphydria wood wasps, were noted inside stromata of d. decipiens. this insect can appear highly suitable as a vector for daldinia (šrutka et al., 2007; pažoutová et al., 2013). probably also the fly noted on stromata of d. childiae preserved in kram f can be considered as a vector for the fungus. parasitic species can be used as aids in the host biogeography (savile, 1975). however, our knowledge of the insects parasitizing daldinia are to scarce to draw any conclusions. acknowledgements i would like to thank m. stadler for his comments, suggestions and important article on secondary metabolites, a. wilczek for collecting daldinia petriniae specimens from the bieszczady zachodnie mts. and t. zatwarnicki and a. palaczyk for suggesting that the fly could be agromyzidae. conflict of interest the authors declare no conflict of interest related to this article. references cesati,v., de notaris, g. 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(2011). importance of secondary metabolites in the xylariaceae as parameters for assessment of their taxonomy, phylogeny, and functional biodiversity. current research in environmental & applied mycology. https://doi.org/5943/cream/1/2/1 stadler, m., læssøe, t., fournier j., decock c., schmieschek b., tichy, h-v., peršoh d. (2014). a polyphasic taxonomy of daldinia (xylariaceae). studies in mycology, 77, 1–143. https://doi.org/10.3114/sim0016 stadler, m., wollweber, h., mühlbauer, asakawa y., hashimoto, t., rogers, j.d., ju, y.m., wetzstein, h.g., tichy, h.v. (2001). molecular chemotaxonomy of daldinia and other xylariaceae. mycological research. 105(10), 1191–1205. https://doi.org/10.1017/s0953756201004890 ju, y.m., rogers, j.d., san martin, f. (1997). a revision of the genus daldinia. mycotaxon 61, 243–293. wojewoda, w., kozak, m., mleczko, p., karsiński, d. (2016). grzyby makroskopijne gorców (karpaty zachodnie) (macroscopic mushrooms of the gorce montains (western carpathians)). instytut botaniki im. w. szafera, polska akademia nauk, ss. 192. [in polish] abstract the article presents the results of the revision of daldinia (ascomycota) specimens preserved in kram f (kraków, poland). the following species were identified: daldinia childiae, d. decipiens, d. loculata, d. loculatoides, d. petriniae and d. vernicosa. three of them were not reported from poland so far: d. childiae, d. loculata, d. loculatoides. aditionaly to polish specimens, also some collections from outside poland are kept in kram f: one specimen of d. concentrica from bulgaria, d. lloydii from ukraine and d. eschscholtzii from cameroon. key words: daldinia, stromatal pigments, wood-inhabiting fungi, fungi-associated insects received: [2022.03.01] accepted: [2022.07.12 rewizja okazów z rodzaju daldinia w kolekcji zielnika kram: daldinia childiae, d. loculata i d. loculatoides, trzy nowe gatunki dla polski artykuł zawiera wyniki rewizji okazów z rodzaju daldinia (ascomycota), przechowywanych w zbiorach kram f. w kolekcjach stwierdzono następujące gatunki: d. childiae, d. decipiens, d. loculata, d. loculatoides, d. petriniae i d. vernicosa. trzy z nich nie były do tej pory podawane z terenu polski: d. childiae, d. loculata, d. loculatoides. w zbiorach kram f znajdują się również kolekcje spoza polski: jeden okaz d. concentrica z bułgarii, d. lloydii z ukrainy i d. eschscholtzii z kamerunu. słowa kluczowe: daldinia, barwniki podkładek, grzyby zasiedlające drewno, owady związane z grzybami information on the authors andrzej chlebicki https://orcid.org/0000-0002-7001-2782 his research to date has focused on fungi found on arctic-alpine plants. in addition, in recent years he has dealt with extremophiles found on rocks, mainly black yeast, and the influence of the electromagnetic field on the development of fungal colonies. 185 annales universitatis paedagogicae cracoviensis studia naturae, 6: 185–202, 2021, issn 2543-8832 doi: 10.24917/25438832.6.11 sabina adamczyk �e osteopathy academy, non-public continuing education institution in krakow, szlak st. 65, 31-153 kraków, poland; sabina04@poczta.fm osteopathic capabilities for headache therapy – a short overview introduction in order to de�ne a headache, one should take into account, �rst of all, its non-speci�city, expressed in subjective perception. �is symptom is most o�en located throughout the head and felt both on the surface of the face skin – in the orbital-temporal area, and deep inside the skull. it is characterised by di�erent intensity and duration. however, headache is not usually a symptom of severe disease, although cluster headaches can lead to severe systemic disturbances (stovner et al., 2006; prusiński, 2012). headache is the most common phrase searched by patients in the google browser around the world (kamiński et al., 2020). it concerns di�erent age groups of society: from children to the elderly (linet et al., 1989; lipton, stewart, 1993; anttila et al., 2006; louw, schmidt, 2015; mrozkowiak et al., 2018). research in recent years has shown that over 90% of people experience at least one headache incident each year. usually, most people experience the so-called random pain, e.g. alcohol syndrome, fatigue, lack of sleep. however, about 15% of people experience more serious ailments in the form of cyclical pains. �ey are characterised by annoyance, but most of all have a negative impact on the quality of life. �ey are a disease phenomenon and may require medical attention. �e most common pains of this type are: tension headaches, migraine and cluster pains (malec-milewska, woroń, 2012; prusiński, 2012). �e most common form of headache is tension type headache (tth), a�ecting approximately 30–80% of cases. �ey are o�en called stress or psychogenic pains and arise from increased muscle tension. patients usually describe tth as dull, compressive, and tightening pains which radiate to the occiput or forehead. �ere is also pain involving the back of the neck. �e cause of tth is, among others, myofascial pain syndrome (mfps), which is characterised by the occurrence of non-speci�c muscle s ab in a a da m cz yk 186 pain in the head and neck; o�en it is not strictly associated with headache. mfps may a�ect up to 55% of head and neck pain patients (stępień, 2003; fumal, schoenem, 2008; chochowska et al., 2015). migraine (lat. migraena) is a type of headache experienced by approximately 15–18% of women and 6–8% of men. it most o�en a�ects young people, under the age of 40, and is usually characterised by unilateral pain lasting from 4 to even 72 hours. migraines may be accompanied by nausea, vomiting, sensitivity to light, noise, odours and vegetative disorders. in its course, migraine may transform into daily headaches or may coexist with a tension headache (gervil et al., 1999; rożniecki et al., 2018). a migraine headache has a speci�c location that usually a�ects the area behind the eyeball, in the forehead, and in the temple. it can have bilateral or alternating symptoms. �is pain is sometimes so strong that it excludes the patient from everyday life. pain of vascular origin is pulsating with the patient’s heart rate and is o�en shooting or excruciating (glaubic-łątka et al., 2004; wójcik-drączkowska et al., 2007). �e classic type of migraine is the so-called migraine with aura. migraine aura is a type of neurological disorder that, in its classic form, a�ects eyesight: it causes visual disturbances – spots in front of the eyes, �ashes of light in the �eld of vision, visual acuity disturbance, and sometimes even loss of vision (domitrz, 2007; wójcik-drączkowska et al., 2007). cluster headache, otherwise known as raeder syndrome or histamine headache, formerly known as horton’s headache syndrome (lat. cephalea hortoni), is the most intense headache. it belongs to the group of primary pains, i.e. those that are not caused by any disease (this group also includes migraine). it most o�en occurs in men between 20 and 50 years of age. its causes are unknown. �is type of pain is described as excruciating, burning, glaring, stinging, drilling, and above all, very intense. it is unilateral pain, located within the trigeminal nerve (sensorial innervating the face). �is pain can also appear in the orbital (behind the eyeball) or temporal area, less commonly in the cheek or jaw (malec-milewska, woroń, 2012; rożniecki et al., 2018). patients su�ering from chronic headache usually undergo pharmacological treatment, which is known to cause many side e�ects over a  prolonged period of time, o�en leading to damage to various internal organs. in recent years, a  phenomenon has been observed that in alternative pain treatment centres, such as physiotherapy, osteopathy or chinese medicine, patients look for other methods of non-pharmacological treatment (stovner et al., 2006; prusiński, 2008; łukasik, et al., 2012; vemuri, got, 2020; healy et al., 2021). it is, among other, a  consequence of returning to herbalism and alternative medicine, as well as promoting an ecological, healthy and hygienic lifestyle. osteopathy is one of the branches of medicine focused on the natural treatment of the whole person (greenman, 2005; speece et al., 2017). diagnostics and treatment in this �eld are based primarily on precise palpation, using manual tests and techniques, o steopathic capabilities for headache therapy – a short overview 187 in order to treat the causes of dysfunction and restore the mobility of each body system. palpation (lat. palpatio), is an examination by touch – a diagnostic method that involves touching the size, shape, hardness or location of a speci�c anatomical structure with the �ngers. �is method is used by doctors in examining the chest and abdominal organs, and also to evaluate the heart rate (bolechowski, 1982; greenman, 2005; wong et al., 2014). osteopathic treatment is based on the interdependence of anatomy and physiology and the perception of each person as a functional unit, capable of recovering on its own, if the structure and physiological functioning of the body are in the right condition (jäkel, hauenschild, 2011). in recent years, this type of therapy has been increasingly used to treat pain, and the basic practices in this area have been developed for a long time (fig. 1. – appendix 1). taking up this type of topic in the review aspect was caused by the desire to present other, bene�cial, and still relatively little known methods of �ghting headaches. �e current level of knowledge on this subject requires systematisation, as well as indication of which osteopathic therapies bring patients the best results in pain therapies. experiments included in the analysis �e bibliography on the e�ects of osteopathic therapies was analysed through search strategies using medical subject headings (mesh) and key words in the text. �is review is methodically based on the review method used by jäkel and hauenschild (2011). �e following databases were searched: jaoa (journal of the american osteopathic association), pubmed.gov. due to the large number of erroneous records, the use of the word “osteopathy” was abandoned in favour of more speci�c phrases, consistent with the chosen topic. �e key words and phrases selected for the search were: “headache osteopathy”, “non-pharmacological migraine treatment”, “tension headache”, “migraine therapy”, “osteopathy and tension headache”. �e “keys words” were used in both polish and english. a selection was made among the publications from 2005–2020. �e search for given phrases resulted in 220 potential publications, of which 132 were from pubmed and 88 from jaoa. by removing duplicate articles, 180 records were obtained. a�er analysing the abstracts, articles not related to the subject were rejected. six publications relating to experiments in the �eld of osteopathic headache treatment were included in the further analysis. �ese were randomized controlled trial (rtc) studies and prospective and retrospective comparative studies: voigt et al. (2011), rolle et al. (2014), adragna et al. (2015), chaibi et al. (2017), d’ippolito et al. (2017), gandol� et al. (2018). �eir short description is presented in table 1. s ab in a a da m cz yk 188 tab. 1. characteristics of the analysed experiments with the use of osteopathic techniques research authors study type subject of study voigt et al. (2011) rct e�cacy of omt in the treatment of migraine rolle et al. (2014) rct studies on the e�ects of om� in patients with frequent episodic tension headaches adragna et al. (2015) comparative study before and a�er the intervention �e in�uence of omt on pain and quality of life in patients with migraines chaibi et al. (2017) rct adverse events in csmt, with single-blind, placebo, and with randomized controlled study for people with migraine d’ippolito et al. (2017) prospective and retrospective comparative studies e�ect of om� on pain and mood disorders in patients with high-frequency migraine gandol� et al. (2018) rct will myofascial treatment and the use of trigger point therapies reduce pain and pain medication intake in patients undergoing onabotulinumtoxina injections for chronic migraine? note: csmt – chiropractic spine manipulation �erapy; omt – osteopathic manipulative treatment; om� – osteopathic manipulative �erapy; rct – randomized controlled trial types of procedures in analysed osteopathic experiments voigt et al. (2011) evaluated the e�ectiveness of osteopathic manipulative treatment (omt), combined with standard care, for women su�ering from migraines, de�ned according to the international classi�cation of diseases (icd-10, g43; migraine with or without aura). �e researchers gathered a clinical group of 42 women, aged 25–65, meeting the de�ned criteria (minimum 3 migraine attacks per month). �irteen women were diagnosed with migraine with aura. �e duration of migraine symptoms ranged from 2 to 45 years; on average 23 years. �e exclusion criteria were as follows: non-medical therapies (e.g. acupuncture, homeopathy) within 8 weeks prior to study initiation, pregnancy, lactation, and neurological diseases (e.g. brain tumours, multiple sclerosis and others). 42 participants were randomly assigned to 2 research groups (21 people each): omt and control. a  licensed physiotherapist performed osteopathic manipulations of each participant in 5 sessions of 50 minutes over a period of 10 weeks in the omt group. osteopathic manipulation involved a variety of techniques at the discretion of the osteopath, tailored to each participant. �e control group did not receive any osteopathic manipulation, sham treatment, or other physical therapy. all women completed standardised health outcomes questionnaires before the study (t1) and 6 months a�er the end of the follow-up (t2). �ese were: health-related quality of life (hrqol), migraine disability assessment (midas), short form-36 (sf-36) and the german “pain questionnaire” (schmerzfragebogen des schmerztherapeutischen kolloquium ev dr. lowendorf ). o steopathic capabilities for headache therapy – a short overview 189 �e italian team of rolle et al. (2014) conducted a  pilot study on the e�ects of osteopathic manipulative �erapy (om�) in patients with frequent episodic tension headaches. �ey carried out randomized (rct), with single-blind, and control of placebo. �e inclusion criterion for the study was the diagnosis of a frequent episodic tension type headache (tth). �e study excluded people: under 18 years or over 65 years of age, taking acute headache medications for 10 or more days a month in the last 3 months, su�ering from pain for less than a year, with mental illnesses and other disorders (e.g. secondary aches of head pains) or with any ongoing prophylaxis during the study period. �is experiment included: a  1-month baseline period, a  1-month treatment period, and a 3-month follow-up period. patients were randomly assigned (coin toss) to the control or experimental group with om�. during 4 weekly treatment sessions, patients from the experimental group underwent corrective om� techniques. �ese techniques were not protocol based but were individually tailored for each patient as described by greenman (2005). patients in the control group underwent a cranial rhythm assessment (sham therapy), considered a placebo – manual techniques were used, but the observed osteopathic disorders were not corrected. at the end of the active treatment period, at 1 and 3 months, patients in both groups were assessed using “headache diaries” and the headache disability inventory (hdi). �ese diaries included: changes in the frequency of headache reported by patients (number of episodes during the period studied), headache intensity (for each episode in the study period; rated from 1 – lowest pain experienced to 5 and above – worst pain experienced), medication use without prescription (total number of drugs used during the period under study). adragna et al. (2015) conducted an experiment on the e�ects of osteopathic medicine (omt) in patients with migraine without aura in the form of a pilot study. �ey examined 8 people: 3 men and 5 women; patients were selected in a private doctor’s o�ce and included in one therapeutic group. �e researchers used four osteopathic treatments in the experiment, carried out over the course of 8 weeks. �e measures of the result were: frequency of attacks, use of analgesics, completed body awareness questionnaires: midas, headache impact test (hit-6), sf-36 and body awareness questionnaire (baq). experimental data was collected at baseline (t0), 1 month a�er the last treatment (t1), and 3 months a�er the last treatment (t2). from the three months prior to t0 and throughout the study, all participants completed a “headache diary” and continued prescribed drug therapy. chaibi et al. (2017) took up the topic related to the occurrence of adverse events in chiropractic spine manipulation �erapy (csmt). investigators conducted a rigorously designed experiment, with single-blind, prospective, randomized clinical trial (rct), using csmt in migraineurs to assess adverse events (aes), a�er manual intervention. �ey recruited a group of 97 migraine patients (83 women and 14 men), aged 18 to 70. s ab in a a da m cz yk 190 participants, who experienced at least 1 migraine attack per month were randomized to the experimental csmt, placebo (non-speci�c manual touch and non-therapeutic pushing manoeuvre), or control (follow-up medication) groups. interventions were administered in twelve 15-minute sessions over a period of 3 months. �e researchers compared the aes of participants who completed the study. �e results were assessed a�er 3, 6 and 12 months. �e initial exclusion criteria were: contraindications for spine manipulation, spinal radiculopathy, pregnancy, depression and csmt in the last 12 months. during the experiment, the exclusion criteria also included: any other manipulative intervention of any physical therapist, or pregnancy. aes were assessed during 703 sessions (355 in the csmt groups and 348 in the placebo groups). d’ippolito et al. (2017) undertook to investigate the e�ect of om� on pain and mood disorders in patients with high-frequency migraine. �ey reviewed the medical records of patients with this type of diagnosis. �e inclusion criteria for the experiment were as follows: diagnosis according to ichd-3b (pain: > 8 and < 15 days per month), participation in the om� program and psychological evaluation, before and a�er the om� program. patients with one of the following criteria were excluded: a di�erent diagnosis of ichd-3b (e.g. tension headache, chronic migraine), somatic or psychiatric disorders (e.g. major depression, psychosis) and the presence of musculo-skeletal disorders, temporomandibular diseases, neurological or rheumatic diseases. 11 people participated in the study. patients included in the experiment were subjected to psychological assessment according to standard psychological assessments. �e following were used to assess the symptoms of depression and anxiety as well as personality patterns: the hamilton depression rating scale (hdrs), state-trait anxiety inventory (stai), forms x-1 (state anxiety), and x-2 (trait anxiety) and the millon clinical multiaxial inventory (mcmi-iii) – is only used during the �rst visit. hdi and hit-6 – used to assess pain and its impact on daily activities. all patients were eligible for osteopathic treatment, but only those who wished to be treated in this way were enrolled in the om� program. four 45-minute sessions were conducted over 8 weeks. �ese procedures focused on correcting the dysfunctions identi�ed at the �rst assessment and were performed using myofascial techniques, balanced ligament tension and osteopathy in the cranial �eld. a�er the om� program, all patients were re-evaluated using the above-mentioned tests. �e last of the analysed experiments by gandol� et al. (2018) aimed to determine, whether myofascial treatment and trigger point therapy will reduce pain and reduce pain medication intake in patients undergoing onabotulinumtoxina injections due to chronic, di�cult-to-treat migraine. adults with persistent migraines took part in the experiment. �is was a single-blind pilot study with two parallel groups (experimental and control). �e osteopathic intervention used here included manipulative treatment, consisting of techniques aimed at improving joint mobility and reducing the sti�ness o steopathic capabilities for headache therapy – a short overview 191 of the so� tissues of the cervico-thoracic spine – the experimental group (12 people). in addition, tens current-healing treatment with a portable device (master 932, elettronica pagani srl, milan, italy) was used – control group (10 people). �e frequency and duration of treatment was the same in both groups: 1 session per week for 4 weeks. non-clinical and demographic data, including personal habits (e.g. co�ee, alcohol and tobacco consumption) were collected during registration using a  questionnaire. an evaluator unaware of treatment allocation recorded the results before treatment (t0), during treatment (t1), and 1 month a�er treatment completion (t2). �e information relating to the detailed characteristics of the individual experiments analysed here related to osteopathic intervention is summarised in table 2. clinical bene�ts of experiments and their analysis in the light of selected literature an experiment conducted by voight et al. (2011) found a signi�cant reduction in total midas; the result was statistically signi�cant in the intervention group as compared to the control. �e number of days with migraines that the patients su�ered while performing their daily duties and activities decreased, but in both the intervention groups with omt and the control group, it was statistically insigni�cant. �e intensity of pain and work disorders and the number of days of incapacity for work caused by migraine showed a statistically signi�cant reduction in the intervention group. �e control group had statistically signi�cant declines only in the functioning of the emotional role. based on the intention-to-treat analysis design, the omt group experienced signi�cant improvements in vitality, mental health, body pain, and physical role functioning (4 of 8 hrqol domains); sf-36 showed statistically signi�cant improvement in the intervention group. overall, this experiment con�rmed the positive e�ects of omt on migraine headaches in terms of: reduced pain intensity and reduced number of days with migraine and disability, and in part improved hrqol. based on “headache diaries” and hdi, no adverse events were recorded throughout the study in any of the groups of the experiment conducted by rolle et al. (2014). �ere was a  signi�cant change in headache frequency in the om� group (approximately 50% reduction from baseline a�er 3 months of follow-up) and an absolute di�erence between the 2 groups at 3 months, with a 33% lower headache frequency in the om� group. over-the-counter medication use only decreased in the om� group at all time points a�er baseline, compared with the mean baseline. pain intensity also slightly decreased over time in the om� group (resulting in an approximately 20% reduction from baseline and a�er 3 months of follow-up). �e overall hdi score showed no signi�cant improvement. however, a comparison between the changes in hdi score in the 2 groups revealed a di�erence in time in the om� group (resulting in an approximately 40% reduction from baseline and a�er 3 months of follow-up). �e s ab in a a da m cz yk 192 ta b. 2 . b as ic in fo rm at io n ab ou t t he a na ly se d ex pe ri m en ts ex pe ri m en t pa tie nt p op ul at io n a ss ig ne d gr ou ps n um be r o f s es si on s/ le ng th o f s es si on pr ac tit io ne r p ro �l e w ay o f r es ul ts c ol le ct in g vo ig t e t a l. (2 01 1) w om en w ith m ig ra in es , ag ed 1 8 – 65 ; 1 3 pe rs on s ha d m ig ra in es w ith a ur a c on tr ol g ro up n = 21 n o in te rv en tio n ph ys io th er ap is t w ith o st eo pa th ic p ow er s c om pa ri so n of re su lts be tw ee n th e 2 gr ou ps o m t g ro up n = 21 5 se ss io ns o f 5 0 m in ut es fo r 10 w ee ks r ol le e t a l. (2 01 4) pe op le a ge d 18 – 6 5 w ith fr eq ue nt e pi so di c pa in c on tr ol g ro up n = 19 sh am th er ap y q ua li� ed o st eo pa th c om pa ri so n of 2 g ro up s: 1m on th b as el in e pe ri od , 1 -m on th tr ea tm en t pe ri od a nd 3 -m on th fo llo w -u p pe ri od o m � g ro up n = 21 4 o m � se ss io ns e ve ry w ee k a dr ag na e t a l. (2 01 5) a du lts w ith m ig ra in es w ith ou t a n au ra o ne th er ap eu tic g ro up n = 8 4 tr ea tm en ts 8 w ee ks q ua li� ed o st eo pa th c om pa ri so n of re su lts at b as el in e (t 0) , 1 m on th a� er la st tr ea tm en t ( t1 ) an d 3 m on th s a �e r l as t tr ea tm en t ( t2 ) c ha ib i e t a l. (2 01 7) w om en a nd m en su �e ri ng fr om m ig ra in es c on tr ol g ro up n = 27 n o in te rv en tio n n ot sp ec i� ed c om pa ri so n of 3 g ro up s, a� er 3 , 6 a nd 1 2 m on th s of in te rv en tio n pl ac eb o gr ou p n = 35 sh am th er ap y c sm t g ro up n = 35 12 se ss io ns o f 1 5 m in ut es fo r 3 m on th s d ’ip po lit o et a l. (2 01 7) a du lts w ith m ig ra in e o ne th er ap eu tic g ro up n = 11 4 se ss io ns o f 4 5 m in ut es fo r 8 w ee ks q ua li� ed o st eo pa th c om pa ri so n of re su lts be fo re (t 1) a nd a �e r in te rv en tio n (t 2) g an do l� e t a l. (2 01 8) a du lts w ith m ig ra in es , a ge d 18 – 6 5 c on tr ol g ro up n = 10 n er ve st im ul at io n (t en s) ; 4 se ss io ns fo r 4 w ee ks o st eo pa th d oc to r c om pa ri so n of re su lts be tw ee n th e 2 gr ou ps b e fo re (t 0) , d ur in g (t 1) a nd a� er tr ea tm en t ( t2 ) o m � g ro up n = 12 1 se ss io n of 3 0 m in ut es p er w ee k fo r 4 w ee ks n ot e: o m t – o st eo pa th ic m an ip ul at iv e tr ea tm en t; o m � – o st eo pa th ic m an ip ul at iv e � er ap y; c sm t – c hi ro pr ac tic s pi ne m an ip ul at io n � er ap y o steopathic capabilities for headache therapy – a short overview 193 obtained results allowed for the conclusion that om� is an interesting alternative in tth therapy. however, it should be remembered that om� is not recommended for everyone and is not completely devoid of side e�ects (rajendran et al., 2012), although this experiment did not �nd any. �e results of the experiment of adragna et al. (2015) showed 100% of somatic dysfunction (sd) in the c1-occipital joint in the �rst session, and only 37% in the same joint in the second session with omt. a reduction in sd was observed between omt sessions, showing a signi�cant reduction in total dysfunction a�er the third and fourth sessions. �e musculoskeletal sd improved with the fourth treatment and the craniosacral system with the second, third and fourth omt treatments. signi�cant changes in the results were observed in the hit-6 scale in the t2 period, midas in the t1 period and the sf-36 scale in the t1 and t2 periods. �e baq, the second midas, and the results of the “headache diary”, despite the reduction of migraine attacks and taking medications, did not produce statistically signi�cant results. however, this experiment generally showed that omt had a positive e�ect in reducing pain and improving the quality of life of migraine patients without aura. �e results of the research team of chaibi et al. (2017) indicated, that the most common adverse event was local tenderness – 11.3% in the cmst group and 6.9% in the placebo group. fatigue on the day of intervention was reported by 8.5% of participants in the csmt group and 1.4% in the placebo group. adverse events were mild and transient, and no serious and other adverse events were reported. �us, this therapy is safe, provided that people with severe contraindications are initially excluded (rajendran et al., 2012; chaibi et al., 2017). d’ippolito et al. (2017) showed that although the number of migraine attacks per month decreased, this was not considered “clinically signi�cant” because the sample size was small and all patients had the same range of headache attacks (> 8 and < 15 days per month). �e results of the stai x-2, hit-6 and hdi questionnaires in the study before treatment (t0) and a�er treatment (t1) showed a statistically signi�cant reduction in means. �e results observed in this small group of patients showed that om� reduced the hit-6 scores. �e change in mean stai x-2 score was not considered “clinically signi�cant” because both the t0 and t1 scores in this test suggested moderate anxiety. �e hdi score showed a statistically signi�cant di�erence between t0 and t1, but it was not considered a “clinically signi�cant improvement” anyway. �ere were no statistically signi�cant changes in the mean hdrs and stai x-1 scores in the period from t0 to t1. in the case of mcmi-iii, the result was inconclusive. however, the analysed retrospective study revealed that patients with high-frequency migraine had statistically signi�cant decreases in hit-6, stai x-2 and hdi scores a�er the om� program. nevertheless, it is impossible to draw de�nitive conclusions about the causal relationship between om� and the changes in score and the clinical e�ect of these s ab in a a da m cz yk 194 changes. one reason is the retrospective, non-randomized selection process, and the other is an equally important, small sample size, which has limited both the generalization of results and the ability to determine the clinical signi�cance of scoring changes. another limitation was the lack of a control group. �e presence of a control group could alleviate potential confounding factors, such as the mechanism of action of therapeutic touch and interactions between therapist and patient, which may have a positive e�ect on headache (keller, bzdek, 1986; autret et al., 2012). however, this experiment showed that om� can have a therapeutic e�ect on pain and mood disorders in patients with high-frequency migraine and could be useful as part of a multidisciplinary treatment program (krause et al., 2017). �e results presented by gandol� et al. (2018) showed no signi�cant di�erences between the groups in the pre-treatment assessment. �ere were also no signi�cant intergroup di�erences in pain intensity in the analysed period. in the post-treatment evaluation, the total consumption of analgesics and non-steroidal anti-in�ammatory drugs (nsaids) was signi�cantly lower in patients treated with osteopathic therapy than in those treated with tens. �e pressure pain thresholds in the muscles of the upper trapezius, occipital and temporal muscles were signi�cantly lower in patients treated with manipulation than those treated with tens. �e total consumption of painkillers, nsaids and triptans was signi�cantly lower a�er a  series of treatments than before treatment in patients undergoing manipulative treatment. no adverse events were reported during and a�er the experiment. �us, according to the authors of this experiment, manipulative techniques aimed at reducing peripheral nociceptive triggers may be of added value in treating chronic migraine symptoms and reducing the use of reliever medications. �e comparison of all the results obtained in the analysed experiments is summarised in table 3. tab. 3. main outcomes and e�ects of analysed experiments, in comparison to control patients or baseline results experiment e�ect/result in comparison to control group and/or baseline results voight et al. (2011) • statistically signi�cant decrease in the total score in the otm group; • pain intensity, work disorders, number of days of inability to work due to migraine, showed a statistically signi�cant reduction in the otm group; in the control group, statistically signi�cant declines in the functioning of the emotional role; • 4 out of 8 spheres tested improved (vitality, mental health, body pain and physical role functioning). rolle et al. (2014) • the frequency of pain and its intensity have decreased; reducing the amount of medications taken; • hdi has not changed signi�cantly. o steopathic capabilities for headache therapy – a short overview 195 adragna et al. (2015) • signi�cant changes expressed in reduced pain; • no signi�cant changes in the evaluation of some tests. chaibi et al. (2017) • local tenderness was greater in the csmt group than in the placebo group; • greater fatigue on the day of intervention in the csmt group than in the placebo group; • the adverse events were mild and transient. d’ippolito et al. (2017) • improvement in general condition but not clinically relevant due to a small sample size. gandol� et al. (2018) • in the experimental group, there was a reduction in the doses of drugs taken and an improvement in the range of motion, especially in �exion and lateral bend. note: hdi – headache disability inventory; csmt – chiropractic spine manipulation �erapy �e method of manual osteopathic therapy considered in all the experiments discussed here belongs to the relatively little-known complementary and alternative medicine treatments, useful in combating pain (vincent, furnham, 1997; vickers, zollman, 1999). �ere are studies containing descriptions of the necessary, more or less complicated, manual techniques in this �eld. an example is the extensive textbook in this area by greenman (2005), which, in addition to the basics, comprehensively and transparently presents osteopathic tests, treatment methods, as well as frequent clinical images and accompanying diagnostic and therapeutic methods. nevertheless, much more popular methods from this group are, for example, acupuncture (e.g. diener et al., 2006; li et al., 2012; linde et al., 2015), methods of brain stimulation, such as biofeedback (nestoriuc et al., 2009) – recently very fashionable, di�erent relaxation methods (evers et al., 2009), or homeopathy (ernst, 1999). �eir e�ectiveness has been tested through numerous medical experiments. in the case of osteopathic treatments, there are still few well-planned experiments con�rming the e�ectiveness of this type of therapy. however, randomly selected experiments for this analysis are an example of a keen interest in this subject. on their basis, it can be concluded that in this type of research it is very important to plan them so that the obtained results are as credible as possible, which was especially emphasized by adragna et al. (2015), rolle et al. (2014) and d’ippolito et al. (2017). according to them, an important role is played here, among others, by sample size, presence of a control group, clear exclusion and inclusion criteria – non-standard selection process, omt tailored to the patient’s needs and a reliable description of the condition before and a�er therapy. as for the selection of appropriate treatments individually for each patient, it usually takes place in the clinical practice of osteopathy, with the use of techniques correcting any observed dysfunctions. �is is because clinical �eld observations suggest that standard osteopathic therapies are less e�ective than those tailored to individual needs (rolle et al., 2014). �e individual relationship of the patient with the therapist-osteopath s ab in a a da m cz yk 196 is also very important, as mentioned earlier (d’ippolito et al., 2017). improving the psychological factors that are believed to aggravate the pain and in�uence the progression of migraine – from episodic to chronic – can be especially important, and a good relationship with the therapist is not neutral in this regard. considering the overall condition of the patient, om� may be an interesting treatment option, for example, episodic pain, characterised by several contraindications and side e�ects. �is therapy is especially indicated for patients not adhering to the treatment regimens and with an increased risk of adverse drugs reactions. manipulation techniques aimed at reducing peripheral nociceptive triggers (pain receptors) may be of great value in the treatment of chronic migraine, especially as their side e�ects are generally minimal or completely unnoticed (rolle et al., 2014; chaibi et al., 2017). an interdisciplinary approach, including pharmacological and non-pharmacological treatment, is also very important (healy et al., 2021). in this regard, the use of omt can reduce acute drug intake and muscular-vascular dysfunction in patients with chronic migraine (gandol� et al., 2018). manipulative osteopathic treatment, for example in the case of somatic dysfunctions of the thoracic spine, may require an interdisciplinary team of specialists. without proper treatment, acute somatic dysfunctions of the thoracic spine can turn into serious chronic problems. although the main moderator of treatment is a quali�ed osteopath or physician, a multidisciplinary and holistic approach, requiring the collaboration of several specialists, is currently preferred (vemuri, got, 2020). �is model emphasizes a multidisciplinary approach to persistent pain due to its intertwined physical and psychological nature (parkin-smith et al., 2015). certainly, combating headaches also requires this kind of interdisciplinary approach. �e articles analysed here supports the hypothesis that osteopathic manipulation has a therapeutic e�ect on pain and mood disorders in patients with high-frequency migraine and may be useful as part of an interdisciplinary therapeutic program. many physicians recognise that the traditional methods of pharmacological pain treatment available to date are o�en unsatisfactory and their social and health costs can be high (jensen, stovner, 2008). positive e�ects of om� on headache control may be due to speci�c neurochemical e�ects, including increases in circulating opioids and serotonin levels, involving the descending serotonergic and noradrenergic pathways (degenhardt et al., 2007). however, the molecular basis of om�’s clinical outcomes is largely unknown and more research is needed to address this issue. conclusion �e experiments presented above con�rm the e�ectiveness of omt manipulative osteopathic therapy in the treatment of headaches of various origins. osteopathic treatment in all its range of possibilities of the techniques used, o�ers great opportunities to reo steopathic capabilities for headache therapy – a short overview 197 duce pain and improve the well-being of su�erers, especially in migraines and tension headaches. �e analysed results also show that the recommended therapy is a  very good supplement to standard drug therapy, thanks to which the use of large doses of drugs can be signi�cantly reduced. researchers of these issues also pay attention to the fact that future experiments should be planned with a su�ciently large sample size. in order to improve the quality of good medical practice, they also encourage osteopaths to use systematic assessment of adverse events in selected manipulative therapy and a holistic, interdisciplinary approach to pain management. con�ict of interest �e author declares no con�ict of interest related to this article. references adragna, v., bertino, a.s., carano, m., soru, a., taranto, g., desideri, r. 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(2011). e�cacy of osteopathic manipulative treatment of female patients with migraine: results of a randomized controlled trial. journal of alternative and complementary medicine, 17(3), 225–230. https://doi.org/10.1089/acm.2009.0673 wójcik-drączkowska, h., bilińska, m., nyka, w. (2007). migrena – rozpoznanie i leczenie (migraine – diagnosis and treatment). forum medycyny rodzinnej, 1(2), 109–114. [in polish] wong, c.k., abraham, t., karimi, p., ow-wing, c. (2014). strain counter strain technique to decrease tender point palpation pain compared to control conditions: a systematic review with meta-analysis. journal of bodywork and movement �erapies, 18(2), 165–173. https://doi.org/10.1016/j.jbmt.2013.09.010 o steopathic capabilities for headache therapy – a short overview 201 appendix 1 fig. 1. classical osteopathic practices in the �eld of headache therapy: a – neck ligament relaxation, b – positional relaxation of the sternocleidomastoid muscle and the inclined muscles, c – stretching of the cervico-thoracic fascia, d – relaxation of the broad neck muscle, e – relaxation of the temporal muscle, f – relaxing the masseter muscle (photo. s. adamczyk) s ab in a a da m cz yk 202 osteopatyczne możliwości terapii bólów głowy – krótki przegląd streszczenie celem przeprowadzonego tu krótkiego przeglądu było usystematyzowanie, a także wykazanie skuteczności technik osteopatycznych w różnego typu terapiach bólu głowy. zestawione powyżej eksperymenty potwierdzają skuteczność manipulacyjnej terapii osteopatycznej omt w tym zakresie. leczenie osteopatyczne w całym swym wachlarzu stosowanych technik, daje ogromne możliwości w redukcji bólu i poprawy samopoczucia osobom cierpiącym, szczególnie na migreny oraz napięciowe bóle głowy. analizowane wyniki wskazują także, że zalecana terapia stanowi bardzo dobre uzupełnienie standardowej terapii lekowej, dzięki czemu zażywanie dużych dawek leków można znacznie ograniczyć. badacze tych zagadnień zwracają jednocześnie uwagę na fakt, aby przyszłe eksperymenty były planowane z odpowiednio dużą wielkością próby. w celu poprawienia jakości dobrej praktyki medycznej, zachęcają również osteopatów do stosowania systematycznej oceny zdarzeń niepożądanych w dobranej odpowiednio terapii manipulacyjnej oraz całościowego, interdyscyplinarnego podejścia do leczenia bólu. key words: alternative medicine, headaches, osteopathic manipulative treatment, om� received: [2021.02.04] accepted: [2021.04.20] 25 annales universitatis paedagogicae cracoviensis studia naturae, 6: 25–47, 2021, issn 2543-8832 doi: 10.24917/25438832.6.2 anna sołtys-lelek1*, wojciech gruszka2 1ojców national park, 32-045 sułoszowa, ojców 9, poland; *ana_soltys@wp.pl 2department of biological sciences, faculty of physical culture in gorzów wlkp., poznań university school of physical education, estkowskiego 13, 66-400 gorzów wlkp., poland roses and hawthorns in an urban area: a case study of gorzów wielkopolski in poland (nw poland) introduction a  record of the �ora in gorzów, poland, has not been coherently compiled. scarce �oristic data are presented in only a few publications. a total of 898 species of vascular plants (518 apophytes and 380 anthropophytes) have been found thus far. studying mainly synanthropic �ora and green areas of the city, the following species were identi�ed: crataegus monogyna jacq., c. laevigata (poir.) dc. (under the synonymic name crataegus oxyacantha l.), c. laevigata ‘paul’s scarlet’ (under the synonymic name c. oxyacantha l. paul’s scarlet), c. punctata jacq., c. crus-galli l., c. coccinea l., and c. ×carrieri fauvel. in the genus rosa, the following species were identi�ed: rosa rugosa �unb., r. canina l. (under the synonymic name r. dumetorum �uill.), r. rubiginosa l., and r. multi�ora �unb. misiewicz (1981, 1986). data on rose and hawthorn species occurring in the study area are also included in zając and zając (2001, 2019), where the following species were listed (generally in atpol squares): crataegus monogyna (ac67 square), c. rhipidophylla (ac57 square) and rosa canina (squares: ac57, ac67). as shown in the publication of sołtys-lelek and barabasz-krasny (2015), areas in the north and east of poland still require research and supplementation in terms of the occurrence of roses and hawthorns. �erefore, the aim of this research was to supplement the state of knowledge on the current distribution of the critical genera crataegus l. and rosa l. in this area, including presenting the full species composition and their distribution around gorzów wielkopolski. a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 26 study area � e city of gorzów wielkopolski, with an area of 86.3 km², is located on the warta river in north-western poland, in lubusz voivodeship. according to the physical and geographic classi� cation, it is situated within two mesoregions “gorzów plain” and “gorzów valley” (kondracki, 2001). � e average annual air temperature is 7.9 °c. � e warmest month is july (17.2 °c), and the coldest month is january (–1.4 °c). � e average annual precipitation is 500–600 mm (studium, 2014). � e length of the growing season is 220 days (misiewicz, 1981). � e city area is varied in terms of plant communities. aquatic and rush communities occur primarily in the warta river valley. meadow-pasture and tall-herb vegetation appears in many places and constitutes an important natural and landscape element in the city. however, most of the area is occupied by anthropogenic segregated and ruderal communities. moreover, there are 10 city parks and 24 larger arranged squares and green areas. � e city area is poor in forest areas. in the eastern areas of the city borders, there is one compact forest complex. moreover, a small fragment of forest complex enters the city area from the north. tree stands in this area are mainly pine, with some oak and birch. one of the most valuable natural areas of the city, where some remnants of naturalness have been preserved, is a mainstay of xerothermic � ora communities, which fig. 1. division of atpol (atlas of poland) squares into squares with a side of 1 km r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 27 has been legally protected as the “gorzowskie murawy” reserve since 2006 (studium, 2014; pyszny et al., 2018). materials and methods floristic materials were collected in 2017–2019 using the �eld cartogram method, based on a  grid of 10 × 10 km atpol squares. each square was divided into 100 smaller squares (1 km side) that were each treated as a single site. details of the atpol grid were adopted in accordance with methodological assumptions of the “distribution atlas of vascular plants in poland – atpol” (zając, 1978; komsta, 2016; verey, 2017 – fig. 1, appendix 1a, b). all herbarium materials were deposited in the herbarium of the university school of physical education, gorzow wielkopolski (poland). data from the literature were also included. �e taxonomic approach and nomenclature were based on the works of zieliński (1985, 1987), christensen (1992, 1997), popek (1996), and henker (2000). �e number of habitats was the basis used to de�ne the frequency of taxa occurrence: 1–4 stands – very rare species, 5–15 – rare species, 15–30 – not so frequent, 31–50 – quite frequent, 51–80 – frequent, 81–100 – very frequent, > 100 – common. abbreviations used in the list of species: (n) – new taxa for the study area, * – anthropophyte, ◊ – cultivated species. results �e list contains 11 hawthorn taxa, including 6 native taxa of the genus crataegus l., belonging to 2 subseries erianthae and crataegus, and 5 classi�ed as cultivated ornamental taxa. moreover, 14 taxa of the genus rosa l. belonged to the sections cinnamomeae dc. (1 species), caninae dc. em. h. christ (11 taxa), and synstylae dc. (1 species). among them, there were 10 native species, including 2 native hybrid forms at the rank of species; 2 anthropophytes; 1 cultivated species; and 1 hybrid form. a total of 13 taxa new to the �ora of the study area were found: 3 hawthorn taxa and 10 rose taxa. a detailed list of localities with gps coordinates is provided in appendix 1 for the genus crataegus and in appendix 2 for the genus rosa. genus crataegus l. ser. crataegus subser. erianthae 1. crataegus laevigata (poiret) dc. synon.: c. oxyacantha l. a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 28 rare species. �is species was reported in the study area by misiewicz (1981) under the synonymic name crataegus oxyacantha l., as generally rare on roadsides and in thickets and parks. it was not found by the authors of this paper. misiewicz (1986) reported at 3 localities: ac6706, ac6715, ac6716 subser. crataegus 2. crataegus rhipidophylla gand. var. rhipipophylla very rare species – 3 records from 3 localities. species previously reported in atpol square ac57 (zając, zając, 2019) 3. crataegus monogyna jacq. var. monogyna quite frequent species – 75 records in 45 stands. species previously reported in atpol square ac67 (zając, zając, 2001) and generally, by misiewicz (1981, 1986). 4. crataegus ×macrocarpa hegetschw nothovar. macrocarpa (n) [c. laevigata (poiret) dc. × c. rhipidophylla gand.var. rhipidophylla] very rare hybrid – 2 records from 2 localities. 5. crataegus ×subsphaericea gand. nothovar. subsphaericea (n) [c. monogyna jacq. × c. rhipidophylla gand. var. rhipidophylla] not so frequent hybrid – 19 records from 16 localities. 6. crataegus ×media bechst. (n) [c. laevigata (poiret) dc. × c. monogyna jacq.] very rare hybrid – 1 record from 1 locality. ornamental, cultivated taxa of hawthorns 7. ◊crataegus laevigata ‘paul’s scarlet’ rare variety reported by misiewicz (1986) as c. oxyacantha l. ‘paul’s scarlet’ – 9 localities: ac5797, ac6704, ac6705, ac6706, ac6707, ac6715, ac6716, ac6723, ac6726, and as element of urban street greenery. 8. *crataegus punctata jacq. very rare species reported by misiewicz (1986) – 1 locality: ac6726 9. *crataegus crus-galli l. very rare species reported by misiewicz (1986) – 1 locality: ac6715 10. *crataegus coccinea l. very rare species reported by misiewicz (1986) – 2 localities: ac6715, ac6716 and as element of urban street greenery. 11. ◊crataegus ×carrieri fauvel. very rare hybrid reported by misiewicz (1986) – 1 locality: ac6715 and as element of urban street greenery. r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 29 fig. 2. rosa dumalis bechst. var. caballicensis (puget) boulenger; a – part of fruiting short shoot, b – thorn, c – glandular fruit, d – fruit, styles of semi-circular head type, e – double-serrated leaf margin, f – part of leaf (underside), hairless. solid bar = 1 cm (photo. 2018, a. sołtys-lelek, specimen from poland, gorzów wielkopolski city, leg. 2017, w. gruszka) genus rosa l. i. sect. cinnamomeae dc. 1. *rosa rugosa �unb. very rare species (anthropophyte) – 5 records in 4 stands. species previously reported by misiewicz (1981, 1986) as occasional on escarpments, roadsides, and embankments. a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 30 ii. sect. caninae dc. em. h. christ. 2. rosa dumalis bechst. (n) syn.: r. afzeliana fr., r. glauca vill, r. caesia sm., r. coriifolia fr. not so frequent species obtained from 25 localities as 4 varieties: a) var. afzeliana (fr.) boulenger rare variety – 10 records from 9 localities. b) var. caballicensis (puget) boulenger (fig. 2) very rare variety – 1 record from 1 locality. c) var. dumalis rare variety – 12 records from 11 localities. d) var. acharii (billb.) boulenger very rare variety – 1 record from 1 locality. e) var. coriifolia (fr.) boulenger very rare variety – 3 records from 3 localities. 3. rosa villosa l. var. villosa (n) very rare species – 1 record from 1 locality. 4. rosa sherardii davies (n) very rare species, obtained from 4 localities as 2 varieties: a) var. sherardii very rare variety – 3 records from 3 localities. b) var. collivaga (cottet) boulenger very rare variety – 1 record from 1 locality. 5. rosa tomentosa sm. var. cinerascens (dumurt.) crépin (n) very rare variety – 1 record from 1 locality. 6. rosa rubiginosa l. rare species, obtained from 10 localities as 2 varieties: a) var. rubiginosa very rare variety – 1 record from 1 locality. b) var. umbellata (leers) dumort. rare variety – 9 records from 5 localities. species previously reported by misiewicz (1981, 1986) as occasional in thickets and on roadside escarpments and buttresses. 7. rosa inodora fr. var. indora (n) very rare species – 4 records from 4 localities. 8. rosa canina l. common species, formerly reported by misiewicz (1981) as r. dumetórum �uill. later, reported by zając, zając, (2001) from atpol squares ac57 and ac67. currently found as �ve varieties at 115 localities: a) var. canina r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 31 rare variety – 9 records from 8 localities. b) var. andegavensis (bastard.) desp. very rare variety – 4 records from 4 localities. c) var. dumalis baker frequent variety – 132 records from 57 localities. d) var. deseglisei (boreau) crépin very rare variety – 2 records from 2 localities. e) var. corymbifera (borkh.) boulenger not so frequent variety – 26 records from 19 localities. 9. rosa jundzillii besler var. jundzillii (n) (fig. 3a – appendix 2) very rare variety – 3 records from 3 localities. 10. rosa ×subcanina (h. christ) vok. (n) hybrid originating from r. dumalis bechst. × r. canina l. with bare leaves that are not hairy. it was reported here following henker (2000) in species rank. not so frequent taxon – 16 records from 16 localities. 11. rosa ×subcollina (h. christ) vok. (n) hybrid originating from r. dumalis bechst. × r. canina l. with hairy leaves. it was reported here following henker (2000) in species rank. very rare taxon – 5 records from 4 localities. 12. rosa canina l. × r. rubiginosa l. (n) very rare hybrid – 1 record from 1 locality. iii. sect. synstylae dc. 13. *rosa multi�ora �unb. very rare species (anthropophyte) – 5 records from 4 stands. species previously reported by misiewicz (1986) as planted in green areas. old garden hybrids occurring as semi-wild 14. ◊rosa ×francofourtana muenchh. (n) (fig. 3b – appendix 2) very rare hybrid – 1 record from 1 locality. discussion within the study area, the list of species in the genus crataegus and rosa comprised a total of 25 taxa, including 11 hawthorn species and 14 rose taxa. among those identi�ed, as many as 13 taxa were new to the �ora of gorzów wielkopolski. �ese were: crataegus ×macrocarpa, c. ×subsphaericea, c. ×media, rosa dumalis, r. villosa, r. sherardii, r. tomentosa, r. inodora, r. jundzillii, r. ×subcanina, r. ×subcollina, an old, cultivated variety of r. ×francofourtana, and the interspeci�c hybrid rosa canina × r. rubiginosa. a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 32 �e study also provided detailed data on both the distribution of localities (appendix 1, 2) and the number of localities of species already reported in the area. �is allowed a  more precise determination of the frequency of occurrence and provided a more accurate picture of their distribution in the study area. �is applies, for example, to rosa rubiginosa or r. canina, which are common species in poland, that were previously reported from a single or few localities (misiewicz, 1981, 1986). �e data obtained during the study con�rmed the poor state of knowledge regarding this area thus far. as indicated previously, some regions, e.g., the northern part of the country, require additional research and supplementation in terms of the occurrence of roses (sołtys-lelek, barabasz-krasny, 2015) and hawthorns. regarding the frequency of rose and hawthorn occurrence in the study area, 17 taxa were classi�ed as very rare and were found in 4 localities, at most. �e rarest taxa were those found at single localities: crataegus ×media, c. crus-galli, rosa tomentosa, r. villosa, r. ×francofourtana, and the hybrid rosa canina × r. rubiginosa. �e most common species were rosa canina (115 localities), crataegus monogyna (45 localities), and rosa dumalis (25 localities). �ese were also the most common species of hawthorns and roses growing wild in poland. during �eld research, hybrid forms were found with characteristics that were intermediate between r. canina and r. dumalis. additionally, forms that di�ered slightly in appearance from typical r. dumalis specimens were identi�ed. according to some authors (henker, 2000), they are attributed to a  hybrid origin (r. dumalis × r. canina) and distinguished as separate taxa; r. ×subcanina had bare, non-hairy leaves, and r. ×subcollina had hairy leaves. according to other authors (zieliński, 1985, 1987), these are only morphologically extreme groups of r. dumalis forms that, when combined with its typical specimens, comprise numerous intermediate individuals. in the present study, the concept by henker (2000) was adopted to distinguish them as separate taxa. �e highest intraspeci�c variation was recorded for rosa canina and r. dumalis, within which 5 varieties each were found. among them, the most noteworthy were rosa canina varieties, which are uncommon in poland (popek, 1996). �ese included: r. canina var. andegavensis with glandular peduncles and bare non-hairy leaves or r. canina var. deseglisei with glandular peduncles and hairy leaves. r. canina var. andegavensis has thus far been known only in the wielkopolska lowland, the sudeten foreland, the płock upland, and piła, and r. canina var. deseglisei from the szczecin, silesian and cracow uplands, the nida basin, the silesian beskid, and piła (popek, 1996; sołtys-lelek, 2011, 2012; sołtys-lelek, gruszka, 2016). other more interesting varieties of roses found in this area undoubtedly include rosa dumalis. r. dumalis var. acharii has glandular peduncles and sepals, and its fruits and leaves are complexly glandular and saw-toothed. in contrast, r. dumalis var. r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 33 caballicensis has glandular peduncles and sepals, but its leaves are singly or doubly saw-toothed. r. dumalis var. acharii has been reported in the miechowska and lubelska uplands and the pieniny mountains. it also occurs sporadically in central and southern europe (popek, 1996). however, r. dumalis var. caballicensis is known only from western pomerania near chojnice (popek, 1996). rosa sherardii var. collivaga, with leaves and peduncles that are not glandular, belongs to the group of rare taxa in poland. it has been reported mainly from western and northern (near kłodzko, wałbrzych, chojnów, zielona góra, szczecin, and piła) and southern poland (pieniny and cracow-częstochowa upland; popek, szeląg, 1993; popek, 1996; sołtys-lelek, 2011; sołtys-lelek, gruszka, 2016). furthermore, an interesting species found in the study area was rosa jundzillii. it is a rare species in poland, occurring mainly in the southern part of the country (sudeten foothills, silesian lowland, and kielce-sandomierska upland), and is distributed in the central part of the country. in the gorzów upland, it has not been reported thus far (zając, zając, 2001; popek, 2002). however, this site is located in a long-urbanized area; thus, its origin is not fully known. it is likely that it may be a remnant of former cultivation. among hawthorns, the most interesting taxa are crataegus ×subsphaericea and crataegus ×macrocarpa, which are mainly reported in the south and east of poland, while information about their occurrence in the northern part of the country is scarce (christensen, 1992; sołtys-lelek, 2011, 2012; oklejewicz, vončina, 2012; oklejewicz et al., 2013, 2014, 2015; sołtys-lelek, gruszka, 2016, 2020; zając, zając, 2019). crategus laevigata, which was previously reported by misiewicz (1981, 1986), could not be found in this study. it is a common species in poland, and the studied area is within its range. �e localities where it occurred may have been transformed due to urban development of the city. however, specimens of this genus may be found in the future. �e species of the studied genus are a valuable element of the city �ora. �e �ora that occurs here is characterised by speci�c conditions in areas that are urbanised to varying degrees. �e �ora of gorzów wielkopolski, on the one hand, is formed spontaneously, because of natural succession, and on the other hand, is also consciously cultivated by man. �e following plants were planted in the study area for their decorative value: rosa rugosa, r. multi�ora, r. ×francofourtana, crataegus laevigata ‘paul’s scarlet’, c. punctata, c. crus-galli, c. coccinea and c. ×carrieri. for rose and hawthorn species, the greatest number of localities was found in habitats described as wastelands. �ey also grow quite numerously on roadsides. �e two most common species in poland – rosa canina and r. dumalis – were found in all distinguished habitat types, such as wastelands, roadsides, forest edges, and urban green areas. additionally, the localities of crataegus monogyna and crataegus ×subsphaericea were recorded in a wide range of habitats, mainly the anthropogenic ones mentioned a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 34 above. only crataegus ×media was found in the natural habitat at the forest edge. �e results obtained in this study were consistent with the literature data, which con�rm the preference of roses and hawthorns in open, thermophilic, and heliophilic habitats (e.g., zieliński, 1987; christensen, 1996; popek, 2002; barabasz-krasny, sołtys-lelek, 2011; sołtys-lelek, 2011, 2012; oklejewicz et al., 2013, 2014, 2015; sołtys-lelek, gruszka, 2016, 2020). in general, a set of native hawthorn species and more than half of the rose species reported in poland (considering only native species) were found within the study area. such a result indicates the high species richness of a small area and human-transformed study site. acknowledgements �e authors of this paper would like to thank mr. krzysztof witków and mr. tadeusz gruszka for their help in �eld research. con�ict of interest �e authors declare no con�ict of interest related to this article. references barabasz-krasny, b., sołtys-lelek, a. 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(1987). rosa l. in: a. jasiewicz (ed.), flora of poland, 5, 48 pp. kraków: institute of botany, polish academy of sciences, 5. r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 37 appendix 1 a) stands of crataegus species crataegus monogyna var. monogyna 1. ac5771 (52°46ʹ02,4ʺ 15°09ʹ43,6ʺ) 2. ac5775 (52°46ʹ04,4ʺ 15°13ʹ23,2ʺ) 3. ac5781 (52°45ʹ41,4ʺ 15°09ʹ45,5ʺ) 4. ac5784 (52°45ʹ29,2ʺ 15°12ʹ41,1ʺ; 52°45ʹ38,7ʺ 15°12ʹ58,5ʺ) 5. ac5785 (52°45ʹ43,5ʺ 15°13ʹ27,7ʺ) 6. ac5786 (52°45ʹ26,1ʺ 15°14ʹ15,3ʺ) 7. ac5788 (52°45ʹ34,9ʺ 15°16ʹ33,0ʺ; 52°45ʹ43,2ʺ 15°16ʹ36,2ʺ; 52°45ʹ56,4ʺ 15°15ʹ58,1ʺ) 8. ac5789 (52°45ʹ31,7ʺ 15°14ʹ19,2ʺ) 9. ac5793 (52°44ʹ58,6ʺ 15°11ʹ42,8ʺ; 52°45ʹ09,6ʺ 15°12ʹ17,3ʺ) 10. ac5794 (52°45ʹ14,8ʺ 15°12ʹ34,8ʺ; 52°45ʹ16,1ʺ 15°12ʹ45,1ʺ) 11. ac5795 (52°45ʹ06,5ʺ 15°13ʹ30,6ʺ; 52°45ʹ23,9ʺ 15°13ʹ42,0ʺ) 12. ac5796 (52°45ʹ05,0ʺ 15°14ʹ17,0ʺ) 13. ac5797 (52°45ʹ12,5ʺ 15°15ʹ10,4ʺ; 52°45ʹ19,2ʺ 15°15ʹ21,3ʺ; 52°45ʹ20,8ʺ 15°15ʹ22,0ʺ) 14. ac5798 (52°45ʹ24,2ʺ 15°15ʹ58,1ʺ; 52°45ʹ25,4ʺ 15°16ʹ40,4ʺ) 15. ac6608 (52°44ʹ34,3ʺ 15°07ʹ56,9ʺ) 16. ac6629 (52°43ʹ20,5ʺ 15°08ʹ21,3ʺ) 17. ac6639 (52°42ʹ40,3ʺ 15°08ʹ46,5ʺ) 18. ac6700 (52°44ʹ17,7ʺ 15°09ʹ44,3ʺ; 52°44ʹ35,6ʺ 15°09ʹ18,2ʺ) 19. ac6703 (52°44ʹ22,0ʺ 15°12ʹ25,5ʺ; 52°44ʹ40,2ʺ 15°12ʹ02,4ʺ; 52°44ʹ44,5ʺ 15°12ʹ15,5ʺ) 20. ac6704 (52°44ʹ32,3ʺ 15°12ʹ39,7ʺ; 52°44ʹ45,3ʺ 15°12ʹ26,3ʺ; 52°44ʹ46,2ʺ 15°13ʹ16,5ʺ; 52°44ʹ46,3ʺ 15°12ʹ32,0ʺ) 21. ac6705 (52°44ʹ42,8ʺ 15°14ʹ07,0ʺ) 21. ac6706 (52°44ʹ47,7ʺ 15°14ʹ24,6ʺ) 22. ac6708 (52°44ʹ45,5ʺ 15°16ʹ24,1ʺ; 52°44ʹ51,6ʺ 15°16ʹ34,1ʺ) 23. ac6711 (52°44ʹ02,6ʺ 15°10ʹ28,9ʺ) 24. ac6712 (52°43ʹ54,1ʺ 15°11ʹ11,3ʺ; 52°43ʹ54,5ʺ 15°10ʹ43,0ʺ; 52°43ʹ55,1ʺ 15°11ʹ23,6ʺ; 52°43ʹ57,2ʺ 15°11ʹ34,6ʺ; 52°44ʹ00,3ʺ 15°11ʹ17,3ʺ) 25. ac6715 (52°44ʹ01,4ʺ 15°13ʹ37,8ʺ; 52°44ʹ04,4ʺ 15°13ʹ35,8ʺ; 52°44ʹ06,6ʺ 15°13ʹ36,6ʺ) 27. ac6716 (52°44ʹ00,0ʺ 15°15ʹ00,4ʺ; 52°44ʹ09,4ʺ 15°14ʹ52,3ʺ; 52°44ʹ19,1ʺ 15°14ʹ47,3ʺ) 28. ac6717 (52°44ʹ03,7ʺ 15°15ʹ57,8ʺ) 29. ac6718 (52°44ʹ20,3ʺ 15°16ʹ50,2ʺ) 30. ac6719 (52°44ʹ05,7ʺ 15°16ʹ57,4ʺ) 31. ac6722 (52°43ʹ24,8ʺ 15°11ʹ04,9ʺ; 52°43ʹ26,8ʺ 15°11ʹ01,0ʺ) a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 38 32. ac6723 (52°43ʹ28,7ʺ 15°11ʹ39,6ʺ; 52°43ʹ42,1ʺ 15°11ʹ39,4ʺ) 33. ac6724 (52°43ʹ40,3ʺ 15°13ʹ07,1ʺ) 34. ac6727 (52°43ʹ22,8ʺ 15°15ʹ13,4ʺ) 35. ac6730 (52°42ʹ49,9ʺ 15°09ʹ06,0ʺ) 36. ac6731 (52°42ʹ41,9ʺ 15°10ʹ06,4ʺ; 52°42ʹ44,0ʺ 15°09ʹ59,4ʺ; 52°42ʹ44,8ʺ 15°10ʹ04,1ʺ) 37. ac6732 (52°43ʹ04,9ʺ 15°10ʹ54,2ʺ) 38. ac6736 (52°42ʹ47,9ʺ 15°14ʹ34,0ʺ; 52°43ʹ00,0ʺ 15°14ʹ37,6ʺ) 39. ac6739 (52°42ʹ50,7ʺ 15°17ʹ52,3ʺ) 40. ac6747 (52°42ʹ22,8ʺ 15°15ʹ37,8ʺ) 41. ac6749 (52°42ʹ40,1ʺ 15°17ʹ26,5ʺ) 42. ac6754 (52°41ʹ58,2ʺ 15°13ʹ14,4ʺ) 43. ac6756 (52°42ʹ00,7ʺ 15°14ʹ46,3ʺ) 44. ac6757 (52°42ʹ00,4ʺ 15°15ʹ47,5ʺ) 45. ac6769 (52°41ʹ40,6ʺ 15°17ʹ14,8ʺ) c. rhipidophylla var. rhipidophylla 46. ac5776 (52°46ʹ13,0ʺ 15°14ʹ07,3ʺ) 47. ac6619 (52°44ʹ09,0ʺ 15°08ʹ36,4ʺ) 48. ac6712 (52°44ʹ08,6ʺ 15°11ʹ25,0ʺ) c. ×media notha var. media 49. ac6729 (52°43ʹ52,8ʺ 15°17ʹ45,5ʺ) c. ×macrocarpa notha var. macrocarpa 50. ac5793 (52°45ʹ09,7ʺ 15°12ʹ13,7ʺ) 51. ac6745 (52°42ʹ13,8ʺ 15°14ʹ06,7ʺ) c. ×subsphaericea notha var. subsphaericea 52. ac5775 (52°46ʹ02,7ʺ 15°13ʹ44,9ʺ; 52°46ʹ16,5ʺ 15°13ʹ57,6ʺ) 53. ac5784 (52°45ʹ26,4ʺ 15°12ʹ45,8ʺ) 54. ac5788 (52°45ʹ57,1ʺ 15°16ʹ07,9ʺ) 55. ac5793 (52°45ʹ09,0ʺ 15°12ʹ19,7ʺ) 56. ac5796 (52°45ʹ02,7ʺ 15°14ʹ41,7ʺ) 57. ac6629 (52°43ʹ32,2ʺ 15°08ʹ33,4ʺ) 58. ac6701 (52°44ʹ16,9ʺ 15°10ʹ38,1ʺ) 59. ac6709 (52°44ʹ36,3ʺ 15°17ʹ03,0ʺ) 60. ac6712 (52°43ʹ54,5ʺ 15°10ʹ55,9ʺ) 61. ac6713 (52°44ʹ03,7ʺ 15°12ʹ07,4ʺ; 52°43ʹ48,7ʺ 15°12ʹ27,4ʺ) r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 39 62. ac6714 (52°43ʹ55,0ʺ 15°13ʹ02,8ʺ; 52°43ʹ52,2ʺ 15°12ʹ44,6ʺ) 63. ac6722 (52°43ʹ24,2ʺ 15°11ʹ11,7ʺ) 64. ac6722 (52°43ʹ26,0ʺ 15°11ʹ23,8ʺ) 65. ac6724 (52°43ʹ40,3ʺ 15°12ʹ35,8ʺ) 66. ac6745 (52°42ʹ14,6ʺ 15°14ʹ07,4ʺ) 67. ac6747 (52°42ʹ30,2ʺ 15°15ʹ20,7ʺ) b) stands of rosa species rosa canina var. andegavensis 1. ac5785 (52°45ʹ34,0ʺ 15°13ʹ43,3ʺ) 2. ac5795 (52°45ʹ06,5ʺ 15°13ʹ30,6ʺ) 3. ac6718 (52°44ʹ09,3ʺ 15°16ʹ13,7ʺ) 4. ac6757 (52°41ʹ54,7ʺ 15°16ʹ08,0ʺ) rosa canina var. canina 5. ac5775 (52°46ʹ04,4ʺ 15°13ʹ23,2ʺ) 6. ac5794 52°45ʹ17,2ʺ 15°12ʹ44,0ʺ) 7. ac5797 (52°45ʹ16,8ʺ 15°15ʹ13,7ʺ) 8. ac6704 (52°44ʹ46,1ʺ 15°12ʹ26,4ʺ) 9. ac6706 (52°44ʹ49,5ʺ 15°14ʹ22,8ʺ) 10. ac6712 (52°43ʹ58,2ʺ 15°11ʹ29,9ʺ) 11. ac6715 (52°43ʹ58,7ʺ 15°13ʹ35,5ʺ) 12. ac6724 (52°43ʹ46,4ʺ 15°13ʹ22,0ʺ; 52°43ʹ43,5ʺ 15°12ʹ32,4ʺ) rosa canina var. corymbifera 13. ac5771 (52°46ʹ02,4ʺ 15°09ʹ43,6ʺ) 14. ac5793 (52°44ʹ53,3ʺ 15°11ʹ31,9ʺ; 52°44ʹ58,6ʺ 15°11ʹ42,8ʺ; 52°45ʹ09,6ʺ 15°12ʹ17,3ʺ) 15. ac5794 (52°44ʹ51,4ʺ 15°13ʹ09,3ʺ) 16. ac5798 (52°45ʹ24,2ʺ 15°15ʹ58,1ʺ) 17. ac6609 (52°44ʹ32,1ʺ 15°08ʹ0,09ʺ) 18. ac6701 (52°44ʹ16,9ʺ 15°10ʹ36,1ʺ; 52°44ʹ16,9ʺ 15°10ʹ38,1ʺ) 19. ac6702 (52°44ʹ18,3ʺ 15°11ʹ24,2ʺ) 20. ac6703 (52°44ʹ40,6ʺ 15°11ʹ48,4ʺ; 52°44ʹ40,4ʺ 15°12ʹ18,3ʺ) 21. ac6705 (52°44ʹ44,2ʺ 15°13ʹ23,9ʺ) 22. ac6707 (52°44ʹ26,7ʺ 15°15ʹ19,1ʺ; 52°44ʹ36,6ʺ 15°15ʹ31,6ʺ) 23. ac6712 (52°43ʹ54,5ʺ 15°10ʹ55,9ʺ; 52°43ʹ58,1ʺ 15°11ʹ18,6ʺ) 24. ac6714 (52°43ʹ45,5ʺ 15°12ʹ30,4ʺ) 25. ac6715 (52°44ʹ03,1ʺ 15°13ʹ25,8ʺ; 52°43ʹ54,2ʺ 15°13ʹ28,4ʺ) a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 40 26. ac6721 (52°43ʹ32,7ʺ 15°10ʹ38,9ʺ) 27. ac6722 (52°43ʹ20,4ʺ 15°11ʹ33,7ʺ) 28. ac6725 (52°43ʹ37,4ʺ 15°13ʹ47,3ʺ) 29. ac6730 (52°42ʹ58,9ʺ 15°09ʹ52,1ʺ) 30. ac6731 (52°42ʹ44,7ʺ 15°10ʹ27,8ʺ) 31. ac6747 (52°42ʹ30,2ʺ 15°15ʹ31,8ʺ) rosa canina var. deseglisei 32. ac6709 (52°44ʹ38,8ʺ 15°17ʹ03,3ʺ) 33. ac6714 (52°43ʹ48,2ʺ 15°13ʹ02,9ʺ) rosa canina var. dumalis 34. ac5699 (52°44ʹ48,1ʺ 15°08ʹ18,1ʺ) 35. ac5776 (52°46ʹ13,0ʺ 15°14ʹ07,3ʺ; 52°46ʹ04,2ʺ 15°14ʹ35,1ʺ) 36. ac5778 (52°46ʹ03,0ʺ 15°15ʹ57,9ʺ) 37. ac5781 (52°45ʹ41,5ʺ 15°09ʹ45,7ʺ) 38. ac5784 (52°45ʹ29,2ʺ 15°12ʹ41,1ʺ; 52°45ʹ26,4ʺ 15°12ʹ45,8ʺ; 52°45ʹ38,7ʺ 15°12ʹ58,5ʺ) 39. ac5785 (52°45ʹ43,5ʺ 15°13ʹ27,7ʺ) 40. ac5786 (52°45ʹ42,7ʺ 15°14ʹ58,0ʺ) 41. ac5788 (52°45ʹ56,4ʺ 15°15ʹ58,1ʺ; 52°45ʹ59,3ʺ 15°16ʹ19,1ʺ; 52°45ʹ35,6ʺ 15°16ʹ45,7ʺ; 52°45ʹ57,1ʺ 15°16ʹ07,9ʺ; 52°45ʹ34,9ʺ 15°16ʹ33,0ʺ) 42. ac5792 (52°44ʹ51,1ʺ 15°10ʹ58,8ʺ) 43. ac5794 (52°44ʹ50,5ʺ 15°12ʹ23,8ʺ; 52°45ʹ14,8ʺ 15°12ʹ34,8ʺ) 44. ac5795 (52°44ʹ59,4ʺ 15°13ʹ43,2ʺ; 52°45ʹ05,1ʺ 15°13ʹ24,0ʺ; 52°45ʹ19,7ʺ 15°13ʹ20,4ʺ; 52°45ʹ20,8ʺ 15°13ʹ14,7ʺ) 45. ac5796 (52°45ʹ08,4ʺ 15°14ʹ18,9ʺ) 46. ac5797 (52°45ʹ17,9ʺ 15°15ʹ49,5ʺ; 52°45ʹ09,6ʺ 15°15ʹ19,8ʺ) 47. ac5798 (52°45ʹ24,5ʺ 15°16ʹ23,6ʺ; 52°45ʹ14,0ʺ 15°16ʹ48,4ʺ; 52°45ʹ24,2ʺ 15°15ʹ58,1ʺ; 52°45ʹ28,8ʺ 15°16ʹ40,8ʺ; 52°45ʹ24,9ʺ 15°16ʹ37,6ʺ) 48. ac6608 (52°44ʹ34,3ʺ 15°07ʹ56,9ʺ) 49. ac6609 (52°44ʹ32,1ʺ 15°08ʹ0,09ʺ) 50. ac6619 (52°43ʹ57,2ʺ 15°08ʹ42,8ʺ) 51. ac6629 (52°43ʹ20,5ʺ 15°08ʹ21,3ʺ; 52°43ʹ32,2ʺ 15°08ʹ33,4ʺ) 52. ac6700 (52°44ʹ35,6ʺ 15°09ʹ18,2ʺ) 53. ac6701 (52°44ʹ16,9ʺ 15°10ʹ36,1ʺ; 52°44ʹ16,9ʺ 15°10ʹ38,1ʺ) 54. ac6702 (52°44ʹ43,0ʺ 15°11ʹ29,4ʺ) 55. ac6703 (52°44ʹ29,9ʺ 15°11ʹ48,2ʺ; 52°44ʹ37,1ʺ 15°12ʹ19,8ʺ; 52°44ʹ18,3ʺ 15°11ʹ38,1ʺ; 52°44ʹ33,8ʺ 15°12ʹ18,3ʺ) r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 41 56. ac6704 (52°44ʹ31,0ʺ 15°12ʹ31,6ʺ) 57. ac6705 (52°44ʹ42,4ʺ 15°14ʹ04,6ʺ; 52°44ʹ44,2ʺ 15°13ʹ26,4ʺ; 52°44ʹ24,0ʺ 15°13ʹ50,9ʺ; 52°44ʹ41,9ʺ 15°13ʹ58,7ʺ; 52°44ʹ42,8ʺ 15°14ʹ07,0ʺ; 52°44ʹ45,8ʺ 15°13ʹ43,4ʺ) 58. ac6706 (52°44ʹ50,1ʺ 15°14ʹ44,9ʺ) 59. ac6707 (52°44ʹ33,4ʺ 15°15ʹ13,8ʺ; 52°44ʹ40,4ʺ 15°15ʹ12,1ʺ) 60. ac6708 (52°44ʹ31,1ʺ 15°16ʹ09,0ʺ; 52°44ʹ35,8ʺ 15°16ʹ29,8ʺ; 52°44ʹ51,6ʺ 15°16ʹ34,1ʺ) 61. ac6710 (52°44ʹ11,0ʺ 15°09ʹ38,8ʺ; 52°44ʹ05,6ʺ 15°09ʹ27,9ʺ) 62. ac6711 (52°44ʹ08,9ʺ 15°10ʹ25,3ʺ) 63. ac6712 (52°43ʹ58,9ʺ 15°10ʹ47,3ʺ; 52°44ʹ00,3ʺ 15°11ʹ17,3ʺ; 52°43ʹ52,7ʺ 15°11ʹ09,1ʺ; 52°43ʹ51,4ʺ 15°11ʹ22,2ʺ) 64. ac6713 (52°44ʹ03,7ʺ 15°12ʹ07,4ʺ; 52°43ʹ58,8ʺ 15°12ʹ21,9ʺ; 52°44ʹ06,1ʺ 15°12ʹ26,5ʺ; 52°44ʹ08,8ʺ 15°12ʹ03,5ʺ; 52°44ʹ03,0ʺ 15°12ʹ02,9ʺ) 65. ac6714 (52°43ʹ55,1ʺ 15°12ʹ59,0ʺ; 52°43ʹ53,6ʺ 15°13ʹ01,0ʺ; 52°43ʹ51,8ʺ 15°12ʹ44,2ʺ; 52°43ʹ51,9ʺ 15°12ʹ44,4ʺ; 52°44ʹ10,9ʺ 15°13ʹ05,3ʺ) 66. ac6715 (52°44ʹ06,6ʺ 15°13ʹ36,6ʺ; 52°44ʹ02,7ʺ 15°13ʹ24,8ʺ; 52°44ʹ04,6ʺ 15°13ʹ34,6ʺ; 52°43ʹ54,4ʺ 15°13ʹ55,4ʺ) 67. ac6716 (52°43ʹ51,0ʺ 15°14ʹ35,5ʺ; 52°44ʹ15,3ʺ 15°14ʹ52,8ʺ) 68. ac6717 (52°44ʹ06,2ʺ 15°15ʹ28,6ʺ) 69. ac6718 (52°44ʹ13,1ʺ 15°16ʹ26,0ʺ; 52°44ʹ05,1ʺ 15°16ʹ11,2ʺ; 52°44ʹ16,4ʺ 15°16ʹ37,1ʺ; 52°44ʹ11,9ʺ 15°16ʹ45,9ʺ) 70. ac6719 (52°44ʹ10,7ʺ 15°16ʹ55,2ʺ) 71. ac6721 (52°43ʹ13,8ʺ 15°10ʹ29,5ʺ; 52°43ʹ37,5ʺ 15°10ʹ36,5ʺ) 72. ac6722 (52°43ʹ26,8ʺ 15°10ʹ56,0ʺ; 52°43ʹ15,7ʺ 15°11ʹ06,5ʺ; 52°43ʹ29,6ʺ 15°11ʹ03,8ʺ; 52°43ʹ21,5ʺ 15°11ʹ08,2ʺ; 52°43ʹ22,6ʺ 15°11ʹ08,8ʺ; 52°43ʹ36,4ʺ 15°11ʹ05,4ʺ; 52°43ʹ36,1ʺ 15°10ʹ57,7ʺ) 73. ac6723 (52°43ʹ28,7ʺ 15°11ʹ39,6ʺ; 52°43ʹ33,7ʺ 15°11ʹ52,3ʺ; 52°43ʹ33,1ʺ 15°12ʹ21,0ʺ; 52°43ʹ39,5ʺ 15°12ʹ12,3ʺ) 74. ac6724 (52°43ʹ40,9ʺ 15°13ʹ06,5ʺ; 52°43ʹ36,1ʺ 15°12ʹ35,5ʺ; 52°43ʹ40,7ʺ 15°12ʹ36,1ʺ) 75. ac6727 (52°43ʹ39,6ʺ 15°15ʹ15,3ʺ; 52°43ʹ26,9ʺ 15°15ʹ34,5ʺ; 52°43ʹ24,1ʺ 15°15ʹ16,4ʺ) 76. ac6729 (52°43ʹ52,8ʺ 15°17ʹ45,5ʺ) 77. ac6730 (52°42ʹ49,9ʺ 15°09ʹ06,0ʺ) 78. ac6731 (52°42ʹ44,7ʺ 15°10ʹ27,8ʺ; 52°42ʹ44,8ʺ 15°10ʹ04,1ʺ; 52°42ʹ44,0ʺ 15°09ʹ59,4ʺ; 52°42ʹ41,9ʺ 15°10ʹ06,2ʺ) 79. ac6732 (52°43ʹ05,4ʺ 15°10ʹ55,1ʺ) 80. ac6733 (52°43ʹ10,5ʺ 15°12ʹ24,5ʺ) a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 42 81. ac6736 (52°43ʹ16,7ʺ 15°14ʹ54,2ʺ; 52°42ʹ51,9ʺ 15°14ʹ39,7ʺ; 52°42ʹ51,9ʺ 15°15ʹ11,3ʺ; 52°42ʹ58,0ʺ 15°15ʹ12,8ʺ; 52°42ʹ47,3ʺ 15°14ʹ25,4ʺ) 82. ac6737 (52°42ʹ57,7ʺ 15°15ʹ17,4ʺ) 83. ac6745 (52°42ʹ15,1ʺ 15°14ʹ08,5ʺ) 84. ac6746 (52°42ʹ29,5ʺ 15°14ʹ22,9ʺ; 52°42ʹ35,5ʺ 15°15ʹ15,1ʺ) 85. ac6747 (52°42ʹ30,2ʺ 15°15ʹ31,8ʺ; 52°42ʹ22,0ʺ 15°15ʹ19,6ʺ; 52°42ʹ30,2ʺ 15°15ʹ20,7ʺ) 86. ac6756 (52°42ʹ00,7ʺ 15°14ʹ46,3ʺ;52°42ʹ08,4ʺ 15°14ʹ42,6ʺ) 87. ac6758 (52°41ʹ44,0ʺ 15°16ʹ44,4ʺ) 88. ac6765 (52°41ʹ36,6ʺ 15°13ʹ41,1ʺ) 89. ac6766 (52°41ʹ27,9ʺ 15°14ʹ59,3ʺ) 91. ac6769 (52°41ʹ40,4ʺ 15°17ʹ21,6ʺ) rosa canina × r. rubiginosa 92. ac6703 (52°44ʹ18,3ʺ 15°11ʹ35,6ʺ) rosa dumalis var. acharii 93. ac6715 (52°44ʹ06,6ʺ 15°13ʹ34,6ʺ) rosa dumalis var. afzeliana 94. ac5793 (52°45ʹ09,0ʺ 15°12ʹ19,7ʺ) 95. ac5794 (52°45ʹ16,7ʺ 15°12ʹ44,4ʺ; 52°45ʹ17,7ʺ 15°12ʹ52,1ʺ) 96. ac6713 (52°43ʹ59,6ʺ 15°11ʹ51,4ʺ) 97. ac6717 (52°44ʹ00,2ʺ 15°15ʹ36,2ʺ) 98. ac6722 (52°43ʹ19,9ʺ 15°11ʹ21,2ʺ) 99. ac6725 (52°43ʹ31,1ʺ 15°14ʹ04,6ʺ) 100. ac6736 (52°43ʹ11,2ʺ 15°14ʹ30,6ʺ) 101. ac6746 (52°42ʹ43,5ʺ 15°14ʹ35,3ʺ) 102. ac6747 (52°42ʹ14,8ʺ 15°15ʹ41,8ʺ) rosa dumalis var. caballicensis 103. ac5793 (52°44ʹ53,3ʺ 15°11ʹ31,9ʺ) rosa dumalis var. coriifolia 104. ac5776 (52°46ʹ03,7ʺ 15°14ʹ40,1ʺ) 105. ac6724 (52°43ʹ13,4ʺ 15°12ʹ46,1ʺ) 106. ac6736 (52°42ʹ57,9ʺ 15°14ʹ41,4ʺ) r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 43 rosa dumalis var. dumalis 107. ac5797 (52°44ʹ58,8ʺ 15°15ʹ18,0ʺ) 108. ac6639 (52°42ʹ39,4ʺ 15°08ʹ47,2ʺ) 109. ac6703 (52°44ʹ30,2ʺ 15°12ʹ12,4ʺ) 110. ac6712 (52°43ʹ48,3ʺ 15°11ʹ21,6ʺ) 111. ac6714 (52°44ʹ13,5ʺ 15°13ʹ05,2ʺ) 112. ac6715 (52°44ʹ06,6ʺ 15°13ʹ36,6ʺ) 113. ac6722 (52°43ʹ26,8ʺ 15°10ʹ56,0ʺ; 52°43ʹ26,8ʺ 15°11ʹ01,0ʺ) 114. ac6723 (52°43ʹ33,1ʺ 15°12ʹ13,4ʺ) 115. ac6731 (52°42ʹ41,5ʺ 15°10ʹ07,1ʺ) 116. ac6749 (52°42ʹ31,9ʺ 15°17ʹ26,4ʺ) 117. ac6756 (52°42ʹ00,7ʺ 15°14ʹ46,3ʺ) rosa inodora var. inodora 118. ac6702 (52°44ʹ17,9ʺ 15°11ʹ23,5ʺ) 119. ac6712 (52°43ʹ44,3ʺ 15°11ʹ33,5ʺ) 120. ac6731 (52°42ʹ41,4ʺ 15°10ʹ08,3ʺ) 121. ac6745 (52°42ʹ14,5ʺ 15°14ʹ06,1ʺ) rosa jundzillii var. jundzillii 122. ac5788 (52°45ʹ35,7ʺ 15°16ʹ29,8ʺ) 123. ac5794 (52°45ʹ16,7ʺ 15°12ʹ44,4ʺ) 124. ac6722 (52°43ʹ20,7ʺ 15°11ʹ02,7ʺ) rosa multi�ora 125. ac5793 (52°44ʹ50,5ʺ 15°12ʹ23,4ʺ) 126. ac5894 (52°45′12,2″ 15°12′27,3″) 127. ac6726 (52°43ʹ40,6ʺ 15°14ʹ42,3ʺ; 52°43′23,4″ 15°14′52,8″) 128. ac6739 (52°42ʹ50,7ʺ 15°17ʹ52,3ʺ) rosa rubiginosa var. rubiginosa 129. ac6702 (52°44ʹ17,7ʺ 15°11ʹ12,8ʺ) rosa rubiginosa var. umbellata 130. ac5795 (52°45ʹ23,7ʺ 15°13ʹ43,2ʺ) 131. ac6703 (52°44ʹ19,8ʺ 15°12ʹ22,6ʺ; 52°44ʹ27,2ʺ 15°12ʹ13,9ʺ; 52°44ʹ27,2ʺ 15°12ʹ13,4ʺ; 52°44ʹ30,2ʺ 15°12ʹ12,4ʺ) 132. ac6704 (52°44ʹ46,3ʺ 15°12ʹ32,0ʺ; 52°44ʹ44,1ʺ 15°12ʹ44,0ʺ) 133. ac6712 (52°43ʹ55,1ʺ 15°11ʹ23,6ʺ) 134. ac6757 (52°41ʹ49,5ʺ 15°15ʹ54,3ʺ) a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 44 rosa rugosa 135. ac5788 (52°45′33,5″ 15°16′41,1; 52°45′39,9″ 15°16′40,7″) 136. ac6716 (52°44′17,4″ 15°14′48,7″) 137. ac6731 (52°42′43,6″ 15°10′02,2″) 138. ac6732 (52°43′03,5″ 15°10′52,4″) rosa sherardii var. collivaga 139. ac6714 (52°43ʹ52,2ʺ 15°12ʹ44,6ʺ) rosa sherardii var. sherardii 140. ac6703 (52°44ʹ18,3ʺ 15°11ʹ38,1ʺ) 141. ac6747 (52°42ʹ32,2ʺ 15°15ʹ30,3ʺ) 142. ac6756 (52°42ʹ05,4ʺ 15°14ʹ32,6ʺ) rosa tomentosa var. cinerscens 143. ac6736 (52°43ʹ07,9ʺ 15°14ʹ31,8ʺ) rosa villosa var. villosa 144. ac5788 (52°45ʹ31,7ʺ 15°16ʹ44,2ʺ) rosa ×francofourtana 145. ac6756 (52°42ʹ08,4ʺ 15°14ʹ42,6ʺ) rosa ×subcollina 146. ac6707 (52°44ʹ32,4ʺ 15°15ʹ30,1ʺ) 147. ac6712 (52°43ʹ52,7ʺ 15°11ʹ9,1ʺ) 148. ac6722 (52°43ʹ20,6ʺ 15°11ʹ34,4ʺ) 149. ac6736 (52°43ʹ07,5ʺ 15°14ʹ31,1ʺ; 52°43ʹ00,7ʺ 15°14ʹ32,8ʺ) rosa ×subcanina 150. ac5795 (52°44ʹ53,7ʺ 15°13ʹ42,3ʺ) 151. ac5796 (52°45ʹ02,5ʺ 15°14ʹ55,2ʺ) 152. ac5797 (52°45ʹ01,6ʺ 15°15ʹ21,6ʺ) 153. ac5798 (52°45ʹ22,9ʺ 15°16ʹ36,8ʺ) 154. ac5799 (52°45ʹ17,2ʺ 15°16ʹ51,4ʺ) 155. ac6629 (52°43ʹ27,9ʺ 15°08ʹ29,3ʺ) 156. ac6703 (52°44ʹ21,0ʺ 15°12ʹ23,3ʺ) 157. ac6705 (52°44ʹ46,6ʺ 15°13ʹ43,4ʺ) 158. ac6708 (52°44ʹ45,5ʺ 15°16ʹ24,1ʺ) r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 45 159. ac6711 (52°44ʹ00,2ʺ 15°10ʹ25,7ʺ) 160. ac6717 (52°44ʹ19,7ʺ 15°15ʹ56,1ʺ) 161. ac6718 (52°44ʹ09,7ʺ 15°16ʹ22,4ʺ) 162. ac6722 (52°43ʹ24,2ʺ 15°11ʹ11,7ʺ) 163. ac6724 (52°43ʹ36,8ʺ 15°12ʹ37,6ʺ) 164. ac6736 (52°42ʹ55,0ʺ 15°14ʹ21,7ʺ) 165. ac6746 (52°42ʹ31,7ʺ 15°14ʹ29,0ʺ) a nn a s oł ty sle le k, w oj ci ec h g ru sz ka 46 appendix 2 fig. 3. rosa jundzillii besler var. jundzillii, specimen from poland, gorzów wielkopolski city, zwiadowców street (a); rosa ×francofourtana muenchh., specimen from poland, gorzów wielkopolski city, podgórna street (b) (photo. 2018, w. gruszka) r oses and haw thorns in an urban area: a case study of g orzów w ielkopolski in p oland (n w p oland) 47 abstract �is paper presents the distribution of species of the genera crataegus and rosa in gorzów wielkopolski, poland. based on literature data and �eld research, 25 taxa were found to occur here, comprising 11 hawthorn species and 14 rose taxa. among the taxa of hawthorns, 6 were native taxa belonging to 2 subseries erianthae and crataegus, and 5 were classi�ed as cultivated ornamental taxa. among the taxa of genus rosa l., there were 10 native species (including 2 native hybrid forms at the rank of species), 2 anthropophytes, 1 cultivated species, and 1 hybrid form. among those identi�ed, as many as 13 taxa were new to the �ora of gorzów wielkopolski. �ese were crataegus ×macrocarpa, c. ×subsphaericea, c. ×media, rosa dumalis, r. villosa, r. sherardii, r. tomentosa, r. inodora, r. jundzillii, r. ×subcanina, r. ×subcollina, the old, cultivated variety r. ×francofourtana, and an interspeci�c hybrid rosa canina × r. rubiginosa. �e occurrence of rosa jundzillii, a species rare in poland and not previously reported in the gorzów upland, was noted during the study. �e following varieties of roses that are rare in poland were also found: r. dumalis var. acharii, r. dumalis var. caballicensis, r. canina var. andegavensis, and r. canina var. deseglisei. �is study showed that although the city area is strongly transformed by man, there is a high diversity of taxa of the studied genera. key words: crataegus, distribution of critical species, list of species, poland, rosa, rosaceae received: [2021.09.02] accepted: [2021.11.02] róże i głogi obszarów zurbanizowanych: na przykładzie gorzowa wielkopolskiego (nw poland) streszczenie w artykule przedstawiono rozmieszczenie gatunków z rodzajów crataegus i rosa na obszarze gorzowa wielkopolskiego. na podstawie danych z bibliogra�i oraz badań własnych stwierdzono tu występowanie 25 taksonów, w tym: 11 gatunków głogów i 14 taksonów róż. spośród głogów, 6 taksonów to gatunki rodzime, należące do subserii erianthae i crataegus, a 5 zaliczanych jest do hodowlanych taksonów ozdobnych. spośród gatunków rodzaju rosa, 10 to rodzime elementy �ory (w tym 2 formy mieszańcowe ujęte w randze gatunku), a wśród pozostałych 2 antropo�ty: 1 tzw. starą odmianę hodowlaną i 1 formę mieszańcową. wśród odnotowanych taksonów, aż 13 jest nowych dla �ory miasta gorzowa wielkopolskiego: crataegus ×macrocarpa, c. ×subsphaericea, c. ×media, rosa dumalis, r. villosa, r. sherardii, r. tomentosa, r. inodora, r. jundzillii, r. ×subcanina, r. ×subcollina oraz stara odmiana hodowlana r. ×francofourtana i mieszaniec międzygatunkowy rosa canina × r. rubiginosa. w trakcie badań odnotowano tutaj występowanie rzadkiego w polsce gatunku rosa jundzillii, który z wyżyny gorzowskiej nie był do tej pory podawany. znaleziono także rzadkie w polsce odmiany róż, jak: dla rosa dumalis: r. dumalis var. acharii i r. dumalis var. caballicensis oraz dla rosa canina: r. canina var. andegavensis i r. canina var. deseglisei. przeprowadzone badania wykazały, iż pomimo że obszar miasta jest terenem silenie przekształconym przez człowieka, to występuje tu duże zróżnicowanie taksonów z analizowanych rodzajów. słowa kluczowe: crataegus, rozmieszczenie gatunków krytycznych, lista gatunków, polska, rosaceae, rosa information on the authors anna soltys-lelek https://orcid.org/0000-0002-9595-3167 author of numerous scienti�c and popular science studies in the �eld of botany and environmental protection. her research interests relate particularly to critical types of roses (rosa) and hawthorns (crataegus). member of the polish and slovak botanical society. wojciech gruszka http://orcid.org/0000-0002-6229-8397 author and co-author of scienti�c and popular science studies in lichenology and botany. his main research interests relate to the ecology and protection of lichens. in addition, he participates in research on the distribution of representatives of rose (rosa) and hawthorn (crataegus) species in poland. iii liceum ogólnokształcące annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x gracjana budzałek*, sylwia śliwińska-wilczewska institute of oceanography, university of gdansk, gdynia, poland, *gbudzalek@gmail.com allelopathic effect of the green macroalga ulva intestinalis (ulvaceae, chlorophyta) on selected baltic cyanobacteria introduction allelopathy is a unique strategy to deter or eliminate organisms coexisting in the same ecosystem (molisch, 1937; śliwińska-wilczewska et al., 2021). in aquatic ecosystems, allelopathic activity depends on the production and secretion of active allelopathic compounds and their effective dispersal in the environment (lewis, 1986). the benthic zone is limited compared to the extensive pelagic zone in the sea, so allelopathic interactions in these coastal ecosystems may be stronger. macroalgae have been found to produce active metabolites that inhibit other organisms that compete with them for light and space (harlin, 1987; jeong et al., 2000), but their allelopathic activity on baltic cyanobacteria is still insufficiently recognised. previous studies have confirmed that green algae of the genus ulva l. (chlorophyta) are capable of allelopathic effects on selected microalgal species (jin, dong, 2003; nan et al., 2004; jin et al., 2005; wang et al., 2007; tang, gobler, 2011). jin and dong (2003) studied the effects of extract, filtrate, and fresh thallus of ulva pertusa kjellman on heterosigma akashiwo (y. hada) y. hada ex y. hara & m. chihara and alexandrium tamarense (lebour) balech. h. akashiwo and a. tamarense were strongly inhibited by aqueous extract and fresh tissue from u. pertusa. in contrast, the authors showed that the green algae-derived filtrate had no significant effect on the cell counts of the test organisms. a year later, nan et al. (2004) investigated the effect of fresh tissue and filtrate from u. pertusa on the tetraselmis subcordiformis (wille) butcher (species form phylum chlorophyta), the heterosigma akashiwo and alexandrium tamarense (representative of miozoa), the skeletonema costatum (greville) cleve, nitzschia closterium (ehrenberg) w. smith, and chaetoceros gracile f. schütt (member of bacillariophyta), the chroomonas placoidea butcher ex g. novarino & i.a.n. lucas (species from phylum cryptophyta), and the isochrysis galbana parke (representative of haptophyta). growth was significantly inhibited for each tested species. additionally, the authors’ results suggest that allelopathic compounds from u. pertusa are highly degradable. the inhibitory effect of extract, filtrate, and fresh thallus from u. lactuca was also demonstrated by tang and gobler (2011). this green macroalgae adversely affected the cell concentrations of aureococcus anophagefferens hargraves & sieburth in sieburth, p.w. johnson & hargreaves and chattonella marina (subrahmanyan) y. hara & m. chihara (member of ochrophyta), the cochlodinium polykrikoides margalef, karlodinium veneficum (d. ballantine) j. larsen in daugbjerg & al., karenia brevis (c.c. davis) gert hansen & moestrup, and prorocentrum minimum (pavillard) j. schiller (species form phylum miozoa), and pseudo-nitzschia multiseries (pavillard) j. schiller (representative of bacillariophyta). wang et al. (2007) investigated the inhibitory effect of extract, filtrate, and fresh thallus of ulva pertusa on the growth of two dinoflagellates h. akashiwo and alexandrium tamarense. on the other hand, jin et al. (2005) investigated the inhibitory effect of fresh thallus and extract of u. pertusa and u. linza l. on the dinoflagellata prorocentrum micans ehrenberg. a recent work indicates that ulva intestinalis l. also has allelopathic properties (budzałek et al., 2021a). it was shown that the addition of the cell-free filtrate obtained from u. intestinalis significantly inhibited growth and photosynthetic efficiency of filamentous cyanobacteria nodularia spumigena mertens ex bornet & flahault and nostoc sp. suprysingly, the authors found that the addition of different concentrations of aqueous extract as well as filtrate stimulated the aphanizomenon sp. there is only one report of allelopathic activity of spirogyra sp. (charophyta) (irfanullah, moss, 2005). in their work, they studied the effects of allelopathic compounds secreted by spirogyra sp. on phytoplankton communities. phytoplankton species dynamics and species composition were apparently not influenced by allelopathy of living or decomposing spirogyra sp. in addition, they investigated the allelopathic effects of filtrate and living thallus of spirogyra sp. on phytoplankton. there was no change in phytoplankton growth and species composition under the allelopathy of living or decomposing spirogyra sp. this study showed that filamentous macroalgae from charophyta phylum probably cannot control phytoplankton biomass in a nutrient-rich environment by secreting allelopathic compounds. allelopathic activity was also confirmed for macroalgae belonging to the genus chara (charophyta) (donk, bund, 2002; berger, schagerl, 2003; mulderij et al., 2003; pakdel et al., 2013; mähnert et al., 2017; złoch et al., 2018). pakdel et al. (2013) studied the effects of extract, filtrate, and live material from the stonewort – chara australis r. brown on the cyanobacteria anabaena variabilis kützing ex bornet & flahault and the green alga scenedesmus quadricauda (turpin) brébisson in brébisson & godey. c. australis had a highly inhibitory effect on the growth of a. variabilis. in contrast, the extract had no significant effect on s. quadricauda. recently, the effect of the extract of baltic c. aspera wild., c. baltica bruzelius, and c. canescens loiseleur on the cyanobacteria synechococcus sp. was demonstrated by złoch et al. (2018). both inhibition and stimulation of growth and photosynthesis of the tested species were demonstrated. c. aspera had both stimulatory and inhibitory effects on the studied cyanobacteria. the other two chara sp. inhibited the growth of cyanobacteria cell numbers. mähnert et al. (2017) showed inhibitory effects of extracts as well as fresh thallus of c. aspera, c. globularis, c. rudis, and c. tomentosa on the green microalgae chlorella vulgaris l., acutodesmus acuminatus (lagerheim) p.m. tsarenko in tsarenko & petlovanny, the cyanobacteria synechococcus elongatus, s. leopoliensis, and the bacterium aliivibrio fischeri beijerinck. strong inhibitory effects of the extract and fresh thallus on cyanobacteria and bacteria were demonstrated. berger and schagerl (2003) demonstrated the inhibitory effect of c. aspera extract on the growth of the filamentous cyanobacteria anabaena cylindrica lemmermann. the allelopathic inhibitory effect of c. aspera was also studied on the green algae scenedesmus acutus meyen (donk, bund, 2002). on the other hand, mulderij et al. (2003) studied the effect of filtrate obtained from chara globularis var. globularis (detharding ex willdenow) r.d. wood and c. contraria var. contraria (a. braun ex kützing) j.a. moore on three green algae. they showed growth stimulation of selenastrum capricornutum printz and chlorella minutissima fott & nováková. however, they showed no significant differences for scenedesmus obliquus (turpin) kützing. toxic cyanobacterial blooms pose a serious threat to the environment and human health, and restoration of affected water bodies can be a challenge. although cyanobacterial blooms are a common phenomenon already, their occurrence and severity are expected to increase in the future due to climate change (reichwaldt, ghadouani, 2012). in fresh and brackish water bodies toxic cyanobacterial blooms have been recorded for at least 2 millennia (zillén, conley, 2010). massive cyanobacterial blooms appear in the baltic sea almost every year during summer, however it is very difficult to predict their location and intensity of occurrence (kahru et al., 2020). the genus aphanizomenon sp. commonly dominates the water column biomass during blooms in the baltic sea, along with nodularia spumigena, which has consistently caused fatalities of both wild and domesticated animals in the baltic sea (wasmund, 1997). a third common species in this basin is nostoc sp., which also exhibits cytotoxic properties (surakka et al., 2005). thus, the aim of this study was to demonstrate the allelopathic effects of baltic macroalga ulva intestinalis l. (syn. enteromorpha intestinalis (l.) nees) thallus on growth and photosynthetic activity of three bloom-forming cyanobacteria aphanizomenon sp., nodularia spumigena, and nostoc sp. these studies help define the role of u. intestinalis allelopathy as a biological factor in the distribution of bloom-forming cyanobacteria in the coastal baltic sea region. fig. 1. ulva intestinalis l. thalli from habitat and light micrographs of tubular thallus (young cell arranged in longitudinal rows). scale bar: 100 mm (a), 60 μm (b), 40 μm (c), 20 μm (d). (photo. g. budzałek) material and methods place of sampling and material cultivation the material used in the experiments consisted of strains of baltic cyanobacteria aphanizomenon sp. (ccba-69), nodularia spumigena (ccba15), and nostoc sp. (ccba81) (so-called target organisms). strains of cyanobacteria were isolated from the natural phytoplankton communities of the coastal waters of the gulf of gdańsk (southern baltic sea) (54°30'53.7"n; 18°54'23.5"e) and are maintained in the culture collection of baltic algae (ccba) at the laboratory of marine plant ecophysiology at the university of gdańsk (latała et al., 2006). the macroalga ulva intestinalis used in the study (donor organism) was collected from the coastal zones of the southern baltic sea region (54°30'08.7"n; 18°33'32.3"e). determination of u. intestinalis (fig. 1) based on the examination of morphological features (such as number of pyrenoids and shape of cells) using an identification keys (starmach, 1972; škaloud et al., 2018). the herbarium sheets (voucher number: ba m50; herbarium website: https://zielnik.ug.edu.pl/en/home/) were prepared in accordance with guidelines in drobnik (2007) and rybak (2018) and deposited at the institute of oceanography, university of gdansk (poland). the studied macroalga and cyanobacteria were cultured on sterile mineral medium f/2 (guillard, 1975) prepared with baltic sea water filtered through glass fiber filters (whatman gf/c) and autoclaved. the salinity was 8 psu as measured with a salinometer (inolab cond level 1, germany). the u. intestinalis and cyanobacterial strains used in the experiments was maintained in 300-ml and 50-ml glass erlenmeyer flasks, respectively. donor and target organisms were cultured at a par intensity of 10 μmol photons m–2s–1 (16:8 h light: dark cycle) and a temperature of 18°c. photosynthetically active irradiance (par) was measured using a quantum meter (li-cor, usa). the light sources used in the experiment were lamps (cool white 40w, usa). the cultures were acclimated to these conditions for 2 days, and these growth conditions were used for the experiments. determination of the allelopathic effect of ulva intestinalis thallus the allelopathic effect of ulva intestinalis thallus was tested according to a method proposed by wang et al. (2007) with modifications. the cyanobacteria monocultures were exposed to three different concentrations of the presence of live thallus of u. intestinalis. target cyanobacterial strains were maintained in 25-ml erlenmeyer flasks. in all experiments, the starting concentration of chlorophyll a in cyanobacterial cultures was 0.4 μg chl a ml–1. the coexistence assays were performed using a mixed culture system of one macroalga and one strain of cyanobacteria. different initial inoculation concentrations (0.01, 0.05, and 0.1 g wet weight ml–1) of fresh algal thallus were inoculated into 25-ml erlenmeyer flasks containing 20 ml of the target cyanobacteria strains. control cultures were prepared analogously, but f/2 medium was added instead of thallus at 0.01, 0.05, and 0.1 ml– 1. the concentration of major nutrients in the control and experimental samples were adjusted to the same level as in the f/2 standard. therefore, the influence of nutrients, micronutrients and vitamins on the experimental result can be excluded. after 7 days of the exposure, before the measurements, the ulva sp. thallus was removed and the cyanobacterial cells concentration as well as chlorophyll a fluorescence parameters were determined. all allelopathic tests were conducted in triplicate. determination of cyanobacterial number of cells the number of cells (n) in aphanizomenon sp., nodularia spumigena, and nostoc sp. cultures was estimated with previously determined linear correlations between cell abundance (n ml– 1) and optical density (od). n was counted using a bürker chamber (48 squares per count) and light microscope following a procedure according to guillard and sieracki (2005), and the od was measured spectrophotometrically at 750 nm with a multiskan go uv-vis spectrophotometer (thermo scientific, massachusetts, usa). the linear correlation between n and od for mentioned cyanobacteria was described by budzałek et al. (2021). od measurements were performed on the 7th days of the experiment and control and converted to cyanobacteria cells. in addition, absorption spectra in the wavelength range from 400 nm to 750 nm with a frequency of 1 nm was determined for cyanobacterial abundances in controls and allelopathic treatments. determination of the chlorophyll a fluorescence in the conducted experiments, the pulsed amplitude modulation (pam) method was used to measure the chlorophyll a fluorescence (fms1, hansatech). this method is widely used to measure chlorophyll a fluorescence in cyanobacteria, both in the laboratory and in the natural environment (schreiber et al., 1995; campbell et al., 1998). samples were taken for chlorophyll fluorescence analysis after the 7th days of the experiment. about 5 ml of target cyanobacteria were filtered through 13-mm glass fiber filters (whatman gf/c). in the next step, the filters were placed in holders. the samples were kept in the dark for 5 min before measurement. in this study, the maximum psii quantum efficiency (fv/fm) and the effective quantum yield of psii photochemistry (φpsii) were determined (campbell et al., 1998). statistical analysis to confirm the allelopathic effect of the extracts obtained from macroalgae on the number of cells and chlorophyll a fluorescence parameters of target cyanobacteria, a one-way anova was performed. data are reported as the means ± standard deviations (sd), where level of significance was p < 0.05. statistical analysis and graphs were performed using statistica® 13.1 software and originpro program, version 2021 (originlab corporation, northampton, ma, usa). fig. 2. the number of cells (n 105 ml–1) of aphanizomenon sp. (a) nodularia spumigena mertens ex bornet & flahault (b), and nostoc sp. (c) in controls (c) and cultures to which were added different concentrations (g ml–1) of fresh thallus (ft) obtained from ulva intestinalis l. after 7 days of the experiment (a) and par absorption spectra determined for this strains at thallus concentrations: 0.01 (b), 0.05 (c), and 0.1 (d). the values shown are mean values (n = 3, mean ± sd). different letters indicate significant differences between the means of the treatments (p < 0.05, one-way anova) results allelopathic effect of ulva intestinalis thallus on cyanobacterial abundances in this study, the number of cells (n 105 ml–1) of aphanizomenon sp., nodularia spumigena, and nostoc sp. in controls and cultures to which were added: 0.01, 0.05, and 0.1 g wet weight ml–1 of fresh thallus obtained from ulva intestinalis after 7 days of the experiment were determined. it was found that thallus obtained from u. intestinalis had no statistically significant effect on the number of cells of the cyanobacterium aphanizomenon sp. at higher concentrations (0.05 and 0.1 g ml–1). on the other hand, it was examined a stimulating effect of 0.01 g ml–1 of the fresh thallus on the number of aphanizomenon sp. cells which constituted 168% (one-way anova, p < 0.05), relative to the control treatment (fig. 2a). on the other hand, it was shown that the fresh thallus addition resulted in a decrease in the number of n. spumigena cells. for this cyanobacterium, the growth was, relative to the control, 45%, 27%, and 46% (one-way anova, p < 0.05, for all), after addition of 0.01, 0.05, and 0.1 g wet weight ml−1 of fresh thallus, respectively (fig. 2b). in experiments with nostoc sp. and u. intestinalis thallus addition, the negative effect on cyanobacterial growth was detected on the 7th day of exposure at 0.1 g wet weight ml−1 and constituted 97% (oneway anova, p < 0.05) of control. in turn, the u. intestinalis thallus addition had no significant effect on the growth of nostoc sp. at lower concentrations (0.01 and 0.05 g ml−1) (fig. 2c). fig. 3. the values of the fluorescence parameter fv/fm (a) and φpsii (b) for aphanizomenon sp. (a) nodularia spumigena mertens ex bornet & flahault (b), and nostoc sp. (c) in controls (c) and cultures to which were added: 0.01, 0.05, and 0.1 (g ml–1) of fresh thallus (ft) obtained from ulva intestinalis l. after 7 days of the experiment. the values shown are mean values (n = 3, mean ± sd). different letters indicate significant differences between the means of the treatments (p < 0.05, one-way anova) allelopathic effect of ulva intestinalis thallus on fluorescence parameters the values of the fluorescence parameter fv/fm (the maximum psii quantum efficiency) and φpsii (the effective quantum yield of psii photochemistry) for aphanizomenon sp., n. spumigena, and nostoc sp. in control and cultures to which were added a different concentrations (0.01, 0.05, and 0.1 g wet weight ml–1) of fresh thallus obtained from u. intestinalis after 7 days of experiment were examined. a stimulating effect of concentrations of 0.01, 0.05, and 0.1 g wet weight ml–1 of u. intestinalis thallus on the values of φpsii of cyanobacteria aphanizomenon sp. were observed and in these conditions these parameters constituted 176%, 129%, and 148% (one-way anova, p < 0.05, for all), respectively (fig. 3ab). surprisingly, addition of thallus of u. intestinalis did not affect the value of fv/fm of aphanizomenon sp. (fig. 3aa). moreover, the addition of fresh thallus obtained from u. intestinalis had no statistically significant effect on the fluorescence parameters fv/fm and φpsii of n. spumigena (fig. 3ba-b). in turn, the fv/fm of nostoc sp. was positively affected after addition of 0.01, 0.05, and 0.1 g wet weight ml−1 of thallus, being, relative to the control, 142%, 153%, and 124% (one-way anova, p < 0.05, for all), respectively (fig. 3ca). moreover, in the fresh thallus addition experiments, the value of φpsii for the mentioned cyanobacterium was significantly different from the control at the concentration of 0.01 g ml−1, when it constituted 202% (fig. 3cb). discussion effects of allelopathic compounds produced by baltic macroalga ulva intestinalis on cyanobacterial growth in the present study, the allelopathic effect of baltic green macroalga ulva intestinalis thallus on growth of three bloom-forming cyanobacteria aphanizomenon sp., nodularia spumigena, and nostoc sp. was investigated. the live thallus had an inhibitory effect on the number of cells of n. spumigena, and nostoc sp. in the first week. whereas cell growth in the aphanizomenon sp. sample was stimulated by u. intestinalis thallus at the lowest concentration (0.01 g wet weight ml–1). in several other works, the authors also showed allelopathic activity of fresh thallus of green algae of the genus ulva on co-occurring phytoplankton species. wang et al. (2007) noted that heterosigma akashiwo and alexandrium tamarense showed different responses when exposed to live u. pertusa thallus. the authors showed that relatively low concentrations had a lethal effect on h. akashiwo, while a. tamarense cells were not completely degraded even at the highest macroalgal concentration. differences in the cell surface structure of h. akashiwo and a. tamarense may account for their different sensitivity to allelopathic compounds (wang et al., 2007). kakisawa et al. (1988) suggested that allelopathic compounds produced by macroalgae may be more active against organisms that lack cell walls. in the present study, three species of baltic cyanobacteria that possess cell walls were used for testing. in addition, cyanobacteria are covered with a mucous envelope of varying thickness, which may also diminish the effect of allelochemicals. this may partly explain the lack of effect of the filtrate and live u. intestinalis mollusks on the cell abundance of aphanizomenon sp. it should be noted here that the composition of allelopathic compounds from green macroalge is still not well known. in a review by budzalek et al. (2021b), known compounds from macroalgae with an active effect on other organisms were listed. among them, it has been shown that the u. intestinalis can produce penostatins a–h, cytochalasans, penochalasins a–h, chaetoglobosin, and communesins a–b with cytotoxic activities. however, more research is needed to unequivocally show which compounds are responsible for inhibiting or stimulating cyanobacteria in the baltic ecosystem. in contrast, uchida et al. (1995) reported that heterocapsa circularisquama horiguchi completely annihilated gyrodinium instriatum freudenthal & j.j.lee by direct cell contact. in the present study, the possibility of inhibition of cyanobacterial growth by direct contact with u. intestinalis was also considered and this possibility was finally ruled out by additionally performing experiments with the addition of the filtrate of this donor green alga alone. since significant changes in the growth of baltic cyanobacteria were also observed under the influence of the filtrate, it can be assumed that the secretion of allelopathic compounds by u. intestinalis is the most probable explanation for the observed growth inhibition of the organisms studied. effects of allelopathic compounds produced by baltic macroalga ulva intestinalis on photosynthesis performance in this work, we also investigated the allelopathic effect of fresh thallus obtained from u. intestinalis on the maximum psii quantum efficiency (fv/fm) and the effective quantum yield of psii photochemistry (φpsii) of some baltic cyanobacteria. it was found that u. intestinalis had no allelopathic effect on fluorescence parameters of tested cyanobacterium n. spumigena. on the other hand, all tested concentrations (0.01, 0.05, and 0.1 g wet weight ml–1) of thallus from mentioned green macroalga stimulated the values of fv/fm or φpsii of cyanobacteria aphanizomenon sp. and nostoc sp. compared to the control. in other works, budzałek et al. (2018, 2021b) and złoch et al. (2018) demonstrated that extract and filtrate of u. intestinalis contains water-soluble allelochemical(s) is able to influence the fluorescence parameters of some baltic cyanobacteria. budzałek et al. (2018) noted that the highest decrease in fv/fm for nostoc sp. was observed after addition of 100 µl ml–1 u. inestinalis extract. on the other hand, in some cases, the extract and filtrate from u. inestinalis caused the stimulation of fluorescence parameters of aphanizomenon sp. and nostoc sp. (budzałek et al., 2021). moreover, extract obtained from chara baltica and c. canescens caused an increase in fluorescence parameters for synechococcus sp. after the seventh day of the exposure (złoch et al., 2018). the low level of these parameters may be evidence of certain disturbances in the photosynthesis process due to allelochemicals (song et al., 2017). on the other hand, stimulating photosynthesis at the low concentration of thallus may indicate the phenomenon of hormesis (stebbing, 1982). it is worth mentioned here, that this is the first report of the allelopathic effect of u. intestinalis thallus on the maximum psii quantum efficiency and the effective quantum yield of psii photochemistry of baltic cyanobacteria. therefore, more studies should be done to further investigate the interactions between macroalgae and cyanobacteria in the aquatic reservoirs. ecological significance of ulva intestinalis allelochemicals and possibilities of their application direct competitive interaction between species is one of the major causes of plant extinction worldwide (jarchow, cook, 2009). competition for available resources is often considered the main competitive mechanism that influences the success of some organisms. studies have shown that the green algae ulva sp., which have a high surface area to volume ratio, exhibit high rates of nutrient uptake. however, ulva sp. have limited ability to concentrate and store nitrogen internally (xu et al., 2012). furthermore, tait and schiel (2011) indicated that light intensity plays an important role in macroalgae productivity. the authors noted that high densities of the brown alga sargassum muticum (yendo) fensholt exclude native species and reduce biodiversity by shading the associated microalgae (white, shurin, 2011). in contrast, svirski et al. (1993) found that growth inhibition of gracilaria sp. cultured in the presence of u. lactuca was not due to shading or nutrient depletion but appeared to be caused by competition for inorganic carbon through the production of allelopathic compounds. nan et al. (2004) found that it is difficult to distinguish competition for available resources from allelopathic interactions in the natural aquatic habitats. therefore, the authors in their study focused on performing experiments with the addition of filtrate obtained from u. pertusa to exclude the effect of competition for available nutrients in the culture and the effect of elevated ph. the authors showed that the growth of h. akashiwo and s. costatum was completely inhibited, even with high nutrient availability. therefore, competition for nutrients cannot explain the growth inhibition of the tested microalgae by u. pertusa. the experiments conducted in the present study were refined to be able to exclude the effect of competition for resources. in the work described above, the experiments were also conducted using mineral medium f/2. hence, the effect of nutrient depletion as a cause of growth inhibition of the cyanobacteria studied in our work can also be excluded. macroalgae are known for their good nutrient uptake capacity (valiela et al., 1997; neori et al., 2004; zertuche-gonzález et al., 2009) and are intentionally used in many parts of the world to reduce nutrient levels in coastal waters (neori et al., 2004; carmona et al., 2006; chopin et al., 2001, 2008). many bloom-forming species are directly or indirectly promoted by nutrients (heisler et al., 2008; anderson et al., 2008) thus macroalgae may reduce the occurrence of these species by decreasing the levels of nutrients available in the ecosystem. however, experiments conducted by tang and gobler (2011) showed that the observed negative effects of u. lactuca on bloom-forming species were not due to nutrient limitation of microalgae or nutrient competition between microand macroalgae. in the present study, the studied cyanobacteria grown on a nutrient-sufficient mineral medium, so it can be concluded that the negative effect of u. intestinalis was the result of the allelopathic compounds released into the medium. macroalgae belonging to the genus ulva are widely distributed. harvesting these macroalgae from shorelines provides an easy and environmentally friendly way to potentially control the species responsible for creating massive blooms (jeong et al., 2000). our results have shown that u. intestinalis can produce and secrete some allelopathic compounds that are able to inhibit the growth of the common, bloom-forming cyanobacteria. acknowledgements the authors would like to thank the editor and two anonymous reviewers for their valuable comments and suggestions to improve the quality of the paper. this research was funded by ugrants-start, grant number 5330c20-gs03-22. references anderson, d.m., burkholder, j.m., cochlan, w.p., glibert, p.m., gobler, c.j., heil, c.a., raphael, m.k., parsons, m.l., rensel, j.e., townsend, d.w., trainer, v.l., vargo, g. a. 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(1997). occurrence of cyanobacterial blooms in the baltic sea in relation to environmental conditions. internationale revue der gesamten hydrobiologie und hydrographie, 82(2), 169–184. https://doi.org/10.1002/iroh.19970820205 white, l. f., shurin, j. b. (2011). density dependent effects of an exotic marine macroalga on native community diversity. journal of experimental marine biology and ecology, 405(1–2), 111–119. http://dx.doi.org/10.1016/j.jembe.2011.05.024 xu, d., gao, z., zhang, x., fan, x., wang, y., li, d., wei, w., zhimeng, z., ye, n. (2012). allelopathic interactions between the opportunistic species ulva prolifera and the native macroalga gracilaria lichvoides. plos one, 7(4). https://doi.org/10.1371/journal.pone.0033648 zertuche-gonzález, j.a., camacho-ibar, v.f., pacheco-ruíz, i., cabello-pasini, a., galindo-bect, l.a., guzmán-calderón, j.m., carranza-macias, v., espinoza-avalos, j. (2009). the role of ulva spp. as a temporary nutrient sink in a coastal lagoon with oyster cultivation and upwelling influence. journal of applied phycology, 21(6), 729. https://doi.org/10.1007/s10811-009-9408-y zillén, l., conley, d.j. (2010). hypoxia and cyanobacterial blooms are not natural features of the baltic sea. biogeosciences discussions, 7(2), 1783–1812. http://dx.doi.org/10.5194/bgd-7-1783-2010 złoch, i., śliwińska-wilczewska, s., kucharska, m., kozłowska, w. (2018). allelopathic effects of chara species (c. aspera, c. baltica, and c. canescens) on the bloom-forming picocyanobacterium synechococcus sp. environmental science and pollution research, 25, 36403–36411. https://doi.org/10.1007/s11356-0183579-5 abstract macroalgae have been found to produce active allelochemicals that inhibit of growth other organisms that compete with them for light and space. however, their allelopathic activity on baltic cyanobacteria is still insufficiently recognised. therefore, this study aimed to demonstrate the allelopathic effects of baltic macroalga thallus (ulva intestinalis) on the growth and photosynthetic activity of three bloom-forming cyanobacteria: aphanizomenon sp., nodularia spumigena, and nostoc sp. this study investigated the cell count of the analysed cyanobacteria (n 105 ml–1), the maximum quantum yield of the second photosystem (psii) in the dark (fv/fm), and the real quantum yield of psii in the light (φpsii) (in the control and the experiments). after 7 days of exposure, the following were added: 0.01, 0.05, and 0.1 g ml–1 of u. intestinalis fresh thallus. it was found that thallus obtained from u. intestinalis had no statistically significant effect on the number of cells of the cyanobacterium aphanizomenon sp. (at 0.05 and 0.1 g ml–1) and nostoc sp. (at concentrations of 0.01 and 0.05 g ml−1). on the other hand, it was examined a stimulating effect of 0.01 g ml–1 of the fresh thallus on the number of aphanizomenon sp. cells which constituted 168%, relative to the control. it was shown that the fresh thallus addition resulted in a decrease in the number of n. spumigena cells (45%, 27%, and 46% after addition of 0.01, 0.05, and 0.1 g wet weight ml−1 of fresh thallus, respectively). in experiments with nostoc sp., the addition of u. intestinalis thallus has been a negative effect on cyanobacterial growth at 0.1 g ml−1 and constituted 97% of control. it was also found, that u. intestinalis had no allelopathic effect on fluorescence parameters of n. spumigena. all tested concentrations of thallus u. intestinalis (0.01, 0.05, and 0.1 g wet weight ml–1) stimulated the values of fv/fm or φpsii of cyanobacteria aphanizomenon sp. and nostoc sp. compared to the control. these studies help define the role of u. intestinalis allelopathy as a biological factor in the distribution of bloomforming cyanobacteria in the coastal baltic sea region. key words: allelopathy, cyanobacteria, green macroalgae, growth, fluorescence, thallus received: [2021.11.05] accepted: [2022.03.22] wpływ allelopatyczny ulva intestinalis na wybrane gatunki bałtyckich sinic streszczenie makroglony mają zdolność do produkowania aktywnych związków allelopatycznych, które wpływają na inne organizmy, konkurujące z nimi o światło i przestrzeń. jednakże ich działanie allelopatyczne na bałtyckie sinice jest wciąż niedostatecznie rozpoznane. dlatego głównym celem niniejszej pracy było wykazanie aktywności allelopatycznej plechy bałtyckiej zielenicy, ulwa (lub taśma czy błonica) kiszkowata – ulva intestinalis, na wzrost i aktywność fotosyntetyczną i wzrost trzech sinic bałtyckich, tworzących masowe zakwity: aphanizomenon sp., nodularia spumigena oraz nostoc sp. w niniejszej pracy badano liczebność komórek analizowanych sinic (n 105 ml–1), maksymalną wydajność kwantową drugiego fotosystemu (psii) w ciemności (fv/fm) oraz rzeczywistą wydajność kwantową psii w świetle (φpsii) (w kontroli oraz w eksperymentach). po 7 dniach ekspozycji, dodawano do nich: 0,01, 0,05, i 0,1 g ml–1 świeżej plechy u. intestinalis. wykazano, że obecność plech u. intestinalis nie miała istotnego wpływu na liczebność komórek aphanizomenon sp. w ilości (0,05 i 0,1 g ml–1) oraz nostoc sp. w ilości plechy (0,01 i 0,05 g ml−1). wykazano także stymulujący wpływ plechy dodawanej w najmniejszej ilości 0,01 g ml–1 na liczebność komórek aphanizomenon sp., która wynosiła 168%, w stosunku do kontroli. dodanie 0,01, 0,05 oraz 0,1 g ml−1 świeżej plechy powodowało obniżenie liczebności komórek n. spumigena (o odpowiednio: 45%, 27%, i 46%, w stosunku do kontroli). dodatkowo, dodanie 0,1 g ml−1 plechy u. intestinalis powodowało zahamowanie wzrostu nostoc sp. o 97%. w pracy wykazano także, że plecha u. intestinalis nie miała wpływu na wartość parametrów fluorescencji u n. spumigena. plecha tego gatunku (w każdym testowanym stężeniu: 0,01, 0,05 oraz 0,1 g ml–1) stymulowała wartość parametrów fv/fm lub φpsii u sinic aphanizomenon sp. i nostoc sp. wyniki uzyskane w niniejszej pracy definiują rolę allelopatii u. intestinalis jako ważnego czynnika biologicznego, wpływającego na występowanie zakwitów sinic w przybrzeżnych rejonach morza bałtyckiego. słowa kluczowe: allelopatia, sinice, zielenice, wzrost, fluorescencja, plecha information on the authors gracjana budzałek the field of her interest is allelopathic interactions between macroalgae and other phytoplankton species. in her research she is focusing mostly on baltic species from puck bay. she is investigating what influence of allelopathic compounds produced by macroalgae have on bloom-forming cyanobacteria and microalgae. sylwia śliwińska-wilczewska https://orcid.org/0000-0002-3147-6605 she is interested in allelopathy of cyanobacteria and microalgae; in particular of picocyanobacteria synechococcus sp. allelopathy plays an important role in interspecific competition and contributes to cyanobacterial bloom maintenance. in her study, the influence of allelochemicals on the growth, chlorophyll fluorescence and photosynthesis irradiance curves of different phytoplankton species was investigated. she is also investigating what influences have environmental factors on produced allelopathic compounds on algae and cyanobacteria. 31 annales universitatis paedagogicae cracoviensis studia naturae, 4: 31–64, 2019, issn 2543-8832 doi: 10.24917/25438832.4.2 krystyna towpasz institute of botany, jagiellonian university, kopernika 31 st., 31-501 kraków, poland; krystyna.towpasz@uj.edu.pl vascular plants of pilzno surroundings (south-eastern poland) introduction an extensive monographic study on the �ora and geobotanical relations of the strzyżów foothills was created at the end of the last century (towpasz, 1987, 1990). �e �ora of the ciężkowice foothills was analysed in a  similar period (kornaś et al., 1996). later, only a  few petty �oristic publications were created to complement the �ora of the strzyżów foothills (oklejewicz et al., 2004; wójcik, 2011; towpasz, 2013). �e natural environment of pilsen surroundings has been recently presented in a monographic study (towpasz, 2018). �e aim of this study is �oristic valorisation of the southern part of the pilzno commune (south-eastern poland). general characteristics of the study area vegetation �e study area belongs to the western outer carpathians and is located within the macroregion of the central beskids foothills and two mesoregions: ciężkowice foothills (pogórze ciężkowickie) and strzyżów foothills (pogórze strzyżowskie) (fig. 1). �e landscape is dominated by arable �elds, locally meadows and pastures (appendix 1a–b). forests occupy a small area and are preserved in places unfavorable to agriculture. rare protected plants grow within them, among others: matteucia struthiopteris, staphyllea pinnata (appendix 1c–d), or scilla bifolia. meadows, arised and maintained on this area as a consequence of constant human intervention, such as: mowing, grazing, fertilisation. depending on the degree of soil moisture, the following types of meadows are distinguished in this area: constantly and periodically moist and fresh meadows. on this area, pastures occupy a  smaller area compared to meadows. for example, the association of water lilies is very rare in local ponds and oxbow lakes. xerothermic vegetation develops here on dry, sunny slopes of hills and river valleys, mainly at southern exposure. synanthropic communik ry st yn a to w pa sz 32 ties, related to human activity, are creating among other as segetal vegetation in �elds and gardens, and as ruderal vegetation accompanying human settlements, communication lines or industrial centers. research methods a  detailed monographic studies of the vascular plant �ora of the strzyżów and ciężkowice foothills dates from the end of the 20th century (towpasz, 1987, 1990; kornaś et al., 1997). in connection with the elaboration of a chapter on the natural environment of pilzno (towpasz, 2018), some of the materials contained in these studies related to the occurrence of vascular plants in the pilzno commune were used, a�er their updating and veri�cation in the �eld in recent years. to complement the �ora of the strzyżów foothills, earlier studies were also used (oklejewicz et al., 2004; wójcik, 2011; towpasz, 2013). fig. 1. localisation of the study area against the regional divisionof the eastern part of the polish carpathians (according kondracki, 1978): 1 – state boundary, 2 – the northern limits of the carpathians, 3 – boundary between the western ana eastern carpathians, 4 – study region v ascular plants of p ilzno surroundings (s outh-eastern p oland) 33 �e order of the species is given in alphabetical sequence, and their nomenclature is consistent with mirek et al. (2002). all localities listed here were located in the atpol – the polish geobotanical grid, 2 km long, in accordance with the methodology proposed by zając (1978). distribution of atpol grid squares in the area of pilzno commune is shown in �gure 2. alien origin plants, mountain species (pawłowski, 1925, 1972) and protected plants (regulation of the minister of the environment of october 9 (…), 2014) are indicated in the list. classi�cation of alien species was adopted according to the atlas zając and zając (2001) and supplemented based on the study by tokarska-guzik et al. (2012). for alien species, found in semi-natural and natural habitats (agriophytes), their belonging to the group of invasive plants in the studied area is given in accordance with the work (zając, zając, 2015). in the prepared �oristic list, the following marks and abbreviations have been used: * – alien species, grn. – górne, dln – dolne, cf – ciężkowice foothills, sf – strzyżów foothills. glossary of geographical names in the pilzno commune �e list includes geographical coordinates for the villages and signatures of the atpol grid squares. fig. 2. distribution of atpol grid squares in the area of pilzno commune k ry st yn a to w pa sz 34 bielowy 49°56′46″n 21°19′36″e (ef 79 33) – village, 6 km se from pilzno (cf) dęborzyn 49°55′07″n 21°18′59″e (ef 79 44) – village, about 7 km se from pilzno (cf) dobrków 49°59′19″n 21°20′01″e (ef 79 04) – village, 1.5 km n from gołęczyna (sf) dulczówka 49°58′43″n 21°17′29″e (ef 79 12) – village, 1 km w from pilzno (cf) gołęczyna 49°58′38″n 21°20′36″e (ef 79 14) – village, about 3 km se from pilzno (sf) jaworze dolne 49°57′18″n 21°21′08″e (ef 79 24) – village in the wisłoka valley, 8 km se from pilzno (sf) jaworze górne 49°55′59″n 21°20′52″e (ef 79 34) – village, 2 km s from jaworze dolne (sf) łęki górne 49°58′26″n 21°10′48″e (ef 78 13) – village, 4 km w from pilzno (cf) mokrzec 49°58′38″n 21°19′36″e (ef 79 13) – village, 1 km e from gołęczyna (sf) parkosz 50°00′13″n  21°18′50″e (ef 69 43) – village in the wisłoka valley, 3 km ne from pilzno (sf) pilzno 49°58′43″n 21°17′29″e (ef 79 02; 79 03; 79 12) – city by the tarnów – dębica road, on the border of the rożnów foothills and the strzyżów foothills (cf, sf) podgrodzie 50°00′42″n 21°20′54″e (ef 69 44) – village, 6 km ne from pilzno (sf) rędziny 49°56′51″n 21°18′08″e (ef 79 22) – hamlet of strzegocice (cf) słotowa 49°56′43″n 21°17′16″e (ef 79 31) – village, 2 km w from strzegocice (cf) strzegocice 49°56′51″n 21°18′08″e (ef 79 22) – village, 5 km s from pilzno (cf) zagórze 49°55′19″n 21°18′05″e (ef 79 33) – village, 1 km nw from dęborzyn (cf) zwiernik 49°56′24″n 21°12′34″e (ef 78 34) – village, 3 km s from łęki górne (cf) results – list of species 1. abies alba mill. – higher montane zone species. in deciduous forests (oak-hornbeam forests, beech forests) and mixed forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), zagórze (ef 79 33). 2. acer campestre l. – in deciduous forests: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 3. a. platanoides l. – in deciduous forests: parkosz (ef 69 43), zwiernik (ef 78 34), gołęczyna (ef 79 14). 4. a. pseudoplatanus l. – higher montane zone species. in deciduous forests: parkosz (ef 69 43), łęki grn. (ef 78 13). 5. achillea millefolium l. – in the meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12), jaworze dln. (ef 79 24). 6. acinos arvensis (lam.) dandy – balks and dry slopes: parkosz (ef 69 43), jaworze dln. (ef 79 24). 7. actaea spicata l. – oak-hornbeam forest over the stream: podgrodzie (ef 69 44). 8. adoxa moschatellina l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 35 9. aegopodium podagraria l. – �ickets over stream, roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12), jaworze dln. (ef 79 24). 10. *aethusa cynapium l. subsp. agrestis (wallr.) dostál – archaeophyte. fields: pilzno (ef 79 02). 11. agrimonia eupatoria l. – roadsides, balks: parkosz (ef 69 43), jaworze dln. (ef 79 24). 12. *agrostemma githago l. – archaeophyte. fields: bielowy (ef 79 33). 13. agrostis capillaris l. – in meadows and �elds: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 02), gołęczyna (ef 79 14), bielowy (ef 79 33). 14. a. gigantea roth – fields: parkosz (ef 69 43), pilzno (ef 79 12), jaworze dln. (ef 79 24), słotowa (ef 79 31). 15. a. stolonifera l. – on stones of the wisłoka river banks: pilzno (ef 79 12), jaworze dln. (ef 79 24). 16. ajuga reptans l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13). 17. alchemilla acutiloba opiz – pastures: łęki grn. (ef 78 13). 18. alisma plantago-aquatica l. – in water reservoir and in drainage ditche: parkosz (ef 69 43). 19. alliaria petiolata (m. bieb.) cavara & grande – in alluvial forests of the wisłoka river: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 02, ef 79 12), jaworze dln. (ef 79 24). 20. allium oleraceum l. – fields: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 21. a. scorodoprassum l. – on edges of thickets: jaworze dln. (ef 79 24). 22. alnus glutinosa (l.) gaertn. – oak-hornbeam forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 23. a. incana (l.) moench – higher montane zone species. in alluvial forests: podgrodzie (ef 69 44). 24. alopecurus geniculatus l. – wet meadows: pilzno (ef 79 02), parkosz (ef 79 03). 25. a. pratensis l. – meadows: parkosz (ef 69 43), łęki grn. (ef 78 13), mokrzec (ef 79 13). 26. alyssum alyssoides (l.) l. – on sandy fallow lands on the wisłoka river: parkosz (ef 69 43), jaworze dln. (ef 79 24). 27. *amaranthus retro�exus l. – epoecophyte. in �elds, in root crops: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 28. *anagallis arvensis l. – archaeophyte. fields: parkosz (ef 69 43), zwiernik (ef 78 34), pilzno (ef 79 02), mokrzec (ef 79 13). 29. *anchusa o�cinalis l. – archaeophyte. on fallow lands and on ruderal habitats: podgrodzie (ef 69 44), mokrzec (ef 79 13). 30. anemone nemorosa l. – in deciduous forests: podgrodzie (ef 69 44), łęki grn. (ef 78 13), parkosz (ef 79 03). k ry st yn a to w pa sz 36 31. angelica sylvestris l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44). 32. *anthemis arvensis l. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34). 33. anthriscus nitida (wahlenb.) hanzl. – wet forest: pilzno (ef 79 12). 34. a. sylvestris (l.) ho�m. – in thickets of the wisłoka river: pilzno (ef 79 12), strzegocice (ef 79 22). 35. anthyllis vulneraria l. – in grasslands on slopes: jaworze dln. (ef 79 24). 36. *antoxanthum aristatum boiss. – holoagiophyte. on sandy dune in pine forest: jaworze dln. (ef 79 24). 37. a. odoratum l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), gołęczyna (ef 79 14). 38. *apera spica-venti (l.) p. beauv. – archaeophyte. fields: parkosz (ef 69 43), zwiernik (ef 78 34), pilzno (ef 79 03). 39. *aphanes arvensis l. – archaeophyte. fields: parkosz (ef 69 43), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 40. aposeris foetida (l.) less. – higher montane zone species. in deciduous forests: bielowy (ef 79 33). 41. arabidopsis thaliana (l.) heynh. – on ruderal habitats: podgrodzie (ef 69 44), łęki grn. (ef 78 13). 42. arabis glabra (l.) bernh. – on dry slopes: parkosz (ef 69 43), jaworze dln. (ef 79 24). 43. arctium lappa l. – roadsides: parkosz (ef 69 43), pilzno (ef 79 02). 44. a. minus (hill) bernh. – roadside and alluvial forests of the wisłoka river: parkosz (ef 69 43), dobrków (ef 79 04). 45. a. tomentosum mill. – roadside: podgrodzie (ef 69 44). 46. arenaria serpyllifolia l. – roadsides: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 47. *armoracia rusticana p. gaertn., b. mey. & schreb. – archaeophyte. roadside: pilzno (ef 79 02). 48. arrhenatherum elatius (l.) p. beauv. ex j. presl. & c. presl. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), bielowy (ef 79 33). 49. artemisia campestris l. – sand dunes and fallow land: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln.(ef 79 24). 50. a. vulgaris l. – roadside: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12). 51. asarum europaeum l. – partially protected species. in deciduous forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 52. asplenium ruta-muraria l. – on walls: pilzno (ef 79 12), strzegocice (ef 79 22). 53. a. trichomanes l. – in oak-hornbeam thickets: jaworze dln. (ef 79 24). 54. *aster novi-belgii l. – hemiagriophyte. invasive species. roadside: podgrodzie (ef 69 44). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 37 55. astragalus glycyphyllos l. – on balks and roadsides: zwiernik (ef 78 34), jaworze dln. (ef 79 24). 56. athyrium �lix-femina (l.) roth – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 57. *atriplex patula l. – archaeophyte. fields: parkosz (ef 69 43), zwiernik (ef 78 34). 58. *avena fatua l. – archaeophyte. fields: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 59. *ballota nigra l. – archaeophyte. roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 60. barbaraea vulgaris r. br. – roadside and meadow: parkosz (ef 69 43). 61. batrachium aquatile (l.) dumort. – in water reservoir: jaworze dln. (ef 79 24). 62. bellis perennis l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13). 63. berteroa incana (l.) dc. – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44). 64. berula erecta (huds.) conville – in drainage ditches: jaworze dln. (ef 79 24). 65. betula obscura kotula – in forests: parkosz (ef 69 43), dobrków (ef 79 04). 66. b. pendula roth – in forests: podgrodzie (ef 69 44), łęki grn. (ef 78 13), gołęczyna (ef 79 14). 67. *bidens frondosa l. – hemiagriophyte. invasive species. on banks of drainage ditches: dobrków (ef 79 04), pilzno (ef 79 12). 68. b. tripartita l. – on banks of rivers and streams: parkosz (ef 69 43), zwiernik (78 34), pilzno (ef 79 12). 69. brachypodium sylvaticum (huds.) p. beauv. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 70. *brassica nigra (l.) w. d. j. koch – epoecophyte. roadside: pilzno (ef 79 02). 71. briza media l. – meadows: słotowa (ef 79 31), jaworze dln. (ef 79 24). 72. *bromus carinatus hook. & arn. – epoecophyte. on lawns and roadsides: pilzno (ef 79 12), jaworze grn. (ef 79 34). 73. b. hordeaceus l. – meadows: parkosz (ef 69 43), łęki górne (ef 78 13), pilzno (ef 79 12), jaworze dln. (ef 79 24). 74. b. inermis leyss. – on dry slopes and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04), pilzno (ef 79 12). 75. *bunias orientalis l. – epoecophyte. on ruderal habitats: jaworze dln. (ef 79 24). 76. calamagrostis arundinacea (l.) roth – in pine forests: jaworze dln. (ef 79 24). 77. c. epigejos (l.) roth – on balks, in thinned forests and on their outskirts: podgrodzie (ef 69 44), słotowa (ef 79 11), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 78. calluna vulgaris (l.) hull – on edges of forests: parkosz (ef 69 43), gołęczyna (ef 79 14), dobrków (ef 79 04), jaworze dln. (ef 79 24). 79. caltha palustris l. – wet meadows, banks of streams: parkosz (ef 69 43), jaworze dln. (ef 79 24), podgrodzie (ef 69 44). k ry st yn a to w pa sz 38 80. calystegia sepium (l.) r. br. – in alluvial forests of the wisłoka river: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 81. camelina microcarpa andrz. – on sandy fallow lands: jaworze dln. (ef 79 24). 82. campanula glomerata l. – dry slopes: jaworze grn. (ef 79 34). 83. c. patula l. – meadows: podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 84. c. persicifolia l. – in thickets of the wisłoka river: podgrodzie (ef 69 44). 85. c. rapunculoides l. – on cereal �elds: parkosz (ef 69 43), pilzno (ef 79 02), gołęczyna (ef 79 14). 86. c. trachelium l. – in thickets: podgrodzie (ef 69 44), gołęczyna (ef 79 14). 87. capsella bursa pastoris (l.) medik. – roadsides: podgrodzie (ef 69 34), parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 02). 88. cardamine amara l. – on the banks of streams: parkosz (ef 69 43), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 89. c. impatiens l. – in deciduous forest: jaworze dln. (ef 79 24). 90. *cardaria draba (l.) desv. – epoecophyte. roadside: mokrzec (ef 79 13). 91. *carduus acanthoides l. – archaeophyte. on balks and roadsides: podgrodzie (ef 69 34), parkosz (ef 69 43), mokrzec (ef 79 13). 92. c. crispus l. – fields: pilzno ef 79 02), dobrków (ef 79 04). 93. carex brizoides l. – oak-hornbeam forests: parkosz (ef 69 43), łęki grn. (ef 78 13), dobrków (ef 79 04), jaworze dln. (ef 79 24). 94. c. caryophyllea latourr. – on lawn at wisłoka’s stone banks: podgrodzie (ef 69 44). 95. c. cuprina (i. sándor ex heu�.) nendtv. ex a. kern. – wet meadows: parkosz (ef 69 43), strzegocice (ef 79 22). 96. c. digitata l. – in forest: podgrodzie (ef 69 44). 97. c. �acca schreb. – pasture: jaworze dln. (ef 79 24). 98. c. gracilis curtis – on banks of drainage ditches: parkosz (ef 69 43), jaworze dln. (ef 79 24). 99. c. hirta l. – pastures: parkosz (ef 69 43), gołęczyna (ef 79 14). 100. c. nigra reichard – wet meadows: łęki grn. (ef 78 13), strzegocice (ef 79 22). 101. c. ornithopoda wild. – on stones of stream banks and on slope grassland: parkosz (ef 69 43), jaworze dln. (ef 79 24). 102. c. ovalis gooden. – meadows: podgrodzie (ef 69 44), pilzno (ef 79 12). 103. c. pallescens l. – meadow: podgrodzie (ef 69 44). 104. c. panicea l. – wet meadows: łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 105. c. paniculata l. – wet meadow: jaworze dln. (ef 79 24). 106. c. pendula huds. – lower montane zone species. in alluvial forest over stream: podgrodzie (ef 69 44). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 39 107. c. pilosa scop. – in forest: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 108. c. remota l. – over stream: dobrków (ef 79 04). 109. c. spicata huds. – on roadside embankment: pilzno (ef 79 12). 110. c. sylvatica huds. – oak-hornbeam forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 111. c. vesicaria l. – wet meadows: parkosz (ef 69 43), dobrków (ef 79 04). 112. c. vulpina l. – wet meadows: parkosz (ef 69 43), zwiernik (ef 78 13), strzegocice (ef 79 22). 113. carlina vulgaris l. – dry meadow: parkosz (ef 69 43). 114. carpinus betulus l. – in deciduous forests, mainly in oak-hornbeam forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), słotowa (ef 79 11), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 115. carum carvi l. – on meadows and balks: podgrodzie (ef 69 44), zwiernik (ef 78 34), słotowa (ef 79 11), jaworze dln. (ef 79 24). 116. * centaurea cyanus l. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 117. c. jacea l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12), jaworze dln. (ef 79 24). 118. c. scabiosa l. – balks: podgrodzie (ef 69 44), słotowa (ef 79 12), jaworze dln. (ef 79 24). 119. centaurium erythraea rafn – partially protected species. dry meadows: parkosz (ef 69 43), jaworze dln. (ef 79 24). 120. cephalanthera longifolia (l.) fritsch – protected species. oak-hornbeam forest: dęborzyn (ef 79 44). 121. cerastium arvense l. – fields: podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 122. c. glomeratum �ull. – field: strzegocice (ef 79 22). 123. c. holosteoides fr. emend. hyl. – meadows: parkosz (ef 69 43), łęki grn. (ef 78 13). 124. c. semidecandrum l. – on sandy dunes: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 125. c. sylvaticum waldst. & kit. – lower montane zone species. oak-hornbeam forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 126. cerasus avium (l.) moench – oak-hornbeam forest: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 127. ceratophyllum demersum l. – in water reservoir: mokrzec (ef 79 13). 128. cerinthe minor l. – on dry slopes: jaworze dln. (ef 79 24). 129. chaenorrhinum minor (l.) lange – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02: ef 79 12). 130. chaerophyllum aromaticum l. – �ickets: pilzno (ef 79 12), jaworze dln. (ef 79 24). k ry st yn a to w pa sz 40 131. ch. hirsutum l. – wet forest: jaworze dln. (ef 79 24). 132. chamaenerion angustifolium (l.) scop. – �ickets edge: zwiernik (ef 78 34). 133. *chamomilla suaveolens (pursh) rydb. – epoecophyte. fields and roadsides: podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 02), parkosz (ef 79 03), dobrków (ef 79 04), słotowa (ef 79 31). 134. chelidonium maius l. – roadsides: podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 03), słotowa (ef 79 11), gołęczyna (ef 79 14). 135. chenopodium album l. – fields: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 02, ef 79 12), jaworze dln. (ef 79 24). 136. ch. glaucum l. – field: podgrodzie (ef 69 44). 137. *ch. hybridum l. – archaeophyte. on ruderal habitats: podgrodzie (ef 69 44), parkosz (ef 69 43). 138. ch. polyspermum l. – fields: podgrodzie (ef 69 44), dobrków (ef 79 04), mokrzec (ef 79 13). 139. * ch. strictum roth – epoecophyte. on ruderal habitat: jaworze dln. (ef 79 24). 140. chimaphila umbellata (l.) w. p. c. barton – in pine forests: jaworze dln. (ef 79 24). 141. chrysosplenium alternifolium l. – in deciduous forests: zwiernik (ef 78 24), jaworze dln. (ef 79 24), podgrodzie (ef 79 44). 142. *cichorium intybus l. – archaeophyte. roadsides: podgrodzie (ef 69 44), parkosz (ef 69 43), słotowa (ef 79 11), jaworze dln. (ef 79 24). 143. circaea lutetiana l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 144. cirsium arvense (l.) scop. – fields: parkosz (ef 69 43), pilzno (ef 79 02, ef 79 12), gołęczyna (ef 79 14). 145. c. oleraceum (l.) scop. – wet meadow: dobrków (ef 79 04). 146. c. palustre (l.) scop. – in balk: podgrodzie (ef 69 44). 147. c. rivulare (jacq.) all. – wet meadows: łęki grn. (ef 78 13), parkosz (ef 69 43), strzegocice (ef 79 22). 148. c. vulgare (savi) ten. – roadsides and pastures: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), gołęczyna (ef 79 14), strzegocice (ef 79 22). 149. clinopodium vulgare l. – on dry slopes and balks: gołęczyna (ef 79 14), jaworze dln. (ef 79 24), zagórze (ef 79 33) ), parkosz (ef 79 43). 150. *consolida regalis gray – archaeophyte. fields: jaworze dln. (ef 79 24), bielowy (ef 79 33). 151. convallaria majalis l. – in oak-hornbeam forests: podgrodzie (ef 69 44), łęki grn. (ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 152. convolvulus arvensis l. – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 02, ef 79 12). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 41 153. *conyza canadensis (l.) cronquist – epoecophyte. roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), mokrzec (ef 79 14). 154. cornus sanguinea l. – �ickets: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 155. coronilla varia l. – in balks and on dry slopes grasslands: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dolne (ef 79 24). 156. corydalis solida (l.) clairv. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 157. corylus avellana l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 158. corynephorus canecens (l.) p. beauv. – on roadside and on sandy fallow lands: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 159. crataegus monogyna jacq. – in thickets of balks: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04), mokrzec (ef 79 13), strzegocice (ef 79 22). 160. c. rhipidophylla gand. var. lindmanii (hrabětová) k. i. chr. – in thickets of balk: jaworze grn.(ef 79 34). 161. c. rhipidophylla gand. var. rhipidophylla – �ickets over the wisłoka river: podgrodzie (ef 69 44). 162. c. subsphaericea gand. – in thickets of balk: jaworze dln. (ef 79 24) (oklejewicz i in. 2004). 163. crepis biennis l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 164. c. capillaris (l.) wallr. – meadow: jaworze dln. (ef 79 24). 165. c. paludosa (l.) moench – in meadow and riparian forest: parkosz (ef 69 43), zwiernik (ef 78 34), dobrków (ef 79 04). 166. c. tectorum l. – on sandy fallow area: jaworze dln. (ef 79 24). 167. cruciata glabra (l.) ehrend. – on balk: jaworze dln. (ef 79 24). 168. cucubalus baccifer l. – in wicker over the wisłoka river: podgrodzie (ef 69 44). 169. cuscuta epithymum (l.) l. – on meadow plants: strzegocice (ef 79 23). 170. c. europaea l. – on meadow plants: parkosz (ef 69 43). 171. cynoglossum o�cinale l. – on gravels of the wisłoka river: parkosz (ef 69 43), jaworze dln. (ef 79 24). 172. cynosurus cristatus l. – meadows: podgrodzie (ef 69 44), dobrków (ef 79 04). 173. cystopteris fragilis (l.) bernh. – on a stone wall: strzegocice (ef 79 22). 174. dactylis glomerata l. – meadows: parkosz (ef 69 43), łęki grn. (ef 78 13). 175. d. polygama horv. – in meadow and oak-hornbeam forest: parkosz (ef 69 43), dobrków (ef 79 04). 176. dactylorhiza majalis (rchb.) p. f. hunt & summerh. – partially protected species. wet meadows: podgrodzie (ef 69 44), łęki grn. (ef 78 13), strzegocice (ef 79 22), jaworze dln. (ef 79 24). k ry st yn a to w pa sz 42 177. daphne mezereum l. – partially protected species. in deciduous forests: parkosz (ef 69 43), dobrków (ef 79 04), dęborzyn (ef 79 43). 178. *datura stramonium l. – epoecophyte. field: gołęczyna (ef 79 14). 179. daucus carota l. – meadows: podgrodzie (ef 69 44), dobrków (ef 79 04). 180. dentaria glandulosa waldst. & kit. – higher montane zone species. oak-hornbeam forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 181. deschampsia caespitosa (l.) p. beauv. – meadows: parkosz (ef 69 43), jaworze dln. (ef 79 24), zagórze (ef 79 33). 182. *descurainia sophia (l.) webb ex prantl – archaeophyte. on ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02). 183. dianthus deltoides l. – on sandy grasslands: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 184. *digitaria ischaemum (schreb). h. l. mühl. – archaeophyte. on fallow area: jaworze dln. (ef 79 24). 185. *d. sanguinalis (l.) scop. – archaeophyte. on ruderal habitat: podgrodzie (ef 69 44). 186. *diplotaxis muralis (l.) dc. – epoecophyte. roadside: pilzno (ef 79 02). 187. dipsacus laciniatus l. – on stones of the wisłoka river bank: pilzno (ef 79 12), strzegocice (ef 79 22). 188. d. sylvestris huds. – on stones of the wisłoka river bank: podgrodzie (ef 69 44), pilzno (ef 79 12), mokrzec (ef 79 13). 189. dryopteris carthusiana (vill.) h. p. fuchs – in forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), dęborzyn (ef 79 44), gołęczyna (ef 79 14). 190. d. dilatata (ho�m.) a. gray – all-mountain species. in forests: podgrodzie (ef 69 44), zwiernik (ef 78 34). 191. d. �lix-mas (l.) schott – in forests: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 192. d. expansa (c. presl) fraser-jenk. & jermy – in forest: jaworze dln. (ef 79 24). 193. *echinochloa crus-galli (l.) p. beauv. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44). 194. *echinocystis lobata (f. michx.) torr. & a. gray – holoagriophyte. invasive species. it grows in alluvial forests: mokrzec (ef 79 13). 195. echium vulgare l. – roadsides: podgrodzie (ef 69 44), zwiernik (ef 78 34), dobrków (ef 79 04), jaworze dln. (ef 79 24). 196. eleocharis palustris (l.) roem. & schult. – on banks of ditches: parkosz (ef 69 43), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 197. elymus caninus (l.) l. – in thickets of rivers and streams: podgrodzie (ef 69 44), pilzno (ef 79 12). 198. e. repens (l.) gould – in �eld communities and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), zwiernik (ef 78 34), gołęczyna (ef 79 14). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 43 199. epilobium hirsutum l. – on edge of ditch: podgrodzie (ef 69 44). 200. e. parvi�orum schreb. – on banks of streams: podgrodzie (ef 69 44). gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 201. e. roseum schreb. – on stones of the wisłoka river bank: pilzno (ef 79 12). 202. epipactis helleborine (l.) crantz – partially protected species. oak-hornbeam forests: jaworze dln. (ef 79 24), jaworze grn. (ef 79 34), dęborzyn (ef 89 04). 203. e. purpurata sm. – protected species. oak-hornbeam forest: dęborzyn (ef 79 44). 204. equisetum arvense l. – in forests, �elds, meadows, roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 02, ef 79 03), jaworze dln. (ef 79 24). 205. e. �uviatile l. – in reeds on water reservoirs banks: parkosz (ef 69 43), łęki grn. (ef 78 13). 206. e. hyemale l. – oak-hornbeam forest: podgrodzie (ef 69 44). 207. e. palustre l. – on banks of ditches: łęki grn. (ef 78 13), dobrków (ef 79 04). 208. e. pratense ehrh. – wet oak-hornbeam forest: łęki grn. (ef 78 13). 209. e. ramosissimum desf. – on stones of river banks: łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 210. e. sylvaticum l. – in pine forests: jaworze dln. (ef 79 24), słotowa (ef 79 31). 211. e. telmateia ehrh. – lower montane zone species. in wet forests and on banks of streams: podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 212. erigeron acris l. – roadsides: parkosz (ef 69 43), zagórze (ef 79 33). 213. eriophorum angustifolium honck.– wet meadows: parkosz (ef 69 43), dęborzyn (ef 79 44). 214. e. latifolium hoppe – wet meadow: łęki grn. (ef 78 13). 215. erodium cicutarium (l.) l’hér. – on ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44). 216. erophila verna (l.) chevall. – on grasslands and dry slopes, �elds: parkosz (ef 69 43), gołęczyna (ef 79 14). 217. erysimum cheiranthoides l. – in �elds and ruderal habitats: pilzno (ef 79 12), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 218. euonymus europaea l. – in forests: zwiernik (ef 78 34), parkosz (ef 69 43), dobrków (ef 79 04), jaworze dln. (ef 79 24). 219. e. verrucosa scop. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki dln. (ef 79 42), jaworze dln. (ef 79 24). 220. eupatorium cannabinum l. – in thickets above streams: podgrodzie (ef 69 44). jaworze dln. (ef 79 24). 221. euphorbia amygdaloides l. – in deciduous forests: parkosz (ef 69 43), dobrków (ef 79 04), jaworze dln. (ef 79 24). 222. e. cyparissias l. – on balks, pastures and dry meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), gołęczyna (ef 79 14). k ry st yn a to w pa sz 44 223. e. dulcis l. – oak-hornbeam forest: zwiernik (ef 78 34). 224. e. esula l. – on dry meadows, balks and pastures: podgrodzie (ef 69 44), pilzno (ef 79 12), jaworze dln. (ef 79 24). 225. * e. helioscopia l. – archaeophyte. fields and roadsides: podgrodzie (ef 69 44), pilzno (ef 79 02), mokrzec (ef 79 13). 226. e. serrulata �ull. – higher montane zone species. in thickets and on stones of rivers: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04), jaworze dln. (ef 79 24). 227. euphrasia rostkoviana hayne – on balk: dobrków (ef 79 04). 228. fagus sylvatica l. – in deciduous forests: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 229. *fallopia convolvulus (l.) á. löve – archaeophyte. fields: parkosz (ef 69 43), pilzno (ef 79 02), dobrków (ef 79 04), jaworze grn. (ef 79 34). 230. f. dumetorum (l.) holub. – in thickets on the wisłoka river: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 231. festuca gigantea (l.) vill. – in forests and thickets above streams: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24), zagórze (ef 79 33). 232. f. ovina l. – meadows: parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 233. f. pratensis huds. – meadows: strzegocice (ef 79 22), jaworze dln. (ef 79 24). 234. f. rubra l. – meadows: parkosz (ef 69 43), słotowa (ef 79 31), jaworze dln. (ef 79 24). 235. ficaria verna huds. – in the forests, on roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 03). 236. filipendula ulmaria (l.) maxim. – wet meadows: parkosz (ef 69 43), dobrków (ef 79 04). 237. fragaria vesca l. – in forests, thickets and dry meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24), bielowy (ef 79 33). 238. frangula alnus mill. – forests and thickets: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 239. fraxinus excelsior l. – in moist and wet forests: parkosz (ef 69 43), dobrków (ef 79 04). 240. gagea lutea (l.) ker gawł. – forests and thickets: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 241. galega o�cinalis l. – on edges of thickets and on meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), strzegocice (ef 79 22). 242. galeobdolon luteum huds. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 243. galeopsis bi�da boenn. – in forests: parkosz (ef 69 43), słotowa (ef 79 31). 244. *g. ladanum l. – archaeophyte. field: parkosz (ef 69 43). 245. g. pubescens besser – in forests and on ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 12), jaworze dln. (ef 79 24). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 45 246. g. speciosa mill. – in forests and thickets: podgrodzie (ef 69 44), dobrków (ef 79 04). 247. g. tetrahit l. – in forests: gołęczyna (ef 79 14), jaworze grn. (ef 79 34). 248. *galinsoga ciliata (raf.) s. f. blake – epoecophyte. in ruderal habitats and on �elds: parkosz (ef 69 43), pilzno (ef 79 12). 249. *g. parvi�ora cav. – epoecophyte. in ruderal habitats and on �elds: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 250. galium aparine l. – in forestss, thickets and on �elds: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 02, ef 79 12). 251. g. mollugo l. – on meadows and balks: łęki grn. (ef 78 13), zagórze (ef 79 43). 252. g. odoratum (l.) scop. – in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 253. g. palustre l. – banks of ponds and wet meadows: parkosz (ef 69 43), bielowy (ef 79 33). 254. g. rivale (sibth. & sm.) – on banks of ditches: parkosz (ef 69 43). 255. g. schultesii vest – in forests and thickets: parkosz (ef 69 43), jaworze dln. (ef 79 24). 256. g. verum l. – meadows and forests: parkosz (ef 69 43), podgrodzie (ef 69 44), strzegocice (ef 79 22). 257. gentianella ciliata (l.) borkh. – in grassland of dry meadow: dobrków (ef 79 04). 258. *geranium dissectum l. – archaeophyte. fields: podgrodzie (ef 69 44), mokrzec (ef 79 13), bielowy (ef 79 33). 259. g. palustre l. – wet meadows: strzegocice (ef 79 22), jaworze dln. (ef 79 24). 260. g. phaeum l. – all-mountain species. in riparian forest over the wisłoka river: parkosz (ef 69 43). 261. g. pratense l. – meadows: podgrodzie (ef 69 44), łęki grn. (ef 78 11), bielowy (ef 79 33). 262. *g. pusillum burm. f. ex l. – archaeophyte. fields: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 263. g. robertianum l. – in forest: jaworze dln. (ef 79 24). 264. g. sanguineum l. – on edge of thickets: jaworze dln. (ef 79 24). 265. geum urbanum l. – wet forests: łęki grn. (ef 78 13), parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 266. glechoma hederacea l. – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), zwiernik (ef 78 34), pilzno (ef 79 12), jaworze dln. (ef 79 24). 267. g. hirsuta waldst. & kit. – in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 268. glyceria maxima (hartm.) holub. – on banks of pond and over the wisłoka river: parkosz (ef 69 43). podgrodzie (ef 69 44). 269. gnaphalium uliginosum l. – wet �elds: parkosz (ef 69 43), jaworze dln. (ef 79 24). 270. gymnocarpium dryopteris (l.) newman – in forest: jaworze dln. (ef 79 24). 271. hedera helix l. – protected species. in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), dobrków (ef 79 04). k ry st yn a to w pa sz 46 272. *helianthus tuberosus l. – holoagriophyte. invasive species. in alluvial forests over the wisłoka river: pilzno (ef 79 12). 273. heracleum sphondylium l. – meadows and roadsides: podgrodzie (ef 69 44), pilzno (ef 79 03), dobrków (ef 79 04). 274. hieracium lachenalii c. c. gmel. – in forests: bielowy (ef 79 33), jaworze dln. (ef 79 24). 275. h. murorum l. – in forests: zwiernik (ef 78 34), gołęczyna (ef 79 14). 276. h. pilosella l. – dry meadows and pastures: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), bielowy (ef 79 33). 277. h. piloselloides vill. – dry meadows: strzegocice (ef 79 22). 278. h. sabaudum l. – in forests: parkosz (ef 69 43), jaworze dln. (ef 79 24). 279. h. umbellatum l. – in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24), słotowa (ef 79 31). 280. holcus lanatus l. – meadows: łęki grn. (ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 281. holosteum umbellatum l. – in slope of grassland: słotowa (ef 79 31). 282. *hordeum murinum l. – archaeophyte. roadside: jaworze dln. (ef 79 24). 283. humulus lupulus l. – in alluvial forests over rivers and streams: podgrodzie (ef 69 44), parkosz (ef 69 43), jaworze dln. (ef 79 24), bielowy (ef 79 33). 284. hypericum hirsutum l. – in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 285. h. humifusum l. – pasture: podgrodzie (ef 69 44). 286. h. maculatum crantz – wet meadow: słotowa (ef 79 31). 287. h. perforatum l. – meadows and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12), gołęczyna (ef 79 14). 288. h. tetrapterum fr. – wet meadows: dobrków (ef 79 04), jaworze dln. (ef 79 24). 289. hypochoeris radicata l. – meadows and pastures: podgrodzie (ef 69 44), parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), słotowa (ef 79 31). 290. *impatiens glandulifera royle – holoagriophyte. invasive species. in alluvial forest: dobrków (ef 79 04). 291. i. noli-tangere l. – wet forests: parkosz (ef 69 43), jaworze dln. (ef 79 24). 292. *i. parvi�ora dc. – holoagriophyte. invasive species. in ruderal habitat: pilzno (ef 79 12). 293. inula britannica l. – meadows: parkosz (ef 69 43), jaworze dln. (ef 79 24). 294. *i. helenium l. – hemiagriophyte. on edges of thickets: bielowy (ef 79 33). 295. iris pseudacorus l. – on banks of ditches: strzegocice (ef 79 22). 296. isopyrum thalictroides l. – in forest: podgrodzie (ef 69 44). 297. jasione motana l. – on sandy dunes and fallow lands: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 47 298. *juglans regia l. – holoagriophyte. currently in study area it settled on balks and in forests: rędziny (ef 79 22), jaworze dln. (ef 79 24). 299. juncus articulatus l. emend. k. richt. – meadows: podgrodzie (ef 69 44), pilzno (ef 79 12). 300. j. bufonius l. – in wet �elds and ditches: podgrodzie (ef 69 44), dobrków (ef 79 04), pilzno (ef 79 12). 301. j. conglomeratus l. emend. leers – meadows: parkosz (ef 69 43), jaworze dln. (ef 79 24). 302. j. in�exus l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 303. *j. tenuis willd. – hemiagriophyte. invasive species. on roadsides and pastures: podgrodzie (ef 69 44), bielowy (ef 79 33). 304. juniperus communis l. – �ickets: parkosz (ef 69 43), jaworze dln. (ef 79 24). 305. knautia arvensis (l.) j. m. coult. – medow: słotowa (ef 79 31). 306. *lactuca serriola l. – archaeophyte. roadside: podgrodzie (ef 69 44). 307. *lamium album l. – archaeophyte. roadsides: łęki grn. (ef 78 13), parkosz (ef 69 43), pilzno (79 12), jaworze dln. (ef 79 24). 308. *l. amplexicaule l. – archaeophyte. fields: jaworze dln. (ef 79 24). 309. l. maculatum l. – on roadsides and ruderal habitats and in thickets above streams: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12), zagórze (ef 79 33). 310. *l. purpureum l. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 03), jaworze dln. (ef 79 24). 311. lapsana communis l. – fields and roadsides: zwiernik (ef 78 34), pilzno (79 12), mokrzec (ef 79 13). 312. lathyrus pratensis l. – meadows: rędziny (ef 79 22), bielowy (ef 79 33). 313. *l. tuberosus l. – archaeophyte. on cereal �elds: parkosz (ef 69 43), pilzno (ef 79 12), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), bielowy (ef 79 33). 314. l. vernus (l.) bernh. – in forests: podgrodzie (ef 69 44), parkosz (ef 69 43 dobrków (ef 79 04). 315. lavathera thuringiaca l. – in thickets over the wisłoka river: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 316. lembotropis nigricans (l.) griseb. – on edge of pine forest: gołęczyna (ef 79 14). 317. lemna minor l. – in pond: parkosz (ef 69 43), jaworze dln. (ef 79 24). 318. l. trisulca l. – in pond: parkosz (ef 69 43). 319. leontodon autumnalis l. – meadows: strzegocice (ef 79 22). 320. l. hispidus l. – roadsides and meadows: podgrodzie (ef 69 44), strzegocice (ef 79 22), jaworze dln. (ef 79 24). 321. *leonurus cardiaca l. – archaeophyte. on ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12), jaworze dln. (ef 79 24). k ry st yn a to w pa sz 48 322. *lepidium campestre (l.) r. br. – archaeophyte. on ruderal habitat: jaworze dln. (ef 79 24). 323. *l. ruderale l. – archaeophyte. on ruderal habitat: pilzno (ef 79 12). 324. leucanthemum vulgare lam. – meadows: parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 325. ligustrum vulgare l. – in xerothermic thickets in balks and in thickets over the wisłoka river: łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 326. lilium martagon l. – protected species. in oak-pine forest: jaworze dln. (ef 79 24). 327. linaria vulgaris mill. – on balks: podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 12), bielowy (ef 79 33). 328. *lithospermum arvense l. – archaeophyte. fields: parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 329. *lolium multi�orum lam. – epoecophyte. in meadows and �elds in cereal crops: parkosz (ef 69 43), pilzno (ef 79 02, ef 79 12), bielowy (ef 79 33). 330. l. perenne l. – meadows and pastures: podgrodzie (ef 69 44), parkosz (ef 69 43), pilzno (ef 79 12). 331. lonicera xylosteum l. – in deciduous forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 332. lotus corniculatus l. – meadows: podgrodzie (ef 69 44), parkosz (ef 69 43), bielowy (ef 79 33). 333. l. uliginosus schkuhr. – wet meadow: parkosz (ef 69 43). 334. luzula campestris (l.) dc. – meadows: podgrodzie (ef 69 44), parkosz (ef 69 43), łęki grn. (ef 78 13), słotowa (ef 79 31). 335. l. multi�ora (retz.) lej. – meadows: gołęczyna (ef 79 14), bielowy (ef 79 33). 336. l. pilosa (l.) willd. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), słotowa (ef 79 31). 337. lychnis �os-cuculi l. – bank of pond: parkosz (ef 69 43), bielowy (ef 79 33). 338. *lycium barbarum l. – epoecophyte. roadsides: parkosz (ef 69 43), pilzno (ef 79 12), strzegocice (ef 79 22). 339. lycopus europaeus l. – on banks of streams: podgrodzie (ef 69 44), zwiernik (ef 78 34), dobrków (ef 79 04). 340. lysimachia nemorum l. – higher montane zone species. in forest: jaworze dln. (ef 79 24). 341. l. nummularia l. – meadows: parkosz (ef 69 43), łęki górne (ef 78 13), jaworze dln. (ef 79 24), słotowa (ef 79 31). 342. l. vulgaris l. – wet meadows and �elds: podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 343. lythrum salicaria l. – over banks of ditches: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 49 344. maianthemum bifolium (l.) f. w. schmidt – in deciduous forests: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04), gołęczyna (ef 79 14). 345. *malva neglecta wallr. – archaeophyte. on ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44). 346. *m. sylvestris l. – archaeophyte. roadsides: podgrodzie (ef 69 44), dobrków (ef 79 04), jaworze dln. (ef 79 24), bielowy (ef 79 33). 347. *matricaria maritima l. subsp. inodora (l.) dostál – archaeophyte. fields and roadsides: parkosz (ef 69 43), mokrzec (ef 79 13), bielowy (ef 79 33). 348. matteucia struthiopteris (l.) tod. – lower montane zone species and partially protected. w olszynie nad potokiem: jaworze dln. (ef 79 24). 349. medicago falcata l. – on balks: parkosz (ef 69 43), gołęczyna (ef 79 14). 350. m. lupulina l. – meadows: parkosz (ef 69 43), pilzno (ef 79 12), strzegocice (ef 79 22). 351. *m. sativa l. – hemiagriophyte. on meadows, balks and on roadsides: parkosz (ef 69 43), pilzno (ef 79 02), jaworze dln. (ef 79 24), bielowy (ef 79 33). 352. *m. ×varia martyn – epoecophyte. on balk: pilzno (ef 79 12). 353. melampyrum arvense l. – on thermophilic meadows, �elds: parkosz (ef 69 43), pilzno (ef 79 12). 354. m. nemorosum l. – in deciduous forests and thickets: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34). 355. *melandrium album (mill.) garcke – archaeophyte. meadows: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 02, ef 79 12), gołęczyna (ef 79 14). 356. melica nutans l. – in deciduous forests: parkosz (ef 69 43), zwiernik (ef 78 34). 357. melilotus alba medik. – on stones of rivers and on roadsides: podgrodzie (ef 69 44), dulczówka (ef 79 12), jaworze grn. (ef 79 34). 358. m. o�cinalis (l.) pall. – on stones of rivers and on balks: podgrodzie (ef 69 44), jaworze grn. (ef 79 34). 359. mentha aquatica l. – on banks of ditches and ponds: parkosz (ef 69 43), dobrków (ef 79 04), jaworze dln.(ef 79 24). 360. m. arvensis l. – fields and roadsides: parkosz (ef 69 43), pilzno (ef 79 02), mokrzec (ef 79 13). 361. m. longifolia (l.) l. – on banks of streams, ditches and in wet forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn.(ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 362. mercurialis perennis l. – in deciduous forests: jaworze dln. (ef 79 24), zagórze (ef 79 43). 363. milium e�usum l. – in deciduous forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln.(ef 79 24). 364. moehringia trinervia (l.) clairv. – in forests and in thickets over the wisłoka river: podgrodzie (ef 69 44), bielowy (ef 79 33). k ry st yn a to w pa sz 50 365. moneses uni�ora (l.) a. gray – in pine forest: jaworze dln. (ef 79 24). 366. monotropa hypopitys l. – in pine forest: jaworze dln. (ef 79 24). 367. mycelis muralis (l.) dumort. – in forest: parkosz (ef 69 43), dobrków (ef 79 04). 368. *myosotis arvensis (l.) hill – archaeophyte. on �elds in cereal and root crops: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 12), strzegocice (ef 79 22). 369. m. palustris (l.) l. emend. rchb. – on wet meadows and on banks of ditches: parkosz (ef 69 43), zwiernik (ef 78 34), dobrków (ef 79 04), bielowy (ef 79 33). 370. m. sparsi�ora pohl. – in wet oak-hornbeam forest: podgrodzie (ef 69 44). 371. m. stricta link ex roem. & schult. – fields: podgrodzie (ef 69 44), bielowy (ef 79 33), jaworze dln. (ef 79 24). 372. m. sylvatica ehrh. ex ho�m. – in wet oak-hornbeam forest: pilzno (ef 79 12). 373. myosoton aquaticum (l.) moench – on stones of the wisłoka river: podgrodzie (ef 69 44). 374. myriophyllum spicatum l. – in water reservoir: jaworze dln. (ef 79 24). 375. *nepeta cataria l. – archaeophyte. pastures: parkosz (ef 69 43), gołęczyna (ef 79 14). 376. nuphar lutea (l.) sibth. & sm. – in pond: parkosz (ef 69 43). 377. nymphaea candida c. presl – partially protected species. in water reservoir: rędziny (ef 79 22). 378. odontites serotina (lam.) rchb. – pastures: parkosz (ef 69 43), jaworze dln. (ef 79 24). 379. * o. verna (bellardi) dumort. – archaeophyte. fields: parkosz (ef 69 43), pilzno (ef 79 02). 380. oenanthe aquatica (l.) poir. – on banks of ponds: parkosz (ef 69 43), podgrodzie (ef 69 44). 381. oenothera biennis l. – on banks of rivers and roadsides: podgrodzie (ef 69 44), mokrzec (ef 79 13), gołęczyna (ef 79 14). 382. ononis arvensis l. – on balks and stones of the wisłoka river: podgrodzie (ef 69 44), dobrków (ef 79 04), słotowa (ef 79 31). 383. *onopordum acanthium l. – archaeophyte. on ruderal habitats: podgrodzie (ef 69 44). 384. origanum vulgare l. – on balks: podgrodzie (ef 69 44), mokrzec (ef 79 13), słotowa (ef 79 31). 385. oxalis acetosella l. – in forest: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 386. *o. fontana bunge – epoecophyte. fields: parkosz (ef 69 43), pilzno (ef 79 02, ef 79 12). 387. padus avium mill. – in moist forests and thickets: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13). 388. *papaver argemone l. – epoecophyte. fields: podgrodzie (ef 69 44), gołęczyna (ef 79 14). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 51 389. *p. rhoeas l. – archaeophyte. fields: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 02), bielowy (ef 79 33). 390. paris quadrifolia l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 391. *parthenocissus inserta (a. kern.) fritsch – holoagriophyte. invasive species. in alluvial forests on rivers and streams: gołęczyna (ef 79 14), jaworze dln.(ef 79 24). 392. *pastinaca sativa l. – archaeophyte. meadows: podgrodzie (ef 69 44). 393. petasites albus (l.) gaertn. – higher montane zone species. in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 394. p. hybridus (l.) p. gaertn., b. mey. & scherb. – over rivers and streams: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 395. peucedanum oreoselinum (l.) moench – on edges of pine forests: gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 396. phalaris arundinacea l. – in reeds over banks of streams and ditches: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 397. phegopteris connectilis (michx.) watt – in forests: jaworze dln. (ef 79 24). 398. phleum pratense l. – meadows: parkosz (ef 69 43), łęki grn. (ef 78 13), jaworze dln. (ef 79 24), bielowy (ef 79 33). 399. phragmites australis (cav.) trin. ex steud. – over banks of drainage ditches: jaworze dln. (ef 79 24), bielowy (ef 79 33). 400. picea abies (l.) h. karst. – in forests, as an admixture: zwiernik (ef 78 34), gołęczyna (ef 79 14), jaworze dln. (ef 79 34). 401. pimpinella maior (l.) huds. – meadow: zwiernik.(ef 78 34). 402. p. saxifraga l. – on balks: podgrodzie (ef 69 44), strzegocice (ef 79 22). 403. pinus sylvestris l. – in forests: parkosz (ef 69 43), słotowa (ef 79 31), jaworze dln. (ef 79 24). 404. plantago intermedia gilib. – fields: parkosz (ef 69 43), pilzno (ef 79 02, ef 79 12). 405. p. lanceolata l. – meadows, pastures and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13). 406. p. major l. – meadows, pastures and roadsides: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 03), jaworze dln. (ef 79 24). 407. p. media l. – on dry meadows and balks: parkosz (ef 69 34), podgrodzie (ef 69 44), dobrków (ef 79 04). 408. platanthera bifolia (l.) rich. – partially protected species. in forests: dobrków (ef 79 04), bielowy (ef 79 33). 409. poa annua l. – pastures and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 410. p. compressa l. – on balks and on dry slopes of grasslands: parkosz (ef 69 43), zwiernik (ef 78 34), jaworze dln. (ef 79 24), bielowy (ef 79 33). k ry st yn a to w pa sz 52 411. p. nemoralis l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 412. p. palustris l. – on alluvial of rivers and over banks of ponds: parkosz (ef 69 43), pilzno (ef 79 12). 413. p. pratensis l. – on meadows, balks and roadsides: podgrodzie (ef 69 44), łęki grn. (ef 78 13), strzegocice (ef 79 22), jaworze dln. (ef 79 24). 414. p. trivialis l. – wet meadows and thickets: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 12), strzegocice (ef 79 22). 415. polygala comosa schkuhr – on dry slopes of grasslands and in balks: latoszyn (ef 69 44), dobrków (ef 79 04). 416. p. oxyptera rchb. – on dry and barren grasslands: zagórze (ef 79 33). 417. p. vulgaris l. – dry meadows: parkosz (ef 69 43), dobrków (ef 79 04). 418. polygonatum multi�orum (l.) all. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 419. polygonum amphibium l. – on wet meadows and over banks of waters: parkosz (ef 69 43), pilzno (ef 79 12), bielowy (ef 79 33). 420. p. aviculare l. – fields and pastures: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 02), bielowy (ef 79 33). 421. p. bistorta l. – wet meadows: parkosz (ef 69 43). 422. p. hydropiper l. – fields: parkosz (ef 69 43), zagórze (ef 79 33). 423. p. lapathifolium l. subsp. brittingeri (opiz) rech. f. – on ruderal habitats: podgrodzie (ef 69 44), pilzno (ef 79 12). 424. p. lapathifolium l. subsp. lapathifolium – fields: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 425. p. lapathifolium l. subsp. pallidum (with.) fr. – fields, ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02), mokrzec (ef 79 13), bielowy (ef 79 33). 426. p. minus huds. – on wet �elds and over banks of ditches: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 427. p. mite schrank – over ditches and on �elds: parkosz (ef 69 43), podgrodzie (ef 69 44). 428. p. persicaria l. – ruderal habitats, �elds: parkosz (ef 69 43), pilzno (ef 79 02), mokrzec (ef 79 13). 429. polypodium vulgare l. – in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24), zagórze (ef 79 33). 430. populus alba l. – in alluvial forests over the wisłoka river: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12). 431. p. nigra l. – in alluvial forests over the wisłoka river: pilzno (ef 79 12), strzegocice (ef 79 22). 432. p. tremula l. – in forests: parkosz (ef 69 43), łęki grn. (ef 78 13), słotowa (ef 79 31). 433. potamogeton crispus l. – in still waters: jaworze dln. (ef 79 24). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 53 434. p. natans l. – in bends of the wisłoka river: jaworze dln. (ef 79 24). 435. p. nodosus poir. – in bends of the wisłoka river: jaworze dln. (ef 79 24). 436. p. pusillus l. – in bends of the wisłoka river: jaworze dln. (ef 79 24). 437. potentilla anserina l. – on �elds, pastures and roadsides: podgrodzie (ef 69 44), parkosz (ef 69 43), pilzno (ef 79 03), bielowy (ef 79 33). 438. p. argentea l. – pastures: podgrodzie (ef 69 44), parkosz (ef 69 43), słotowa (ef 79 11). 439. p. erecta (l.) raeusch. – on edges of forests: bielowy (ef 79 33). 440. p. pusilla host – higher montane zone species. on stones of the wisłoka river: parkosz (ef 69 43). 441. p. reptans l. – meadows and roadsides: parkosz (ef 69 43), pilzno (ef 79 02, ef 79 12), jaworze dln. (ef 79 24). 442. p. supina l. – pastures: parkosz (ef 69 43). 443. primula elatior (l.) hill – partially protected species. in wet forests and meadows: parkosz (ef 69 43), jaworze dln. (ef 79 24). 444. prunella vulgaris l. – meadows and pastures: parkosz (ef 69 43), dobrków (ef 79 04). 445. prunus spinosa l. – in thickets and on balks: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 446. pteridium aquilinum (l.) kuhn – in mixed forests, in cutting places and balks: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 447. pulmonaria obscura dumort. – in deciduous forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 448. pyrola chlorantha sw. – partially protected species. in pine forest: jaworze dln. (ef 79 24). 449. p. minor l. – partially protected species. in pine forest: jaworze dln. (ef 79 24). 450. p. rotundifolia l. – partially protected species. in pine forest: jaworze dln. (ef 79 24). 451. pyrus communis l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24), bielowy (ef 79 33). 452. p. pyraster (l.) burgsd. – on balks and in thickets: parkosz (ef 69 43), dobrków (ef 79 04), mokrzec (ef 79 13). 453. quercus petraea (matt.) liebl. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 454. q. robur l. – in forests: łęki grn. (ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 455. *q. rubra l. – holoagriophyte. invasive species. in forests: dobrków (ef 79 04), mokrzec (ef 79 13). 456. ranunculus acris l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), zwiernik (ef 78 34), pilzno (ef 79 03), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). k ry st yn a to w pa sz 54 457. *r. arvensis l. – archaeophyte. in cereal crops: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 458. r. auricomus l. – wet meadows: parkosz (ef 69 43), strzegocice (ef 79 22). 459. r. cassubicus l. – in forest: parkosz (ef 69 43). 460. r. �ammula l. – on banks of water reservoirs: parkosz (ef 69 43). 461. r. lanuginosus l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44). 462. r. polyanthemos l. – dry meadows: gołęczyna (ef 79 14), jaworze dln. (ef 79 24), bielowy (ef 79 33). 463. r. repens l. – in meadows and ruderal habitats: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 12). 464. r. sardous cranz – pastures: parkosz (ef 69 43), dobrków (ef 79 04). 465. *raphanus raphanistrum l. – archaeophyte. fields: parkosz (ef 69 43). 466. reseda lutea l. – roadsides: podgrodzie (ef 69 44), pilzno (ef 79 02), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 467. *reynotria japonica houtt. – holoagriophyte. invasive species. in forests and on ru deral habitats: gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 468. rhamnus cathartica l. – in thermophilic thickets on balks: parkosz (ef 69 43), podgrodzie (ef 69 44). 469. rhinantus serotinus (schönh.) oborný – meadows and �elds: parkosz (ef 69 43). 470. ribes spicatum e. robson – in forests: jaworze dln. (ef 79 24), gołęczyna (ef 79 14). 471. *robinia pseudoacacia l. – holoagriophyte. invasive species. in forests: parkosz (ef 69 43), mokrzec (ef 79 13). 472. rorippa ×armoracioides (tausch) fuss – on edge of thickets: jaworze dln. (ef 79 24). 473. r. austriaca (cranz) besser – over the wisłoka river: pilzno (ef 79 12). 474. r. palustris (l.) besser – over the wisłoka river: pilzno (ef 79 12). 475. r. sylvestris (l.) besser – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02), gołęczyna (ef 79 14). 476. rosa canina l. – in thickets, on balks and on edges of forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24), bielowy (ef 79 33), jaworze grn. (ef 79 34). 477. r. rubiginosa l. – in thickets over the wisłoka river: jaworze grn. (ef 79 34). 478. rubus bifrons vest – on edge of thickets: podgrodzie (ef 69 44). 479. r. caesius l. – �ickets: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), słotowa (ef 79 31). 480. r. hirtus waldst. & kit. – forests and thickets: pilzno (ef 79 12), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 481. r. idaeus l. – forests and thickets: podgrodzie (ef 69 44), łęki grn. (ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), bielowy (ef 79 33). 482. r. montanus lib. ex lej. – �ickets: parkosz (ef 69 43), jaworze dln. (ef 79 24). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 55 483. r. plicatus weihe & nees – in thickets over the wisłoka river: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 484. *rudbeckia laciniata l. – holoagriophyte. invasive species. in alluvial forests: łęki grn. (ef 78 13), dulczówka (ef 79 12). 485. rumex acetosa l. – meadows: podgrodzie (ef 69 44), słotowa (ef 79 31), jaworze dln. (ef 79 24). 486. r. acetosella l. – fields: podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24), bielowy (ef 79 33). 487. r. aquaticus l. – on bank of ditch: strzegocice (ef 79 22). 488. r. conglomeratus murray – in ditches and on wet meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04), jaworze dln. (ef 79 24). 489. r. crispus l. – meadows: łęki grn. (ef 78 13), dobrków (ef 79 04). 490. r. obtusifolius l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), słotowa (ef 79 31). 491. r. sanguineus l. – wet forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 492. r. thyrsi�orus fingerh. – on meadows and on roadside slopes: parkosz (ef 69 43), pilzno (ef 79 12). 493. sagina procumbens l. – wet meadows: jaworze dln. (ef 79 24). 494. salix alba l. – in alluvial forests over the wisłoka river: podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12), słotowa (ef 79 31). 495. s. caprea l. – �ickets: podgrodzie (ef 69 44), strzegocice (ef 79 22). 496. s. cinerea l. – in riparian forest over stream: strzegocice (ef 79 22). 497. s. fragilis l. – in riparian forests over streams: łęki grn. (ef 78 13), parkosz (ef 69 43). 498. s. purpurea l. – in riparian forests over streams: podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 499. s. triandra l. – in alluvial forests over the wisłoka river: parkosz (ef 69 43), pilzno (ef 79 12), strzegocice (ef 79 22). 500. s. viminalis l. – in riparian forests over rivers: łęki grn. (ef 78 13), parkosz (ef 69 43), jaworze dln. (ef 79 24). 501. salvia glutinosa l. – higher montane zone species. in forests: podgrodzie (ef 69 34), parkosz (ef 69 43), gołęczyna (ef 79 14). 502. sambucus ebulus l. – on balks: dobrków (ef 79 04), bielowy (ef 79 33). 503. s. nigra l. – forests and thickets: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), bielowy (ef 79 33), jaworze grn. (ef 79 34). 504. s. racemosa l. – higher montane zone species. in forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 505. sanguisorba minor scop. – on dry slope: jaworze dln. (ef 79 24). k ry st yn a to w pa sz 56 506. sanicula europaea l. – in forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze grn. (ef 79 34). 507. saponaria o�cinalis l. – roadsides: podgrodzie (ef 69 44), pilzno (ef 79 12), gołęczyna (ef 79 14). 508. sarothamnus scoparius (l.) w. d. j. koch – on edge of forest: jaworze dln. (ef 79 24). 509. scilla bifolia l. – all-mountain species, partially protected. in forests: parkosz (ef 69 43), podgrodzie (ef 69 44). 510. scirpus sylvaticus l. – on wet meadows and over banks of ditches: podgrodzie (ef 69 44), dobrków (ef 79 04), bielowy (ef 79 33). 511. *scleranthus annuus l. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), mokrzec (ef 79 13), gołęczyna (ef 79 14), zagórze (ef 79 33). 512. s. perennis l. – on sandy dunes and fallow lands: podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 513. s. polycarpos l. – on sandy fallow land: gołęczyna (ef 79 14). 514. scrophularia nodosa l. – �ickets: parkosz (ef 69 43). 515. s. umbrosa dumort. – over banks of pond: jaworze dln. (ef 79 24). 516. sedum acre l. – on stones of the wisłoka river: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 517. s. maximum (l.) ho�m. – on roadside slopes and balks: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 518. s. sexangulare l. – on stones of the wisłoka river: podgrodzie (ef 69 44), parkosz (ef 69 43). 519. selinum carvifolia (l.) l. – on edge of forest: strzegocice (ef 79 22). 520. senecio barbaraeifolius (krock.) wimm. & grab. – wet meadows: jaworze dln. (ef 79 24), dęborzyn (ef 79 44). 521. s. �uviatilis wallr. – in thickets on the wisłoka river: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 12), mokrzec (ef 79 13). 522. s. jacobaea l. – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04). 523. s. ovatus (p. gaertn., b. mey. & schreb.) willd. – higher montane zone species. in forests and on cutting places: zwiernik (ef 78 34), jaworze grn. (ef 79 34). 524. s. sylvaticus l. – on cutting places: jaworze dln. (ef 79 24). 525. *s. vernalis waldst. & kit. – epoecophyte. roadside: jaworze dln. (ef 79 24). 526. s. viscosus l. – on stones of the wisłoka river: podgrodzie (ef 69 34). 527. *s. vulgaris l. – archaeophyte. on ruderal habitats: pilzno (ef 79 12). 528. seseli annuum l. – in pine forests: jaworze dln. (ef 79 24). 529. *setaria pumila (poir.) roem. & schult. – archaeophyte. fields: parkosz (ef 69 43) pilzno (ef 79 02). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 57 530. *s. viridis (l.) p. beauv. – archaeophyte. fields: parkosz (ef 69 43). podgrodzie (ef 69 44), dobrków (ef 79 04), pilzno (ef 79 12), jaworze dln. (ef 79 24). 531. *sherardia arvensis l. – archaeophyte. fields: parkosz (ef 69 43), pilzno (ef 79 02), dobrków (ef 79 04), bielowy (ef 79 33). 532. silene nutans l. subsp. nutans – dry meadow: jaworze dln. (ef 79 24). 533. s. vulgaris (moench) garcke – meadows and roadsides: podgrodzie (ef 69 44), zwiernik (ef 78 34), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 534. *sinapis arvensis l. – archaeophyte. fields: parkosz (ef 69 43), pilzno (ef 79 02, ef 79 12). 535. *sisymbrium o�cinale (l.) scop. – archaeophyte. in �elds and ruderal habitats: parkosz (ef 69 43), zwiernik (ef 78 34), pilzno (ef 79 12). 536. solanum dulcamara l. – wet forests: dobrków (ef 79 04), jaworze dln. (ef 79 24). 537. *s. nigrum l. emend. mill. – archaeophyte. roadsides: parkosz (ef 69 43), pilzno (ef 79 12), mokrzec (ef 79 13). 538. *solidago gigantea aiton – holoagriophyte. invasive species. in alluvial forests over rivers and on roadsides: podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12), mokrzec (ef 79 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 539. s. virgaurea l. – in mixed forests: jaworze dln. (ef 79 24), bielowy (ef 79 33). 540. sonchus arvensis l. – fields and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02), dobrków (ef 79 04), słotowa (ef 79 31). 541. *s. asper (l.) hill – archaeophyte. in �elds of root crops and in ruderal habitats: parkosz (ef 69 43), dobrków (ef 79 04). 542. *s. oleraceus l. – archaeophyte. in �elds of root crops: dobrków (ef 79 04). 543. sorbus aucuparia l. emend. hedl. – forests and thickets: parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 544. *spergula arvensis l. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), słotowa (ef 79 31), jaworze dln. (ef 79 24). 545. s. morisonii boreau – on sand dune: jaworze dln. (ef 79 24). 546. spergularia rubra (l.) j. presl & c. presl – on sandy fallow lands: gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 547. spirodela polyrhiza (l.) schleid. – in pond: mokrzec (ef 79 14). 548. stachys palustris l. – on bank of pond: parkosz (ef 69 43). 549. s. sylvatica l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24), bielowy (ef 79 33). 550. staphyllea pinnata l. – protected species. in oak-hornbeam forest: podgrodzie (ef 69 44). 551. stellaria graminea l. – meadows: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), dobrków (ef 79 04), pilzno (ef 79 12). 552. s. holostea l. – in oak-hornbeam forests: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04), bielowy (ef 79 33). k ry st yn a to w pa sz 58 553. s. media (l.) vill. – in forests and thickets and in �elds and ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 02), słotowa (ef 79 31). 554. s. nemorum l. – in forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 555. s. uliginosa murray – wet meadows: zwiernik (ef 78 34). 556. symphytum o�cinale l. – on moist meadows, banks of ditches and on roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02). 557. s. tuberosum l. – in oak-hornbeam forests and alluvial forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24), słotowa (ef 79 31). 558. *syringa vulgaris l. – holoagriophyte. in thickets: parkosz (ef 69 43). 559. *tanacetum parthenium (l.) sch. bip. – epoecophyte. roadsides: dobrków (ef 79 04), mokrzec (ef 79 13). 560. t. vulgare l. – on pastures, roadsides and in ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 03), strzegocice (ef 79 22). 561. taraxacum o�cinale f. h. wigg. – meadows, �elds and roadsides: podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 02, ef 79 03), dobrków (ef 79 04), gołęczyna (ef 79 14). 562. teesdalea nudicaulis (l.) r. br. – on sandy �elds: parkosz (ef 69 43), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 563. *telekia speciosa (schreb.) baumg. – holoagriophyte. in alder forest above the stream: jaworze dln. (ef 79 24). 564.*�laspi arvense l. – archaeophyte. in �elds and in ruderal habitats: parkosz (ef 69 43), pilzno (ef 79 03, ef 79 12). 565. �ymus pulegioides l. – on dry meadows and balks: parkosz (ef 69 43), pilzno (ef 79 03), dobrków (ef 79 04). 566. t. serpyllum l. emend. fr. – in pine forests: gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 567. torilis japonica (houtt.) dc. – in thickets over the wisłoka river: podgrodzie (ef 69 44), mokrzec (ef 79 13). 568. tragopogon orientalis l. – meadows: jaworze grn. (ef 79 34). 569. trientalis europaea l. – in pine forests: gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 570. trifolium arvense l. – roadsides: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 571. t. aureum pollich – on balks: parkosz (ef 69 43), dobrków (ef 79 04). 572. t. campestre schreb. – in grasslands on dry slopes and in meadows: podgrodzie (ef 69 44), łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 573. t. dubium sibth. – meadows: podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 12). 574. t. fragiferum l. – on wet meadows and pastures: podgrodzie (ef 69 44), dobrków (ef 79 04). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 59 575. t. hybridum l. subsp. hybridum – wet meadows. parkosz (ef 69 43), dobrków (ef 79 04), słotowa (ef 79 31). 576. t. medium l. – on balks and dry meadows: parkosz (ef 69 43), dobrków (ef 79 04). 577. t. pratense l. – on stones of the wisłoka river and on meadows: podgrodzie (ef 69 44), łęki grn. (ef 78 13), dulczówka (ef 79 12). 578. t. repens l. – meadows: parkosz (ef 69 43), łęki grn. (ef 78 13), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 579. trisetum �avescens (l.) p. beauv. – meadows: łęki grn. (ef 78 13), jaworze dln. (ef 79 24). 580. tussilago farfara l. – in �elds, balks and ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02, ef 79 03), jaworze dln. (ef 79 24), bielowy (ef 79 33). 581. typha latifolia l. – in water reservoirs: parkosz (ef 69 43), dobrków (ef 79 04), dulczówka (ef 79 12), mokrzec (ef 79 13), jaworze dln. (ef 79 24). 582. ulmus glabra huds. – oak-hornbeam forests: podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 583. u. laevis pall. – oak-hornbeam forests: parkosz (ef 69 43), słotowa (ef 79 31). 584. urtica dioica l. – in moist deciduous forests and ruderal habitats: parkosz (ef 69 43), łęki grn. (ef 78 13), pilzno (ef 79 03), jaworze dln. (ef 79 24). 585. * u. urens l. – archaeophyte. on ruderal habitats: parkosz (ef 69 43), mokrzec (ef 79 13), gołęczyna (ef 79 14). 586. vaccinium myrtillus l. – in mixed forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), gołęczyna (ef 79 14). 587. v. vitis-idaea l. – in mixed forests: podgrodzie (ef 69 44), dobrków (ef 79 04), gołęczyna (ef 79 14). 588. valeriana simplicifolia kabath – wet meadows: parkosz (ef 69 43), dobrków (ef 79 04), bielowy (ef 79 33). 589. *valerianella dentata (l.) pollich – archaeophyte. fields: pilzno (ef 79 02), dobrków (ef 79 04). 590. *v. rimosa bastard – archaeophyte. fields: parkosz (ef 69 43). 591. verbascum densi�orum bertol. – on gravels of the wisłoka river: parkosz (ef 69 43), jaworze dln. (ef 79 24). 592. v. phlomoides l. – roadsides: podgrodzie (ef 69 44). 593. v. thapsus l. – on dry slope: parkosz (ef 69 43). 594. *verbena o�cinalis l. – archaeophyte. on ruderal habitats: mokrzec (ef 79 13). 595. veronica anagallis-aquatica l. – on banks of streams and drainage ditches: podgrodzie (ef 69 44), dobrków (ef 79 04). 596. *v. arvensis l. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), strzegocice (ef 79 22), jaworze dln. (ef 79 24). k ry st yn a to w pa sz 60 597. v. beccabunga l. – on banks of streams: podgrodzie (ef 69 44), jaworze dln. (ef 79 24), bielowy (ef 79 33). 598. v. chamaedrys l. – roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 03), gołęczyna (ef 79 14). 599. v. hederifolia l. – fields: parkosz (ef 69 43), podgrodzie (ef 69 44), gołęczyna (ef 79 14). 600. v. montana l. – oak-hornbeam forests: podgrodzie (ef 69 44), zwiernik (ef 78 34), jaworze dln. (ef 79 24). 601. v. o�cinalis l. – in forests, on their outskirts and in fallow areas: zwiernik (ef 78 34), gołęczyna (ef 79 14), jaworze dln. (ef 79 24), jaworze grn. (ef 79 34). 602. * v. persica poir. – epoecophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 02), strzegocice (ef 79 22). 603. * v. polita fr. – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), dobrków (ef 79 04). 604. v. serpyllifolia l. – fields: podgrodzie (ef 69 44), strzegocice (ef 79 22). 605. v. spicata l. – on sand dune: podgrodzie (ef 69 44). 606. *v. triphyllos l. – archaeophyte. fields: podgrodzie (ef 69 44), gołęczyna (ef 79 14). 607. viburnum opulus l. – in alluvial forests: podgrodzie (ef 69 44), parkosz (ef 69 43), łęki grn. (ef 78 13), dobrków (ef 79 04). 608. *vicia angustifolia l. – archaeophyte. in �eld communities: parkosz (ef 69 43), podgrodzie ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 12), gołęczyna (ef 79 14). 609. v. cracca l. – meadows, �elds and roadsides: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13), pilzno (ef 79 12), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 610. *v. dasycarpa ten. – epecophyte. fields: parkosz (ef 69 43), pilzno (ef 79 02), dobrków (ef 79 04), bielowy (ef 79 33). 611. *v. hirsuta (l.) gray – archaeophyte. on cereal �elds. parkosz (ef 69 43), podgrodzie (ef 69 44), zwiernik (ef 78 34), pilzno (ef 79 12), gołęczyna (ef 79 14). 612. v. sepium l. – �ickets: parkosz (ef 69 43), zwiernik (ef 78 34), pilzno (ef 79 12). 613. *v. tetrasperma (l.) schreb. – archaeophyte. on cereal �elds: łęki grn. (ef 78 13), gołęczyna (ef 79 14), słotowa (ef 79 31). 614. *v. villosa roth – archaeophyte. on cereal �elds: parkosz (ef 69 43), mokrzec (ef 79 13). 615. vinca minor l. – oak-hornbeam forests over the wisłoka river: podgrodzie (ef 69 44). 616. *viola arvensis murray – archaeophyte. fields: parkosz (ef 69 43), podgrodzie (ef 69 44), pilzno (ef 79 02), gołęczyna (ef 79 14), jaworze dln. (ef 79 24). 617. v. canina l. – on edges of forests: gołęczyna (ef 79 14), jaworze dln. (ef 79 24), bielowy (ef 79 33). 618. v. hirta l. – in thickets over the wisłoka river: podgrodzie (ef 69 44). v ascular plants of p ilzno surroundings (s outh-eastern p oland) 61 619. v. odorata l. – in riparian forests and wet oak-hornbeam forests: parkosz (ef 69 43), podgrodzie (ef 69 44), łęki grn. (ef 78 13). 620. v. reichenbachiana jord. ex boreau – oak-hornbeam forests: parkosz (ef 69 43), podgrodzie (ef 69 44), jaworze dln. (ef 79 24). 621. v. riviniana rchb. – oak-hornbeam forest: parkosz (ef 69 43). 622. v. rupestris f. w. schmidt – on sandy fallow area: jaworze dln. (ef 79 24). 623. v. tricolor l. – in cereals on sandy �elds: gołęczyna (ef 79 14), bielowy (ef 79 33). 624. *xanthium albinum (widder) h. scholz – hemiagriophyte. on the wisłoka river alluvials: parkosz (ef 69 43). 625. *x. strumarium l. – epoecophyte. on ruderal habitats: parkosz (ef 69 43), podgrodzie (ef 69 44), mokrzec (ef 79 13), jaworze dln. (ef 79 24). summary �e list of vascular plants presented above has 625 taxa, of which 514 are native and the remaining 111 are alien taxa. within them 70 are archaeophytes, 21 are epoecophytes, 6 are classi�ed as hemiagriophytes, and 14 are holoagriophytes. (appendix 1e–g). �irteen alien species have the status of invasive plants. among native plants, 19 mountain species were found: 4 lower montane zone species, 12 higher montane zone species and 3 all-mountainous. eleven protected species have been reported on study area, among others orchids: cephalanthera longifolia, epipactis purpurata (complete protection), or dactylorhiza majalis (partial protection). con�ict of interest �e author declares no con�ict of interest related to this article. references kondracki, j. (1978). geogra�a �zyczna polski. warszawa: wydawnictwo pwn. [in polish] kornaś, j., medwecka-kornaś, a., towpasz, k. (1997). rośliny naczyniowe pogórza ciężkowickiego. zeszyty naukowe uniwersytetu jagiellońskiego, prace botaniczne, 28, 1–170. [in polish] mirek, z., piękoś-mirkowa, h., zając, a., zając, m. (2002). flowering plants and pteridophytes of poland – a checklist. in: z. mirek (ed.), biodiversity of poland, 1–442. 1. kraków: w. szafer institute of botany, polish academy of sciences. oklejewicz, k., jóźwiak, a., skrzypek, b., wolak, n. (2004). uzupełnienie do �ory pogórza strzyżowskiego (se polska). fragmenta floristica et geobotanica polonica, 11, 199–202. [in polish] pawłowski, b. (1925). geobotaniczne stosunki sądeczyzny. prace monogra�czne komisji fizjogra�cznej. kraków: wyd. polska akademia umiejętności. [in polish] pawłowski, b. (1972). szata roślinna gór polskich. in: w. szafer, k. zarzycki (eds.), szata roślinna polski, 1–342. 2. warsaw: polish scienti�c publishers pwn. [in polish] regulation of the minister of the environment of october 9, 2014 on the protection of plant species (rozporządzenie ministra środowiska z dnia 9 października 2014 r. w sprawie ochrony gatunkowej roślin) dz.u. z 2014 r. poz. 1409. [in polish] k ry st yn a to w pa sz 62 tokarska-guzik, b., dajdok, z., zając, m., zając, a., urbisz a., danielewicz, w., hołdyński, c. (2012). rośliny obcego pochodzenia ze szczególnym uwzględnieniem gatunków inwazyjnych. warszawa: generalna dyrekcja ochrony środowiska. [in polish] towpasz, k. (1987). rośliny naczyniowe pogórza strzyżowskiego. zeszyty naukowe uniwersytetu jagiellońskiego, prace botaniczne, 16, 1–160. [in polish] towpasz, k. (1990). charakterystyka geobotaniczna pogórza strzyżowskiego. zeszyty naukowe uniwersytetu jagiellońskiego, rozprawy habilitacyjne, 178, 1–242. [in polish] towpasz, k. (2013). uzupełnienia do �ory roślin naczyniowych pogórza strzyżowskiego (karpaty zachodnie). część ii. fragmenta floristica et geobotanica polonica, 20(1), 41–50. [in polish] towpasz, k. (2018). środowisko naturalne okolic pilzna. in: b. stanaszek (ed.), pilzno, monogra�a miasta do 1945 roku, 11–30. pilzno: towarzystwo przyjaciół pilzna i ziemi pilźnieńskiej [in polish] wójcik, t. (2011). notatki �orystyczne ze strzyżowa i okolic (pogórze strzyżowskie). fragmenta floristica et geobotanica polonica, 18(1), 83–90. [in polish] zając, a. (1978). założenia metodyczne „atlasu rozmieszczenia roślin naczyniowych w polsce”. wiadomości botaniczne, 22(3), 145–155. [in polish] zając, a., zając, m. (2001). distribution atlas of vascular plants in poland. nakładem pracowni chorologii komputerowej instytutu botaniki uj, kraków s. xii + 716. [in polish/english] zając, a., zając, m. (2015). distribution of kenophytes in the polish carpatians and their foreland. kraków: nakładem instytutu botaniki uniwersytetu jagiellońskiego. [in polish/english] v ascular plants of p ilzno surroundings (s outh-eastern p oland) 63 appendix 1 dulczówka near pilzno, agricultural landscape – a; alluvial forest over the wisłoka river – b; protected species: staphyllea pinnata l. – c, matteucia struthiopteris (l.) tod. – d; invasive holoagriophytes: impatiens glandulifera royle – e, solidago gigantea aiton – f, echinocystis lobata (f. michx.) torr. & a. gray – g, (photo. k. towpasz) k ry st yn a to w pa sz 64 abstract �e paper presents the occurrence of vascular plant species in the southern part of the pilzno commune based on monographic studies from the area of ciężkowice and strzyżów foothills (western carpathians). �e study contains a list of plant species, both native and of alien origin. for each species its habitat and sites in the atpol network were given. key words: vascular plants, distribution, pilzno commune, ciężkowice foothills, strzyżów foothills received: [2019.09.05] accepted: [2019.11.22] rośliny naczyniowe okolic pilzna (południowo-wschodnia polska) streszczenie w  pracy przedstawiono występowanie gatunków roślin naczyniowych na obszarze południowej części gminy pilzno w oparciu o opracowania monogra�czne z terenu pogórzy ciężkowickiego i strzyżowskiego (karpaty zachodnie). artykuł zawiera wykaz roślin, zarówno rodzimych, jak i  obcego pochodzenia. dla każdego gatunku podano jego siedlisko i lokalizację w sieci atpol. słowa kluczowe: rośliny naczyniowe, rozmieszczenie, gmina pilzno, pogórze ciężkowickie, pogórze strzyżowskie information about author krystyna towpasz she is retired professor at the institute ob botany of the jagiellonian university in kraków. she is interested in geobotany, plant ecology and plant protection. 203 annales universitatis paedagogicae cracoviensis studia naturae, 6: 203–225, 2021, issn 2543-8832 doi: 10.24917/25438832.6.12 angelika kliszcz department of agroecology and plant production, faculty of agriculture and economics, university of agriculture in kraków, al. mickiewicza 21, 31-120 kraków, poland; angelika.kliszcz@urk.edu.pl phenological growth stages and bbch-identification keys of jerusalem artichoke (helianthus tuberosus l.) introduction �e growth stages of development of particular plant species were constructed and proposed by many authors around the world since decades. chen (2013) indicates that the tradition of observation and recording the phenological events of many cultivated and ornamental plants in ancient times were occurred. �e interesting plant species that is being rediscovered today is jerusalem artichoke, topinambour (helianthus tuberosus l.). both, the growing attention of scientists in the context of its interesting physiological, biochemical and genetic predispositions invasive plant (balogh, 2008; tokarska-guzik et al., 2012), resistant to salt stress (zhang et al., 2016), quite di�cult to correctly identify h. tuberosus and h. strumosus l. and their natural hybrids – they both produce tubers from all the other species of the helianthus genus (kays, nottingham, 2008), as well as the growing interest in this plant as a raw material in the food industry (high inulin content, an easily hydrolysable fructan (barhatova et al., 2015), component of sophisticated alcoholic beverages (rossini et al., 2016), health-promoting properties of tubers (cieślik, filipiak-florkiewicz, 2000; kulczyński, gramza-michałowska, 2016; radovanovic et al., 2014), make this plant nowadays more and more noticeable. additionally, the energy sector approves this plant as an energetic crop (bioethanol and biofuel production, pellets production due to a large amount of biomass produced per unit area with a  satisfactory caloric value (kowalczyk-juśko et al., 2012; johansson et al., 2015; sawicka et al., 2019; bedzo et al., 2020). �ere are also other applications (such as blasting in forest frames as an alternative food for wild animals (dreszczyk, brzezowska, 2008), especially wild boars, a  substrate medium for the production of mushrooms and shunts (đorđević et al., 2010), or for industry processes with the use of microorganisms laboratory cultures (meng et al., 2021) make this plant more and more famous. a ng el ik a k lis zc z 204 however, each multi-purpose plant has its limitations, among which it should be noted: invasive nature (high inulin content, thanks to which the tubers winter in the soil down to –30°c, rapid growth and early growth of the aboveground parts shading the surface, vegetative reproduction mostly climatic zones, remains in the position despite herbicidal fallow (balogh, 2008; tokarska-guzik et al., 2012; pacanoski, mehmeti, 2020). �e increasing number of papers according jerusalem artichoke in web of science (from 1910 to 2000: 615 articles, 2001–2010: 221 articles, 2011–2015: 292 articles, 2016–2020: 480 articles) and agricultural events focusing on jerusalem artichoke (stapor, 2020) is gathering more and more people interested in this plant around the world. more information about origin, history of discover and various naming of this plant can be found in �e biology and chemistry of jerusalem artichoke (kays, nottingham, 2008). detailed information about probiotic and pharmaceutical properties of this plant was published by mystkowska et al. (2015), its multipurpose use was provided by sawicka et al. (2012), and energy properties of the plant announced gao et al. (2016). recently, the very valuable review, with a whole view on this plant appeared (rossini et al., 2019). �e aim of the study is to focus on the growth and development of jerusalem artichoke (h. tuberosus) plants grown from the tubers in temperate climate zone and propose a bbch (biologische bundesanstalt, bundessortenamt and chemicsche industrie) identi�cation key, which is expected for uni�cation academic and practical discussion about this plant. due to the fact that in our climate the seeds are not fully developed and their role in propagation of the plant is negligible, the proposed bbch key describes all development phases except the ripening of seeds phase (is omitted). �e developmental biology of jerusalem artichoke – state of the art �e general strategy of jerusalem artichoke (ja) is to invests actual carbon and nutrients early in its development into stem (incoll, neales, 1970), branch, and leaf growth, facilitating the exploitation of aboveground resources. later in the developmental cycle carbon and nutrients are allocated to rhizomes and tubers (mclaurin et al., 1999), enabling the species to spread alongside, i.e. colonising new areas. �e success of its allocation patterns is in simultaneous synchronization of the belowand aboveground biomass growth and development (fig. 1). �e plants produce considerable amounts of aboveground biomass, which acts for carbohydrates reservoir although it has c3 photosynthetic mode (podlaski et al., 2017). helianthus tuberosus plants have a strong photoperiod response (short-day plant) (terzić et al., 2012). one of the �rst research exploring photoperiod phenomenon in plants was based on jerusalem artichoke species (garner, allard, 1923). shortly, photoperiodic plants identify day length in the leaves and then transfer the signal (�origen) to the shoot apex for the onset of the formation of in�orescence. �e day length also a�ects the start of formation tubers (i.e. tuberisation). typically initiation of tuberisation 205 begins from 5 to 13 weeks a�er emergence (swanton, cavers, 1989; hay, o�er, 1992; mclaurin et al., 1999). generally, the development of aboveground biomass goes through successive phases, from the sprouting, full side branching (bbch 39n), to full drying o� the plant (bbch 98). �e belowground development starts with roots development (bbch 04), then rhizome development (bbch 40-49), and tuber development (bbch 70-79). �e bbch system of monitoring physiological scales in plants any lively discussion between people from di�erent regions needs a common language. except for the known need to communicate in the same language, it is nonetheless necessary to think about the same phenomena in the same way, especially when the discussion concerns a dynamic process in biological systems, additionally modi�ed by environmental processes in�uenced in various intensity in every point on the world map. scientists’ passion, farmer’s needs, and entrepreneurs’ interests underlie the universal bbch scale (biologische bundesanstalt, bundessortenamt and chemische industrie) currently in force (meier, 2018). �e monograph did not appear at once (meier et al., 2009). many interesting works on this topic (troitzky, 1925; fleckinger, 1948; feekes, 1941; large, 1954) were published through the 20th century, to meet the demands of 1) fig. 1. general view on developmental stages of jerusalem artichoke (helianthus tuberosus l.) p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 206 uni�cation and clari�cation of de�nitions of concepts in botanical scienti�c discussion, 2) simpli�cation the decision making in process of plant protection by farmers and avoid the misunderstanding between farmers and agrochemical companies or agricultural insurance agents, and 3) development of agrometeorology. however, a real acceleration of universal concepts describing phenological stages during plant development occurred a�er a publication scale by zadoks et al. (1974). �e authors presented an adjusted and re�ned numeric decimal scale for such plants like cereals and rice, which gave the direct base for currently wide-spread used bbch scale. �e bbch coding system is an improvement of the zadoks et al. (1974) coding system, it includes also the dicotyledoneous plants and more monotyledoneous plants species. �e �rst publication of the bbch codes of some crops (bleiholder et al., 1989) (appears in working group consisted of sta� members from four chemical companies) was the �rst step to join the forces in 1991 with german scientists (from �e federal biological research centre for agriculture and forestry, bba), who published booklets describing phenological stages of particular crops (meier, 1985). �e �rst outcome of this cooperation was the principles of the enhanced general bbch scale (hack et al., 1992), which was the base for members of this group for publishing (with experts in each crop) the “extended bbch scales for speci�c crops” in various branch journals. �e �rst bbch monograph edited by meier (1997) was published in four languages and describes the phenological development stages of 27 crops and wild plants. adamczewski and matysia (2005) published the bbch scale in polish, a�er earlier published work (e.g. gąsowski, ostrowska, 1993). milestone for the international acceptance of the bbch codes used in plant protection management process was the decision for establishing the bbch scale mandatory for all o�cial plant protection trials made in 2004 and 2006 by eppo (european and mediterranean plant protection organization) (meier et al., 2009). nowadays the bbch monograph (meier, 2018) includes 48 identi�cation keys for crops and additional key for weeds (dicotyledons, graminae, monocotyledons, perennial plants). recently, the developmental biology of many other crops has been described in a key on the bbch scale, e.g. to harmonize production processes (rajan et al., 2011; zhao et al., 2019; singh et al., 2021). although, despite of multi-purpose use and increased awareness of h. tuberosus species, there is a lack of o�cial bbch identi�cation key describing phenological growth stages of h. tuberosus. in this article, developmental biology of jerusalem artichoke in temperate climate was presented. �e phenological stages of this plant were proposed in the obligatory bbch identi�cation key. 207 material and methods experimental site and plant material �e plants were grown in experimental �eld located in experimental station in krakow – mydlniki (50°05′08.5″n; 19°51′08.3″e; university of agriculture in kraków, poland) in 2020. during the 7 months (mid-april to mid-november 2020) growth and development of helianthus tuberosus plants (cv. rubik) were monitored. �e seed tubers were intentionally le� in the �eld previous autumn (november 2019) as an irregular population close to the natural one. in spring, plants were chosen and monitored during the vegetation season (completely random assignment was applied). �e soil was unfertilized and characterised a well moisture content during whole vegetation period (due to impact of an neighbourhood of underground watercourse). �e bbch scale �e bbch principal growth stages were the basis for these considerations (tab. 1). tab. 1. principal growth stages of helianthus tuberosus l. according to bbch scale (a�er meier et al., 2009) stage (number 0–9) description 0 germination / sprouting / bud development 1 leaf development (main shoot) 2 formation of side shoots/tillering 3 stem elongation or rosette growth/shoot development (main shoot) 4 development of harvestable vegetative plant parts or vegetatively propagated organs / booting (main shoot) 5 in�orescence emergence (main shoot) / heading 6 flowering (main shoot) 7 development of fruit 8 ripening or maturity of fruit and seed 9 senescence, beginning of dormancy meteorological data �e observations were carried out with the background of weather conditions in temperate climate (poland). �e meteorological data was obtained from climatedata.org (2020). �e graphical relation of mean temperatures [°c] and total precipitation values [mm] for each month were reported as a gaussen-walter climatogram with łukasiewicz modi�cation (walter, 1976; łukasiewicz, 2006) (fig. 2). �e rule for the construction of such a graph is that the values of mean temperature and total precipitation are plotted with maintaining a  ratio of 1°c to 4 mm of precipitation. �is balance determines the di�erence between precipitation and evapotranspiration. it helps to read o� the amount of evapotranspiration and thus estimate the excess or shortage of precipitation for plants on a local scale (treder et al., 2018). p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 208 results �e stages of phenological phases of helianthus tuberosus was constructed for temperate climate zone (tab. 2 – appendix 1). principal growth stage 0: sprouting helianthus tuberosus tubers have endodormancy, which means that an internal mechanism prevents sprouting even though the environmental conditions may be suitable. it is linked with the conditions (winter months) of their region of origin (temperate zone, continental climate). �e main shoot develops from apical bud located on the belowground seed tuber (fig. 3a–b). �ere are possible other lateral sprouts growing from the same tuber at the same time (from axillary buds), but the plant seems to develop lateral sprouts a little bit later so as not to inhibit the growth of the main shoot. because some tubers are located fairly deep in the ground, the length of the �rst sprout may achieve even 30 cm (bbch 08, fig. 3b), due to high vitality and aèuence of substances in tubers. sprouts a�er reaching a suitable cumulative temperatures grow in the soil quite fast regardless of weather conditions. fig. 2. gaussen-walter climatogram for the studied area (in months january–november 2020) 209 principal growth stage 1: leaf development (main shoot) �e main shoot (stem) grows quite fast and tends to shade the stand. �e development of aboveground leafy biomass realizes quickly. when the plant meets favourable weather, it generates next leaves pair in each new week. leaves are numerous, with the opposite arrangement in the lower third, alternate above. in the studies about the dynamics of this stage, it will be worth to mark with an asterix the last pair of the leaves with opposite arrangement (e.g. bbch16*). generally, the number of leaves pairs depend on biotope richness, and to a lesser extent on the weather conditions. once the side branches appear from the apical buds in any leaves pair, the development of the plant shall be determined in accordance with appropriate next phase (bbch 3_). principal growth stage 2: formation of other sprouts �is stage occurs not always here. generally, the plant skips with its development from leaf development phase (bbch 1_) to the next phase (bbch 3_). �e strategy of forfig. 3. �e examples of some phenological stages of proposed bbch coding system for jerusalem artichoke (helianthus tuberosus l.) (photo. a. kliszcz) p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 210 mation of other stems (lateral sprouts) from the underground parts of the plant (tuber, rhizomes, underground part of the main stem) seems to be linked with the need for more photosynthetic area within the plant, what constitutes their carbohydrate supply for new tubers or it is a result of mechanically injured belowground stem. �e number of shoots that emerge are linked with e.g. shape of tuber and it is variety depended. �e tuber more branched produces more shoots (tubers branched vs. tuber with apical dominance) (kays, nottingham, 2008). �e formation of other sprouts from the tuber also depends on the depth at which the tuber is located, tuber size and number of its axillary buds. in general, since the delineation of the sources of additional stems is unclear, all shoots that came above the ground surface a�er the main shoot drop into one category. it is worth to notice that the emergence of other sprouts (stems) may appear during the whole vegetative phase. �e �nal number of stems is constituted at the end of the growing season, when the plant enters the senescence phase. principal growth stage 3: stem elongation and development of lateral branches on main shoot (side branching) in this stage the development of the stem and upper leaves continues. side branching appears simultaneously on the main shoot from the bottom of the plant. �e presence and degree of branching depend on the variety, plant population density, and other factors (like branch location on the plant, photosynthetic potential, environmental factors). �e lateral branches are formed in the axils of the leaves, starting at the base of the plant. at each node, there is commonly two opposite-located lateral branches. very rare is the triple, when three leaves emerge from one node. rapid growth of branches on the plant diminishes in the middle of growth cycle and again increases when axillary buds start to developing into �owering branches. vertical development of the plant is terminated by �ower bud formation at the apex of the stems. not all bbch codes may occur during the development of the plant in this stage. as soon as the next axillary buds begin to develop in the next (upper) pair of leaves on the plant, the bbch code is counted there (e.g. bbch 32-1 move to bbch 33-0, even if the leaf development on the lower branch continues). �e size of axillary buds should be at least 2 cm to be considered as the next level of this development phase (fig. 3d). typically, every next node with leaves pair generates the twin opposite branches, but it depends on the plant’s current needs (e.g. shading, processes of translocating carbohydrates). �e plant may omit some pairs of leaves without developing any branches there. if this is the case for the third pair of leaves, the bbch 33-0 remains until a fourth pair of leaves has developed and the axillary buds in the axils of this leaves appear. another case may arise when fourth pair of leaves has already hosted emerging twin branches (e.g. already with two leaves each), then the bbch 33-_ is valid until in the axis of the ��h pair of leaves appear new axillary buds. 211 principal growth stage 4: rhizomes booting �e plant starts to form rhizomes in the early stages of biomass acquisition and accumulation (from 1.5 to 8 weeks a�er emergence; fig. 3e). �e underground portion of the stem (4 to 5 cm below the soil surface) is the basis for the emergence of rhizomes. �ey grow in a slight downward angle (fig. 3f) with internodes length varying substantially among clones (kays, nottingham, 2008). principal growth stage 5: in�orescence emergence �e shi� between vegetative and generative phase in jerusalem artichoke (short-day clones) is strongly dependent on photoperiod. �e in�orescence appears in the speci�c location on the plant of ja (i.e., the top of the stem and later – on branch apices) and their formation involves temporal order. because of that the bbch coding system for this phase was focused on �rst appeared examples of top in�orescence, and then on branched-located ones (e.g. bbch 51 -> bbch 52, 53). long-cycle varieties (ca. 9 months vegetation) o�en produce buds but no �owers (denoroy, 1996), and their aerial parts are moving then straight on to senescence phase (bbch 9_). principal growth stage 6: �owering �e �owering is starting from the top of the main shoot (bbch 61-65) and will proceed to the bottom (along with side shoots embedded on the main shoot, i.e. bbch 66-69). flower stalks have frequently between 10 and 15 cm for rubik cultivar, depending on closeness to the plant axis. �is paper focus on domesticated clone (cv. rubik), and it was observed that in temperate climate (fig. 2) it takes ca. 21 days for the plants to proceed �owering (from the tight bud stage (bbch 51) to senescence stage (bbch 90)). �e detailed visualisation of chronological sequence of �owering phase of single �ower was presented in the book concerning the biological and chemical issues of ja plants (kays, nottingham, 2008). �e bbch scale required the �owering sequence of all �owers on the plant due to the fact that ja plants produce many �owers (not a  single one as the sun�ower produces). and for that reason presented bbch scale includes the whole �owering biology of this plant with the background of single �ower development steps. principal growth stage 7: tuber bulking while tuber initiation appears to be in part controlled by carbohydrate supply, tuber bulking is strongly modulated by photoperiod, even in clones that are day neutral for �owering (kays, nottingham, 2008). cumulative temperature is linearly correlated with tuber number, and cumulative degree days (≥ 520 degree days) can be used for predicting the onset of tuberisation (spitters et al., 1988). in the cross section tuber p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 212 could be divided (from exterior to interior) into: epidermis, cortex, outer medulla, inner medulla, and pith (mazza, 1985). shortly a�er the beginning of �owering, remobilization of nutrients from canopy into the developing tubers begins. photoperiod and carbohydrate supply are a critical factors in the tuberisation response. simultaneously with the proceeding bulking stages of the tuber, the dormancy onset occurs. �e onset of dormancy is progressing gradually. firstly, into dormancy enter rhizomes and small, young tubers, with more and more areas of the tuber establishing dormant. �e larger, more mature tubers enter into dormancy at the end within the whole plant. although, the initiation of dormancy in large tuber may occur even the tuber is not fully �lled (kays, nottingham, 2008). principal growth stage 8: ripening seed flowers are o�en sterile (domesticated clones). swanton et al. (1992) stated that most of the ja plants have no more than 5 seeds per �ower head. according to westley (1993) only a  44 % of mature seeds are capable of germination, whereas only 33% are able for reproduction – during the �rst season (wild clones). in this phase, the increased translocation of the assimilates to the tubers takes place, and this coincides with and a�ect the seed ripening as well. because the propagation of the plant by the seed is of marginal importance, and it occurs when the plant is drying o� (transfer of the symbionts to the tubers, i.e. tuber bulking, fig. 3g–h), no special phase was assessed for process regarding seeds ripening. principal growth stage 9: senescence �e drying o� of the whole plant occurs when the belowground part of the plant enters dormancy. �e process of senescence accompanies the plant virtually throughout the entire growing season. �e �rst leaves on the main stem senesced �rstly, and it happens well before it starts blooming. some authors argue this fact that the plant shades the lower leaves as it grows (zubr, 1988). �e meteorological data (fig. 2) shows no shortages of water in the studied period, i.e. the precipitation line is above the line representing the temperature, such ratio (1°c to 4 mm precipitation) is proposed by łukasiewicz (2006) for climate conditions in poland (temperate). �erefore, the plants had favourable conditions for development during whole vegetation season. discussion �e jerusalem artichoke (helianthus tuberosus l.) is an aged tuber crop with a lately aroused attraction following its multipurpose usage (cao et al., 2008; van wyk, wink, 2008; ma et al., 2011; maj et al., 2013; mystkowska, zarzecka, 2013). �e growth of 213 plants and certain developmental phenomena are governed by a few main factors: the total amount of heat a plant received during a certain period, the portion of various wavelengths from sunlight, number of days with sunlight and the duration of the day, richness of soil and their su�cient moisture (biggs et al., 2007; kocsis et al., 2007; puangbut et al., 2012; ruttanaprasert et al., 2014). however, the developmental biology of wild, domesticated, and intermediate clones di�er signi�cantly, but general overview of phenological stages of jerusalem artichoke seems to be constant (fig. 1). �e plant have to go through all stages to produce tubers (fig. 3a), a main propagules for next growing season. however, the development of this species is realised in two parallel directions: vegetative and generative (pawłowski, jasiewicz, 1971; vaughan, geissler, 2001). �is complicates the naming of its developmental stage. dual naming can be a  solution. when the side branching phase continues, the rhizomes booting occurs, and then the proper description could be e.g. bbch 38-2/46. similarly, as the tubers �ll, the aboveground biomass withers (it could be written as bbch 78/97). of course, the codes can be used individually to indicate only the state of the aerial vegetative biomass (respectively, bbch 38-2, or bbch 97). but for detailed ecological studies of this plant it is bene�cial to use dual nomenclature. �is paper focuses on domesticated clone (cv. rubik), which belongs to the group of short-day clones. �e �rst attempt to name and standardise jerusalem artichoke (ja) developmental biology stages was made by paungbut et al. (2015). however, they designed their own system without using the bbch codes. �ey arrange all development of ja into three main groups of stages: vegetative stages (v), reproductive stages (r), and tuberisation stages (t). �e authors cultivated the plants in �ailand (tropical area) in 2011–2012, and observed developmental biology of ja during opposite seasons, both, the early-rainy season and the drier post-rainy season. �ere are 15 phases in their concept (adding to this number of pairs of leaves on the main stem, depending on the nodes produced; shortly, if the main stem develops 12 leaves pairs, there will be 25 phases in total). �e bbch codes allow for a more accurate description of each phase through careful observation of plant morphology. �erefore, in presented concept according to bbch codes there are at least 100 various phases, which precisely de�ne every moment of development (e.g. bbch 3_ is describing side branching). more detailed descriptions of the ja plants are already included in the phenotype studies of the various clones of this species (diederichsen, 2010), which is no necessary to include it in bbch system formation. it is worth to note that with modi�ed biotope features (unfavourable conditions), the length of developmental stages could be shorten (e.g. the time of bulking tubers) or abandoned (the �owering phase does not always occur). additionally, the genetic features play a pivotal role as well (kays, kultur, 2005; skiba, sawicka, 2016) and also wild vs. domesticated clones develop with di�erent dynamics (feher et al., 1999; breton p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 214 et al., 2017). for example, serieys et al. (2010) examined 142 clones of ja deposited in inra library and stated that 80% of them perform the �owering phase between september and october, and 10% of them did not enter this phase at all. on the other hand, the biomass enhancement and its dynamics could be induced with richness of site or fertilisation level (bogucka et al., 2021). interesting research on the dynamics of nutrient uptake was carried out by izsaki and németh (2013). �e authors examined two varieties of ja and stated that in both cases the maximum nutrient uptake was recorded on the 155th day of vegetation, which corresponds with beginning of 7th phase (bbch 70-79), i.e. tubers bulking. manipulation of duration of development processes could be intentionally forced in agricultural practice, which is targeted on high-quality, plentiful tubers yield. in literature the manipulation of planting date (puangbut et al., 2012), mowing date of plant top (acar et al., 2011), dates of pruning radius (gao et al., 2018), harvest time and storage (saengthongpinit, sajjaanantakul, 2005) or another agrotechnical treatments are known (puttha et al., 2013; dias et al., 2016; gao et al., 2019). �erefore, the proper managing of the plantation of topinambur plants seems to be an important factor for high yield of tubers in many agroecosystems. conclusion �e scale for coding the phenological growth stages of helianthus tuberosus l. species is needed. �is plant becomes more and more popular because of its multi-purpose use and ease of cultivation in all types of soil all over the world. �e latest edition of bbch monograph (meier, 2018) does not cover this need. jerusalem artichoke has many desirable growing traits such as cold and drought tolerance, wind and sand resistance, saline tolerance, strong fecundity and high pest and disease resistant. generally, the development of aboveground biomass goes through successive phases, from the sprouting, full side branching (bbch 39n), to full drying o� of the plant (bbch 98). �e belowground development starts with roots development (bbch 04), then rhizome development (bbch 40-49), and tuber development (bbch 70-79). it is worth to notice that the development of the plant from a point (bbch 49) realises in two parallel directions: vegetative and generative, i.e. when at least one rhizome starts to thicken at its end, it means that the plant begun a generative phase. �e concern about their developmental biology is also essential for managing the termination strategies for this genus, as the ja is an invasive plant in most ecosystems. �erefore growing attraction of this plant force their key in bbch coding system to harmonize discussion about this plant in the future. 215 acknowledgements �e research was �nanced by the ministry of science and higher education of poland. con�ict of interest �e author declares no con�ict of interest. references acar, r., ada, r., özköse, a. 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(1988). jerusalem artichoke as a �eld crop in northern europe, in topinambour (jerusalem artichoke). report eur 11855. luxembourg: commission of the european communities. 221 appendix 1 tab. 2. phenological growth stages of helianthus tuberosus l. according to the bbch scale bbch code (2-digit) description of the development from the tuber principal growth stage 0: sprouting 00 innate or enforced dormancy, tuber not sprouted 01 beginning of sprouting: �rst sprout visible (< 1 mm) 02 end of dormancy: sprout 2–3 mm 03 first (main) sprout further growth (< 1 cm) 04 beginning of root formation; �rst sprout further growth (< 5 cm) 05 first sprout further growth (< 15 cm) 06 first sprout further growth (< 15 cm) 07 first sprout further growth (< 20 cm) 08 first sprout further growth (< 30 cm and more) 09 breakthrough sprouts on the ground principal growth stage 1: leaf development (main shoot) 10 �e �rst pair of leaves begins photosynthesis (leaves are still rolled up, but green) 11 first pair of leaves fully developed 12 second pair of leaves fully developed 13 �ird pair of leaves fully developed 14 fi�h pair of leaves fully developed 15 seventh pair of leaves fully developed 16 ninth pair of leaves fully developed 17 eleventh pair of leaves fully developed 18 �irteenth pair of leaves fully developed 19 fi�eenth pair of leaves fully developed 19n sixteenth (and further) pair of leaves fully developed principal growth stage 2: formation of other sprouts 20 only the main shoot is visible above the ground 21 �e �rst side shoot appears above the ground 22 �e second side shoot appears above the ground 2.. �e next side shoots appear above the ground principal growth stage 3: development of lateral branches on main shoot (side branching) 30 beginning of developing lateral branches: the �rst (twin) axillary buds appear in the axils of the �rst pair of leaves on the main shoot 31-1 �e �rst pair of developed leaves growing out from the axils of the �rst pair of leaves on the main shoot 31-2 �e second pair of developed leaves growing out from the axils of the �rst pair of leaves on the main shoot 31-… successive pairs of developed leaves growing out from the axils of the �rst pair of leaves on the main shoot p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 222 32-0 �e �rst (twin) axillary buds appear in the axils of the second pair of leaves on the main shoot 32-1 �e �rst pair of developed leaves growing out from the axils of the second pair of leaves on the main shoot 32-.. successive pairs of developed leaves growing out from the axils of the second pair of leaves on the main shoot 33-0 �e �rst (twin) axillary buds appear in the axils of the third pair of leaves on the main shoot 33-1 �e �rst pair of developed leaves growing out from the axils of the third pair of leaves on the main shoot 33-.. successive pairs of developed leaves growing out from the axils of the third pair of leaves on the main shoot 34-0 �e �rst (twin) axillary buds appear in the axils of the ��h pair of leaves on the main shoot 34-1 �e �rst pair of developed leaves growing out from the axils of the ��h pair of leaves on the main shoot 34-.. successive pairs of developed leaves growing out from the axils of the ��h pair of leaves on the main shoot 35-0 �e �rst (twin) axillary buds appear in the axils of the seventh pair of leaves on the main shoot 35-1 �e �rst pair of developed leaves growing out from the axils of the seventh pair of leaves on the main shoot 35-.. successive pairs of developed leaves growing out from the axils of the seventh pair of leaves on the main shoot 36-0 �e �rst (twin) axillary buds appear in the axils of the ninth pair of leaves on the main shoot 36-1 �e �rst pair of developed leaves growing out from the axils of the ninth pair of leaves on the main shoot 36-.. successive pairs of developed leaves growing out from the axils of the ninth pair of leaves on the main shoot 37-0 �e �rst (twin) axillary buds appear in the axils of the eleventh pair of leaves on the main shoot 37-1 �e �rst pair of developed leaves growing out from the axils of the eleventh pair of leaves on the main shoot 37-.. successive pairs of developed leaves growing out from the axils of the eleventh pair of leaves on the main shoot 38-0 �e �rst (twin) axillary buds appear in the axils of the thirteenth pair of leaves on the main shoot 38-1 �e �rst pair of developed leaves growing out from the axils of the thirteenth pair of leaves on the main shoot 38-.. successive pairs of developed leaves growing out from the axils of the thirteenth pair of leaves on the main shoot 39-0 �e �rst (twin) axillary buds appear in the axils of the ��eenth pair of leaves on the main shoot 39-1 �e �rst pair of developed leaves growing out from the axils of the ��eenth pair of leaves on the main shoot 39-.. successive pairs of developed leaves growing out from the axils of the ��eenth pair of leaves on the main shoot 39n-0 �e �rst (twin) axillary buds appear in the axils of the sixteenth (or successive) pair of leaves on the main shoot 223 principal growth stage 4: rhizomes booting 40 �e �rst rhizome starts to grow 41 �e other rhizomes start to develop 42 �e rhizomes elongate 43 �e rhizomes still elongate and start to branch 44 �e rhizomes elongate and branch still (< 30 cm from plant) 45 �e rhizomes elongate and branch still (< 50 cm from plant) 46 �e rhizomes elongate and branch still (< 80 cm from plant) 47 �e rhizomes elongate and branch still (< 100 cm from plant) 48 rhizomes are developed and most of them have not thickened ends yet 49 �e ends of most rhizomes start to thicken (rhizome tips to twice the diameter of subtending rhizome) principal growth stage 5: in�orescence emergence 50 in�orescence not visible 51 in�orescence just visible between youngest leaves on the main shoot (tight bud stage) (peduncles elongate) in�orescence just visible on the upper branches (50% branches on the top have in�orescence just visible); tight bud stage (peduncles elongate) in�orescence just visible on the lower branches; tight bud stage (peduncles elongate) 52 first top ligules (ray �ower corollas) exposed and green (peduncles elongate) 53 most top ligules exposed and green (peduncles elongate) 54 ligules on the upper branches exposed and green (peduncles elongate) 55 most top ligules exposed and green (peduncles elongate) 56 ligules on the upper branches exposed and green (peduncles elongate) 57 ligules on the lower branches exposed and green (peduncles elongate) 58 ligules on the top and upper branches yellow-green (in�orescence still closed) 59 most ligules on the plant are yellow-green (in�orescence still closed) principal growth stage 6: floweringi 60 top ligules beginning to unroll (disk �ower corollas yellow and closed) 61 first top ligules open 62 first top ligules with emerging anthers from the corolla 63 additional anthers and �rst stigmas emerging on outer whorls on the top ligules 64 about half of top disk �owers open with stigmas emerged 65 all of the top disk �owers open with stigmas emerged (in bloom) 66 �ird part of disk �owers in lateral branches (from the top) are in bloom (outer whorl �owers on the top displaying initial stigma senescence) 67 two thirds of disk �owers on lateral branches (from the top) are in bloom (top ligules wilting and initial drying) 68 80% of disk �owers on lateral branches (from the top) are in bloom 69 end of �owering: almost all disc �ower have �nished �owering on the plant, ray �orets dried principal growth stage 7: tubers bulking 70 tubers bulking (10% of all) 71 tubers bulking (20% of all) 72 tubers bulking (30% of all) 73 tubers bulking (40% of all) p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) a ng el ik a k lis zc z 224 74 tubers bulking (50% of all) 75 tubers bulking (60% of all) 76 tubers bulking (70% of all) 77 tubers bulking (80% of all) 78 tubers bulking (90% of all) 79 tubers are full (100%), maximum of the total tuber mass reached principal growth stage 8: ripening seedii principal growth stage 9: senescence 90 40% of aboveground green parts of the plant has dried up 91 50% of aboveground green parts of the plant has dried up 92 60% of aboveground green parts of the plant has dried up 93 70% of aboveground green parts of the plant has dried up 94 80% of aboveground green parts of the plant has dried up 95 90% of aboveground green parts of the plant has dried up 96 100% of aboveground green parts of the plant has dried up 97 aboveground biomass has > 20% moisture w/w 98 aboveground biomass has < 20% moisture w/w 99 tuber harvested, dormancy i a synthesis of an in�orescence development a�er kays and nottingham (2008) (changed) ii seed is developing along the tuber bulking 225 fazy rozwojowe słonecznika bulwiastego (helianthus tuberosus l.) w propozycji oznaczeń skali bbch streszczenie celem pracy było zbadanie i standaryzacja faz rozwojowych słonecznika bulwiastego (helianthus tuberosus l.), rosnącego w klimacie umiarkowanym, na podstawie klucza oznaczeń bbch. tego rodzaju analizę wykonano po raz pierwszy, co było oczekiwane w dyskursie naukowym, jak i praktycznym. rosnące zainteresowanie tym gatunkiem, zarówno z punktu widzenia naukowego, jak i utylitarnego, stawia potrzebę nazwania jego poszczególnych stadiów rozwojowych oraz ich standaryzacji w zakresie nomenklatury. słonecznik bulwiasty jest rośliną o wielokierunkowym wykorzystaniu w różnych gałęziach przemysłu. surowcem w przemyśle spożywczym są bulwy, które gromadzą znaczne ilości inuliny – łańcuchowego polimeru fruktozy, o istotnych właściwościach probiotycznych. dzięki temu bulwy są cennym składnikiem żywności funkcjonalnej, substratem w produkcji farmaceutyków, czy napojów alkoholowych, a także pozwalają na przetrwanie gatunku w środowisku w okresie zimowym. formowanie się bulw zachodzi przez znaczną część rozwoju ontologicznego gatunku (bbch 49) i związane jest głównie z fotoperiodem, sumą temperatur efektywnych oraz obecnością nadziemnej biomasy rośliny, z której zachodzi alokacja asymilatów do bulw w okresie rozwoju generatywnego. spośród wielu innych zastosowań, roślina ta jest wykorzystywana jako surowiec energetyczny, gdyż naturalnie wyschnięta biomasa nadziemna, pod koniec sezonu wegetacyjnego, zawiera niską zawartość wody i plasuje ten gatunek w środku listy roślin energetycznych. key words: topinambour, bbch scale, phenological stages received: [2021.06.02] accepted: [2021.09.17] p henological grow th stages and b b c h -identification keys of jerusalem artichoke (h elianthus tuberosus l.) annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x aleksandra izdebska, natalia malejky-kłusek, małgorzata kłyś department of ecology and environmental protection, institute of biology, pedagogical university of cracow, podchorążych 2, 30-084 kraków, poland, *email natalia.malejky@tlen.pl, aleksandra.izdebska2@student.up.krakow.pl, malgorzata.klys@up.krakow.pl composting as a “golden method” to solve the organic household waste problem? – short revision introduction according to the act (dz. u. 2013 poz. 21), waste is defined as each substance or object that the owner disposes of, intends to dispose of, or is obligated to dispose of. the regulation of the minister of environment (dz.u. z 2014 r., poz. 1923) divides the waste into 20 groups according to the source of its origin, including, among others litter from the leather and fur industry, agricultural, gardening, hydroponic farming, fishery, forestry, game shooting, food processing as well as a household with fractions collected selectively. the waste catalogue divides such waste into subgroups and types, assigns their codes, and specifies hazardous. this classification distinguishes 3 basic refuse selection criteria – by its source of origin, properties, and components. each one of us is a daily “producer” of refuse. waste produced by private individuals has been classified into the code 20 group – household with fractions collected selectively. household litter includes waste from private homes as well as from other sources (except for hazardous) whose nature and composition are similar to a household. such refuse is the main source of pollution in all parts of the environment and is a major nuisance contributing to environmental degradation (czop, hatlapa, 2007; sklamowski, 2009). based on the regulation of the minister of environment (dz.u. z 2014 r., poz. 1923), the household waste group is divided into three subgroups: segregated household and selectively collected household refuse (e.g., paper and cardboard, glass, clothes), litter from garden and park (biodegradable, soil and earth, including rocks and other non-biodegradable) and other household litter (e.g., sink basin deposits, large-size waste). mailto:natalia.malejky@tlen.pl according to information provided by statistics poland, in 2017, the quantity of household waste produced in poland reached 11.969 thousand tons. this figure is 2.7% higher than in the previous year, hence we are seeing an increase in the quantity of produced household litter (https://stat.gov.pl/obszary-tematyczne/srodowisko-energia/srodowisko/zmiana-systemugospodarki-odpadami-komunalnymi-w-polsce-w-latach-2012-2016,6,1.html). the highest quantity of household refuse produced in 2020 per one resident was recorded in dolnośląskie province (400 kg), and the lowest quantity in podkarpackie province (236 kg) (https://bdl.stat.gov.pl/bdl/dane/podgrup/tablica). the act of 14 december 2012 outlines the hierarchy of waste management methods. first and foremost, it is necessary to minimize the production of waste (1), next prepare it to be reused (2), recycle it (3), then put it through other recovery processes (4), and finally neutralize it (5). compliance with those rules is of utmost importance to decrease the adverse impact of litter on human health and the environment. moreover, it will contribute to optimizing the utilisation of refuse-derived materials. in poland, mixed waste is subjected to the following processes according to their management methods hierarchy: recycling, thermal transformation, other processing methods, and depositing in landfills. the fact that the quantity of selectively collected refuse in poland is growing every year is the cause for optimism (fig. 1). fig. 1. the comparison of selective waste collection in poland in 2012–2020 (source: https://bdl.stat.gov.pl/bdl/dane/podgrup/tablica) the increase in the quantity of selectively collected litter is undoubtedly attributable to the introduction of an amendment to the act of 13 september 1996 on maintaining cleanliness and order in municipalities (dz.u. z 1996 nr 132 poz. 622), which went into effect as of 1 january 2012. under article 6k. sec. 1 item 3 of that act, if waste is not collected and collected selectively, the municipal council shall introduce higher rates of fees. it is also the responsibility of the commune to obtain an appropriate level of recycling and to limit biodegradable municipal refuse deposited. the main component of household waste is organic matter. its share amounts to 40%, therefore it is important to implement the procedures aiming at reintroducing the organic components into the environment. the processing of household rubbish by biological methods, including composting, can be employed to recover the matter in the form of fertiliser or gas, and, at the same time, to reduce the quantities of refuse sent to landfills. composting is a method that is based on natural processes occurring in the environment through the activity of microorganisms, whose effectiveness is augmented by artificially optimized conditions. the composting process may involve the following types of waste: selectively collected kitchen and garden, refuse from green areas (city parks and green spaces), household-like litter generated by the industry and artisan shops and organic from the food industry as well as sediments from sewage treatment plants (czop, hatlapa, 2007; wengierek, 2014; czop, żydek, 2017). compost is an organic fertiliser. it contains decomposed remnants of plant-based materials. may contain faecal sludge and natural fertilisers. after the humification process is finished, the compost looks like hummus. it is dark and its smell can be compared to that of damp garden soil. depending on the type of substrate, it may contain nitrogen, potassium oxide, teraphosphorus decoxide as well as macro and microelements (krysztoforski, 2011). the analysis of the quantities of refuse subjected to the composting processes in poland in 2012–2020 shows that the largest volume of waste was composted in 2018, and the smallest – in 2020, when it was more than 3 times less than in 2018 (fig. 2). recently, i.e. in 2016–2020, the largest volumes of rubbish were composted in świętokrzyskie province. it should be noted that in 2018, that province recorded a significant, more than six-fold increase in the quantity of composted waste as compared to the previous year. on the other hand, the smallest quantities of composted household waste were recorded in 2016 in łódzkie province. it turns out that between 2014 and 2020, lubelskie and lubuskie provinces did not at all utilise composting as a refuse disposal method (fig. 3). fig. 2. quantities of waste recycled through composting in poland in 2012–2020 (source: https://bdl.stat.gov.pl/bdl/dane/podgrup/tablica) fig. 3. amount of waste subjected to recovery composting in individual provinces in poland in 2012–2020 (source: https://bdl.stat.gov.pl/bdl/dane/podgrup/tablica) certain local governments have assigned a high priority to composting through implementation special programs. one of the examples is the environment and agriculture department of the wrocław city hall (south-west poland), which has been realizing a program of compost bins distribution among wrocław residents as well as educational institutions located throughout the city. the purpose of the program is to educate the residents on correct composting of household refuse, promote composting of waste for the residents’ own needs, increase the quantities of recycled biodegradable rubbish and, at the same time, reduce the quantities of household waste generated in the city of wrocław that are being sent to landfills. beneficiaries of the program receive a compost bin free-of-charge for 36 months, and after that time the bin becomes the property of the user. the compost bin can last for 12 years and can recycle up to 1274 kg of biological waste per year provided that it is used according to the instructions and it is fully utilised (http://bip.um.wroc.pl/artykul/305/5331/programudostepniania-kompostownikow). the advantages of employing the composting method in refuse management processes include recirculation of significant quantities of biodegradable litter, sanitary and epidemiological hygienisation treatment of rubbish, reduction of the quantities of refuse sent to landfills by 30-50%, technical availability and easiness of use, economic availability, simultaneous disposal of several types of organic waste, and the potential for utilising the products of the composting process (drozd et al., 1996; manczarski, 2007; skalmowski, 2009; olczyk, 2012; łabętowicz et al., 2019). this paper attempts to evaluate the waste composting process as partial solution to the issue of management of refuse, both mixed and segregated. in addition, the article evaluates the potential for utilising the products of the household rubbish composting process. composting systems composting is a biothermal process that is divided into two main phases: i – thermophilic composting – a high-temperature phase during which the temperature may exceed 70°c; this phase is very important to the hygienisation treatment process; ii – mesophilic composting – the maturity phase which is characterized by a drop in temperature and the creation of humins (manczerski, 2007; skalmowski, 2009). some authors distinguish as many as 4 phases of this process: i – initial composting, in which the composting process is initiated and the multiplication of microorganisms, ii – thermophilic or intensive composting phase, iii – mesophilic phase known as proper composting, iv – compost maturation (temperature drop, humins formation) (manczerski, 2007). the overall course of the composting process is affected by several factors, including organic matter content, temperature of the compost mass, ambient temperature, structure, humidity, carbon/nitrogen (c/n) ratio and volatile compounds (sasaki et al., 2003; ozimek, kopeć, 2012; shimizu 2017). the duration of those phases depends on the applied technology http://bip.um.wroc.pl/artykul/305/5331/program-udostepniania-kompostownikow http://bip.um.wroc.pl/artykul/305/5331/program-udostepniania-kompostownikow and the composition of biomass to be composted. the intensity of exothermic decomposition of organic substances depends on their susceptibility to decomposition. lignin and scleroproteins such as e.g. keratin are very resistant to decomposition. hemicellulose and cellulose also do not decompose well. on the other hand, fats as well as most of sugars and proteins decompose very well (jędrczak, haziak, 2005). there are several ways of classifying the composting systems. selectively collected organic waste may be composted in open air on prisms under natural conditions (fig. 4). fig. 4. scheme of composting in prisms with active aeration (source: public domain, changed) this process is preceded by pre-treatment of refuse. it involves removing materials that may be harmful and grinding the compost to obtain the appropriate granulation. in addition, composting processes may take place in artificial conditions, in chambers or on concrete slabs; in that case, the waste is also pre-treated. composting prisms may be set up in open air, under the roof or in an enclosed space. they are usually placed on concrete slabs, which make it easier to drain leachate and artificially aerate the prisms. from time to time, the prisms may be flipped using specialized machinery. composting chambers may have different forms (fig. 5). in most of the cases, they are enclosed, may be movable or immovable, and may take various shapes (drums, tunnels, towers, rectangular chambers). composting chambers intensify the composting processes and create optimal conditions for their course. they make it easier to drain leachate and neutralise odours (manczarski, 2007; juda-rezler, manczarski, 2010; gawała-widera et al., 2011). fig. 5. scheme of an exemplary gazebo composter: a – cover, b1, b2 – chambers for bio-waste and earthworms, c – lower chamber for liquid fertiliser (source: public domain, changed) depending on various factors such as quantity and quality of refuse, location and conditions of the factory, the biomass composting processes are carried out in different systems. in most of the cases, the refuse is shredded, any contaminations (mostly metals) are separated from it, and then the waste is mixed with a mature compost, sawdust and other wood byproducts which act as structural material and keep moisture at an adequate level. organic waste collected separately is usually composted in open air, sometimes after installing an external insulation layer. the prisms are periodically flipped, and they are fitted with aeration and leachate discharge systems. sometimes this process is shielded from the external environment by installing a total or partial roof over the prisms. the best way of composting kitchen organic waste is storage in chambers due to odours neutralization, while green waste should be composted in prisms (manczarski, 2007; manczarski, 2013). potential for utilising of household waste composting process products biological processing of household waste has many advantages. products created through the household refuse composting processes are a source of thermal energy and biogas, and, as such, they may be practically utilised. some composted material, e.g. coffee grounds, lipid fraction from swine manure are used as biofuel feedstocks (liu, price, 2011; cho et al., 2021). compost, as a final product of that process, is used as fertiliser and soil additive for growing plants (mylavarapu, zinati, 2009). it is used in reclamation of degraded areas, to improve the soil structure, and as a component of fodders and mulches (czop, żydek, 2017). how the compost will be ultimately used depends on its characteristics and physical properties. it should not contain any “hard” contaminants such as e.g. glass or plastics. it should be “rich” in organic matter as well as substances that can be absorbed by plants. in addition to that, important characteristics that are taken into consideration include not presence of diseasecausing microorganisms, degree of its maturity as well as type of raw material from which it was made (baran et al., 2011; czop, żydek, 2017). composting process as a source of energy the composting process turns organic waste into fertiliser. this process is accompanied by production of large quantities of thermal energy, which could be utilised in practice e.g. in agriculture and horticulture. according to rosik-dulewska and grabda’s (2001) research, increase of soil temperature in foil tunnels accelerates growth and development of plants and boosts the crop yields. sołowiej (2007a) used the compost-generated thermal energy for optimizing the conditions for vegetable cultivation in foil tunnels. utilisation of that thermal energy resulted in shorter plant development period and increased crop yields. please note that such energy doesn’t cost anything, so there is also an economic incentive to use it. moreover, in his work devoted to the concept of using a compost prism as a low-temperature thermal energy source in vegetable cultivation, he (sołowiej, 2007b) proposed a design of an installation which transfers the heat from the compost prism via the pipe collector which is connected to the soil-heating pipes in the foil tunnel. such installation would have to be fitted with a circulating pump, which would force the circulation of water through the system. such system would allow the transfer of heat under certain conditions, and it would not have an adverse impact on the composting process. sołowiej (2007b) emphasized that usage of compost as a low-temperature thermal energy source would improve the profitability of vegetable cultivation in foil tunnels through significant acceleration of plant growth and increased vegetable crop yields, it would save conventional energy carriers, and, at the same time, reduce the quantities of pollutants discharged into atmosphere which usually accompany the traditional heating methods. compost can be used as a renewable sour of energy due to its organic carbon content. however, the problem is the moisture content of the compost, which can be reduced e.g. by adding cardboard. moreover, it was found that adding cardboard after the completion of the composting process increases the energy content of the compost (raclavská et at., 2011). lignocellulosic biomass is also one of the renewable energy sources. it is a source of biofuels, including bio-fuel, biohydrogen, and biogas. the use of lignocellulosic biomass will contribute to the reduction of fossil fuel consumption (koul et al., 2022). compost used as fertiliser and soil additive for growing plants compost is used for fertilising the soil in orchards arable fields, forestry, and green areas. its composition may be enriched depending on the needs of the soil in which it is supposed to be used. to enrich the compost, the following products are added: fertilisers, alkalising agents (to increase ph) as well as peat/lignite (to increase the number of carious substances) (czop, żydek, 2017). compost rich in micro-elements, e.g. zinc, and copper, can be used for the production of fertiliser mixtures (baran et al., 2011). one of the compost’s properties is nutrient retention in the soil and improvement of soil structure due to the carious substance content. in addition, compost reduces the erosion of phosphorus, making it easier to absorb it by plants and microorganisms (czop, żydek, 2017). in the study on composting process with the addition of animal-derived refuse, anders and nowak (2008) concluded that adding meat waste during the composting process yielded a very good quality compost that could be used as organic fertiliser. to be used as fertiliser, the compost must meet priority quality requirements that are prescribed by the regulations. therefore, it is very important to know the origin of the material which will be used in the composting process. composts obtained from mixed household waste are of much lower quality than those produced from selectively collected waste, mostly because mixed household waste is contaminated with heavy metals (for example zinc, chromium, copper) plastics, and glass (manczarski, 2007; olczyk, 2012). in the study on the impact of selective collection of household waste on the properties of composts, olczyk (2012) proved that the biodegradable fraction selected from the household waste had met the requirements stipulated in the regulation of the minister of agriculture and rural development of 18 june 2008 in the matter of implementation of selected provisions of the act on fertilisers and fertilisation (dz.u. 2008 nr 119 poz. 765), and, therefore, after processing, it could be used as fertiliser. after analysing the composition of the compost obtained from mixed and not segregated at the source waste, olczyk (2012) noted that it contained large quantities of heavy metals which disqualified it from being used in agriculture. compost used as a soil supplement significantly enhanced strawberry plant (fragaria ×ananassa duchesne ex rozier) growth and fruit quality. in experiments, peter nutrient solution (50% fertilizer) was added to a mixture of 50% soil plus 50% compost. this increased the dry weight of the plants twice, fruit yield by more than 70%, and fruit size by 15% as compared to control. compost with fertilizer also enhanced leaf chlorophyll content (wang, lin, 2002). the addition of compost resulted in improvement of both physical and chemical properties of the soils as well as increased parsley (petroselinum crispum (mill.) fuss.) yields in research provided by mylavarapu and zinati (2009). poultry manure used in combination with tillage increased grain yield by about 40% compared with tillage only (agbede, ojeniyi, 2009). compost from municipal solid waste (msw) not source separated and bark and sewage sludge (bs) had a positive effect on soil softness, porosity, availability of nutrients for plants, and water holding capacity. both materials influenced the growth of apple (malus sp.) and vine (vitis vinifera l.) cuttings and prevented the development of weeds (pinamonti, zorzi, 1996). but inadequate processing of farmyard manure, compost or slurry can be a reason for increased weeds. to kill the weed seeds, the temperature must be sufficiently high during the composting phase (bàrberi, 2002). compost that meets the relevant standards can be used in ecological agriculture. as reported by illera-vives et al. (2013) compost with seaweed, fish refuse (their skin, heads and spine) and pine bark are used in such agriculture. they found that fish waste contained a lot of nitrogen and phosphorus. the sea wood contained high amounts of potassium, sodium and moisture. on the other hand, pine bark had a high c/n ratio. due to the low metal content of all types of waste, this compost is suitable for cultivation and it can be used in ecological agriculture. compost used in reclamation of degraded areas research conducted by kujawska et al. (2016) has shown that mixing a mineral component (containing mining waste) with an organic component (in the form of compost originating from an organic fraction of household waste) creates a mineral and organic base that can be used to create a reclamation layer for degraded areas. this research evaluated the following parameters of mixes: the content of biogenic elements, macroelements, and microelements as well as the toxicity of the obtained base to an experimental plant – common wheat (triticum aestivum l.). despite increased contents of heavy metals in the obtained base, they did not accumulate in the plant biomass. the obtained organic base had physical and chemical properties which were similar to the properties of the soil. compost bs is a valid alternative to the soil and organic soil conditioners on the market in the recovery of the degraded area. in the studies conducted by pinamonti and zorzi (1996) compost secured a rapid emergence and regular growth of herbaceous species covering degraded areas. compost used to improve the structure of the soil and as a component of fodders and mulches compost has a broad application in terms of improvement of the soil structure. it decreases erosion processes through the reduction of rainwater surface runoff; it increases the durability of the surface of park pathways as well as recreation and sports amenities; it helps reduce the creation of puddles when mixed with sand, clay, or small gravel (czop, żydek, 2017). to improve the structure of the soil and increase the amount of carbon in it, manure is used, appropriate irrigation, cover, and energetic plants are planted, as well as agroforestry (lal, 2004). some crop residues were used in soil mulching and producing animal fodder (koul et al. 2022). as a component of fodders and mulches, compost contributes to the development and growth of pigs and chickens. due to the content of vitamin b12, natural antibiotics, and iron ions, compost was observed to have a positive impact on poultry which is less susceptible to diarrhoea and contagious diseases (czop, żydek, 2017). utilisation of compost and legal regulations according to the requirements stipulated in the national waste management plan 2022 (m.p. 2016 poz. 784), by 2020, no more than 35% of the mass of biodegradable household waste should be sent to landfills, taking as a base the quantity of waste produced in 1995 (4.38 million megatons). under the regulation of the minister of environment of 25 may 2012 in the matter of limiting the mass of biodegradable household waste sent to landfills and the method of estimating the limits of such waste (dz.u. z 2012 r., poz. 676), in 2012, the limit of the mass of biodegradable household waste sent to landfills was 70%, in 2013–2015 – 50%, in 2016– 2017 – 45%, and 2018–2019 – 40%. based on the aforementioned regulations, the priority objective of biodegradable household waste management is to reduce the quantities of waste that is being sent to landfills. the implementation of that plan may be significantly aided by composting, which contributes to the reduction of quantities of household waste sent to landfills by as much as 40%. in addition, composting allows recovering the organic matter in the form of fertiliser which may be used in agriculture or orchards. however, it must meet the quality requirements which are evaluated based on the following three criteria: composition, physical properties, physicochemical properties, and chemical properties; sanitary and hygienic condition; and degree of maturity or stabilization. such criteria for organic fertilisers and soil additives for growing plants are defined in the regulation of the minister of agriculture and rural development of 18 june 2008 in the matter of implementation of selected provisions of the act on fertilisers and fertilisation (tab. 1–2). tab. 1. quality requirements for solid fertilisers (based on the regulation of the minister of agriculture and rural development of 18 june 2008 in the matter of implementation of selected provisions of the act on fertilisers and fertilisation) parameter unit organic fertiliser organic and mineral fertilises mineral fertiliser organic substance [% dry mass] ≥ 30 ≥ 20 potassium, k2o [% mass] ≥ 0.2 ≥ 1 ≥ 2 phosphorus, p4o10 ≥ 0.2 ≥ 0.5 ≥ 2 total nitrogen, n ≥ 0.3 ≥ 1 ≥ 2 tab. 2. permissible quantities of contaminants in fertilisers (based on the regulation of the minister of agriculture and rural development of 18 june 2008 in the matter of implementation of selected provisions of the act on fertilisers and fertilisation) contamination organic fertiliser organic and mineral fertiliser mineral fertiliser unit [mg/kg dry mass] arsenic (as) ≤ 50 chromium (cr) ≤ 100 ≤ 100 cadmium (cd) ≤ 5 ≤ 5 ≤ 50 nickel (ni) ≤ 60 ≤ 60 lead (pb) ≤ 140 ≤ 140 ≤ 140 mercury (hg) ≤ 2 ≤ 2 ≤ 2 fertilisers and soil additives for growing plants must meet certain sanitary-hygienic requirements in terms that they must not contain any live eggs of intestinal parasites ascaris sp., trichuris sp., toxocara sp. or salmonella bacteria. an equally important quality requirement that the compost must meet so that it could be used as a fertiliser is its maturity. compost maturity is taken into consideration when evaluating the compost’s usefulness as a fertilizer and an additive for growing plants. according to the regulation of the minister of agriculture and rural development of 18 june 2008 in the matter of implementation of selected provisions of the act on fertilisers and fertilisation, a mature compost meets the following criteria: at4 (transpiration) – below 10 mg o2/g of dry mass, loi (loss on ignition) – below 35% of dry mass, and toc (total organic carbon) – below 20% of dry mass. conclusions the organic matter from household waste may be recovered through composting in the form of a fertiliser, which can be broadly used, as well as in the form of thermal energy and biogas. in addition, less waste will be sent to landfills thanks to composting. therefore, this process can benefit us in two ways: it protects the environment and helps agriculture. when it comes to environmental protection, there is less organic matter that is being sent to landfills, and, therefore, the landfills will occupy a smaller area. on top of that, composting eliminates sanitary hazards. when it comes to aiding agriculture, the products of the composting process can be used as fertiliser. to be used as fertiliser, the compost must meet priority quality requirements that are prescribed by the regulations. composts obtained from mixed household waste are of much lower quality than those produced from selectively collected waste. however, due to growing quantities of selectively collected household waste, including biodegradable refuse, and due to the increasing percentage of its volumes being biologically transformed, we can be optimistic that one day we will see widespread use of waste-derived fertilisers. thus, composting can be the “golden method” of solving household waste problems, but it still requires additional financial outlays and regulations, as well as an appropriate incentive for society. references agbede, t. m., ojeniyi, s. o. 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[in polish] kompostowanie – „złota metoda” na rozwiązanie problemu organicznych odpadów domowych? – krótki przegląd streszczenie w polsce ilość odpadów poddanych recyklingowi organicznemu – kompostowaniu począwszy od roku 2015 wzrasta. obserwuje się także rosnącą ilość selektywnie zbieranych odpadów komunalnych, w tym biodegradowalnych. recykling organiczny bioodpadów daje możliwość odzyskania materii w formie kompostu lub stabilizatu, przy jednoczesnym zmniejszeniu ilości odpadów kierowanych na składowiska. kompost przeznaczony na nawóz musi spełnić określone wymagania jakościowe zgodnie z ustawą o nawozach i nawożeniu. otrzymany nawóz znajduje szerokie zastosowanie m.in., jako: środek wspomagający uprawę roślin, podłoże w rekultywacji terenów zdegradowanych, dodatek do pasz i ściółek. słowa kluczowe: materia organiczna, unieszkodliwianie odpadów, nawozy, wysypiska śmieci received: [2022.03.18] accepted: [2022.07.27] annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832 doi: 10.24917/25438832.7.x weed communities of jerusalem artichoke (helianthus tuberosus l.) cultivation alina stachurska-swakoń*, rakowska rita, sabina klich department of plant ecology, institute of botany, jagiellonian university, gronostajowa 3, 30-387 kraków, poland, *alina.stachurska-swakon@uj.edu.pl introduction the european initiative “clean energy for all europeans” facilitates the transition away from fossil fuels toward cleaner energy. the changed directive from december 2018 points to 32% energy consumption from renewable sources (eu directive, 2018/2001). it causes demand to search for various sources to gain the eu target. one of the sources of clean energy is organic matter and the interest in plants as the source of renewable energy has increased in the last twenty years. the plants could be used in two ways in the context of renewable energy: the plant biomass can be burned to produce electricity from the heat or can be transformed and converted into liquid fuels (e.g., tuck et al., 2006; tilman et al., 2009; araújo et al., 2017). to achieve high yields from biomass production, the use of additional water, fertilizers, and pesticides are usually needed. additionally, the competition for land between food and fuel uses was recently recognised (haberl, 2015). the choice of cultivated plant species for energy crops depends mainly on purpose; however, the environmental conditions are also taken into consideration. to the group of species for biomass combustion belong among others: miscantus ×giganteus j. m. greef & deuter ex hodk. & renvoize, reynoutria sachalinensis (f. schmidt) nakai, rosa multiflora thunb., salix viminalis l., sida hermaphrodita (l.) rusby (e.g., anioł-kwiatkowska et al., 2009; wróbel et al., 2011; tokarska-guzik et al., 2012). plant species used to receive liquid fuels are corn, and sugar beet (rossini et al., 2019). jerusalem artichoke (ja) (helianthus tuberosus l.) can be used both for above-ground biomass for burning and undergrowth tubers for ethanol production (swanton et al., 1992; rossini et al., 2019). there are several characteristics that make ja worthy of cultivation for an energy crop: rapid growth, a high carbohydrates content, high dry mass (monti et al., 2005; baldini et al., 2006; rossini et al., 2019), pathogen tolerance, minimal external production costs (monti et al., 2005), ability to grow in marginal lands (kays, nottingham, 2007). total dry matter production ranges from 6 to 9 t/ha under limiting conditions and from 20 to 30 t/ha under favourable conditions, or even to 35 t/ha in the case of some genetic lineages (denoroy, 1996; kays, nottingham, 2007; liu et al., 2011). environmental conditions influence the biomass allocation strategy (swanton et al., 1992; gao et al., 2011), and assimilation activities also vary between developmental stages and genotypic lines (swanton, cavers, 1989; kocsis et al., 2007; gao et al., 2011). the species is tolerant to drought, frost, and salinity; it can easily grow in different types of soil (kosaric et al., 1984; swanton et al., 1992; denoroy, 1996; baldini et al., 2006). the ja is a perennial plant native to central north america (swanton et al., 1992). the original use of this plant was for food for humans and livestock. the north american indians appreciated ja as a readily available source of food. the species was introduced to europe as early as the beginning of the xvii century (balogh, 2008). it has escaped cultivation and started to invade natural plant communities. in poland, it was recorded for the first time in 1872 (sudnik-wójcikowska, 1987). its spread into natural and semi-natural habitat began in 1960, today it could be found along rivers, ponds and at the edges of forests (tokarska-guzik, 2005; towpasz, stachurska-swakoń, 2011, 2018; popiela et al., 2015; zając, zając, 2015; jarek, stachurska-swakoń, 2016). nowadays, the species has the status of invasive in europe: poland, austria, italy, germany, france, and hungary (wittenberg, 2005; balogh, 2008; tokarskaguzik et al., 2012; filep et al., 2018). h. tuberosus belongs to the asteraceae family and is one of the sixty-six species of the genus helianthus l. native to america (balogh, 2008). it creates coarse stems reaching above 3 m with numerous ovate leaves up to 25 cm long (swanton et al., 1992; kays, nottingam, 2007). it produces yellow inflorescences (capitula) that are 5–10 cm in diameter (fig. 1, appendix 1). the plant reproduces by seed and spreads by tuber-bearing rhizomes. the species is known to be highly polymorphous (kays, nottingham, 2007). the cultivation of energy crops is still a little researched subject in terms of their impact on biodiversity. the energy crops are a specific type of plantation, the crops are harvested usually in the autumn. in syntaxonomy, weed communities constitute a separate group of ecosystems that arise spontaneously under conditions of a specific, but extreme anthropopressure. the aim of the research was to study the weed species that could occur in the plantation of the h. tuberosus. we hypothesize that in the ja plantations develop communities from the polygono-chenopodietalia (r.tx. et lohm. 1950) j.tx. 1961 order, as the ja has long development during the vegetation season and the crops are harvested in autumn. fig. 2. the locality of the polanka haller (wielickie foothills, western carpathians) (https://pl.m.wikibooks.org/ wiki/map.png, modified) study area, data collection, and data analyses the research was carried out in the pogórze wielickie foothills (west carpathians, s poland) in the experimental plantation in the area of “polanka haller” belonging to the jagiellonian university (fig. 2). the experimental farm is located in an area with a mean annual temperature of 7.8 c. the gray-brown podzolic soils dominated here. the detailed characteristic of the area is given by drużkowski (1998). the phytosociological relevés were made in the first year of plantation of helianthus tuberosus – (ja) using the braun-blanquet (1964) approach, in 2007. the time of august and september was chosen as the best to make relevés in the plantation according to the developmental stage of ja. the area of 100 m2 was chosen as the appropriate for the aim of the study and the seventeen stands were chosen as representative of the plantation area in accordance with the ja cover. the whole area of the ja plantation on the jagiellonian university farm was 3.25 ha at that time. additionally, soil ph was measured in the field in each plot using the hellige colorimetric method. the phytosociological table was prepared based on the results of upgma with the ruzicka coefficient (mvsp package). the syntaxonomical affinity of the species follows matuszkiewicz (2005). the names of vascular plants follow mirek et al. (2002). the correlations between plant cover were examined using the kendall coefficient (statistica 13.0 software). habitat characteristics were performed using the phytoindication method with indicator values according to ellenberg (ellenberg et al., 1992). weighted average values of indicators were calculated for all relevés: light – l, soil moisture – f, soil acidity – r, and nitrogen – n, taking into account the number of species. the mann–whitney non-parametric test was used to check the influence of soil requirement of weeds (statistica 13.0 software). results the cover of helianthus tuberosus – (ja), as a crop plant, was differentiated in the study area and the relevés were made in representative plots with its cover between 35 and 100%. the average cover of the plants in the relevés was 83%. vascular plants were noticed in every plot, however, their number and cover varied between plots. the cover of the plants was highly correlated with the abundance of ja and ranged between 5 and 70% with a mean of 30%. in plots with the cover of 100% ja cover, weeds covered 3–20% of the plot area (appendix 2 – tab. 1). a total of 82 species of vascular plants were noticed in h. tuberosus with the range of 5 to 36 in one relevé with the area of 100 m (appendix 2 – tab. 1). more than half (45 species, 55%) were noticed only sporadically. the group of plants that achieved the iv degree of constancy included: agrostis stolonifera, cirsium arvense, elymus repens, and rumex obtusifolius. the listed species were also more numerous: they occurred with a coverabundance value between 1 and 3. they represent the ruderal communities of the artemisietea class and all of them are perennial plants. field weeds belonging to the stellarietea mediae class were represented by the largest group and consisted of 26 species (31.7% of all species); however, only 17 occurred in more than 20% of plots. furthermore, the group was not homogenous and contained the characteristic species of both weed crop types: cereal of the order centauretalia cyani (9) and root crops of the order polygono-chenopodietalia (8). the most common species in the data set, reaching the iii degree of constancy, were: apera spica-venti, matricaria maritima subsp. inodora, and vicia tetrasperma characteristic of centauretalia cyani and chenopodium album, and sonchus oleraceus characteristic of polygono-bidentetalia. the group of meadow species that occurred in the plots with ja consisted of 24 species (29.3%); however, only 8 species were abundant in more than 20% of the plots. they represent molinio-arrhenatheretea class, plants with a wide ecological spectrum. additionally, a few sporadic species were moisture preferred like mentha longifolia and scirpus sylvaticus. meadow species were often noted in the plots. fig. 3. coverage of weed species (a) and a number of weed species (b) in relation to jerusalem artichoke cover in jerusalem artichoke plantations (p < 0.005) a mean number of 20 vascular plant species was observed in one relevé. this number differentiated between plots and ranged between 8 and 36. the number of weed species was significantly negatively correlated with the cover of ja (fig. 3; y=-1.19x+ 129, r2=0.91; p<0.0001), the high land cover of ja resulted in a smaller number of vascular plants that cooccurred with the crop-plant. similarly, a negative correlation between the cover of ja and the cover of other vascular plants (fig. 3; y=-0.44x+56.67, r2=0.8; p<0.0001). fig. 4. mean weighted values of the ellenberg’s indicators for l – light, f – soil moisture (a) and r – acidity, n – soil nitrogen content (b), calculated for all relevés with jerusalem artichoke; sequence of phytosociological relevés according to tab. 1 – appendix 2 most of the plants accompanying ja cultivation are perennial, mainly hemicryptophytes (42 species, 51.2%) with a smaller number of geophytes (14 species, 17%). therophytes were represented by 21 species (25.6%). considering the plant origin, 12 species were archaeophytes (14.6%), while 7 species (8.5%) represented kenophytes such as solidago canadensis, erigeron annuus, e. canadensis. the species richness and composition were connected with the ja cover, which was also reflected in the mean ellenberg values. the statistical significance (u mann – whitney test) difference of indicators between stands with relatively low (3-4) and high (5) ja cover was detected for the moisture and nitrogen indicator (fig. 4). the moisture indicator was higher with the closed canopy of ja (respectively 5.48 for ja=3-4 and 5.89 for ja=5, p< 0.001). the nitrogen indicator was also lower, with an average value of 6.0 when the ja cover was lower and 7.19 for ja cover 5 (p<0.001). discussion the impact on the biodiversity of perennial crops intended for energy purposes is poorly understood, as it is a relatively new direction of production introduced on agricultural land. some authors emphasize the need for the monitoring of energy crops in terms of their impact on biodiversity at various levels (e.g., rowe et al., 2007; feledyn-szewczyk et al., 2011; klima et al., 2019). usually, energy plantations are less intensively managed and less disturbed than arable fields. in this aspect, some benefits for local biodiversity could be perceived when we compare them with arable fields (rowe et al., 2007; dauber et al., 2010; stanley, stout, 2013) and a general decline in european agriculture landscapes. the presented research contributes to the understanding of biocenotic and floristic relationships in energy crops, since the studies on the infestation of energy crops are carried out rarely. the energy crops are usually composed with the alien species to the local habitat, and the weed composition is not yet stable. from the syntaxonomical point, it could be interesting the affinity of the weed composition in energy crops to the syntaxonomical classification. in the literature, most attention is paid to weed infestation in willow plantations, as this species is cultivated more often. our research indicates that the weeds in the jerusalem artichoke (ja) plantation do not create a stable composition. the occurred species represent various communities, the spontaneous vegetation consists of species that are characteristic for arable fields, ruderal, and meadow habitats. most species appeared sporadically with low importance for species composition. as ja plantation could persist for a few years, perennial cultivation promotes the appearance of perennial species, the appearance of troublesome weeds, and the presence of meadow species. the difficulties to identify weed communities were also indicated by other studies on weed infestation in energy crops (korniak, 2007; rola et al., 2007; sekutowski and badowski, 2007; trąba et al., 2009; anioł-kwiatkowska et al., 2009). as in the case of our studies, the composition of species representing different syntaxonomical units: forest, meadow, ruderal, and segetal plants were reported. the authors indicate that the species richness of the energy crops depends on the age of cultivation and the previous state of the habitat. plantations established in meadow habitats should be more diverse in weed composition than those established on former agricultural lands. koncekova et al. (2014) assumed an influence by both the impact of prior land use and the impact of crops from the surrounding arable fields. in their studies on spontaneous vegetation of miscanthus plantation, they found a higher share of species included in the group of species occurring in root crops and cereals. in the third year of their observation, the share of perennial species increased. the authors explained it with little land disturbance that creates good conditions for perennial species. the life form and some morphological traits of the cultivated species could also influence the weed community. sobisz and ratuszniak (2009), comparing species of weed companions in willow, rose, and ja plantations, indicated that the smallest number of weeds was recorded in helianthus cultivation, under dense and shadow conditions, on the other side the highest in salix cultivation. in his study, there were no species with high degrees of constancy in helianthus cultivation, and he did not find a group that would positively distinguish ja crops. ziaja and wnuk (2009) pointed to the impact of crop duration on the composition and diversity of herbaceous plants accompanied energy crops. baum et al. (2012) found the highest similarity in plant composition of willow plantations with marginal grassland strips, grasslands, or even mixed forests. janicka et al. (2020) indicated the significance of soil condition that influences the plant diversity in salix crops. similarly, the diversity of weed species in the miscanthus crop was dependent on soil type (feledyn-szewczyk et al., 2011). there is a group of species that we could consider generalist perennial weeds that occur in various energy crops. to this group, generalist energy crop weeds, belong: convolvulus arvensis, elymus repens, equisetum arvense, ranunculus repens, and rumex obtusifolius. we expected more species of archaeophytes in our plots, as ja crops were established in the formerly cultivated fields, and therefore seeds of field weeds could persist in the soil bank. the low number of archaeophytes recorded during the conducted research coincides with observations from other energy plantations (e.g., anioł-kwiatkowska et al., 2009). the decline of local flora in archaeophytes is often documented (e.g., meyer et al., 2013; stachurskaswakoń, trzcińska-tacik, 2014; towpasz and stachurska-swakoń, 2018). introducing new crop plants may intensify this trend. the low number and frequency of the accompanying plant species could be related to the allelopathic potential of ja. the published experiments showed that wild and cultivated decomposing ja residues, particularly leaves and stems, have a phytotoxic potential (tesio et al., 2011). the allelopathic potential of this plant was studied for several crops and weeds. according to vidotto et al. (2008), the species showed a varied sensitivity to aqueous extracts from ja. the species group including amaranthus retroflexus and echinochloa crus-galli belonged to the sensitive species, and chenopodium album, the common weed species belonged to the group poorly sensitive. in another experiment, the germination rates of elymus repens, solidago gigantea, and trifolium vulgare were not influenced by extracts of ja (filep et al., 2016). the allelopathic potential is also known for the other species of the helianthus genus (kliszcz, 2018; puła et al., 2019). in addition, invasive species often reveal the allelopathic potential for the germination process of accompanying species (e.g., zandi et al., 2020). according to kompała-bąba and błońska (2008), ja creates its own communities with different species depending on place history and soil types. it grows together with dominants of nitrophilous communities of semi-shaded margins (aegopodium podagraria) or perennials from the artemisietea class such as urtica dioica, artemisia vulgaris, cirsium arvense, equisetum arvense, or with other alien species such as solidago canadensis or s. gigantea. in his natural range, the species build communities along with a wide ecological tolerance species such as cirsium arvense and elymus repens. these two species were also noticed in our research. conclusions the spontaneous vegetation of jerusalem artichoke (ja) crops consists of varied groups of species, and we did not notice the repeatable composition of plants. the list contains species with low cover and low frequency across our dataset of relevés. in general, the associated vegetation of ja is similar to other perennial energy crops. it can be proposed the group of generalist energy crop weeds with convolvulus arvensis, elymus repens, equisetum arvense, ranunculus repens, and rumex obtusifolius. the history of the plantation, the vicinity of plant associations, and the local habitat condition could be major drivers of the diversity of weeds in the plantations. as ja is also an invasive species and it is difficult to eradicate, in that case, the monitoring of ja plantation is needed. conflict of interest the authors declare no conflict of interest related to this article. references anioł-kwiatkowska, j., kącki, z. śliwiński, m. 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[in polish] appendix 1 fig. 1. jerusalem artichoke helianthus tuberosus l.; capitulum-type inflorescences (a), compact aggregation of the study species (b) (photo a. stachurska-swakoń) appendix 2 tab. 1. helianthus tuberorus l. crops on the wielickie foothills (western carpathians) successive number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 c o n st a n c y field number 18 20 21 22 23 25 26 27 28 29 30 31 32 57 59 61 63 date [2007 year] 3.08 3.08 3.08 3.08 3.08 4.08 4.08 7.08 7.08 7.08 7.08 7.08 7.08 14.09 14.09 14.09 14.09 area [m2] 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 soil ph 7 6 6 5.5 7 6 7 7 6 6 7 6 6 6 6 6 6 cover of h. tuberosus [%] 60 60 40 35 60 85 85 97 99 85 100 100 100 100 100 100 100 cover of weeds [%] 70 60 70 80 70 40 30 10 7 15 15 15 20 3 3 3 3 exposure s n s n sse sse ne se s slope [degree] 5 5 5 3 3 5 3 3 3 number of weed species 36 37 31 36 35 26 23 21 8 17 11 11 15 9 9 10 10 crop plant helianthus tuberosus 4 4 4 3 4 5 5 5 5 5 5 5 5 5 5 5 5 v weeds: agrostis stolonifera 3 3 3 3 3 2 1 1 + 1 . . 1 . . + . iv cl: artemisietea elymus repens 2 3 3 3 3 3 3 . 1 2 2 2 2 . . . . iv cirsium arvense 1 2 1 1 1 2 . 1 1 + 1 . + + . + . iv rumex obtusifolius 1 . . . + 1 + + + . + 1 2 + . + + iv tanacetum vulgare 3 + 2 3 3 1 1 . . 1 . . . . . . . iii artemisia vulgaris 1 . 1 2 . . + + . . + . . . . + . ii galeopsis pubescens + 1 . . + . . . + . . + . . + + + ii urtica dioica . . . . . . . . + 1 + 1 . . . 3 ii o: centauretalia cyani matricaria maritima subsp. inodorum 2 1 2 2 2 2 1 + . 1 . + . . . . . iii apera spica-venti + 1 1 1 1 + + . + + . . . . . . . iii vicia tetrasperma + + 1 1 2 1 + . + . . . . . . . iii matricaria chamomilla 1 1 1 1 1 . . . . . . . . . . . . ii papaver rhoeas + 2 + + . . + . . . . . . . . . . ii vicia hirsuta 1 2 + 1 1 . . . . . . . . . . . . ii o: polygono-bidentetalia sonchus arvensis 3 + 3 2 2 . 1 + . . . + . . + . + iii chenopodium album 1 1 . 1 1 + + . . . . . + + . . . iii oxalis stricta + + + 1 1 + . . . . . . . . + . . ii capsella bursa-pastoris 1 1 + + + . . . . . . . . + . + . ii cl: stellarietea mediae galeopsis tetrahit . 2 2 1 1 . + . . . . . . . + + . ii polygonum aviculare + . + . . . . . + . . . . . . . + ii tussilago farfara 1 + 1 1 1 . . . . . . . . . . . . ii viola arvensis + + + + + . . . . . . . . . + . . ii cl: molinioarrhenatheretea ranunculus repens + 2 2 . 1 1 . 1 . + 1 + 1 . . . . iii stachys palustris 1 . 1 2 2 1 . . . . . + . 1 1 . . iii achillea millefolium 2 2 . 2 . . + . . . . . . . . . . ii crepis capillaris 1 . . 1 1 . + + . . . . . . . . . ii daucus carota 2 . 2 2 3 . 1 . . . . . . . . . . ii phleum pratense . . . + . . . + . . . . 1 . . + . ii poa pratensis . . . . . + . + . + + + + . . . . ii vicia cracca + 1 . + + . . . . . . . . . . . . ii companions erigeron annuus 1 + . . 2 2 2 1 . + . . . . + . 1 iii erigeron canadensis 1 1 2 1 1 2 + + . + . . . . . . . iii equisetum arvense . + + . + 2 + + . + . . . . . . . iii epilobium sp. . . . . . + . + . 1 . . . + + + . ii mentha arvensis 1 + . . 1 . + + . . + . . . . . . ii galeopsis bifida 1 + . + . . . . . . . . + . . . . ii taraxacum officinale . + + . + . + . . . . . . . . . . ii sporadic species: centauretalia cyani: centaurea cyanus 1, 2; polygonum tomentosum 4, 5; polygono-bidentetalia: raphanus raphanistrum 5, 14; rumex crispus 1: 1, 2, 4; setaria pumila 4; veronica persica 3, 7; stellarietea mediae: geranium robertianum 3, 14; lactuca serriola 5; myosotis arvensis 3, 6; sinapis arvensis 5; molinioarrhenatheretea: cerastium holosteoides 6; cirsium oleraceum 11; deschampsia caespitosa 13; festuca pratensis 2; geranium palustre 17: 1; heracleum sphondylium 2: 2, 6; juncus effusus 6; lysimachia nummularia 10; l. vulgaris 4; mentha longifolia 4; plantago major 6; potentilla anserina 2: 1, 13:1; scirpus sylvaticus 13; trifolium hybridum 1; trifolium pratense 2; veronica chamaedrys 17: 1; artemisietea: chaerophyllum aromaticum 11; cirsium vulgare 8; torilis japonica 6, 10: 1; vicia angustifolia 2; vicia sepium 3, 5; companions: betula pendula (c) 3: 1, 4; calamagrostis epigejos 4, 7; carex brizoides 17: 2; epilobium hirsutum 1; geranium dissectum 4; gnaphalium sylvaticum 1, 6:1, 8; g. uliginosum 6; medicago lupulina 5; myosoton aquaticum 8, 12 13; plantago intermedia 3: 1; rubus caesius 8; solidago canadensis 3, 4, 5; stellaria graminea 2; veronica arvensis 2. abstract jerusalem artichoke could be used as a source of renewable energy in the meaning of biomass combustion or liquid fuels production. the presented study concerned on the impact of ja plantation for biomass combustion on plant diversity. the spontaneous vegetation of ja crops studied on the basis of phytosociological methods consisted of varied groups of species that contain weeds (32%), meadow (29%), and ruderal (13%) species. most of the species occurred sporadically (55%) with low frequency. most of the plants accompanying ja cultivation were perennial, mainly hemicryptophytes (51%) with a smaller number of geophytes (17%). therophytes constituted 25% of spontaneous flora of ja crops. it can be proposed the group of generalist energy crop weeds with convolvulus arvensis, elymus repens, equisetum arvense, ranunculus repens, and rumex obtusifolius. keywords: energy crops, invasive species, renewable energy, topinambour, weeds received: [2022.02.16] accepted: [2022.05.18] zbiorowiska chwastów w uprawach topinamburu (helianthus tuberosus l.) streszczenie uprawy roślin energetycznych stanowią wciąż mało zbadany obiekt pod kątem wpływu na różnorodność biologiczną. celem prezentowanej pracy była charakterystyka spontanicznej roślinności towarzyszącej uprawie topinamburu (helianthus tuberosus l.), jednego z gatunków uprawianych na cele bioenergetyczne wykorzystywanego, zarówno w postaci biomasy, jak i biopaliw. gatunki współwystępujące z topinamburem nie tworzyły trwałych zbiorowisk chwastów polnych, reprezentowały różne grupy syntaksonomiczne: chwasty polne (32% gatunków), gatunki łąkowe (29%), ruderalne (13%). terofity stanowiły 25% zanotowanych gatunków. większość roślin (55% gatunków) występowała sporadycznie z niskim pokryciem. zarówno liczba towarzyszących gatunków jak i ogólne pokrycie roślin towarzyszących było ujemnie skorelowane z pokryciem topinamburu. do gatunków, które można uznać za pospolicie występujące w uprawach energetycznych można zaliczyć: convolvulus arvensis, elymus repens, equisetum arvense, ranunculus repens, rumex obtusifolius. słowa kluczowe: rośliny energetyczne, gatunki inwazyjne, energia odnawialna, topinambur, chwasty information on the authors alina stachurska-swakoń https://orcid.org/0000-0003-0381-4520 department of plant ecology she is interested in the vegetation ecology, particularly in the mechanisms of plant communities changes and flora under natural and anthropogenic influence, plant ecology, and plant protection. rakowska rita https://orcid.org/0000-0002-4255-4576 she is interested in beaver influence on vegetation, ias, nature management and conservation. sabina klich https://orcid.org/0000-0001-5412-8390 she is interested in the development of water and marsh vegetation within dam reservoirs, plant ecology, invasive species, vegetation in protected areas and habitats protection. 194 annales universitatis paedagogicae cracoviensis studia naturae, 5: 194–219, 2020, issn 2543-8832 doi: 10.24917/25438832.5.13 mohamad hesam shahrajabian1, wenli sun1, qi cheng1,2* 1biotechnology research institute, chinese academy of agricultural sciences, beijing 100081, china 2college of life sciences, hebei agricultural university, baoding, hebei, 071000, china; global alliance of hebau-cls&heqis for bioal-manufacturing, baoding, hebei 071000, china; *chengqi@caas.cn chinese jujube (ziziphus jujuba mill.) – a promising fruit from traditional chinese medicine with an understanding of traditional chinese medicine (tcm) comes a unique knowledge of chinese herbs and fruits, as both typical ethnic foods and traditional health-promoting foods. additionally, the exploration of asian food culture provides historical information related to traditional chinese medicine and the use of chinese herbs and fruits in dietary applications (e.g. khoshkharam et al., 2020; shahrajabian et al., 2019 a,b, 2020 a,b; sun et al., 2019 a,b, 2020). �e chinese jujube (ziziphus jujuba mill. or z. jujuba), which originated in china, has a history that spans more than 4,000 years and is recognised as the most important fruit species belonging to the rhamnaceae juss. family (mardare et al., 2016), especially in asia. it was �rst described scienti�cally by carolus linnaeus as rhamnus ziziphus l., in “species plantarum” in 1753. later in 1768, philip miller concluded it was su�ciently distinct from rhamnus genus to merit separation into a new ziziphus genus. miller named it z. jujuba, using linnaeus’s species as a name for the genus (bean, 1988). chinese jujube and the similar indian jujube (z. mauritiana lamk.) are largely used in traditional asian medicine as super fruits (krishna et al., 2016) – fig. 1. indian jujube (also belonging to family rhamnaceae l.) is a tropical/subtropical fruit native to the northern hemisphere (pareek, 2013). for example, li et al. (2018) discovered that the chinese jujube originates from sour jujube (z. acidojujuba mill.) and is an economically very important genus (zhang et al., 2015). �ey also concluded that most jujube cultivars have a certain correlation with their origin, and there are obvious gene exchanges between sour jujube, chinese and indian jujube cultivars. its pulp is eaten mostly fresh but may be dried or processed into confectionary recipes in bread, cakes, compotes and candy (krška, mishra, 2008; mishra, krška, 2017). 195 c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine wang and hu (2016) reported that jujube is an important fruit in china and has attracted signi�cant interest because of its common consumption as food, a food additive, as a �avouring and in tcm (mahajan, chopda, 2009). in china, a wine made from jujubes, called “hong zao jiu” is also produced. in a chinese book on herbal medicine, huangdi neijing (475-221 bc), jujube was described as one of the �ve most valuable fruits in china. in shennong bencao jing (300 bc-200 ad), an earlier book recording medicinal herbs, jujube was considered to be one of the superior herbal medicine that prolonged life-span by nourishing blood, improving quality of sleep, and regulating the digestive system (chen et al., 2017). according to gupta (2004), jujube, along with date palms and grapes, started to be domesticated on the indian subcontinent around the year 9,000 bc, together with wheat and barley, which were cultivated from the very beginning of agriculture. �e aim of this article was to review the most important information about the chinese jujube’s (1) botanical characteristics, (2) cultivation in china and around the world, (3) variety of cultivars, (4) crop pests and (5) importance in traditional cuisine and chinese medicine. fig. 1. chinese jujube’s dry fruit – drupe (ziziphus jujuba mill.) (photo. m.h. shahrajabian) m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 196 jujube classification and species characteristic ziziphus jujuba, commonly known as jujube, red date and chinese date, is a species that has many scienti�c names. �ese names are used as synonyms – a list of synonyms and a detailed botanical description (�ower, fruits and trunk, branches, culture and other characteristics) of this species can be found in the appendix (tab. 1, points 1–2 – appendix 1). as a species found on �ve continents, it is characterised by a very large variety of cultivars (tab. 1, points 3–7 – appendix 1), which proves it is a plant that has been used by humans for centuries. similarly, z. mauritiana is characterised by a very diverse local (customary) nomenclature, which proves that it is a plant commonly known for cultivation in various regions, especially in asia (tab. 2, points 1–2 – appendix 1). from an economic point of view, fruits are the most important raw material derived from species of the ziziphus genus. �ere are three phases of fruit maturation of the chinese jujube, which are based on colour, �esh �rmness and composition, such as starch, sugar, acid and water. �e phases are 1) white mature: the fruit is near full size and shape – the skin of the fruit is thin and changes from green to greenish white in colour, and the �esh becomes white and loose with less juice and sugar and more starch; 2) crisp mature: the fruit skin is half to fully red in colour, becomes thicker, harder and easily separated from the �esh which becomes crisp, juicy and sweet, containing more sugar and acid; and 3) fully mature: sugar content of the �esh increases rapidly and water content begins to decrease, while the �esh near the stone and fruit stalk becomes yellow and so� and the skin changes to a dark red and the fruit becomes wrinkled. �ese three stages of ripening determine the speci�c use and properties of the fruits of not only chinese jujube but all species of this genus. jujubes adapt to a wide range of elevation, from 0 to 2,000 m, between latitude 18o14ʹ to 45o and longitude 76° to 124° and in soil ph 5.5 to 8.5. jujubes may tolerate −30°c in the winter and 49°c in the summer (miri, 2018). eight major phenological stages have been observed for chinese jujube: bud growth; leaf formation; leaf growth; shoot growth; formation of in�orescence; �owering; fruit formation, growth and development; fruit maturation and winter dormancy (sapkota et al., 2020). jujube trees adapt to drought conditions and produce su�cient yield under severe conditions (sülüşoğlu et al., 2014). �e tree can survive with an annual rainfall of only 200 mm but, for better fruit set and fruit quality, more precipitation or supplemental irrigation is needed. shahin et al. (2011) found that increasing water stress signi�cantly reduced rate of stem length, number of leaves per plant, leaf area and fresh and dry weight of di�erent plant organs. �ey also concluded that the date genotype of the chinese jujube had the highest statistically signi�cant performance in increasing vegetative growth compared with other genotypes, when under water stress. additionally, 197 this species is autogamic, so only one tree is needed for fruit-set. additionally, jujubes are also tolerant of high salinity, alkalinity and root exposure from erosion or roots being deeply covered by blowing sand, but chinese jujube should not be planted in the shade of other trees (yao, 2013). jujube, as a nutritious fruit, is important especially for people in low-income groups because of its relatively inexpensive cost for value, compared to other available fruits. it is also a relatively less perishable fruit and its cultivation by large-scale famers can be economically bene�cial and help to maintain national economic levels (islam et al., 2016). jujube cultivation in china chinese jujube is considered an ideal economic crop for arid and semiarid areas of the temperate zone where common fruit trees do not grow well (liu, 2010). chinese jujube has become a leading fruit tree in the northern part of china with a total production of 4,250,000 t in 2009 with a planting area of 1,500,000 ha (ping et al., 2012). china produces and exports more jujube than any other country in the world, and it has been estimated that more than 90 percent of the world’s products made from jujube are provided by china (zhang et al., 2012). according to the xinjiang statistical yearbook 2015, there was a total planting area of 483,628 ha and yearly production of more than 2.5 million t (chen et al., 2017). in china, the jujube tree is popular as a woody species that reduces soil erosion while producing an economic crop in response to the government policy of changing from small grain production to conservation forestry on the loess plateau (wu et al., 2013). �e henan province used to be one of the most important jujube production regions in china (wang, hu, 2016) – fig. 2. �e planting area for jujube in shanxi province exceeds 100 acres, which are mostly planted in the mountains where dry farming is the primary mode of cultivation due the severe scarcity of water for irrigation (chen et al., 2014). guo et al. (2017) found that wild jujube (ziziphus acidojujuba mill.) is highly tolerant to alkaline, saline and drought stress. liu et al. (2016) also recommended that, in order to fully utilise the limited arable land and achieve fast economical returns, jujube/wheat intercropping be employed in xinjiang uygur autonomous region in china by planting wheat between the lines of jujube tree when jujube trees are young. because of its positive ecological, economic and social bene�ts, the jujube/wheat intercropping system became one of the most widely applied agro-forestry systems in xinjiang, northwest china. jujube production has developed at a rapid and signi�cant pace over the past 39 years in china. its annual production has increased more than 15 times from 1980 to 2015 from 376,000 t to over 6,000,000 t, on a fresh weight basis. ninety percent of c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 198 this production is concentrated in six northern provinces, namely hebei, xinjiang, shandong, shanxi, shaanxi and henan (fig. 2). moreover, the current total growing area in china is approximately 2 million hectares. xinjiang provinces in western china have seen a massive increase in production in the last 10 years with both the introduction of new cultivars and favourable growing conditions for producing premium quality fruit (lin et al., 2013). �e 10 leading cultivars of chinese jujube in china and other imported cultivars are shown in the appendix (tab. 1, points 3–4 – appendix 1). jujube cultivation in different parts of the world china is undoubtedly one of the largest producers of ziziphus jujuba in the world (chen et al., 2017; višnjevec et al., 2019; shahrajabian et al., 2019c). over 40% of this plant’s crop is grown in china (fig. 3). however, other countries have shown a growing interest in producing this species, such as egypt, iran, saudi arabia, algeria and iraq, primarily due to the health-promoting properties of the fruit and ease of growth (tab. 1, point 5 – appendix 1). in lebanon, jordan and other middle eastern countries, the fruit is eaten as snacks or alongside a dessert a�er a meal. in persian cuisine, the dried drupes are known as annab, while in azerbaijan, it is commonly eaten as a snack and known as innab. �e fig. 2. �e major cultivation areas of jujube (ziziphus jujuba mill.) in china (chen et al. 2017 – changed) 199 turks use a similar name, hunnap. z. jujuba grows in northern pakistan and is known as innab, commonly used in the tibb unani system of medicine. in india, the fruits are dried in the sun and the hard nuts are removed, then it is pounded with tamarind, red chilies, salt and jiggery. in both china and korea, a sweetened tea syrup containing jujube fruits is available in glass jars, and canned jujube tea or jujube tea in the form of teabags is also available. jujube has been introduced worldwide because of its high nutritional and economic values (gao et al., 2013; zhang, li, 2018). ghazaeian (2015) concluded that climate is very important for the qualitative and quantitative characteristics of jujube. a demand for fresh jujubes continues to outweigh supply on the local markets (johnstone, shan, 2016). western australia is close to south-east asia, thus its counter-seasonal production versus the northern hemisphere may provide an opportunity to market australian grown fresh chinese jujube for the increasing o�-season demand in these countries. �e jujube industry in western australia has an exciting future, as it grows well in many areas of the country. �ere are also many cultivars, which con�rms the signi�cant interest of local growers (tab. 1, point 6 – appendix 1) ciocarlan (2000) reported that the chinese jujube could be found in a semi-spontaneous status in the dobrogea region, which is located between the danube and the black sea in romania; there are several places where the chinese jujube exists in naturalised populations. in romania, the tree was brought “via the silk road” 2,000 years ago (stănică, vasile, 2008; stănică, 2009). it is cultivated in europe, but its primarifig. 3. world production of jujube (ziziphus jujuba mill.); additionally about 6% other countries together – azerbaijan, usa, korea, australia and others (data source: johnstone, 2017) c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 200 ly researches have been conducted in romania, italy and macedonia (cossio, bassi, 2013; johnstone, 2014; markovski, velkoska-markovska, 2015; višnjevec et al., 2019). in north america, jujubes were found to be cultivated mainly in the southwest, southern and south-eastern states – from north and south carolina to florida and from georgia to california, as well as in pennsylvania (yao et al., 2015). �e plantation of jujube is becoming increasingly popular in this part of the world, as evidenced by its numerous cultivars (tab. 1, point 7 – appendix 1). jujube pests and other problems with fruit production balikai et al. (2013) described almost 130 pest species that were recorded on ziziphus crops in india and speci�ed that 177 species of insect (some of them are listed in table 3) and non-insect were recorded to be jujube pests around the world. in india, in an ipm governmental meeting in 2015, 10 pests were cited as pests of national signi�cance: fruit �ies carpomyia vesuviana costa (diptera: tephritidae), fruit borers meridarchis scyrodes meyr (lepidoptera: carposinidae), green slug caterpillars �osea sp. (lepidoptera: limacodidae), grey caterpillars �iacidas postica walker (lepidoptera: noctuidae), mites larvacarus transitans ewing (tetranychoidea: tenuipalpidae), ber beetles adoretus pallens banchard (coleoptera: scarabaeoidae), grape mealybugs maconellicoccus hirsutus green (hemiptera: pseudococcidae), ber mealybugs perissopneumon tamarindus green (hemiptera: pseudococcidae), thrips scirtothrips dorsalis hood (�ysanoptera: �ripidae) and termites odontotermes obesus rambur (isoptera: termatidae). tab. 3. taxonomical position and nature of various insects observed on ecosystems with jujube (nizamani et al., 2015) no. common name technical name family order major pests 1. jujube lead roller ancylis sativa liu tortricidae lepidoptera 2. jujube hairy caterpillar euproctis fraternal moore lymentriidae lepidoptera 3. jujube beetle adretus pallens blanchard scarabaeidae coleoptera minor pests 4. cutworm agrotis biconica kollar notuidae lepidoptera 5. thrip scirtothrips dorsalis hood rambutanae thysanoptera 6. jujube looper achaea janata linn. notuidae lepidoptera 7. jassid amrasca biguttula biguttula ishida cicadilidae 8. aphid aphis gossipie glover aphididae hemiptera 9. jujube gray weevil myllocerus discolor boheman curculionidae coleoptera 201 sporadic pest 10. dusky cotton bug oxycareous hyalinipennis costa lygaediae hemiptera 11. green grasshopper dichromorpha viridis scudder acrididae orthoptera 12. jujube butter �y tarucus balkanicus freyer lycaenidae lepidoptera 13. moth orgyia postica wlk lymantriidae lepidoptera in europe, the following jujube pests were noted: carpomyia vesuviana, c. incomplete becker, bactrocera zonata saunders, ceratitis capitata wiedemann and hispa sp., grammadera clara brunner von wattenwyl (balikai et al., 2013). in romania, the most signi�cant pests of z. jujuba are carpomya vesuviana and c. incomplete – dipterous that lay their eggs in july under the fruit epiderma, and carposina sasakii mats. – a lepidopterous which, in china, destroys 15–20% of fruits; with other minor pests being ceratitis capitata, cydia molesta busck and polycrosis botrana schu�ermuller (stănică, 1997). in �eld experiments, while monitoring pest incidence, several species were identi�ed that may become a threat: halymorpha halys stal (heteroptera: pentatomidae), metcalfa pruinosa say (homoptera: flatidae), ceratitis capitata and nezara viridula l. (heteroptera: pentatomidae); in the same experiment scientists reported other potential polyphagous pests, like weevils, fruit borers and moths that insigni�cantly damaged the fruits (ciceoi et al., 2017). hua et al. (2015) found that jujube fruit cracking has become a major concern in jujube production and it can a�ect fruit quality and yield and crop productivity, resulting in signi�cant economic loss. zeraatgar et al. (2018) concluded that salicylic acid and calcium nitrate play an important role in maintaining and extending post-harvest quality of fresh jujube fruit. �ese substances could cause at least a 10-day delay in the reduction of some the fruit’s bene�cial attributes. disease and cracking resistant cultivars are shown in table 1, point 8 – appendix 1. jujube nutritional composition and chemical constituents kader et al. (1982) stated that, relative to most other fresh fruits, chinese jujubes are lower in water content and titratable acidity and higher in total sugars (mostly reducing sugars) and phenolics. �e major minerals in jujube are phosphorus, potassium, calcium and manganese. in addition, there are also high amounts of sodium, zinc, copper and iron. chinese jujubes are very rich in ascorbic acid (vitamin c) content which increase with maturation to 559 mg/100 g fresh weight. �is was also con�rmed in other studies (e.g. pareek, 2013). �e fruits are nutritious, being high in �avonoids and vitamins b1 and b2, thus it can be considered a so-called functional food – having nutritional as well as medicinal uses (huang et al., 2008). c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 202 rahman et al. (2018) noted that 51.99–71.75% of the chinese jujube is edible, with the edible part containing 82.35–89.63% carbohydrates, 4.43–6.01% protein, 0.48–0.63% lipids, 2.80–4.80% polysaccharides, 45.64–88.97 mg/100 g ascorbic acid, 132.16–196.58 mg/100 g phenolics and 101.17–132.04 mg/100 g �avonoids in its dry matter. �e jujube fruit is rich in mineral content and �bre, a good source of food for direct consumption and it may be a useful food additive when dried (hendek ertop, atasoy, 2018; višnjevec et al., 2019). jujube fruit, especially in dried and powder form, can be valorised as a fortifying and hydrocolloid ingredient, due to its high carbohydrate content. however, the content of di�erent substances may vary depending on the breeding cultivar. li et al. (2007) provided the proximate composition of �ve cultivars of chinese jujube (‘jinsixiaozao’, ‘yazao’, ‘jianzao’, ‘junzao’, ‘sanbianhong’). in their experiment, total phenols, minerals and vitamins were determined for the fruits of these cultivars; signi�cant variation was found for content of water (17.38–22.52%), carbohydrate (80.86–85.63%), proteins (4.75%–6.86%), lipids (0.37–1.02%), soluble �bre (0.57–2.79%), insoluble �bre (5.24–7.18%), reducing sugar (57.61–77.93%) and ash (2.26–3.01%). glucose and fructose were identi�ed as major soluble sugars in all �ve cultivars, while rhamnose, sorbitol and sucrose were also present in lower amounts. chen et al. (2018) reported the ‘junzao’ cultivar contained relatively low levels of total dietary �bre, protein, total sugar and total titratable acids. �e ‘huizao’ cultivar possessed an intermediate level sugar-to-acid ratio and intermediate levels of ascorbic acid. �e ‘dazao’ cultivar showed high levels of total dietary �bre, protein, sugar and total acids. in their experiment, principal components analysis indicated that the parameters that di�erentiated these jujube cultivars appeared to be the total dietary �bre, protein, total sugar, fructose, glucose, sucrose and total titratable acids. chen et al. (2017) noted that jujube exerts neuroprotective activity, including protecting neuronal cells against neurotoxin stress, stimulating neuronal di�erentiation, increasing expression of neurotrophic factors and promoting memory and learning. flavonoid, cyclic adenosine monophosphate (camp) and jujuboside may potentially be the bioactive ingredients accounting for the aforesaid biological activities. �ese �ndings imply that jujube is a potential candidate for use in the development of health supplements for prevention and/or treatment of neurological diseases. seven chemical markers found in jujube, including kaempferol 3-o-rutinoside, quercetin 3-o-rutinoside, (-)-catechin, (-)-epicatechin, swertish, spinosin and camp, have also been associated with neuroprotection (fig. 4 – appendix 2). �e structures of the triterpenic acids, nucleosides and nucleobases, as well as saccharides, in jujube fruits are presented in �gure 5 – appendix 2. �e appendix also contains a list of all of the most important chemical ingredients contained in jujube fruit (tab. 1, point 9 – appendix 1). 203 traditional medicinal uses and potential health benefits of jujube jujube (zisiphus jujuba) has been used as a tcm plant for many years for its various and numerous health bene�ts, including anti-in�ammatory (yu et al., 2012), anti-cancer (plastina et al., 2012), gastro-intestinal protective (huang et al., 2008), anti-oxidant (cheng et al., 2012), anti-insomnia and neuro-protective (yoo et al., 2010) properties. jujube fruits and seeds are still used in chinese and korean traditional medicine and are believed to alleviate stress, according to the modern medicine industry (mill goetz, 2009). among other things, jujube causes a decrease in the blood levels of glucose and lipids and causes a signi�cant decline in triglyceride, ldl and cholesterol levels (hemmati et al., 2015). tahergorabi et al. (2015) reported that different parts of the jujube have been used for curing di�erent kinds of illness such as diabetes, diarrhoea, skin infections, liver complaints, urinary disorders, obesity, fever, pharyngitis, bronchitis, anaemia, cancer, insomnia and, of course, for blood puri�cation and toni�cation of the gastro-intestinal tract. �e jujube leaf, which is the main by-product of the jujube, has been used in tcm for thousands of year, to improve sleep, to nourish the heart and soothe the nerves and to reduce haemorrhaging and diarrhoea (zhang et al., 2014; damiano et al., 2017). modern studies found that the jujube leaves were rich in bioactive components and have various physiological and pharmacological functions (mahajan, chopda, 2009; damiano et al., 2017). �e aqueous ethanol extract of the jujube leaf is used as an energetic constituent for hepatosis and wound healing in animal trials (hovanet et al., 2016; bai et al., 2017). jujube-leaf green tea extracts can inhibit human hepatocellular carcinoma cells by activating amp-activated protein kinase (ampk) (huang et al., 2009; liu et al., 2017); additionally, an unidenti�ed β-d-glucosidase inhibitor has been found in jujube leaf extract (jo et al., 2016). based on iranian traditional medicine, local healers have used powders made of the stem bark and leaves of jujube to cure wounds on di�erent parts of the body, including oral wounds such as aphthous (hamedi et al., 2016). roots and bark of jujube have been used to treat dysentery; the bark has been reported to cure boils and found helpful for the treatment of diarrhoea (mahajan, chopda, 2009) and jujube seeds were used to cure eye diseases and help with leucorrhoea. currently, fruits of jujube are widely used in iranian folk medicine as an antitussive, laxative agent and blood pressure reducer (hamedi et al., 2016). in persian traditional medicine, jujube fruit is also used in combination with other herbal medicines to treat colds, �u and coughing. jujube fruit contains �avonoids, vitamins, amino acids, organic acids, polysaccharides, and microelements (li et al., 2007, 2016), which have been found useful in spleen diseases and supporting body systems (shen et al., 2009). hamedi et al. (2016) found that the fruit of the jujube is digestible; is tonic; works like an aphrodisiac; is c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 204 laxative; removes biliousness, burning sensations, thirst and vomiting and is also used in curing tuberculosis and blood diseases. beavo and brunton (2002) found that jujube fruits contain a certain amount of camp, which has a positive e�ect on the heart muscle, nutritional myocardium and diastolic blood vessels, as well as having anti-arrhythmia and anti-platelet aggregation e�ects. recent phytochemical research on jujube fruits has revealed anti-cancer, anti-in�ammatory, anti-obesity, immuno-stimulating, anti-oxidant, hepato-protective and gastro-intestinal protective properties, as well as inhibition of foam cell formation in macrophages (abedini et al., 2016; keerthi et al., 2016; rajopadhye, upadhye, 2016; alhassan et al., 2019). cosmulescu et al. (2017) mentioned that jujube fruits are rich in bioactive compounds (�avonoids, carotenoids and anthocyanin) and can complement a healthy human diet. shi et al. (2018) concluded that changes in jujube fruit colour are associated with changes in antioxidant activity. flavonoids and polysaccharides could contribute to the antioxidative e�ect of jujube (choi et al., 2011). hoshyar et al. (2015) indicated that, among other things, z. jujuba ameliorates the adverse e�ects of nmu carcinogenesis and could be useful, for example, in treating mammary tumours in humans. taechakulwanijya et al. (2013) found that jujube seed extracts were not toxic to control vero cell lines but induced cell death in jurkat leukemia t cell lines, making them promising candidates for more elaborate studies of their anticancer mechanisms. �e triterpene acids from jujube are considered active ingredients, having anti-in�ammatory and anti-cancer properties (tahergorabi et al., 2015). betulinic acid and jujuboside b might be the active components underlying bene�cial e�ects on the cardiovascular system (seo et al., 2013). vafaei and abdollahzadeh (2015) reported that jujube fruit extract could accelerate burn wound healing. jujube polysaccharides are thought to be the primary active ingredient contributing to jujube’s immune-modulating and hemato-poietic functions (zhao et al., 2008). �e jujube polysaccharides, composed of glucose (23%), xylose (31.3%), mannose (12.9%) and fructose (21.6%), possess antioxidant e�ects that may have contributed to observed positive e�ects (wang, 2011). gao et al. (2013) mentioned that jujube polysaccharides are reported to be useful in ameliorating intestinal oxidative injury resulting from ischemia and reperfusion. additionally, it was found that taking a combination of jujube fruit and low doses of routine pharmaceutical drugs can improve and cure ulcerative colitis disease (gheibi et al., 2018). generally, the vitamin and mineral content of jujube fruit helps to support cardiovascular health and enhance metabolism. �e most important pharmacological properties of jujube are anti-diabetic e�ects, hypnotic-sedative and anxiolytic e�ects, neuroprotective activity, inhibition of sweetness, anti-cancer activity, antimicrobial 205 activity, anti-ulcer activity, anti-in�ammatory and anti-spastic e�ects, anti-allergic activity, permeability-enhancing activity, cognitive activity, anti-fertility/contraceptive properties, hypotensive and anti-nephritic e�ects, cardiovascular activity, immunostimulant e�ects, anti-oxidant e�ects and wound healing activity (anbarasi, brindha, 2013; pareek, 2013). additionally, varghese and patil (2005) revealed that jujube leaves exert an insecticide e�ect against helicoverpa armigera hübner and triboliun confusum jacquelin du val. it has been shown that jujube leaf insecticide activity occurs through inhibition of digestive and mitochondrial enzymes which lead to retarded larval growth. conclusion jujube or chinese dactyl (zizyphus jujuba) is a popular cultivated plant in many parts of the world, especially in china and iran. its fruit is an edible oval drupe, which has a wide range of applications, especially in food and medicine. jujube trees have the ability to adapt to various biotic and abiotic stresses such as salinity and drought. chinese jujube is grown in temperate regions, while the similar indian jujube (z. mauritiana) is cultivated in hot arid regions of india. china is still the largest producer of jujube fruit in the world, although recently there has been a notable increase in interest in jujube fruit production in other parts of the world. jujube fruit contains 23 types of amino acids that are not found in most other kinds of fruits. it also contains vitamin c, ribo�avin and thiamine. �e major components of the jujube fruit are used, in combination with other medicinal plants and fruits, in tcm. �e jujube leaf, which is the main by-product, has also been used in tcm for thousands of years. jujube has numerous important pharmacological activities and is considered a valuable source of nutraceuticals. it is a plant with great healing potential in diseases of civilisation, including various types of cancer, and is an important source of nutrients, easily available in the poorer regions of the world. con�ict of interests �e authors declare that there is no con�ict of interests regarding the publication of this paper. references abedini, m.r., erfanian, n., nazem, h., jamali, s., hoshyar, r. 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(2008). characterization of water soluble polysaccharides from organs of chinese jujube (ziziphus jujuba mill. cv. dongzao). european food research and technology, 226, 985–989. https://doi.org/10.1007/s00217-007-0620-1 c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 212 appendix 1 tab. 1. detailed characteristics of chinese jujube (ziziphus jujuba mill.) 1) scienti�c classi�cation and synonyms (liu, 2010) kingdom: plantae; unranked: angiosperms; unranked: eudicots; unranked: rosids; order: rosales; family: rhamnaceae; genus: ziziphus; species: z. jujuba mill. synonyms: paliurus mairei h. lev; rhamnus jujuba l., r. soporifera lour., r. zizyphus l. ziziphus jujuba (l.) lam., z. jujube (l.) gaertn, z. mairei (h. lev.) browicz & lauener, z. nitida roxb., z. orthacantha dc., z. poiretii g.don nom. illeg., z. rotundata dc., z. sativa gaertn., z. soporifera (lour.) stokes, z. spinosa (bunge) hu ex f.h. chen, z. tomentosa poir., z. trinervia roth nom illeg., z. vulgaris var. inermis bunge, z. vulgaris var. spinosa bunge, z. zizyphus (l.) h. karst., z. zizyphus (l.) meikle., z. jujubum st.-lag. 2) description of all plant: �ower, fruit and trunk branches, culture and other characteristics (liu, 2010; markovski, velkoska-markovska, 2015; ivanišová et al., 2017) height: 457 to 1070 cm; spread: 305 to 915 cm; crown uniformity: irregular outline or silhouette; crown shape: oval, round; crown density: open; growth rate: medium; texture: �ne. foliage – leaf arrangement: alternate; leaf type: simple; leaf margin: crenate, serrulate; leaf shape: lanceolate, ovate; leaf venation: bowed; leaf type and persistence: deciduous; leaf blade length: 5 to 10.5 cm, less than 5.5 cm; leaf colour: green; fall colour: yellow; fall characteristic: showy. flower – colour: yellow; flower characteristics: inconspicuous and not showy; spring �ower. fruit – shape: oval, round; length: 2.5 to 12.5 cm; fruit covering: �eshy; colour: black, red; fruit characteristics: it attracts squirrels and other mammals; fruit, twigs, or foliar cause signi�cant litter, showy. trunk and branches – grow mostly upright and will not droop, not particularly showy, should be grown with a single leader, thorns are present on the trunk or branches. breakage: resistant; current year twig colour: brown; current year twig thickness: medium. other characteristics – roots: surface roots are usually not a problem. culture – light requirement: tree grows in part shade/part sun; and also grows in full sun; soil tolerances: clay, loam, sand, slightly alkaline, acidic, well-drained; drought tolerance: high. 3) �e 10 leading cultivars in china (liu, 2010) cultivar name use main producing area ‘dongzao’ fresh hebei, shandong ‘linyilizao’ fresh shanxi ‘pozao’ dry hebei ‘changhongzao’ dry shandong ‘yuanlingzao’ dry shandong, hebei ‘muzao’ dry shanxi, shannxi ‘bianhesuan’ dry henan ‘jinsxiaozao’ dry, fresh hebei, shandong ‘huizao’ multipurpose henan, xinjiang ‘zanhuangdazao’ multipurpose hebei, northwest 4) potential jujube cultivars for importation from china (liu, 2010) cultivar name province overview ‘baodeyouzao’ (oil jujube) shanxi, shaanxi, yellow river • high yielding, early maturing, • average fruit weight 11.6g, 3.5 × 2.1cm, • deep red skin, dense, juicy �esh with sweet and sour �avour, • low fruit cracking, low fruit drop; 213 ‘dongzao’ (winter jujube, winter date, lubei) shandon, hebei • large tree, high yield, late fruiting, • large fruit (11.5g ave, max 35g, 2.7-3.4cm × 2.6 × 3.4cm), • �in peel, �esh crisp, white, juicy, sweet and rich, • high yield and large, late fruit; ‘guantanzao’ shanxi • medium sized tree, weak vigour, productive and stable yield, mid-late maturity, • med-large fruit, oblong shaped, (3.5 × 2.5cm), ave fruit weight 10g (max 12g), • �ick peel, sweet �esh, less juice, • low fruit cracking, high edible rate and dry quality, adaptability to drought and disease; ‘huizao’ (ash date, zinzhuang) xinzheng city, henan province • medium sized tree, oval shaped fruit (3.2 × 2.3cm), average fruit weight 12.3g, max 13.3g, • crisp, sweet and juicy fruit, • fruit is susceptible to cracking/splitting. suitable for dried fresh and processed or candied dates; ‘jinsixiaozao’ hebei • small fruit, average weight of 5g, skin is bright red, �esh is white, dense and crisp texture, medium juiciness, sweet and slightly sour, • mid-late maturing fruit, • good for storage and transportation, good quality for fresh or dried; ‘kongfusucuizao’ qufu, shandong • tree is talk and strong, • fruit is long oval shaped, average fruit weight 12g, max 20 g, • crisp, juicy, sweet fruit, slightly sour, • early maturing variety, drought tolerant and resistant to rust and anthracnose, fruit shrink disease; ‘linyilizao’ (li) shanxi, yuncheng, linyi • small tree, dense foliage, large fruit, diameter 4.2 × 4.0cm, average fruit weight 30g, max 40g, • �e fruit is thin and ochre-red with white �esh that is crisp, juicy and sweet, • late maturing fruit, • �e fruit is big and beautiful with a high edible rate, adaptable tree; ‘ningyangliuyuexian’ (june fresh) • smaller tree, dense branches, no thorns, • fruit long tube shape, average fruit weight 13.6g, • peel medium-thick, light purple, �esh green and white. fine quality fruit that is crisp, sweet and slightly acidic, • resistant to cracking, good quality fresh jujube however adaptability is poor and the tree requires deep and fertile soil conditions. early maturing; ‘zanhuangdazao’ (gold silk jujube) hebei, zanhuang • tree is tall and upright, fruit oblong or obovate, diameter 4.1 × 3.1cm, average fruit weight 17.3g, max 29g. peel is thick and dark reddish brown. flesh nearly white, dense, medium juice, sweet and sour, • only natural triploid, fruit suitable for dry, candied and fresh. versatile, adaptable, resistant to drought, suitable for warm climate; ‘zaocuiwang’ hebei • strong tree vigour and easy to manage, early maturing, • high yield, 2-year-old gra�ed plants produce about 5kg, • strong resistance to drought, waterlogging and salinity. very lows cracking and rust; c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 214 ‘zaofengzui’ shandong • shorter growth period of 85d. fruit nearly round, diameter 3.0 × 2.9 cm, average fruit weight 12.1 g, max 16.3 g, fruit size uniform, • peel think and crisp, �eshy texture and delicate, juicy, • wide adaptability and strong resistance to anthracnose, ulcer disease, black spot disease rate of less than 3%, rot disease rate of 6%; ‘zaoshulizao’ hebei • larger oval or pear-shaped fruit, diameter 3.5 × 3.2 cm, average fruit weight 17.8 g, max 27 g, • peel is thin, reddish brown, �esh thick, green white or milky white, crisp, juicy, sweet, slightly sour, • high and stable yield, no cracking and good quality fruit, • early results of plastic greenhouse cultivation show average yields of 1.7 kg in the second year, 3.9 kg in the third year and 5.0 kg in the fourth year. 5) world distribution (liu, 2010; johnstone, 2017) region country asia afghanistan, armenia, azerbaijan, bengla, burma, china, cyprus, india, iraq, iran, israel, japan, kyrgyzstan, lebanon, malaysia, mongolia, pakistan, palestine, south korea, syria, �ailand, turkey, turkmenistan, uzbekistan europe bulgaria, england, france, germany, greece, italy, czech, macedonia, portugal, romania, russia, slovenia, spain, ukraine, yugoslavia africa egypt, tanzania, tunisia north america canada, usa oceania australia, new zealand 6) cultivars in australia (johnstone, 2014) cultivar name use characteristics ‘li’ fresh large, mid-season ripening ‘chico’ fresh fruit is round but �attened, excellent fresh fruit with a apple-like sweet-acid taste, good either fresh or dried, mid-late season ripening ‘shanxi-li’ fresh very large fruit ‘ga866’ fresh sweet fruit with a higher sugar level, large and elongated fruit ‘redlands’ fresh very large, sweet, round fruit and mid-season ripening ‘silverhill’ fresh med-large, elongated fruit, very sweet, very late season ripening, good for humid areas ‘sherwood’ fresh, dried fruit is very dense and excellent quality, late season ‘honeyjar’ fresh, dried small but very unique, sweet tasting, crunchy, juicy fruit, very thinskinned, early ripening, considered a fresh eating jujube but also good dried ‘sihong’ fresh, dried, processed excellent, large, round fruit, mid-season ripening, one of the best fresh varieties, also good processed or dried ‘suimen’/’shuimen’ fresh, dried, processed fruit is elongated, fairly good fresh but better dried or processed, mid-season ripening ‘lang’ dried, processed large, pear-shaped fruit, mid-late ripening, good variety for dried fruit, very good for processing, not suited for humid areas, �e tree is very upright and virtually thorn-less ‘don-polenski’ dried similar to lang with a better, crisper �avour ‘�ornless’ dried fruit similar to lang ‘admiral wilkes’ dried, processed elongated and very late ripening, best processed or dried 215 7) current cultivars in the usa (yao, 2013) sources cultivar names f.n. meyer ,s collection ‘li’, ‘lanf’, ‘shui men’ (shuimen, sui men or sui), ‘mu/mu shing hong’, ‘so’, ‘yu’ chico breeding program ‘ga866’, ‘gi-7-62/chico’, ‘gi-1183’, ‘thornless’ cultivars from across the united states alabama ‘silverhil’l, ‘ed hegard’, ‘swobada’ california ‘don polenski’, ‘jin’, ‘porterville’, ‘redland #4’, ‘sugarcane’ georgia ‘fitzgerald’, ‘leon burk’, ‘prine’ florida ‘geant’ kansas ‘topeka’ kentucky ‘priest’ louisiana ‘abbeville’, ‘sherwood’ pennsylvania ‘tsao’ tennessee ‘r3t1’ texas ‘texas tart’ washington, dc ‘admiral wilkes’ imports after the 1990s r. meyer ,s import ‘shanxi li’, ‘honeyjar’, ‘globe’, ‘september late’, ‘ant admire’, ‘sihong’ j. gilbert ,s import ‘qiyuexian’/’autumn beauty’, ‘mango dong zho’/’winter delighttm’, ‘black seatm’, ‘cocotm’, etc. nmsu alcalde ,s import ‘jinsi #2’, ‘jinsi #3’, ‘pitless’, ‘junzao’, ‘teapot’, etc., with total of 30 cultivars. 8) disease resistant cultivars (liu et al., 2013) fruit cracking resistant: ‘chuanlingzao’, ‘chenwudongzao’, ‘chahuzao’, ‘mopanzao’, ‘jianzao’, ‘guantanzao’, ‘xuezao’, ‘huluchanghong’, ‘hebeilongxuzao’, ‘baodeyouzao’; fruit shrink resistant: ‘chuanlingzao’, ‘chengwudongzao’, ‘changjixinzao’, ‘suyuanling’, ‘ningyangliuyuexian’, ‘kongfusucuizao’, ‘pingguozao’. 9) �e major chemical components of jujube (liu, 2010; gao, et al., 2013; chen et al., 2017; cosmulescu et al., 2017; shahrajabian et al., 2019c) a) carbohydrate such as: glucose, fructose, sucrose, rhamnose, and sorbitol; b) vitamin, especially vitamin c and vitamin b complex; c) fatty acids such as: oleic, linoleic (omega-6), palmitic and palmitoleic acids; d) alkaloids, the cyclic peptide alkaloids, mauritine a, mucronine d, amphibine h, nummularine a-b, sativanine a-h, and –k, frangulanine, jubanine a-c, scutianine c-d and ziziphine a (were isolated from stem bark of jujube), the alkaloids coclaurine, isoboldine, norisoboldine, asimilobine, iusiphine and iusirine (were isolated from leaves), e) glycosides: �avonoid glycosides/spinosins, glycosides/saponins; f ) terpenoids, such as colubrinic acid, alphitolic acid, 3-ocis-p-coumaroylalphitolic acid, 3-o-transpcoumaroylalphitolic acid, 3-o-cis-p-coumaroylmaslinic acid, 3-o-trans-pcoumaroylmaslinic acid, oleanolic acid, betulonic acid, oleanonic acid, zizyberenalic acid, betulinic acid, zizyberanal acid, zizyberanone, zizyberanalic acid and ursolic acid. c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 216 tab. 2. selected characteristics of indian jujube (ziziphus mauritiana lamk.) (shahrajabian et al., 2019c) 1) local names of indian jujube (morton, 1987; ved et al., 2016) amharid: kurkura; arabic: nabak (fruit), sidr; bengali: ber, boroi, kool, ber, boroi; burmese: zee-pen, zizidaw, eng-si; english: dunks, jujube, indian cherry, indian jujube, indian plum, geb, ber, common jujube, chinese date, chinese apple, bear tree, desert apple; filipino: manzanita; french: jujube, jujubier, jujubier commun, le jujubier, le jujubier sauvage, liane croc-chien; german: indischer jujubenstrauch; gujarati: bordi; hindi: baer, badari, elladu, ber, khati, jelachi; indonesian: widara, dara, bidara; khmer: putrea; lao: sino-tibetan; malay: bidara, jujub, epal siam; mandinka: tomborongo, tomboron moussana, toboro; nepali: bayer; sanskrit: kuvala, karkandhu, badara, ajapriya, madhuraphala; somali: geb, gub; spanish: yuuba, ponsere, perita haitiana; swahili: mkunazi; tamil: elandai, yellande; �ai: ma thong, ma tan, phutsan; tigrigna: geva; trade name: jujube; urdu: ber; vietnamese: tao nhuc, tao. 2) list and origin of 37 cultivars in india – cultivar name and state (singh et al., 2017) 1.‘bc-1’ – rajasthan, 2.‘umran’ – maharashtra, 3. ‘seb’ – rajasthan, 4. ‘illaichi’ – punjab, 5. ‘tikadi’ – rajasthan, 6. ‘gola’ – haryana, 7.‘reshmi’ – haryana, 8.‘cazri-gola’ – rajasthan, 9.‘banarasi karaka’ – uttar pradesh, 10.‘aliganj’ – uttar pradesh, 11.‘katha’ – rajasthan, 12.‘z-g-3’ – punjab, 13.‘mundia’ – haryana, 14.‘bagwadi’ – punjab, 15.‘maharwali’ – rajasthan, 16.‘banarasi pebandi’ – uttar pradesh, 17.‘s x k hybrid’ – rajasthan, 18.‘sanaur-5’ – haryana, 19.‘�oronless’ – punjab, 20.‘kali’ – rajasthan, 21.‘jogia’ – rajasthan, 22.‘kaithli’ – haryana, 23.‘chhuhara’ – maharashtra, 24.‘dandan’ – punjab, 25.‘gola gurgaon’ – haryana, 26.‘chonchal’ – haryana, 27.‘popular gols’ – haryana, 28.‘akrota’ – haryana, 29.‘laddu’ – uttar pradesh, 30.‘�ar bhubraj’ – rajasthan, 31.‘ponda’ – haryana, 32.‘wilayati’ – punjab, 33.‘�ar sevika’ – rajasthan, 34.‘narikeli’ – west bengal, 35.‘sua’ – haryana, 36.‘vikas’ – gujarat, 37.‘babu’ – gujarat. 217 appendix 2 fig. 4. structures for chemical compounds in jujube having potential neuroprotection e�ect. seven chemical markers found in jujube including kaempferol 3-o-rutinoside, quercetin 3-o-rutinoside, (-)-catechin, (-)-epicatechin, swertish, spinosin, and camp were reported to possess e�ect on neuroprotection (chen et al., 2017) c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine m oh am ad h es am s ha hr aj ab ia n, w en li s un , q i c he ng 218 fig. 5. structures of triterpenic acids, nucleosides and nucleobases as well as saccharides in jujube fruits. (a) – triterpenic acids; (b) – nucleosides and nucleobases; (c) – saccharides (guo et al., 2015) 219 jujuba pospolita (ziziphus jujuba mill.) – obiecujący owoc z tradycyjnej medycyny chińskiej streszczenie głożyna pospolita, jujuba pospolita lub chiński daktyl (zizyphus jujuba mill.; rhamnaceae l.), to popularna w wielu częściach świata roślina uprawna. jej owocem jest jadalny, owalny pestkowiec, który ma szerokie zastosowanie w  kuchni i  tradycyjnej medycynie. jujuba jest uprawiana w  regionach o  klimacie umiarkowanym, podczas gdy podobna do niej głożyna omszona (z. mauritiana lam.) jest uprawiana w gorących i suchych regionach indii. największym producentem owoców jujuby na świecie są chiny, choć w ostatnim czasie zauważalny jest wzrost zainteresowania produkcją tego owocu w innych częściach świata. owoc jujuby zawiera 23 rodzaje aminokwasów, których nie ma w większości innych owoców. zawiera również witaminę c, rybo�awinę i tiaminę. główne składniki owoców jujuby są używane w połączeniu z innymi roślinami i owocami leczniczymi w tradycyjnej medycynie chińskiej. liść jujuby, który jest głównym produktem ubocznym, był również używany w celach leczniczych od tysięcy lat. jujuba ma wiele ważnych właściwości farmakologicznych i można ją uznać za cenne źródło nutraceutyków. jest rośliną o dużym potencjale leczniczym w  leczeniu chorób cywilizacyjnych, takich jak nowotwory, a  także ważnym źródłem składników odżywczych, łatwo dostępnych w biednych regionach świata. keywords: chinese dactyl, fruit, jujube, pharmacological science, traditional asian medicine received: [2020.07.23] accepted: [2020.09.14] c hinese jujube (ziziphus jujuba m ill.) – a prom ising fruit from traditional c hinese m edicine annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x wojciech w. a. kowalski 1 , katarzyna komarzewska 2 1 department of botany and nature conservation, western pomerania university of technology in szczecin, j. słowackiego 17, 71–434 szczecin, poland, e-mail: wojciech.wakowalski@wp.pl 2 chief inspectorate of environmental protection, wały chrobrego 4, 70–502 szczecin, poland water trophy assessment of the cooling system of the “dolna odra” power plant on the basis of algae indicator organisms introduction cooling waters are characterised by specific features with changed not only thermals, but very often they have also different hydro-chemical parameters. the ecological factors of such habitats develop the quantity and quality of phytoplankton structure. many indicators are used to determine water quality. one of them is saprobity. sladeček and sládečková (1996) describe it as the content in the aquatic ecosystem of organic matter capable of biochemical decomposition. different levels of organic pollution are accompanied by specific biocenosis developing in the water column on its surface among the littoral vegetation or on the bottom of the reservoir. the theoretical basis of the saprobes system are ecological relationships between the biocenosis and environmental factors. the system of indicator organisms was developed, proposed by cohn (1853, 1875) and mez (1898) and elaborated in full by kolkwitz and marson (1908, 1909) with subsequent multiple modifications. with this system, in a revised form by liebmann (1951, 1962), the degree of water pollution is assessed on the basis of decomposable organic compounds (turoboyski, 1979). the studies of phytoplankton in heated waters were carried out e.g. in the ecosystems of lakes of the konin-pątnów power plant complex (central poland). they were carried out by the department of hydrobiology of the adam mickiewicz university in poznań (burchardt , 1977). however, they are concerned mainly with the structure of phytoplankton and the qualitative and quantitative changes taking place in it. they did not include determining the trophic status of the lake’s waters. research on contaminated waters, including heated ones, was also carried out in poland by turoboyski (1967, 1969). in this study, the analyses of algae diversity and the assessment of the degree of water trophy based on the indicator organisms of planktonic algae were performed within the hydrological system used for the cooling process of the technical infrastructure of the “dolna odra” power plant. study area the research material was collected from four sites located in different parts of the odra river (western poland) ecosystem. all sites are located in the międzyodrze zone and are associated with the waters of the eastern odra – known as the regalica – 53°21′20″n 14°33′18″e (fig. 1). fig. 1. location of test stands of the cooling system of the „dolna odra”; a – test benches, b – state border, c – cities, settlements, d – railway lines, e – expensive, f – canals, oxbow lakes, rivers, f – “dolna odra” power plant, g – power station site 1 – canal waters with natural thermals supplied to the power plant. the site is located in the area of the cold canal, adjacent to the river current, which supplies water with natural thermals to the “dolna odra” cooling system intake; site 2 – the waters of the east odra river above the mouth of the warm canal. the station is located in the river’s waters above the point of the warm channel estuary to the mainstream of the eastern odra flowing along the main riverbed, characterised by unchanged thermals of discharge waters; site 3 – the waters of the warm canal discharging the cooling water. the station is situated in the channel which discharges the discharge water with increased thermal temperature from the cooling system of the “dolna odra” power plant. plankton samples were collected in the area of the experimental fishing station wrmitż zut in szczecin, which is the place of ichthyological research. site 4 – the waters of the east odra river downstream of the warm canal with elevated temperature due to post-cooling waters. the site is located in the międzyodrze area slightly below the junction of the warm canal with the current of the east odra river. in this part of the main current, the rivers with unchanged thermals mix with the waters flowing from the warm water discharge canal. materials and methods the study focuses primarily on the assessment of water trophies based on the identified algae species. from each of the four sites located in different parts of the hydrological system, 4 samples with a volume of 18 litres of water were collected during the year and concentrated with a plankton mesh to 1 litre. from this volume, after thorough mixing of the sample, 3 microscopic slides were prepared to determine the qualitative composition and the value of the frequency of the organism occurrence (h) expressed by the number of specimens of individual species in the field of view (starmach, 1955). the observed species were determined with the use of taxonomic literature. the method of pantle and buck (1955) used here introduces the saprobic index (sw) determined based on the so-called saprobial values of individual species (s) and frequency of the organism occurrence (h). the saprobic value for various types of water was assumed according to the criteria presented in table (1). the frequency of the organism occurrence (h) is determined based on the percentage of individuals about all individuals of all species (tab. 2). tab. 1. saprobic value [s] for various types of water zone designation s xenosaprobity x 0 oligosaprobity o 1 β–mesosaprobity β 2 α–mesosaprobity α 3 polisaprobity p 4 tab. 2. the frequency of the organism occurrence [h] for different percentage ranges percentage of a taxa h up to 1 % 1 1 – 3 % 2 4 – 10 % 3 10 – 20 % 4 20 – 40 % 5 40 – 100 % 6 the basis for the calculation of the saprobic index (s) for individual samples was the floristic analysis and determination of the frequency of taxa in the samples. the index of the saprobic value (si) and the indication weight of the taxon (ii) was obtained based on data from the literature (sladeček, sládečková, 1996). the abundance of a taxon found (h) in the field of view was examined based on a scale according to starmach (1989): 1 – rarely, when the species is present in about 1% of the fields of view; 3 – often, when the species is present in 5 – 30 fields of view; 5 – mass, when the species is present in 60 – 100 fields of view. when determining the trophic status of ecosystem waters, only taxa with known saprobity were taken into account, the value and indicator values of which were presented and taken into account when determining the trophic status of ecosystem waters in the literature (sladeček, sládečková, 1996). the pantle and buck (1955) saprobic index (sw) is calculated according to the formula:   h sh s i w    (1) where: sw – saprobic index, si – saprobic value of the species, h – the frequency of the species occurrence. the product is calculated for each marked species. the pattern of panle and buck (1955) with the modification of marvan (after sladeček, sládečková, 1996) introduces the indication weight of individual species ii:     ii iii ih ihs s    (2) where: s – saprobic index, si – saprobic value of the species, ii – the indication weight of the individual taxon (range from 1 to 5, where 1 – oligosaprobity, 2 – β-mezosaprobity, 3 – α-mezosaprobity, 4 – polisaprobity, 5 – izosaprobity), h – the abundance of a taxon found. the values of the saprobic index inform about the degree of water clarity. these values are classified in the following ranges: oligosaprobity s = 1.00, between oligoand βmezosaprobity s = 1.50, β-mezosaprobity s = 2.00, between β-mezoand αmezosaprobity s = 2.50, αmezosaprobity s = 3.00, polisaprobity s = 3.50. results and discussion any aquatic organism can be used as an indicator. if we know at least estimate their living environment, we can use its presence to determine the water quality (sladeček, sládečková, 1996; ostrowska, 2012). according to burchardt et al. (1994) a bioindicator is any taxon whose presence or item of number is associated with a specific set of physicochemical conditions that define the framework for the functioning of a specific state of the biocenosis. saprobic systems contain lists of species with their indicating value and saprobial values. most systems takes into account different systematic groups of organisms e. g. bacteria, algae or invertebrate animals. there are also those in which there is only one selected group e. g. diatoms, euglenophytes or benthic invertebrate. for the saprobial system, many lists of species have been developed, together with their saprobicity determination for the use of water quality research institutions. the expression of the use of algae as indicators of environmental quality are systems of organisms based on practical observations of their occurrence in specific environmental conditions (kawecka, eloranta, 1994). another measure of water quality is the trophy of the reservoir, determined both by the content of nutrients, characteristic organisms, chlorophyll concentration, biomass size, etc. many researchers have tried to improve and complete the kolkwitz and marsson system (1908, 1909); there have been many studies comprehensively developing the issue of biological assessment of pollutants, as well as attempts to modify the saprobes system and introduce other methods. in poland, it was mainly turoboyski (1970a, 1970b, 1973, 1976) who worked on it, which determined the index value of individual species, indicating the ranges of variability of their quantitative occurrence in individual pollution zones, which contributed to the refinement of the results of studies of waters contaminated with the classical method. starmach (1955) divided the waters on the basis of the presence of certain species of algae into: spring waters – cataract, clean – oligosaprobic, slightly contaminated – β–mesosaprobic, heavily polluted – α–mesosaprobic, extremely contaminated – polysaprobial, partially poisoned, poisoned. the classification of algae clearly differs, especially in the polysaprobic, oligosaprobic and cataract zones, the latter including waters not contaminated with any sewage. the zones of αand β–mesosaprobic waters, due to their significant similarity, differ in more difficult to perceive features in the occurrence of individual algae taxa. this is due to the fact that eutrophic water species need a higher concentration of nutrients to achieve maximum growth compared to oligotrophic waters, where cells reach their maximum growth rate even at low nutrient concentrations. various species of algae are used to assess the environment. the most useful in bioindication are diatoms (ostrowska, 2012). other algae used as indicators of the characteristics of the aquatic environment are: green algae, including charophytes, desmidiales, and golden algae. when determining water pollution zones on the basis of biological analyses performed, it is advisable to compare these results with chemical analyses, as the complete assessment of water consists of a complex of physical, chemical and biological factors. the chemical definition of water pollution zones expressed by the value of bod5 and dissolved oxygen (turoboyski, 1979) is as follows: polisaprobic zone: bod5 >15 mg/l o2, dissolved oxygen < 2 mg/l o2, α–mezosaprobic zone: bod5 5–15 mg/l o2, dissolved oxygen 2–4 mg/l o2, β–mezosaprobic zone: bod5 3–5 mg/l o2, dissolved oxygen 5–9 mg/l o2, oligosaprobic zone: bod5 < 3 mg/l o2, dissolved oxygen > 9 mg/l o2. species composition and saprobic value of the taxon altogether 101 algae taxa were identified in 16 samples collected from the cooling system of the „dolna odra” power plant. representatives of the following phyla were distinguished: cyanoprocaryota, dinophyceae, chrysophyceae, bacillariophyceae, euglenophyceae, and chlorophyceae (tab. 3 – appendix 1; fig. 2). fig. 2. percentage share of all organisms (a) and indicator species (b) at studied sites: a – cyanoprocaryota, b – dinophyceae, c – chrysophyceae, d – bacillariophyceae, e – euglenophyceae, f – chlorophyceae 1. species frequently found in the studied waters (fig. 3–6 – appendix 2) the most numerous group in the studied waters are green algae of the order chlorococcales. common planktonic species are pseudopediastrum boryanum, pediastrum duplex var. duplex, lacunastrum gracillimum, tetradesmus lagerheimii, desmodesmus communis, desmodesmus opoliensis and lemmermannia triangularis. cyanobacteria are represented by: aphanocapsa delicatissima, planktothrix agardhii, limnothrix planctonica, microcystis aeruginosa, microcystis wesenbergii, pseudanabaena mucicola, dolichospermum flosaquae, aphanizomenon flexuosum. another group of algae abundant in these waters are diatoms: aulacoseira granulata var. granulata, aulacoseira granulata var. angustissima, fragilaria crotonensis, stephanodiscus hantzschii, asterionella formosa, cyclotella meneghiniana, actinocyclus normanii, synedra ulna, melosira varians, nitzschia sigmoidea, ulnaria danica. 2. indicator species in the studied waters the qualitative taxonomic composition of the studied sites shows that the most numerous group of indicator species allowing for the calculation of the saprobic index (s) and determination of belonging to the β–mesosaprobic zone were the chlorophyceae taxa (tab. 4). green algae represent approximately 42% of the total number of species. at the same time, among this group of algae, as much as 45.6% are bioindicators. the largest group is chlorococcales. these data are consistent with previous studies (sládeček, sládečková, 1996). the small share of bioindication species from the order volvocales is astonishing. of the 57 taxa considered to be bioindication organisms of this order, only 2 were recorded. an important role in the assessment of the saprobic of these waters is also played by representatives of bacillariophyceae (19 bioindicators) and cyanoprocaryota (14 bioindicators). tab. 4. share of indicator species at studied sites (1–4). taxonomy number of taxa/number of site total number of species 1 2 3 4 d u ri n g t h e re se a rc h p e ri o d [%] in d ic a to r sp e c ie s [%] t o ta l in d ic a to r t o ta l in d ic a to r t o ta l in d ic a to r t o ta l in d ic a to r cyanoprocaryota 16 8 15 8 16 9 14 10 23 22.7 14 20.6 dinophyceae 2 1 1 2 2 2.,0 1 1.5 chrysophyceae 1 1 2 1 2 2 2.0 1 1.5 bacillariophyceae 15 11 16 10 19 12 18 14 29 28.7 19 27.9 euglenophyceae 1 3 2 3 3.0 2 2.9 chlorophyceae: 35 23 23 17 29 21 24 18 42 41.6 31 45.6 volvocales chlorococcales desmidiales 1 29 5 1 21 1 2 16 5 1 14 2 2 21 6 2 16 3 2 19 3 1 16 1 2 33 7 2.0 32.7 6.9 2 27 2 2.9 39.7 2.9 total 66 42 56 37 68 43 61 48 101 100 68 100 3. saprobic index (s) species with a wide ecological spectrum, occurring in waters with different trophies and not having a specific saprobe value and indicator value in the calculations were not taken into account. for taxa occurring sparse in the analysed samples and having an indicator value assume value 1 for the frequency of the species occurrence. the degree of water pollution of individual sites was determined on the basis of bioindicators, i.e. species of algae with an indicator value. during the research cycle, the saprobic index fluctuated at individual sites (fig. 7), reaching a value from 1.613 to 2.024 (tab. 5). its average values in waters with different thermal temperatures were on a similar level. the value of the water saprobic index value of all sites indicates their belonging to the waters of the β–mesosaprobic. according to sladeček and sládečková (1996), the saprobic index value ranging from 1.51 to 2.50 (mean s = 2.0) indicates the waters of the β– mesosaprobic. this zone is characterised by the course of biochemical processes under aerobic conditions as a result of which complete oxidation of the intermediate products of decomposition of organic compounds takes place. fig. 7. saprobic index [s] at four sites during the months july – december 2004; 1-4 – numbers of site (see in the text), 5 – min.-max. values of ß–mesosaprobic zone tab. 5. saprobic index value in the test waters; 1-4 – study sites date site /saprobic index 1 2 3 4 09.07.2004 1.951 1.613 1.952 2.024 10.08.2004 1.871 1.857 1.832 1.828 06.09.2004 2.024 1.919 1.877 1.978 07.12.2004 1.832 1.813 1.898 1.945 average saprobic index 1.919 1.800 1.889 1.944 a characteristic feature of the waters of the β–mesosaprobic zone is the domination of chemosynthetic and photosynthetic autotrophs, nitrifying bacteria, and algae, mainly bacillariophyceae and chlorophyceae. the waters of the studied sites have a similar species composition of algae. 4. influence of changes in water thermals on the quality composition of phytoplankton the temperature of the aquatic environment is not only a factor influencing the development of phytoplankton but also its composition (sládeček, sládečková, 1996; ostrowska, 2012). the analyses of the qualitative composition of phytoplankton showed slight differences between individual sites (tab. 3 – appendix 1). the structure of phytoplankton as well as the percentage share of individual systematic groups during the research cycle remain similar and have nonsignificant differences in the quality composition of phytoplankton. this is visible not only in its structure but also in the percentage share between the individual systematic groups of algae. such a structure of phytoplankton results from the presence of species with a wide ecological amplitude about the environmental thermals. an important factor here is also a more stable water temperature in large rivers in which there is a tendency to equalize it due to its constant mixing. conclusions the obtained results allow the following conclusions to be drawn: average value of the saprobic index allows to classify of the analysed waters of the β– mesosaprobic zone; the highest saprobic index, as well as the average index, are found in the waters below the warm channel (site 4). this is surely a consequence of the increase in the temperature of the waters of this site by warm discharge waters from the power plant, as well as the probably changed chemical composition; the phytoplankton of the studied waters is typical for slowly flowing, medium fertile waters; planktonic species are the absolute dominant taxa. benthic forms appear sporadically and their presence should be considered accidental; the material does not contain species that are rare for algae and the taxa that are present belong to eurobionts. conflict of interest the authors declare no conflict of interest related to this article. references algaebase database. 2022. https://www.algaebase.org/ burchardt, l. 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[in polish] appendix 1 tab. 3. floristic spectrum of alage. the valid species names of the identified taxa were given on the basis of literature data and the algaebase database (2022); si – saprobic value of the taxon site 1 – canal water with natural thermals supplied to the power plant no. taxonomy/taxon si research terms 0 9 .0 7 .2 0 0 4 1 0 .0 8 .2 0 0 4 0 6 .0 9 .2 0 0 4 0 7 .1 2 .2 0 0 4 cyanoprocaryota 1. aphanocapsa delicatissima w. et g. s. west . + . + . 2. planktothrix agardhii (gomont) anagnostidis et kom. 1.6 + . + . 3. limnothrix planctonica (wolosz.) meffert . + . . + 4. oscillatoria sp. . + . . . 5. microcystis aeruginosa (kütz.) kütz. 1.8 + + + + 6. dolichospermum sigmoideum (nygaard) wacklin, l.hoffmann & kom. . + . . . 7. cf. anabaenopsis arnoldii apterarj 1.6 + . . . 8. cf. aphanizomenon flosaquae ralfs ex bornet & flahault 2.2 + + . . 9. chroococcus turgidus (kütz.) nägeli 1.3 . + . . 10. microcystis viridis (a. braun) lemm. 1.8 . + . . 11. microcysris wesenbregii (kom.) kom. ex kom. 1.8 . + + + 12. pseudanabaena mucicola (naumann & huber-pestalozzi) schwabe . . + + . 13. aphanizomenon flexuosum kom. & kováčik 2.2 . + + . 14. woronichinia compacta (lemm.) kom. et hindák . . . + . 15. merismopedia teniussima lemm. 2.5 . . + . 16. lyngbya attenuata f. e. fritsch . . . . + bacillariophyceae 17. aulacoseira granulata var. granulata (ehr.) simonsen 1.8 + + + + 18. aulacoseira granulata var. angustissima (o.f.müller) simonsen 1.8 + + + + 19. fragilaria crotonensis kitton 1.4 + + + + 20. stephanodiscus hantzschii grunow 2.7 + + + . 21. asterionella formosa hassall 1.4 + + . + 22. cyclotella meneghiniana kütz. 2.7 + + + + 23. actinocyclus normanii (gregory) hustedt . + + + + 24. ulnaria danica (kütz.) compère & bukhtiyarova . + + + + 25. melosira varians agardh 1.6 + + + + 26. nitzschia sigmoidea (nitzsch) w. smith 2.5 + + . + 27. synedra ulna (nitzsch) ehr. . + . . + 28. diatoma tenuis agardh 1.5 + + . . 29. nitzschia acicularis (kütz.) w. smith 2.4 + . . . 30. diatoma vulgaris bory de saint-vincent 2.2 . . + . 31. fragilaria sp. . . + . . chlorophyceae 32. micractinium pusillum fres. 2.5 + . + . 33. mucidosphaerium cf. pulchellum (h.c.wood) c.bock, proschold & krienitz . + . . . 34. coelastrum astroideum de-not de notaris 2.0 + + + + 35. stauridium tetras (ehr.) e. hegewald 1.8 + + + . 36. pseudopediastrum boryanum (turp.) e.hegewald 1.9 + + + + 37. lacunastrum gracillimum (west & g.s. west) h.mcmanus 1.9 + + + + 38. pediastrum duplex meyen 1.9 + + + + 39. monactinus simplex (meyen) corda 1.5 . + + + 40. tetradesmus lagerheimii m.j.wynne & guiry 2.2 + + + . 41. desmodesmus denticulatus (lagerh.) s.s.an, t.friedl & e.hegewald . + . + . 42. desmodesmus subspicatus (chod.) e.hegewald & a.w.f.schmidt . + + . . 43. verrucodesmus verrucosus (y.v.roll) e.hegewald 1.7 + . + . 44. desmodesmus communis (e.hegewald) e.hegewald 2.1 + . + . 45. desmodesmus opoliensis (p.g.richt.) e.hegewald 2.2 + + + . 46. tetradesmus obliquus (turp.) m.j.wynne 2.8 . . + . 47. desmodesmus armatus (chod.) e.hegewald . . + + . 48. desmodesmus intermedius (chod.) e.hegewald . . + + + 49. desmodesmus spinosus (chod.) e.hegewald . . . + . 50. lemmermannia triangularis (chod.) c.bock & krienitz 2.0 + . + . 52. tetrastrum staurogeniiforme (schröd.) lemm. 2.2 + . + . 53. tetraedron caudatum (corda) hansg. 2.0 + . + . 54. tetraedrom minimum (a. br.) hansg. 2.0 . . + . 55. tetrastrum elegans playf. 1.5 + . + . 56. actinastrum hantzschii var. subtile lagerh. wolosz. 2.3 + + + . 57. actinastrum hantzschii var. hantzschii lagerh. 2.3 + . + . 58. pandorina smithii chod. 2.0 + + + . 59. monoraphidium griffithii (barkeley) komárkova-lagnerová 2.2 . + . . 60. golenkinia radiata chod. 1.8 . + . . 61. quadricoccus ellipticus hortob. . . . + . 62. closterium nordstedti var polystictum (nygaard) rúźička . + + . . 63. closterium acerosum ehr. ex ralfs . . + . . 64. closterium cf. moliniferum ehr. ex ralfs 2.1 . + . . 65. staurastrum gracile ralfs ex ralfs . + + . . 66. staurastrum paradoxum meyen ex ralfs . . . + . number of taxa total 66 43 38 43 20 percentage share [%] 100 65.2 57.6 65.2 30.3 number of indicators total 42 31 28 32 14 percentage share [%] 63.6 47.0 42.4 48.5 21.2 dominant aphanizomenon flexuosum . . + . diatoms centricae . . + . continued: site 2 – the waters of the east odra river above the mouth of the warm channel no taxonomy/taxon si research terms 0 9 .0 7 .2 0 0 4 1 0 .0 8 .2 0 0 4 0 6 .0 9 .2 0 0 4 0 7 .1 2 .2 0 0 4 cyanoprocaryota 1. aphanocapsa delicatissima w. et g. s. west . . . + . 2. planktothrix agardhii (gomont) anagnostidis et kom. 1.6 + . + . 3. limnothrix planctonica (wolosz.) meffert . + . . . 4. microcystis aeruginosa (kütz.) kütz. 1.8 + + + + 5. dolichospermum affine (lemm.) wacklin, l.hoffmann & kom. 2.0 . + . . 6. dolichospermum flosaquae (bréb. ex bornet & flahault) p.wacklin, l.hoffmann & j.kom. 2.0 . . + . 7. microcystis marginata (menegh.) kütz. . . . + . 8. microcystis wesenbergii (kom.) kom. ex kom. 1.8 + + + + 9. pseudanabaena mucicola (naumann & huber-pestalozzi) schwabe . . + + . 10. aphanizomenon flexuosum kom. & kováčik 2.2 . + + . 11. merismopedia tenuissima lemm. 2.5 . . + . 12. woronichinia compacta (lemm.) kom. et hindák . . + . . 13. chroococcus turgidus (kütz.) nägeli 1.3 . + . . 14. anabaena sp. . . . + . 15. snowella lacustris (chod.) kom. & hindák 1.5 . . + . bacillariophyceae 16. aulacoseira granulata var. granulata (ehr.) simonsen 1.8 + + + + 17. aulacoseira granulata var. angustissima (o.f.müller) simonsen 1.8 + + + + 18. fragilaria crotonensis kitton 1.4 + + + + 19. stephanodiscus hantzschii grunow 2.7 + + + . 20. asterionella formosa hassall 1.4 + + + + 21. cyclotella meneghiniana kütz. 2.7 + + + + 22. actinocyclus normanii (gregory) hustedt . + . + + 23. ulnaria danica (kütz.) compère & bukhtiyarova . + + + + 24. melosira varians agardh 1.6 . + . + 25. nitzschia sigmoidea (nitzsch) w. smith 2.5 + . . + 26. synedra ulna (nitzsch) ehr. . + . + + 27. diatoma tenuis agardh 1.5 . + . . 28. amphora ovalis (kütz.) kütz. 1.5 . . + . 29. gomphonema acuminatum ehr. 0.9 . . . + 30. fragilaria sp . . . . + 31. cymbella sp. . . . . + chrysophyceae 32. dinobryon divergens imhof 1.8 + . . . euglenophyta 33. colacium vesiculosus ehr. 1.9 . . + . chlorophyceae 34. micractinium pusillum fres. 2.5 + . . . 35. schroederia setigera (schröd.)lemm 1.7 + . . . 36. coelastrum astroideum de-not de notaris 2.0 . . + . 37. pseudopediastrum boryanum (turp.) e.hegewald 1.9 + + + + 38. lacunastrum gracillimum (west & g.s. west) h.mcmanus 1.8 + + . . 39. pediastrum duplex meyen 1.8 + + + + 40 monactinus simplex (meyen) corda 1.5 + + + . 41. tetradesmus lagerheimii m.j.wynne & guiry 2.2 + . . + 42. desmodesmus communis (e.hegewald) e.hegewald 2.1 + + + . 43. desmodesmus opoliensis (p.g.richt.) e.hegewald 2.2 + + . + 44. desmodesmus armatus (chod.) e.hegewald . + . . . 45. desmodesmus spinosus (chod.) e.hegewald . + . . . 46. verrucodesmus verrucosus (y.v.roll) e.hegewald 1.7 + . . . 47. lemmermannia triangularis (chod.) c.bock & krienitz 2.0 + . . . 48. tetraedron caudatum (corda) hansg. 2.0 . + . . 49. actinastrum hantzschii var. subtile lagerh. wolosz. 2.3 . . + . 50. pandorina smithii chod. 2.0 + + . . 51. eudorina elegans ehr. 2.2 . + . . 52. closterium nordstedtii var. polystictum (nygaard) rúźička . + + + . 53. closterium strigosum var. strigosum bréb. 2.2 + . . . 54. closterium moliniferum ehr. ex ralfs 2.1 . + . . 55. staurastrum gracile ralfs ex ralfs . + . . . 56. staurastrum paradoxum meyen ex ralfs . . + . . number of taxa total 56 32 28 29 19 percentage share [%] 100 57.1 50.0 51.8 33.9 number of indicators total 40 24 23 21 14 percentage share [%] 71.4 42.9 41.1 37.5 25.0 dominant microcystis sp. div., aphanizomenon flexuosum + diatoms centricae + continued: site 3 – water of the warm channel draining cooling water no. taxonomy/taxon si research terms 0 9 .0 7 .2 0 0 4 1 0 .0 8 .2 0 0 4 0 6 .0 9 .2 0 0 4 0 7 .1 2 .2 0 0 4 cyanoprocaryota 1. aphanocapsa delicatissima w. et g. s. west . . + . 2. planktothrix agardhii (gomont) anagnostidis et kom. 1.6 + . + + 3. limnothrix planctonica (wolosz.) meffert + . . . 4. microcystis aeruginosa (kütz.) kütz. 1.8 + . + + 5. dolichospermum cf.affine (lemm.) wacklin, l.hoffmann & kom. 2.0 . + . . 6. dolichospermum flosaquae (bréb. ex bornet & flahault) p.wacklin, l.hoffmann & j.kom. 2.0 . + . . 7. microcystis viridis (a. braun) lemm. 1.8 . + . . 8. microcystis marginata (menegh.) kütz. . + + . 9. microcystis wesenbergii (kom.) kom. ex kom. 1.8 + + + + 10. pseudanabaena mucicola (naumann & huber-pestalozzi) schwabe . + + + 11. aphanizomenon flexuosum kom. & kováčik 2.2 . + + . 12. aphanizomenon cf. gracile (lemm.) lemm. 1.5 . . + . 13. lyngbya attenuata f. e. fritsch + . . . 14. anabaena sp. + . + . 15. hapalosiphon sp. . + . . 16. snowella lacustris (chod.) kom. & hindák 1.5 . . + . bacillariophyceae 17. aulacoseira granulata var. granulate (ehr.) simonsen 1.8 + + + + 18. aulacoseira granulata var. angustissima (ehr.) simonsen (o. müller) simonsen 1.8 + . + . 19. fragilaria crotonensis kitton 1.4 + + + + 20. stephanodiscus hantzschii grunow (in cleve & grunow) 2.7 + + . . 21. asterionella formosa hassall 1.4 + + + + 22. cyclotella meneghiniana kütz. 2.7 + + + + 23. actinocyclus normanii (gregory) hustedt . + + + . 24. synedra ulna var. danica (nitzsch) ehr. (kütz.) hustedt . + + + + 25. melosira varians agardh 1.6 . . + + 26. nitzschia sigmoidea (nitzsch) w. smith 2.5 + . + 27. synedra ulna (nitzsch) ehr. . . . + + 28. diatoma tenuis agardh 1.5 + . + . 29. nitzschia acicularis (kütz.) w. smith 2.4 + . . . 30. nitzschia cf. littoralis grunow . . + . . 31. navicula cincta (ehr.) ralfs in pritchard . + . . . 32. pinnularia nobilis (ehr.) ehr. 1.1 . + . . 33. gyrosigma attenuatum (kütz.) rabenhorst 2.2 . . . + 34. fragilaria sp. . . + + + 35. navicula sp. . + . . . chrysophyceae 36. dinobryon sociale ehr. . + . . . 37. dinobryon divergens imhof 1.8 . . . + dinophyceae 38. ceratium hirundinella (o.f. müller) schrank 1.2 + . . . 39. gymnodinium sp . . . + . chlorophyceae 40. mucidosphaerium pulchellum (h.c.wood) c.bock, proschold & krienitz 2.3 . . . 41. coelastrum astroideum de-not de notaris 2.0 + + + . 42. coelastrum microporum näg. 2.1 . . . + 43. stauridium tetras (ehr.) e.hegewald 1.8 . . + . 44. pseudopediastrum boryanum (turp.) e.hegewald 1.9 + + + + 45. lacunastrum gracillimum (west & g.s. west) h.mcmanus 1.8 + + . 46. pediastrum duplex meyen 1.8 + . + + 47. monactinus simplex (meyen) corda 1.5 + + + . 48. tetradesmus lagerheimii m.j.wynne & guiry 2.2 + . + . 49. desmodesmus denticulatus (lagerh.) s.s.an, t.friedl & e.hegewald . . . + . 50. desmodesmus subspicatus (chod.) e.hegewald & a.w.f.schmidt . + . + . 51. desmodesmus communis (e.hegewald) e.hegewald 2.1 + . + + 52. desmodesmus opoliensis (p.g.richt.) e.hegewald 2.2 + + + + 53. desmodesmus armatus (chod.) e.hegewald . + + . . 54. lemmermannia triangularis (chod.) c.bock & krienitz . . . + . 55. tetraedron caudatum (corda) hansg. 2.0 + . . . 56. actinastrum hantzschii var. subtile lagerh. wolosz. 2.0 + . . . 57. actinastrum hantzschii var. hantzschii lagerh. 2.3 + . + . 58. pandorina smithii chod. 2.0 + . . . 59. eudorina elegans ehr. 2.0 + + . + 60. golenkinia radiata chod. 2.2 + . . . 61. lagerheimia ciliata (lgerh.) chod. 1.8 + . . . 62. closterium nordstedti var. polystictum (nygaard) rúźička 2.0 + . . . 63. closterium acutum bréb. . + . + . 64. closterium srtigosum bréb. . + . . . 65. closterium moliniferum ehr. ex ralfs 2.2 + . . . 66. staurastrum gracile ralfs ex ralfs 2.1 . . + . 67. staurastrum paradoxum meyen ex ralfs . + . . . 68. mucidosphaerium pulchellum (h.c.wood) c.bock, proschold & krienitz . . + . . number of taxa total 68 43 26 36 21 percentage share [%] 100 63.2 38.2 52.9 30.9 number of indicators total 44 30 16 23 16 percentage share [%] 64.7 44.1 23.5 33.8 23.5 dominant microcystis sp. div., aphanizomenon flexuosum + continued: site 4 – the waters of the east odra river below the warm channel having increased thermals by cooling waters no. taxonomy/taxon si research terms 0 9 .0 7 .2 0 0 4 1 0 .0 8 .2 0 0 4 0 6 .0 9 .2 0 0 4 0 7 .1 2 .2 0 0 4 cyanoprocaryota 1. aphanocapsa delicatissima w. et g. s. west . . + . 2. aphanocapsa grevillei (berkeley) rabenhorst 1.4 . . + . 3. merismopedia tenuissima lemm. 2.5 . . + . 4. chroococcus turgidus (kütz.) nägeli 1.3 . + . . 5. planktothrix agardhii (gomont) anagnostidis et kom. 1.6 + . . . 6. limnothrix planctonica (wolosz.) meffert + . + . 7. microcystis aeruginosa (kütz.) kütz. 1.8 + . + + 8. dolichospermum flosaquae (bréb. ex bornet & flahault) p.wacklin, l.hoffmann & j.kom. 2.0 . . + . 9. microcystis viridis (a. braun) lemm. 1.8 . + . . 10. microcystis marginata (menegh.) kütz. . + + . 11. microcystis wesenbergii (kom.) kom. ex kom. 1.8 + + + + 12. pseudanabaena mucicola (naumann & huber-pestalozzi) schwabe . + + . 13. aphanizomenon flexuosum kom. & kováčik 2.2 + + + . 14. snowella lacustris (chod.) kom. & hindák 1.5 . + . . bacillariophyceae 15. aulacoseira granulata var. granulata (ehr.) simonsen 1.8 + + + + 16. aulacoseira granulata var. angustissima (o.f.müller) simonsen 1.8 + + + + 17. fragilaria crotonensis kitton 1.4 + + + + 18. stephanodiscus hantzschii grunow 2.7 + + + + 19. asterionella formosa hassall 1.4 + + + + 20. cyclotella meneghiniana kütz. 2.7 + + + + 21. actinocyclus normanii (gregory) hustedt . . + . . 22. ulnaria danica (kütz.) compère & bukhtiyarova . + + . . 23. melosira varians agardh 1.6 . + + + 24. nitzschia sigmoidea (nitzsch) w. smith 2.5 + + + + 25. synedra ulna (nitzsch) ehr. + + . + 26. diatoma tenuis agardh 1.5 + + + . 27. nitzschia acicularis (kütz.) w. smith 2.4 . + . . 28. brebissonia lanceolata (agardh) r.k.mahoney & reimer 1.5 . + . . 29. cymatopleura solea (bréb.) w. smith 2.3 . . + + 30. iconella biseriata (bréb.) ruck & nakov 1.5 . . . + 31. surirella elegans ehr. 1.3 . . . + 32. fragilaria sp. . + . + chrysophyceae 33. dinobryon divergens imhof 1.8 . + . . dinophyceae 34. ceratium hirundinella (o.f.müller) dujardin 1.2 . + . . euglenophyceae 35. lepocinclis acus (o.f.müller) marin & melkonian . + + . . 36. lepocinclis oxyuris (schmarda) marin & melkonian 2.5 . . + . 37. colacium vesiculosum ehr. 1.9 . + . . chlorophyceae 38. micractinium pusillum fres. 2.5 + . + . 39. mucidosphaerium pulchellum (h.c.wood) c.bock, proschold & krienitz 2.3 + . . . 40. coelastrum astroideum de-not de notaris 2.0 . + + + 41. stauridium tetras (ehr.) e.hegewald 1.8 . . + . 42. parapediastrum biradiatum (meyen) e.hegewald 1.8 . . + . 43. pseudopediastrum boryanum (turp.) e.hegewald 1.9 + . + + 44. lacunastrum gracillimum (west & g.s. west) h.mcmanus 1.8 + + . . 45. pediastrum duplex meyen 1.8 + + + + 46. monactinus simplex (meyen) corda 1.5 . . + . 47. tetradesmus lagerheimii m.j.wynne & guiry 2.2 + . + . 48. desmodesmus denticulatus (lagerh.) s.s.an, t.friedl & e.hegewald + . . . 49. desmodesmus communis (e.hegewald) e.hegewald 2.1 + + + . 50. desmodesmus opoliensis (p.g.richt.) e.hegewald 2.2 + + + . 51. desmodesmus armatus (chod.) e.hegewald + + . . 52. desmodesmus subspicatus (chod.) e.hegewald & a.w.f.schmidt . . . + 53. lemmermannia triangularis (chod.) c.bock & krienitz 2.0 . . . + 54. tetrastrum staurogeniiforme (schröd.) lemm. 2.2 + . . . 55. actinastrum hantzschii lagerh. var. subtile wolosz. 2.3 + . + + 56. crucigenia fenestrata (schmidle) schmidle 2.1 + . . . 57. pandorina smithii chod. 2.0 + . + . 58. eudorina elegans ehr. 2.2 + . . . 59. closterium nordstedtii var. polystictum (nygaard) rúźička + + . . 60. closterium moliniferum ehr. ex ralfs 2.1 . + . . 61. staurastrum gracile ralfs ex ralfs + + . . number of taxa total 61 32 36 33 21 percentage share [%] 100 52.5 59.0 54.1 34.4 number of indicators total 47 25 25 29 18 percentage share [%] 77.0 41.0 41.0 47.5 29.5 dominant microcystis sp. div., aphanizomenon flexuosum + appendix 2 fig. 3. parapediastrum biradiatum (meyen) e.hegewald – a, microcystis wesenbergii (kom.) kom. ex kom. – b, microcystis aeruginosa (kütz.) kütz. – c, pediastrum duplex meyen – d, stauridium tetras (ehr.) e. hegewald – e, monactinus simplex (meyen) corda – f fig. 4. monactinius simplex (meyen) corda – f1, lacunastrum gracillimum (west & g.s. west) h.mcmanus – g, actinastrum hantzschii lagerh. – h, asterionella formosa hassall – i, aulacoseira granulata var. granulata (ehr.) simonsen – j fig. 5. ceratium hirundinella (o.f. müller) schrank – l, coelastrum asteroideum de-not de notaris – m, fragilaria crotonensis kitton – n, tetradesmus lagerheimii m.j.wynne & guiry – o fig. 6. desmodesmus communis (e.hegewald) e.hegewald – r, desmodesmus opoliensis (p.g.richt.) e.hegewald – s, mucidosphaerium pulchellum (h.c.wood) c.bock, proschold & krienitz – t, nitzschia acicularis (kütz.) w. smith – u, planktotrix agardhii (gomont) anagnostidis et kom. – w, snowella lacustris (chod.) kom. & hindák – z abstract the paper presents the results of water trophy assessment from various stations of the cooling system of the “dolna odra” (poland) power plant. the assessment was made based on the indicator phytoplankton species collected four times at four sites located in different parts of the hydrological system cooling the technical infrastructure of the power plant. the sites were characterised by water with a natural temperature for individual seasons, as well as by thermals changed as a result of the discharge of cooling waters. for each site, the saprobic index was calculated based on indicator species. the analysis showed that the changes in trophic conditions, both in waters with changed thermals and in waters not subject to the influence of after-cool waters, are not subject to significant changes. the waters of all studied sites in the analysed periods of the year should be classified according to sladeček and sládečková (1996) to the waters of the β–mesosaprobic zone, with a saprobic index value ranging from 1.51 to 2.50 (average s = 2.0). this zone is characterised by the course of biochemical processes under aerobic conditions, as a result of which complete oxidation of the intermediate products of decomposition of organic compounds takes place. key words: after-cooling water, indicator species, phytoplankton, saprobic index, water trophy of the powerplant’s hydrological infrastructure received: [2022.10.24] accepted: [2022.11.12] ocena trofii wód układu chłodzącego elektowni „dolna odra” na podstawie składu jakościowego organizmów wskaźnikowych glonów streszczenie w pracy przedstawiono wyniki oceny trofii wód z różnych stanowisk układu chłodniczego elektrowni „dolna odra” (polska). oceny dokonano w oparciu o wskaźnikowe gatunki fitoplanktonu zebranego w czterech terminach roku na czterech stanowiskach, zlokalizowanych w różnych częściach systemu hydrologicznego schładzającego infrastrukturę techniczną elektrowni. stanowiska charakteryzowały się wodami o temperaturze naturalnej dla poszczególnych pór roku, jak i zmienną termiką w wyniku zrzutu wód pochłodniczych. dla każdego stanowiska obliczono indeks saprobowy na podstawie gatunków wskaźnikowych. analiza wykazała, że zmiany w zakresie trofii, zarówno w wodach o zmienionej termice, jak i wodach nie podlegających oddziaływaniu wód pochodniczych, nie ulegają istotnym zmianom. wody wszystkich badanych stanowisk w analizowanych okresach roku należy zaklasyfikować wg sladeček i sládečková (1996) do wód strefy β–mezosaprobowej, mających wartość indeksu saprobowości w granicach od 1,51 do 2,50 (średnio s = 2,0). strefa ta charakteryzuje się przebiegiem procesów biochemicznych w warunkach aerobowych, w wyniku których zachodzi całkowite utlenienie pośrednich produktów rozkładu związków organicznych. słowa kluczowe: wody pochłodnicze, gatunki wskaźnikowe, fitoplankton, wskaźnik saprobowy, trofia wodna infrastruktury hydrologicznej elektrowni information on the authors wojciech w. a. kowalski the author is a specialist in the field of algology. his research interests concern both single species of algae and whole groups of marine and freshwater algae, with particular emphasis on rare and endangered taxa. a special taxonomic group of interest is the taxa associated with the ecosystems of bog bogs, as well as freshwater red algae. katarzyna komarzewska she is a specialist in the field of algology. her research is focused on the various aquatic ecosystems, especially those transformed anthropogenically. she is professionally involved in phytoplankton research as part of monitoring carried out in western pomerania in the central research laboratory in szczecin (north poland). 附表8 annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832, e-issn 2545-0999 doi: 10.24917/25438832.7.x peiman zandi 1,3 *, aminu darma 2 , agnieszka tatoj 3 , xue zhou 1 , ayşe çalık 4 , mohamad hesam shahrajabian 5 , alminda magbalot-fernandez 6 , ewald schnug 7 1 international faculty of applied technology, yibin university, yibin 644000, china, *peiman.zandi@yibinu.edu.cn 2 institute of environment and sustainable development in agriculture, chinese academy of agricultural sciences, beijing 100081, china 3 department of botany, institute of biology, pedagogical university of krakow, 30-084 kraków poland; 4 department of field crops, faculty of agriculture, harran university, 63300 şanlıurfa, turkey 5 biotechnology research institute, chinese academy of agricultural sciences, beijing 100081, p.r. china 6 college of agriculture, rizal memorial colleges inc., davao city 8000 philippines 7 department of life sciences, institute for plant biology, technical university of braunschweig, 38106, braunschweig, germany perspective on the influence of ultraviolet radiations in plant growth and development introduction light is one of the most critical environmental parameters regulating plant growth and development. photomorphogenesis and photosynthesis are subjected to light intensity, photoperiod, and spectral composition (paradiso, proietti, 2021). compared with light intensity and photoperiod, the impacts of light spectra on modulating plant growth and development are more intricate. among these different light spectra, blue and red lights are highly absorbed by chlorophyll in leaves, thus driving photosynthesis (taiz et al., 2014). however, increasing studies proved that, besides blue and red light, other light spectrums such as ultraviolet (uv) radiation with a wavelength of 200-400 nm critically influence terrestrial plants as well (verdaguer et al., 2017). in nature, uv radiation comprises three wavebands (uv-a: 315-400 nm, uv-b: 280-315 nm, uv-c: 200-280 nm) (blaustein, searle, 2013; cie, 2020) – fig. 1. fig. 1. the penetration of uv-a and uv-b rays to the earth (source: www.uvdi.com; modified) many studies have centred on plant responses to uv-b and extensive data are now available on the regulatory responses mediated by the uv-b photoreceptor uvr8 (bernula et al., 2017; coffey et al., 2017), as well as the retarding impact of uv-b on plant growth (liu et al., 2011; huché-thélier et al., 2016; vanhaelewyn et al., 2020). therefore, changes in plant chemistry, such as increased accumulation of flavonoids with high affinity to uv-b and altered epicuticular wax composition, exemplify plants’ acclimation responses to uv-b exposure (gonzalez et al., 1996; kakani et al., 2003; bassman, 2004). despite the widespread distribution of uv-a on the earth and its significant effects on terrestrial plants, consolidated information on plant responses to uv-a is limited (chen et al., 2019; rai et al., 2021). additionally, no definite evidence is available regarding its photoreceptor (verdaguer et al., 2017), and insofar as the ongoing experiments’ results have shown the blue photoreceptors phototropins and cryptochromes are potential candidates for uv-a absorption (casal, 2013; wang et al., 2020). plants adjust their physiological and morphological processes in response to altering light conditions, which may suppress or stimulate plant growth (hogewoning et al., 2010, 2012). many studies have shown the acceleration of plant growth and modification of plant morphology in response to uv-a radiation (tezuka et al., 1993; victório et al., 2011; chen et al., 2019). for example, plants grown under supplemental uv-a exhibited a larger leaf area index (antonelli et al., 1997), increased rosette diameter (biswas, jansen, 2012; schulz et al., 2021), and a higher biomass accumulation (lee et al., 2014; bernal et al., 2015). the uv-a-induced morphological changes may be attributed to blue/uv-a absorbing cryptochrome, which plays a pivotal role in shade avoidance (keller et al., 2011). by contrast, some studies reported no significant responses or adverse effects (e.g., prevention of biomass accumulation, leaf expansion, and stem elongation) under the presence of uv-a radiation (tsormpatsidis et al., 2008; baroniya et al., 2013); this is likely due to uv-a induced stress (kreslavski et al., 2013a; vanhaelewyn et al., 2020). in this context, the direction of plant growth in response to uv-a is not constant. furthermore, available data do not allow the effect of the uv-a dosage to be determined because of uncertain information on the fluency rates of uv-a radiation involved (verdaguer et al., 2017). therefore, it is still unknown whether the growth of plants exhibits a dosage response under uv-a radiation. photosynthesis is the ultimate basis for plant growth (taize et al., 2014). conventionally, the uv-a component of sunlight has been considered to be detrimental to photosynthesis, as indicated by a decrease in maximum quantum efficiency of psii photochemistry (fv/fm), gas exchange rates, and electron transport rate (turcsányi, vass, 2000; nayak et al., 2003; kreslavski et al., 2013b). on the other hand, uv-a inclusion generally escalated the photosynthesis rate (tezuka et al., 1994; turnbull et al., 2013). also, under non-saturating (low) light conditions, uv-a can enhance photosynthesis rates (palma et al., 2021). for instance, uv-a increased the photosynthesis rate by 8-10% in poa annua l., sorghum halepense (l.) pers., and nerium oleander l. under the background photosynthetic active radiation (par) intensity of 500 μmol m -2 s -1 (mantha et al., 2001). in this aspect, supplemental uv-a radiation is expected to enhance carbon assimilation in indoor cultivation systems where the background par is often far below the light saturation point. nevertheless, it is still unknown how uv-a affects the photosynthesis system of individual plants in a controlled environment. recently, the rapid development of plant factories with artificial light (pfal) indicates that various kinds of artificial light procedures are useful during crop cultivation and could affect human development along the feeding pathways (kato et al., 2010). on the other hand, the vast majority of research on lighting in plant factories has been published with only red and blue radiations (hernández, kubota, 2016; reference therein; trouwborst et al., 2016). moreover, the ratio of par–uv-a, which plays an essential role in the responses of plants to uv-a exposure (krizek, 2004), is prone to differ to a great extent according to how a plant factory was designed. in retrospect, it remained unknown whether additional uv-a radiation would benefit this indoor cultivation system. as a result, we attempted to provide information on the effect of supplemental uv-a radiation on plant growth and physiology in a controlled environment in this review. ultraviolet (uv) radiations ultraviolet radiations have been spotlighted because of their vital role in depleting the stratospheric ozone layer, which increases its level on the ground (rowland, 2006; bernhard et al., 2010). consequently, tremendous research efforts have been focused on elucidating the effect of the depleted ozone layer (i.e., increased uv-b level) on plant physiology and agriculture. primarily, studies were conducted in a chamber, in vitro, and in a location that can irradiate high uv-b levels to determine uv-b’s damage to plants. accordingly, it was demonstrated that uv radiations reduce the growth and yield (teramura, sullivan, 1994; caldwell et al., 1998; kakani et al., 2003a; gao et al., 2004), destroying cell microstructure (hollósy, 2002) causing damage, and impairment to photosynthesis processes (kataria et al., 2014). however, many preceding studies overlooked whether uv-b dosage in nature is harmful enough to underlie the undesirable consequences shown in the chamber studies. as a case in point, it was revealed that the reduction in plant growth by increased uv-b radiation in areas affected by ozone decline, which is the leading cause of an increase in uv level, since 1980 it is unlikely to have gone beyond 6% (ballaré et al., 2011). moreover, an increasing number of researchers are arguing that uv cannot be classified in the category of detrimental plant rays (jansen, bornman, 2012), and uv can be exploited in agricultural lands (wargent, jordan, 2013; thomas, puthur 2017). although montreal protocol effectively suppressed the emission of ozone-depleting substances and several factors such as cloud and aerosol changes decreased the uv level on the surface these days (neale et al., 2021), living organisms on the earth are still exposed to higher levels of uv than before. hence, a need to examine the influence of uv on plants and their coping strategy under uv-induced stresses is required. effects of uv on plant growth biomass accumulation usually, uv-b exposure negatively affects plant growth, developmental processes, and morphology (zuk-golaszewska et al., 2003; cechin et al., 2007; salama et al., 2011; chaudhary, agrawal, 2015; dotto, casati, 2017). for example, it was found that the total dry weight of four cucumber cultivars (krizek et al., 1997), the fresh and dry weight of soybean plants (zhang et al., 2014; skórska et al., 2019), and aerial dry mass of lettuce (tsormpatsidis et al., 2008) were significantly decreased by uv-b in field attenuation studies. an apparent disparity in growth parameters was observed among rice varieties after exposure to uv-b (hidema, kumagai, 2006; faseela, puthur, 2019). field enhancement studies have shown that increased uv-b dose can decrease the total dry weight of all wild-type arabidopsis sp. examined (cooley et al., 2001) and the total biomass of coleus forskohlii briq (takshak, agrawal, 2015), regardless of sampling stages. the same results were observed in soybeans cultivated in a controlled environment with ambient and overall lesser uv-b doses (biggs et al., 1981). in addition, an examination of the effect of uv dose on the above-ground dry weight of lettuce revealed that as the uv wavelength decreases, the synthesis of biomass is strongly inhibited (tsormpatsidis et al., 2008). similarly, longer exposure to some uv-a wavelengths has been linked to an increase in biomass in red-leaf lettuces (samuoliene et al., 2020). contrary to uv-b, which strongly interferes with the accumulation of photosynthates, uv-a tends to be weaker and equivocal; some plants had decreased biomass due to uv-a (xiong et al., 2001; kataria, guruprasad, 2012a), but others remained the same (zhang et al., 2014) or increased (kataria et al., 2013; lee et al., 2014). although contrasting results limit interpretation, several literature sources confirmed that genetic aspects such as cultivar and ecotype play a critical role in a plant’s sensitivity. in a study where parameters are compared by monocots and dicots or by c3 and c4 species, the effects of uv-a were by and large driven by whether a plant is a monocot or dicot, rather than the photosynthetic mechanisms (kataria et al., 2013). in a study by häder (1996) who compared the german wheat cultivar and chilean cultivars grown at high altitudes where uv level is escalated, it turned out that changes in light conditions led to a considerable reduction in the root and leaf dry weights of the german cultivar, implying the presence of a genetic factor that enables uv resistance. leaf area/size there are already abundant reports showing that leaf area, size, or structure is decreased by uv-b (barnes et al., 1996; nogues et al., 1998; cooley et al., 2001; kakani et al., 2003b; wargent et al., 2009; kataria, guruprasad, 2012b; klem et al., 2012; zhang et al., 2012; fina et al., 2017; rai, agrawal, 2017; rana, abdulkarim, 2021). with the addition of uv-b rays, reduction in leaf size was mainly associated with the reduced epidermal cell size (yang, yao, 2008; jacque et al., 2011), increased cell wall peroxidase activity, and collateral decrease in the number of cells (wargent et al., 2009). the effect of uv-b on the leaves of arabidopsis sp. plants was highly attributed to light status after uv treatment (sztatelman et al., 2015). these authors showed that leaves left in darkness after uv-b treatment had substantially higher chlorophyll content and photosynthetic efficiency than those kept in constant light. another study, also observed that irradiation of light-emitting diodes substantially increased chlorophyll a and b content, leaf area, and overall growth in lettuce (miliauskienė et al., 2021). lately, it was reported that the reduction in leaf expansion in uv-b-irradiated leaves is imputed to the decline in cell production and shortened growth zone (fina et al., 2017). recent studies revealed that uv-a has frequently increased the area and size of leaves (victório et al., 2011; zhang et al., 2012; kataria et al., 2013; khoshimkhujaev et al., 2014; zhang et al., 2014), as opposed to uv-b that inhibited leaf expansion. in bean plants, supplemental uv-a stimulated the elongation of leaves and, consequently, resulted in a larger leaf area (antonelli et al., 1997). this phenomenon is thought to be the upregulation of cryptochrome implicated in shade avoidance, as the absence of blue light led to the reduction in leaf area and the ratio of the lamina to petiole in arabidopsis (keller et al., 2011). in a study by brazaitytė et al. (2009), it was identified that uv-a light influenced an enlargement of leaf area and the amount of secondary metabolites such as phenols, α-tocopherol, and β-carotene in mustard microgreens. nevertheless, some researchers also reported that the differences in species or genotypes within the same species resulted in various responses such as reduction (krizek et al., 1997; qian et al., 2021) and no changes in leaf size (cooley et al., 2001; sullivan et al., 2003). thus, deeper insight into the relationship between cryptochrome and uv-a-induced leaf expansion is needed, which would enable the better exploitation of uv-a by the crop. effects of uv on plant physiology it’s not hard to find reports of uv-b radiations resulting in reduced photosynthesis rate (teramura, sullivan, 1994; nogues et al., 1998; joshi et al., 2007; kataria, guruprasad, 2012a; takshak, agrawal, 2015; zhang et al., 2016). other deleterious effects of uv-b radiation are oxidative stress-induced photosynthetic pigments impairment (kataria et al., 2014; kataria, 2017), secondary metabolite variations (escobar-bravo et al., 2017), changes in cuticular wax deposition and anatomical characteristics (nascimento et al., 2015; willick et al., 2018), extensive harm to photosystem i (psi) and ii (psii) proteins and disturbance in the electron transport chain (zhang et al., 2016; rai et al., 2018). notably, uv-b has the potential to reduce the maximum quantum yield of psii (φpsii max: fv/fm), an indicator of the health of the photosynthesis system in higher plants (keiller et al., 2003; pfündel, 2003; lee et al., 2014; zhang et al., 2016). this is thought as the consequence of photosynthetic components such as photosystem ii (tyystjärvi, 2008; dobrikova et al., 2013; kataria et al., 2014) and enzymes involved (imbrie, murphy, 1982; yu et al., 2013; köhler et al., 2017). additionally, an appreciable decline in stomatal conductance (gs) (nogués et al., 1999; tossi et al., 2014) and production of reactive oxygen species (ros) if exposure to uv-b acted as a distress factor in plants (hideg et al., 2013) were reported as well. the increase in ros generation in response to uv radiation has been attributed to the disruption of metabolic activities and escalated activity of membrane-localized nadph-oxidase (nawkar et al., 2013). teramur and salliva (1994) and kataria et al. (2014) have provided more detailed information about uv-b-caused deterioration of photosynthesis processes. there are also some indications that augmented co2 may alleviate the damaging consequences of uv-b-exposed plants by providing photosynthate and other photoprotective benefits (qaderi et al., 2005; koti et al., 2007; wargent, jordan, 2013). although some preceding in vitro uv-a research indicated that uv-a enhanced photosynthetic activities, there are many opposite cases in that uv-a hindered photosynthesis (mantha et al., 2001; joshi et al., 2013; bernal et al., 2015). the mechanisms underlying increased stimulation of photosynthesis are: a) direct absorption of uv-a by chlorophylls and carotenoids (buschmann, lichtenthaler, 1998), b) indirect absorption of uv-a-induced fluorescence emitted by phenolic compounds (mantha et al., 2001; johnson, day, 2002), c) stomatal opening by cryptochromes that perceived the fluorescence (mantha et al., 2001). other alterations corresponding to uv-a-enhanced photosynthesis are increased rubisco activity in wheat, cotton, and sorghum (kataria et al., 2013), the higher performance index of photosynthesis (piabs) of red lettuce (lollo rosso) (tsormpatsidis et al., 2008), and better yield of electron transport per trapped exciton (et0/tr0) and piabs in maize plants (shine, guruprasad, 2012). interestingly, uv-a was found to have the potential to protect plants from uv-b; when uv-a and uv-b were irradiated together, barley seedlings cultured in weak photosynthetically active radiation exhibited better fv/fm and quantum yield of regulated light-induced thermal energy dissipation (φnpq) than ones with uv-b only (štroch et al., 2015). in cluster bean plants (cyamopsis tetragonoloba (l.) taub), the ratio of fv/fm and oxygen evolution reaction activity that uv-b impaired was restored when uv-a was simultaneously irradiated with uv-b (gartia et al., 2003; joshi et al., 2007). the interaction of other stress factors with uv as a regulator as the perspective for uv radiation often varies from a stressor to a regulator of plant development, extensive attempts are being made to manipulate uv (wargent, jordan, 2013; kataria et al., 2017; thomas, puthur, 2017). one of these attempts, for example, is to ‘vaccinate’ plants with uv radiation to induce cross-resistance to other stress factors (loconsole, santamaria, 2021). the interaction between uv and three -representative abiotic stresses – high light, temperature, and waterwill be discussed in the following. high light although uv implemented in studies mentioned below appears to have restrained quantitative growth and retarded photosynthetic capacities in plants, uv radiations were proven to be a successful primer to high light stress (huang et al., 2019). one of the most effective approaches to efficiently use energy by excess light is the xanthophyll cycle (demmig-adams, adams, 1996; latowski et al., 2011). in barley plants, a bigger pool of xanthophyll cycle pigments and higher flavonoid content diminished the decreases in light-saturated co2 assimilation rate (amaxmaximum rate of photosynthesis) and fv/fm (klem et al., 2015). nevertheless, the xanthophyll cycle barely contributed to the photoprotection in rice upon exposure to high light (zhao et al., 2017); instead, d1 protein turnover in uv-b-treated leaves was proven as the protective mechanism the plants exhibited due to higher fv/fm values (olsson et al., 2000). in their study on tomato plants, tao et al. (2021) showed that d1 turnover drives a more critical function than the xanthophyll cycle in photoprotection under sub-high temperature and high light conditions. apart from the protein turnover, uv-b also increased amax, which is believed to function as the combination of elevated concentrations of chlorophyll and uv-absorbing compounds (uac) (xu, qiu, 2007). poulson et al. (2006) found that arabidopsis plants cultivated under uv-b light effectively coped with high light stress; uv-b reduced the loss of photosynthetic capacities expressed by amax, gs, fv/fm, and transpiration rate (e). interestingly, this was because uv-b-irradiated seedlings had lower фpsii (an indicator of photoinhibition) and higher non-photochemical quenching (npq), which lessened non-recoverable photoinhibition (lal, edwards, 1996; niyogi, 1999). water compared to plants subjected to single stress treatments, the combination of two stresses resulted in less reduction in plant biomass (bernal et al., 2013, 2015), cuticle thickness (drilas et al., 1997), needle losses in conifers (petropoulou et al., 1995), and also helped to sustain photosynthetic apparatuses and needle water relations (manetas et al., 1997). in several research projects, concurrent uv treatments increased water use efficiency (wue) and water contents under drought stress. as a case in point, uv-a functioned a critical role in increasing leaf relative water content (rwc) and leaf wue when laurus nobilis l. plants were subjected to escalated uv-a dose and reduced water supply (bernal et al., 2015). plant photomorphogenic responses to destructive uv-b radiation might be assumed as an adaptative mechanism under conditions akin to high-light environments, such as water stress (gitz, liu-gits, 2003). not to mention that uv-b’s presence under drought helped maintain a better water economy in various species as well (manetas et al., 1997; zhao et al., 2009; bandurska, cieślak, 2013). this phenomenon was primarily accompanied by thicker leaves (drilias et al., 1997), lower gs (poulson et al., 2002), and increased production of substances such as osmolytes (schmidt et al., 2000), hormones (bandurska, cieślak, 2013), and antioxidant enzymes (balakumar et al., 1993) which facilitated the adaptation of plants to the extreme environmental conditions. temperature despite the limited number of research performed, some molecular-based results have been obtained revealing that uv-b robustly weakens thermomorphogenesis (i.e., stem elongation), which shows symptoms similar to shade avoidance signaling, elicited by high temperature (favory et al., 2009; hayes et al., 2017; tavridou et al., 2020). apart from these molecular proofs, some reports prove amelioration in temperature damage in seedlings receiving uv-b. for instance, young conifer seedlings grown under additional uv-b were more tolerant both to chilling and heating stresses and maintained higher fv/fm values compared to those exposed to ambient level uv-b (l’hirondelle, binder, 2005). a similar increase in fv/fm was also observed in lettuce seedlings in an experiment simulating the heat and high par radiation stressors caused by nursery-to-greenhouse transfer (wargent et al., 2011). moreover, seedlings pre-acclimatized to uv-b inclusive environment showed relatively higher photosynthetic and growth rates than those pre-acclimatized to uv-b depleted environments (wargent et al., 2011; müller et al., 2013). when cucumber seedlings were cultivated in a growth chamber supplemented with uv-b radiation, more seedlings survived heat stress and were taller than those that did not receive uv-b. such a phenomenon was followed by no alterations in the level of malonaldehyde (mda) and by a lower membrane injury index in cucumber plants under uv-b exposure (teklemariam, blake, 2003). in a study by neugart et al. (2014), it was found that the combined effect of moderate uv-b (0.75 kj m–2 d-1) and temperature (15°c) substantially increased the mrna expression of flavonol 3′hydroxylase in kale (brassica oleracea var. sabellica). lastly, in soybean plants grown under high temperatures, electrolyte leakage, expressed as relative injury to tissue in this research, was also alleviated by additional uv-b radiation (koti et al., 2007). furthermore, this uv-b supplementation reduced the net co2 assimilation (an) by heat stress. conclusion as a crucial constituent of light, ultraviolet radiation (uv) has a significant impact on plant general growth and development in many different ways. uv-b, a component of uv light, has been strongly linked to plant growth interference through decreasing biomass accumulation, photosynthesis impairment, and plant physiology disruption (tab. 1). on the other hand, another light component (uv-a – tab. 2) has been shown to upregulate the plant’s photosynthetic process, increase its resistance to several environmental stresses and have the potential of protecting the plant against the harmful effect of uv-b when illuminated together. furthermore, it can be concluded that cross-talking these two important light components can enhance water efficiency and alleviate problems linked to thermal stress. thus, utilizing the importance of ultraviolet radiations, particularly uv-a and uv-b, could assist in promoting plant growth and development, particularly in an enclosed environment. conflict of interest the authors declare no conflict of interest related to this article. references antonelli, f., grifoni, d., sabatini, f., zipoli, g. 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(2003). the effect of uv-b radiation on plant growth and development. plant, soil and environment, 49(3), 135–140. https://doi.org/10.17221/4103-pse perspektywy wpływu promieniowania ultrafioletowego na wzrost i rozwój roślin streszczenie coraz więcej dowodów wskazuje na wielowymiarową rolę promieniowania ultrafioletowego we wzroście i rozwoju roślin. ultrafiolet-a (uv-a) jako główny składnik promieniowania uv w przyrodzie, ma szeroki zakres wpływu na rośliny. ponieważ istnieją pewne dowody wskazujące na jego pozytywne działanie jako regulatora rozwoju roślin w kontrolowanym środowisku, od dawna obserwuje się zainteresowanie wykorzystaniem uv-a w rolnictwie. jednak rola uv-a we wzroście roślin pozostaje w dużej mierze nieznana, a liczba badań zajmujących się wpływem uv-a w kontrolowanym środowisku jest nadal zbyt mała. przeprowadzony tu przegląd ilustruje również wpływ innych składników uv na wzrost, morfologię i fizjologię roślin, takich jak: uv-b, a także wzajemne oddziaływanie innych stresów abiotycznych i uv-a, jako regulatorów wzrostu. na podstawie przedstawionej tu syntezy można stwierdzić m.in., że niższe dawki promieniowania uv-a mogą stymulować wzrost roślin w kontrolowanym środowisku. być może przyczyni się to już wkrótce do optymalizacji receptury uprawy roślin w pomieszczeniach. słowa kluczowe: akumulacja biomasy, fizjologia roślin, czynniki stresowe, promieniowanie słoneczne received: [2022.08.01] accepted: [2022.11.05] abbreviations amax – maximum rate of photosynthesis; e – transportation rate; uv – ultraviolet radiations; ps – photosystem, par – photosynthetic active radiation; ros – reactive oxygen species, nadph – nicotinamide adenine dinucleotide phosphate; uac – uv-absorbing compounds, wue – water use efficiency, rwc – relative water content, mda – malonaldehyde, mrna – messenger ribonucleic acid. appendix 1 tab. 1. comparison of the neutral, positive and negative effects of ultraviolet light (uv-a: 315–400 nm) on plant development; the table presents sample studies no. succ. plant parameters the way of impact neutral positive negative 1. leaf area indicators cooley et al., 2001; sullivan et al., 2003; tsormpatsidis et al., 2008; baroniya et al., 2013 antonelli et al., 1997; victório et al., 2011; zhang et al., 2012; kataria et al., 2013; khoshimkhujaev et al., 2014; zhang et al., 2014 krizek et al., 1997; qian et al., 2021 2. leaves rosette diameter biswas, jansen, 2012, schulz et al., 2021 3. biomass tsormpatsidis et al.., 2008, baroniya et al., 2013; zhang et al., 2014 kataria et al., 2013; lee et al., 2014; bernal et al., 2015 xiong et al., 2001; kataria, guruprasad, 2012a 4. stem elongation tsormpatsidis et al., 2008; baroniya et al. 2013 5. photosynthesis efficiency tezuka et al., 1994; buschmann, lichtenthaler, 1998; mantha et al., 2001; johnson, day, 2002; kataria et al., 2013; turnbull et al., 2013; palma et al., 2021 turcsányi, vass, 2000; mantha et al., 2001; nayak et al., 2003; joshi et al., 2013; kreslavski et al., 2013b; bernal et al., 2015 a. maximum psii quantum efficiency (fv/fm) gartia et al., 2003; joshi et al., 2007; štroch et al., 2015 b. gas exchange rate nayak et al., 2003 c. electron transport rate shine, guruprasad, 2012 6. water use efficiency (wue) bernal et al., 2015 7. relative leaf water content (rwc) bernal et al., 2015 tab. 2. comparison of the positive and negative effects of ultraviolet light (uv-b: 280–315 nm) on plant development; the table presents sample studies no. succ. plant parameters positive under water or temperature stress negative 1. leaf area indicators barnes et al., 1996, nogues et al., 1998, cooley et al., 2001, kakani et al., 2003b, yang, yao, 2008, wargent et al., 2009, jacque et al., 2011, kataria, guruprasad, 2012b, klem et al., 2012, zhang et al., 2012, fina et al., 2017, rai, agrawal, 2017, rana, abdulkarim, 2021 2. stem elongation favory et al., 2009, hayes et al., 2017, tavridou et al., 2020 3. biomass biggs et al., 1981, krizek et al., 1997, cooley et al., 2001, hidema, kumagai, 2006, tsormpatsidis et al., 2008, zhang et al., 2014, takshak, agrawal, 2015, faseela, puthur, 2019, skórska et al., 2019, 4. photosynthesis efficiency wargent et al., 2011; müller et al., 2013, sztatelman et al., 2015; teramura, sullivan, 1994, nogues et al., 1998, joshi et al., 2007, kataria, guruprasad, 2012a, kataria et al., 2014, takshak, agrawal, 2015, zhang et al., 2016 a. chlorophyll content sztatelman et al., 2015 kataria et al., 2014, kataria, 2017 b. maximum psii quantum efficiency (fv/fm) l’hirondelle, binder, 2005; wargent et al., 2011; müller et al., 2013 keiller et al., 2003, pfündel, 2003, tyystjärvi, 2008, dobrikova et al., 2013, lee et al., 2014 kataria et al., 2014, zhang et al., 2016, rai et al., 2018 c. electron transport rate zhang et al., 2016, rai et al., 2018 5. stomatal conductance nogués et al., 1999, tossi et al., 2014 6. water management manetas et al., 1997; zhao et al., 2009; bandurska, cieślak, 2013 226 annales universitatis paedagogicae cracoviensis studia naturae, 6: 226–250, 2021, issn 2543-8832 doi: 10.24917/25438832.6.13 peiman zandi1,2*, joanna puła3, xing xia2, elke bloem4, aminu darma2, yaosheng wang2, ingrid turisová5, qian li6, luu ngoc sinh7, na li2 1international faculty of applied technology, yibin university, yibin 644000, p.r. china; z_rice_b@yahoo.com 2institute of environment and sustainable development in agriculture, chinese academy of agricultural sciences, beijing 100081, p.r. china 3department of agroecology and crop production, faculty of agriculture and economics, university of agriculture, mickiewicza 21 ave, 31-120 krakow, poland 4institute for crop and soil science julius kühn-institut, federal research centre for cultivated plants, bundesallee 69, 38116 braunschweig, germany 5department of biology and ecology, faculty of natural sciences, matej bel university in banská bystrica, tajovského 40, banská bystrica 974 01, slovakia 6institute of vegetables and flowers, chinese academy of agricultural sciences, beijing 100081, p.r. china 7faculty of sciences and technology, hanoi metropolitan university. 98 duong quang ham, cau giay, hanoi, vietnam more insight into the concept of iron plaque formation and its characteristics in rice (oryza sativa l.) introduction rice (oryza sativa l.) is the second most widely cultivated cereal crop, and around half of the world’s population depends on rice consumption (bazrkar khatibani et al., 2019). however, the enhanced concentration of toxic metal(loids) in paddy soil due to anthropogenic activities has stimulated their increased accumulation in rice (norton et al., 2014; clemens, ma, 2016), posing a signi�cant negative impact on human health via the food chain pollution. as a typical semi-wetland plant, rice is characterised by releasing oxygen (o2) from the root surface in the rhizosphere when growing in a saturated or anaerobic environment; this induces the oxidation of ferrous (fe2+) to ferric (fe3+) and subsequently their precipitates as fe oxides on the outermost cell layers of the root (sundby et al., 1998; hu et al., 2014). iron oxide deposits are known as irregular porous coatings on the roots of hydrophytes (tripathi et al., 2014). according to the literature reviewed by khan et al. (2016), several studies have analysed the iron plaque (ip) deposits by employing various techniques, including x-ray di�raction and energy-dispersive x-ray microanalysis. �ese deposits consisted mainly of a mixture of lepidocrocite [g-feo(oh)], goethite [α-feo(oh)], and ferric phosphate (fepo4), in which manganese (mn) and fe usually 227 co-occur (co-precipitate/adsorb). under laboratory conditions, bacha and hossner (1977) reported a ratio of fe (43): mn (1) in the ip of rice roots. either amorphous or crystalline in structure, ip is composed dominantly of 63% ferrihydrite [(fe3+)2o3 × 0.5 h2o], 32% goethite, and 5% of siderite (feco3) (hansel et al., 2001). �e latter is only formed on microsites where co2 concentration from respiration is high due to insu�cient eèux (hansel et al., 2001). sey�erth et al. (2010) showed that ips are not evenly distributed on rice roots. predominantly mature roots show a high level of ip, while thin immature roots show minimum ip formation (sey�erth et al., 2010, 2011). in contrast, the lowest and highest incorporation rate of rice root sections into ip coating from (oxyhydr) oxides (ferrihydrite 81–100%) sources were attributed earlier to the root base and root tips, respectively (liu et al., 2006). fe hydroxides’ mineralogy in ip of rice root showed that ferrihydrite was the main constituent of ip (sey�erth et al., 2011). frommer et al. (2011) showed that the accumulation of mn and fe highly depends on the root thickness. based on their observation, fe accumulation near �ne immature roots (< 100 µm) was more discernible than at tick mature roots (⁓500 µm) compared with mn. a  recent study on primary roots by zandi et al. (2020) showed that root surface plaque deposition was predominantly built of fe, regardless of root sections. �e unique accumulation pattern of ips was later attributed to fe’s kinetics and thermodynamics, mn redox transformations, and the extent of root aeration (khan et al., 2016). �e disparities in observations may emanate from genotypic di�erences in the roots’ oxidising capacities and the biogeochemical conditions used in these studies. root ip may also contain a variety of other metals and metalloids such as aluminium (al), arsenic (as), cadmium (cd), chromium (cr), mercury (hg), nickel (ni) and lead (pb) (tripathi et al., 2014). �is feature can indicate the involvement of ip in the adsorption of both anions and cations (liu et al., 2004a; yang et al., 2020) and thus introduce it as an essential barrier for heavy metal uptake and accumulation (hossain et al., 2009a, b; hu et al., 2014; sun et al., 2016; xia et al., 2020; zandi et al., 2020). it should be pointed out that sequestration mechanisms of heavy metals on ip remain mostly unknown. growing evidence suggested that dissolved organic carbon (doc) and sulphur (s) could participate in the precipitation of fe hydroxides on the root surface (liu, huang, 2003; sun et al., 2016; yang et al., 2016). �ey probably modify fe hydroxide’s surface properties and molecular structures in ip, thus in�uencing the binding mechanisms of heavy metals to ip. �e speciation of s has also been shown to a�ect and improve the barrier feature of ip against cr uptake and accumulation in rice roots (zandi et al., 2020). furthermore, a�er being reduced in upper plant parts, s by-products can chelate cr and cd, and thereby actively reduce their mobility and toxicity in roots and shoots (zhang et al., 2013; cao et al., 2018; yamazaki et al., 2018; zandi et al., 2021). among di�erent biotic or abiotic factors involved in ip formation and development, root-released o2 is speculated to show a strong involvement (khan m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 228 et al., 2016; huang et al., 2020). �e physical barrier of ip has been ascribed to the proportion of roots coated by iron oxides (xia et al., 2020), the composition type of iron oxides (amaral et al., 2017) and the ability to develop new roots (zandi et al., 2021). �is review primarily aimed at providing more insight into the intrinsic features of iron oxide plaques, the in�uencing factors on their root surface build-up, and their key role in growth parameters and heavy metal sequestration and immobilisation in rice as a semi-wetland plant. characterisation of iron plaque wetlands sediments are composed of iron (fe) elements. ferrous ion (fe2+) is a unique species in anoxic sediment pore water capable of donating electrons (tripathi et al., 2014). ferric ion (fe3+) is an electron acceptor fe species available dominantly in oxic pore water. in addition to the fe plaque, mn plaque may also be formed on hydrophyte roots through oxidisation of mn2+ by root-released o2 (hu et al., 2007). �e formation process of ip (fig. 1) primarily refers to the following chemical reactions (amaral et al., 2017): fe3++ e− (reducing factors in the bulk/rhizosphere soil) fe2+ (1) fe2+ + o2 (released/di�used from root) fe3+ (iron plaque) (2) mn2++ o2 (released/di�used from root) mn4+ (manganese plaque) (3) �e concentration gradient is an essential factor in triggering the depletion of fe2+ from bulk sediment and its further movement into the hydrophyte rhizosphere. in the rhizosphere, root-released o2 oxidises fe 2+ to fe3+ that subsequently precipitates as feooh (or iron oxyhydroxide) and accumulates on the root surface of hydrophyte and soil particles as ip (fig. 1) (hu et al., 2007; yang et al., 2014). �e interior root penetration of iron deposition (plaque distribution) varies among species. �ese di�erences in penetration may result from the dissimilar potential of various species in oxidising rhizosphere because of their variant characteristics of root radial oxygen loss (rol), root respiration and/or variance in the extent of soil oxygen demands (st-cyr, crowder, 1989). inner parts of rice roots viz. epidermis, hypodermis and exodermis coated with iron spots/stains, were detected so far (pereira et al., 2014). internal iron hydroxide precipitation is possible at low rol. in addition to the typical root plaque formation, shoot bases and rhizomes of hydrophytes also can partially be covered by ip (povidisa et al., 2009). 229 �e conception of iron plaque formation: in�uencing factors �e formation of ip occurs during the oxidation of ferrous (fe2+), formed during periods of anaerobic soil conditions, to ferric (fe3+) iron (or iron oxyhydroxides) and the precipitation of resultant ferric oxide on the outer surface of roots (khan et al., 2016). many physicochemical characteristics of soils and/or sediments including organic matter, soil texture, soil redox potential (eh) (syu et al., 2013), soil ph (zhang et al., 2019a), soil water management (zhang et al., 2019b), fe/ mn availability (shi et al., 2004), phosphorus (p) (hu et al., 2005), selenium (se) (huang et al., 2020), as (lee et al., 2013) and s supply (hu et al., 2007), root-released o2 (radial oxygen loss-rol) (huang et al., 2020), root exudates (becker, asch, 2005; wu et al., 2014), root enzymatic activates (lee fig. 1. iron plaque formation in oryza sativa l.; a − roots (cv. xiangzaoxian 31) and rhizosphere; b1–3 – roots of di�erent rice cultivars (b1 – cv. hashemi, b2 – cv. lemont, b3 – cv. xiangzaoxian 31); c1–2 – roots of rice seedlings cv. xiangzaoxian 31 (upper row) and cv. hashemi (lower row) before and a�er iron plaque induction, respectively, under laboratory conditions; d1–3 − roots of rice seedlings cv. kazemi (d1), cv. hashemi (d2) and cv. xiangzaoxian 31 (d3) a�er harvest (photo. p. zandi and x. xia) m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 230 et al., 2007), microbial activities (neubauer et al., 2007; references therein; huang et al., 2012a), plant genotypes (syu et al., 2014) and plant age a�ect ip formation (fig. 2). �e formation of ip may arise (or result) from neutrophilic fe2+ oxidising bacteria (neubauer et al., 2007, 2008). �e discovery of both lithotrophic fe-oxidising bacteria (feob) and fe-reducing bacteria (ferb) (king, garey, 1999) on the root surface of many wetland plants indicate that plaque-associated microbes may directly in�uence ip formation. �e feob could contribute substantially to the formation of ip (neubauer et al., 2007, 2008). such bacteria have been shown to account for over 40–60% of fe (iii) mineral precipitation on roots of wetland hydrophytes under microoxic (⁓5–30 μm o2) conditions (neubauer et al., 2002; maisch et al., 2019). �ere is a general agreement that the reductive dissolution of ip results from rice root decay mediated by ferb (wang et al., 2009; huang et al., 2012a). root surface microbial fe (iii) reduction o�en results in rapid decomposition and subsequent release of crystalline iron minerals and their associated toxic metal(loids) to the immediate rhizosphere (weiss et al., 2004, 2005). �is event usually leads to some notable environmental problems. while biological oxidation of fe2+ may occur naturally, its biotic oxidation from rol is likely to be of greater signi�cance (tripathi et al., 2014; huang et al., 2020). �e local conditions dramatically impact the extent and importance of biological oxidation at the root interface. based on the heterogeneous distribution of soil oxygen due to microbial behaviour, root distribution, and soil morphology, paddy soils’ eh and/or conditions vary (yamaguchi et al., 2014). maisch et al. (2019) indicated that rol central role in ferric iron mineral formation is not only limited to the root surface but also to the whole soil matrix, where it expands potential microoxic living niches for microaerophilic fe(ii) oxidisers (microfeox) throughout the entire rhizosphere. soil moisture regime has been suggested to a�ect the amount of iron plaque formed on rice roots (liu et al., 2010). �is amount was found much lower under lower soil moisture contents than under submerged conditions (chen et al., 2008). continuous �ooding conditions in paddy �elds signi�cantly increase the abundance of iron-reducing bacteria (e.g., latescibacteria, desulfuromonadales, and geobacteraceae) under anaerobic rhizospheric conditions, leading to less ip formation around rice roots (zhang et al., 2019b). however, because the availability of cd, zinc (zn), copper (cu), and other metals in the rhizosphere has decreased, the absence or reduced formation of root plaques cannot promote their presence in rice grains (xu et al., 2013; eduardo et al., 2014; zhang et al., 2019b). in a continuously �ooded culture, changes in the soil ph (increase) and eh (decrease) tend to increase the formation of hydrous oxides of fe, mn, aluminium (al) and other metals in �ooded soils with a high a�nity for metal adsorption on them (more immobilisation), leaving less soil-available metal concentration for uptake in plant roots (eduardo et al., 2014; li et al., 2015). 231 fig. 2. comparison of factors a�ecting iron plaque formation in rhizosphere where rice roots environment is located; +, – and +/– are respectively indicative of positive, negative or conditional impacts on iron plaque formation; more explanations are in the text (courtesy of word diagram: p. zandi) elemental s (s0) could induce the formation of fe and mn plaques on the root surface and mineral particles in the rhizosphere (hue et al., 2007; yang et al., 2014). on the other hand, sulphate (so4 2-) minerals signi�cantly impacted ip’s formation and barrier function compared to s0 minerals (hu et al., 2007). di�erent concentrations of fe plaque were formed on rice roots depending on the amount of s content in soil m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 232 solution (fan et al., 2010; sun et al., 2016). a su�cient s supply has been suggested to play a central role in iron oxide formation and suppression of potentially toxic metals uptake and bioavailability in rice (hu et al., 2007; khan et al., 2016; sun et al., 2016; yang et al., 2016; zandi et al., 2021). excessive addition of s supply reduced the proportion of oxidised iron minerals through extra rhizosphere fe2+ depletion via binding with monosul�de (s2-) to form pyrite (fes/fes2) minerals. �e resultant precipitates of fes/fes2 in bulk soil have been reported to diminish the mass �ow of fe 2+ (and/or mn2+) ions from bulk soil to the rhizosphere zone, leading to less ip formation on root surfaces (hu et al., 2007; fun et al., 2010; sun et al., 2016). moreover, the abundance of s2– ions in rhizosphere soil could also deplete more oxygen owing to s2oxidisation. figure (3) brie�y illustrates s cycling and its relationship with ip formation in submerged conditions. in several studies, authors have demonstrated that the absence of p in the rhizosphere (p de�ciency) enhances the oxidising capacity of roots and consequently leads to more ip formation (chen et al., 2008; hussain et al., 2009b; fu et al., 2010, 2014). se-mediated enhancement of ip formation was attributed to its function in elevating the rol of rice roots (huang et al., 2020). rol is assumed to be the most important driving force in the formation of plaques (stcyr, crowder, 1989; xu, yu, 2013; yang et al., 2014). wetland plant species, including rice cultivars, are di�ered based on their rol oxidising capability (deng et al., 2009; yao et al., 2011). rice roots porosities, aerenchyma structure and exodermis layer are three main controlling factors for molecular oxygen transfer and radial oxygen loss (yamada et al., 2005; wu et al., 2011; yamauchi et al., 2013). among these, aerenchyma tissues enable the transport of oxygen to the roots under hypoxia conditions, and high root porosity tends to rol from root to rhizoplane (kirk et al., 2014; huang et al., 2020). �ere is a general agreement that rice seedlings with their weak aerenchyma structure in their primary roots are more susceptible to paddy �elds’ anaerobic conditions than older rice plants. �ere were signi�cant disparities in rol, ip formation and as/cd accumulation in roots of rice plants among plant growth stages studied, the lowest occurred at the grain-�lling period (wang et al., 2013). some studies reported a signi�cant di�erence among various rice cultivars in ip formation, regardless of whether they were grown in hydroponic, glass beads or soil culture (liu et al., 2006; wu et al., 2012; lee et al., 2013; syu et al., 2014). in a  comparison study by zhang et al. (2021), it was demonstrated that nano-fe3o4-modi�ed biochar (bc–fe) was more e�cient in ip formation and cd immobilisation in rice roots than ferrous sulfate (feso4), or chelated iron (edta-fe). �ey have also shown that biochar loaded with nanoparticles of fe could signi�cantly raise the proportion of crystallised ip on rice roots by up to 31.8%–35.9%. it has been reported that as feeding can accelerate the formation of plaque on hydrophilic plants’ 233 roots (lee et al., 2013). �e underlying mechanism in accelerated ip formation under as feeding conditions has been earlier attributed to as stress-induced formation of superoxide anion (h2o2, o2 −) in di�erent plant parts, which later tend to form more plaque by di�using free radicals and o2 − out of the root (mishra et al., 2011). plaque formation has been generally reported to have negative associations with organic matter, inorganic carbonates (co3) and eh, and a positive association with extractable fe in the substrate (batty et al., 2000). iron immobilisation in the presence of inorganic carbonates (e.g., feco3) is a typical phenomenon in wetlands. high organic matter content in soil matrix has been found to bind iron and make it less available for cycling (perret et al., 2000; weiss et al., 2003). �e relationship between the root plaques and the concentrations of organic matter in sediments has been linked to the degree of organic matter decomposition and the creation of anoxia (reducing) conditions around the root surface. syu et al. (2013) suggested that the eh of the soil and the extent of fe2+ released into soil solution are determined by two factors, the content of fig. 3. conceptual model illustrating sulphur cycling and iron plaque formation under the in�uence of submerged conditions. (courtesy of conceptual model: p. zandi) m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 234 iron oxides and organic matter in �ooding soils. �ey further concluded that soil iron budget alone is not su�cient to form ip on roots of hydrophytes in �ooding conditions but when coincides with high contents of organic matter elevates the extent of fe2+ ions release into soil solutions which further are oxidised to form fe3+ on root surfaces (syu et al., 2013, 2014). amongst the leading factors a�ecting the extent of ip formation are organic matter and clay composition of soil texture. soils with low clay contents are better candidates for ip deposition on roots than soils with high clay contents. �is report rests on the speculation that the low clay soils boundless iron, making it more available in solution than the higher clay soils. it is generally accepted that organic matter contents in clay soils with iron reservoir characteristics (bacha, hossner, 1977) are more than in sandy soils (dou et al., 2016). st-cyr, crowder (1989) stated that soil organic matter usually counterbalances the e�ect of low/high clay contents on ip formation. in other words, clay minerals compete with organic matter for iron adsorption. organic matters can instead adsorb iron desorbed from clay colloids to bound carbonates in paddy soils rich in organic matter. �e binding of iron and carbonates has been proposed as a prerequisite step in the formation of plaques. overall, it may be concluded that the e�ect of clay content on ip formation depends signi�cantly on the amount of soil organic matter and the positive correlation between clay and carbonate-bound iron, which can be achieved on the condition that organic matter proportion in the soil is constant. a su�cient amount of fe2+ (or mn2+) present in the soil sediment is required for plaque formation (yang et al., 2011; jia et al., 2018; singha et al., 2019). �e degree of iron deposition on roots has been related to iron concentration in solution (taylor et al., 1984) and to the dominant form of iron. taylor et al. (1984) demonstrated that chelated iron, whether it is in oxidised (fe-eddha-ferric-ethylenediamine di (o-hydroxy-pheny1 acetic acid), fe-edt (a-ferric-ethylenediaminetetraacetic acid) or reduced (febpds-ferrous-4,7-di[4-pheny1sulfonate] 1,10-phenanthroline) states, had little chance to produce root plaques compared with fe2+ in solution. �eir �nding supported the contention that supplying the soil culture medium with fe3+, fe3+ chelates, or fe2+ chelates results in less extensive plaque formation. however, at lower solution culture ph values (<4), ferric iron could produce more plaques compared with ferrous iron. at these lower ph values, signi�cant concentrations of fe3+ ions were suggested to maintain in a soluble/ colloidal state and plants had utilised their reduction at the solution-root interface for plaque generation. of di�erent soil iron fractions, the fraction at which iron is bound to carbonates has been central for the formation of root iron plaques. in contrast to hydroponic studies, showing a positive association between ip formation and fe2+ in solution (bacha, hossner, 1977; taylor et al., 1984), the exchangeable iron fraction in soil samples was not necessarily related to ip accumulation. �e underlying reason for this might be that iron oxyhydroxide plaques are not directly produced from 235 fe2+ a�er oxidation but rather from iron carbonate siderite (feco3) that resulted from exchangeable iron transformation (fig. 4). siderites are subsequently oxidised by the oxidation potential of rhizosphere and rhizospheric co2 concentration to goethite (α-feooh) or lepidocrocite (ƴ-feooh). �e extent of fe and/or mn plaque formed on roots increases in proportion to the concentration of organic matter responsible for creating an anoxic environment in which these elements (a�er being reduced) are oxidised to form fe and mn oxides on root surfaces (syu et al., 2013, 2014). mn and fe plaques are positively correlated to each other (ye et al., 2003). in an earlier study by crowder and coltman (1993), it was shown that the amount of ip formed on the root surface was fully consistent with the amount of ph and mn2+ in the rhizosphere. similar to the results of taylor et al. (1984), the formation level of fe oxide plaque on rice roots remained higher upon increasing the ph value, as indicated by zhang et al. (2019a). soil ph impact on ip deposition on root surfaces in the rhizosphere can be considered from two aspects: direct control of iron concentration in solution and dissolution of precipitated root iron and the indirect in�uence of root oxidising capacity. in other words, the ph of the soil can control the net oxidising capacity of the roots and hence cause the precipitation of iron in the rhizosphere. earlier reports showing that the ip formation elevated linearly as the soil ph ranged between 3 and 4.6 in typha latifolia l. (taylor et al., 1984) or 3 and 5.3 in oryza sativa l. probably, the given ph ranges diminished the solubility rate of fe3+ on the roots, leaving more root iron deposition. as with iron plaque reduction (lower ip accumulation) at ph above 5, it is speculated to have been occurred by soluble iron depletion due to fe2+ oxidation in the culture solution (taylor et al., 1984; st-cyr, crowder, 1989). �ese researchers found a positive correlation between lower ip accumulation and lower carbonate-bound iron fractions under rhizosphere ph of more than 5, suggesting the iron-bound to carbonate fraction to be the precursor to ip formation on the roots. �e relative quantities and reactivities of the reductants and oxidants in the rhizosphere portray the extent to which the net oxidative and reductive nature of the rhizosphere is determined. as the soil ph decreases, phenolic compounds as fe3+ reductants are released into the immediate rhizosphere as a result of suberin and lignin decomposifig. 4. conceptual word diagram illustrating the relationship between iron-bound-to-carbonates (siderite) and iron plaque formation in the rhizosphere (courtesy of conceptual word diagram: p. zandi) m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 236 tion/ hydrolysis in root outer cell layers (endodermis and exodermis). in addition, at low ph, rice roots lose much of their oxidising power that will tend to lower the oxidation of fe2+ in the solution, leading to negligible ip deposition. on the other side, at higher soil ph values, the oxidative capacity of the rhizosphere increases the possibility of ip accumulation to a degree (of ph) at which there is no iron available in the solution. in addition to the concentration and form of fe, the soil eh was also characterised as a driving component for plaque formation. �e impact of eh on plaque formation depends primarily on its in�uence on iron concentration in soil solution (taylor et al., 1984). an increase in the soil eh may not restrict the plaque formation process until the soil solution iron is not decreased. on the other side, very negative eh has been found to initiate an increase in soil oxygen demand more than the oxidising power of roots, resulting in the mitigation or absence of plaques. �e eh value decreases with depth in the submerged soil pro�le (schmidt et al., 2011). plaques are mostly found on roots near the surface of submerged soil where the eh value is neither too high nor too low to prevent plaque formation. christensen et al. (1998) proved that ip is formed in sediment with relatively low eh because high concentrations of reduced mn and fe di�use towards the surface of roots where they eventually were oxidised due to root oxygen release. �e oxidising capacity of plant roots, as a biotic factor, is an essential factor in controlling ip formation (xu, yu, 2013; cheng et al., 2014; huang et al., 2020). in addition to this, root plaque formation may also be in�uenced, as outlined above, by root exudates, root enzymatic activities and hydrophyte genotypes (tripathi et al., 2014). based on the studies conducted so far, there has been a general belief over the possible implication of root-released o2 on fe oxidation in the rhizosphere (khan et al., 2016; huang et al., 2020). although root-released o2 has been introduced as the primary cause of iron oxide plaque formation in wetland plants, in some exceptions (e.g., menyanthes trifoliata l. and molinia caerulea (l.) moench), root enzymatic activity could play a decisive role in oxidising ferrous iron in the rhizosphere. root exudates (organic acids, phytosiderophores, etc.) have a high capacity in the reductive dissolution of iron oxides (lee et al., 2007) and mobilisation of iron in the rhizosphere (wu et al., 2014). �e �ow rate of oxygen through the aerenchyma system is substantially associated with plant characteristics and environmental conditions (temperature and humidity) (colmer, 2003; yamauchi et al., 2013; amaral et al., 2017). rice cultivars with higher rol rates showed a higher potential in root plaque formation (cheng et al., 2014). however, in some cases, there has been strong evidence of the more appreciable e�ect of root enzymatic activity on oxidising fe(ii) for rol scavenging (becker, asch, 2005; wu et al., 2014). �e occurrence of fe oxide plaque, which is believed to be liable for the sequestration and immobilisation of various metal(loids) (hansel et al., 2001; bailey-serres et al., 2012), is common in wetland and aquatic plant species such as typha latifolia, phragmites australis (cav.) trin. ex steud, aster tripolium l. (= tripolium pannonicum 237 subsp. tripolium (l.) greuter), and spartina alterni�ora loisel.; root and rhizome surface of the cymodocea serrulata (r. brown) ascherson & magnus; mangrove seedlings viz., avicennia schaueriana stapf & leechman ex moldenke, laguncularia racemosa (l.) c.f. gaertn., rhizophora mangle l. and oryza sativa (tripathi et al., 2014). �ese species are usually grown under partially or all-time �ooded conditions. �is suggests that they possess a versatile development of internal aeration (air lacunae) systems that facilitate oxygen di�usion from the atmosphere to the plant roots for root respiration by basic dispersion through the air spaces within the cortical tissue (bedford et al., 2001). because some parts of roots are ‘leaky’ (to some extent suberised), oxygen di�uses outside of roots’ parts where it encounters fe2+ and precipitates (mostly abiotically) as fe3+ (�omas et al., 2005). �erefore, the plant’s ability to oxidise the rhizosphere is one of the critical factors underlying root iron oxide formation (chen et al., 2007). some wetlands’ seasonal impacts on ip accumulation were exclusively attributed to their closeness to the free-�owing water source containing carbonates. in addition to this, other factors which include ecotypic abilities in biomass and photosynthesis production for providing oxygen for rol and iron oxidation, as well as site-speci�c characteristics in ph and eh, were proposed as principals for site di�erences in root fe deposition (st-cyr, crowder 1989). e�ect of rice origin on iron plaque formation recent studies have pointed to the signi�cant di�erences among various rice cultivars and genotypes in ip formation (wu et al., 2012; lee et al., 2013; cheng et al., 2014; syu et al., 2014; zandi et al., 2020). �e amount of fe content in root plaques of lowland rice cultivars was reported to be higher than upland rice cultivars (pereira et al., 2014). moreover, rice cultivars’ di�erence with respect to ips was attributed to exodermis selectivity, fe nutrition solutions and variations in rol (khan et al., 2016). lee et al. (2013) reported a signi�cant variation in the amount of root plaque among various rice cultivars grown in hydroponic culture. similarly, syu et al. (2014) demonstrated that the degree of root plaque formation in 14 indica rice genotypes was remarkably lower than that of 14 japonica rice genotypes. �e di�erence was ascribed to the lower oxidation capability of indica rice roots compared with japonica rice roots. according to the result by weiss et al. (2005), plaque accumulation in di�erent plant species is characterised by their ability to release oxygen into the rhizosphere. such potential in a speci�c plant species can also be related to the age of plants (chen et al., 2008). �e duration between tillering and the ear emergence stage in rice coincided with a dramatic increase in rol and fe plaque (wang et al., 2013). in another experiment on ten rice genotypes a�er iron treatment, it was found that the di�erence between genotypes in root and plaque fe content was in close association with iron in�ux regulatory mechanisms in these genotypes (asch et al., 2007). additionally, asch et al. (2007) demonstrated that sensitive m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 238 rice genotypes had more iron content in the root and root iron plaque than resistance rice genotypes. �e amount and order of ip formation among rice cultivars studied so far were found di�erent for hydroponic and soil cultures; thereby indicating the multilateral controlling capability of root plaques (lee et al., 2013; syu et al., 2013). in a glasshouse study conducted to assess rice genotypic tolerance against as-rich irrigation water, liu et al. (2006) found the concentration of ip formation o�en varied among genotypes. �ese authors observed a similar pattern of ip formation (tip> middle> base) in the roots of di�erent rice genotypes. in soil and hydroponic study under various aeration conditions, wu et al. (2012) similarly found an additional mass of root plaque at di�erent root zones among various genotypes studied. despite higher discharge of oxygen from root tips (nishiuchi et al., 2012), the occurrence of root plaque was more apparent in older root parts of rice plants (williams et al., 2014). �is could be explained by rapid development of immature root parts (growing root tips) (sharma, kaur, 2020). iron plaque: bu�er or barrier for metal translocation? great endeavours towards scrutinising ip function in metal sequestration and translocation have yielded con�icting results (tripathi et al., 2014; khan et al., 2016; amaral et al., 2017). earlier experiments suggest that ip’s barrier feature suppresses the uptake of elements (christensen et al., 1998; batty et al., 2000), while others estimate ip as a bu�er/reservoir allowing the uptake of elements (ye et al., 2001). apparently, the entry of potentially phytotoxic metal(loids) into rice roots can be either decreased or increased based on the degree of ip formation (tripathi et al., 2014). �e heavy coating of ip may impede the uptake and concentration of zn in sea aster and rice, cd (liu et al., 2007; xu et al., 2018) and cr (xia et al., 2020; zandi et al., 2020) in rice and as in rice (liu et al., 2006). zhou et al. (2007) concluded a direct association between the quantities of se accumulated in plaque and the amount of ip encircled root surface. �us, their studies demonstrated that se concentration in shoots and roots of rice plants decreased upon increasing ip formation. according to batty et al. (2000), under ph conditions of 6.0, ip presence lowers the content of cu in the shoots of a robust perennial grass, common reed (phragmites australis). moreover, spartina densi�ora brong showed a higher potential to maintain essential metals around the root surface and control metal absorption by increasing root plaques’ formation (cambrollé et al., 2008). �ere have been reports of ip mediated reduction in cd concentration in response to fe fertiliser application to cd-contaminated paddy soil (liu et al., 2008). �e extended formation of ip might be used as a trapping strategy to immobilise cd and cr from the rhizosphere through adsorption; however, it may not a�ect their uptake and translocation (khan et al., 2016). it was found that the presence of ip constitutes a barrier to entry (acquisition) and circulation (uptake and accumulation) of both cr(iii) and cr(vi) species in rice (yu et al., 2017). �is result was later contradicted 239 by zandi et al. (2020) who reported a greater inhibitory function of ips against cr(vi) compared to cr(iii). sorption experiments have shown higher net as enrichment in ip for rice seedlings treated with monomethylarsenate (mma) compared to monomethylthioarsenite (mmta), and no enrichment for dimethylmonothioarsenate (dmmta) treatment (kerl et al., 2019). �eir results also showed that methylated thioarsenates (mta) have very little binding/sorption capacity to amorphous ferrihydrite, indicating a low barrier capacity of ip for mta. �e role of ip as a nutrient reservoir/bu�er is recommended, particularly when decreasing the supply of nutrients (khan et al., 2016). its direct impact on exerting changes corresponding to the translocation of nutrients or toxic metal(loids) is o�en plant-species and element-speci�c (tripathi et al., 2014). accordingly, it has been demonstrated that ip exhibits greater a�nity for selenite se (iv) than for ni or cu in typha latifolia; this indicates deferential a�nity and sequestration capacity of ip for di�erent ions. ye et al. (2001) reported that root plaque could act as a cu reservoir. �is result was violated by a recent study by peng et al. (2018) who concluded that root surface iron oxides are strong barriers against cu oxide uptake and translocation in rice plants. otte et al. (1989) stated that ip increased zn and as uptake by aster tripolium. besides, p and zn concentrations in rice plants were lower in the absence of ip than in those with ip, suggesting that ip acted as a zn and p reservoir. ye et al. (1997) reported that the root surface of typha latifolia without ip had lower concentrations of cu than those with ip and that the presence of ip enhances ni translocation. as in hydroponic cultures, the degree of ip formation on root surfaces was positively correlated with net p enrichment in ip both in the �eld (dwivedi et al., 2010) and pot (liang et al., 2006) cultures. unfortunately, there is no report in the literature on the positive/negative impact of ips on nutrient transporters. �e disparity in uptake patterns (ascending/descending) of metal(loids) in the presence of ip was strongly attributed to the amount of ip formed, the mineral composition of ips and the type and availability of metallic ions (including anions and cations) in the growth media (tripathi et al., 2014; khan et al. 2016). role of iron plaque in sequestration and uptake of heavy metals root ips have been predominately found associated with metal(loid)s sequestration (or deposition), uptake and translocation, including pb, cu, zn, cd, as, cr, al, sb (chen et al., 2006; jiang et al., 2009; huang et al., 2012b; xu, yu, 2013; xu et al., 2015; xia et al., 2020). �is is because a noticeable amount of metals bind to the fe plaque by complex formation due to fe hydroxide’s high a�nity for di�erent metals (machado et al., 2005; khan et al., 2016). in most studies, root ip reduced metal translocation to root and shoot (xu et al., 2015). �e ip that forms on the roots of aquatic plants is an indispensable sorbent of both cations and anions metal(loid)s (chen et al., 2006; cheng et al., 2014) and is liable for the attenuation of metal(loid) uptake into wetland plants m ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 240 (jiang et al., 2014). reduction in ip formation due to continuous �ooding conditions did not necessarily contribute to an increase in cd and other metals accumulation in rice grains (zhang et al., 2019b). �is was ascribed to the �ood-mediated reduction in root-to-shoot translocation and availability of metals in the rhizosphere (xu et al., 2013; zhang et al., 2019b). what is probably the most determinative factor in controlling plaque formation is the presence of soluble soil iron and plant species capable of creating an oxidised rhizosphere (chen et al., 2006). �ese fe plaques are precipitated at the oxic-anoxic interface of the aquatic rhizosphere where oxygen from radial oxygen loss and fe(ii) from the reductive dissolution of fe (oxyhydr)oxides intercept (liu et al., 2006). �is process may be biotic or abiotic (sey�erth et al., 2011; xu, yu, 2013; khan et al., 2016 – fig. 2), and the type of fe phase that forms and its potential to sorb metal(loid)s is dependent upon the solution chemistry and rate of oxidation (yamauchi et al., 2013; amaral et al., 2017). taylor, crowder (1963) reported about the possible involvement of ip in the immobilisation of ni and cu in the rhizosphere of typha latifolia. zhang et al. (1998) suggested that zn uptake might be in�uenced both by the presence and amount of ip precipitates on the root surface. liu et al. (2004a) reported that about ⁓90% of total as was concentrated in ip on the rice root surface. �e formation of ip under p de�ciency conditions had a signi�cant e�ect on as sequestration in ip and reduction in root to shoot translocation of as (liu et al., 2004b; hu et al., 2005). cu accumulation in roots and shoots of rice plants was negatively associated with the extent of ip development (peng et al., 2018). in other words, the physical presence of ip could dramatically reduce the amount of cu in the abovementioned rice tissues. in a study of three rice cultivars using cr (iii) treatments, hu et al. (2014) concluded that the degree of ip formation and cr concentration in ip increased in the treatment with no p application. in addition to the inevitable function of root plaques in suppressing the transfer of toxic metal(loids) in wetland-living species, the importance of those additional factors a�ecting the uptake process of elements, namely, plant species, ionic species and their concentrations, ph of the rhizosphere, particular fe phases and the development of new lateral roots, should not be overlooked (khan et al., 2016). for instance, zandi et al. (2021) showed how newly developed lateral roots negate the bene�t of ip by providing barrier-free absorption conditions. given the fact that the whole root axis is not covered by ip, an absorption gap may be available around root tips for penetration of nutrients (batty, younger, 2003; yamaguchi et al., 2014). not to mention that the redox (eh) state and amount of fe2+ produced, as well as the speci�c reactive surface area, may also be involved in the uptake of metal/ loids (tripathi et al., 2014). it should be pointed out that high-ph induced formation of ip and expression of metal transporter genes were responsible for absorption and uptake of toxic metal elements, such as cd, mn and zn (zhang et al., 2019a). 241 role of iron plaque in plant growth �ere are con�icting reports regarding the variable in�uence of ip on the growth of di�erent plant parts during ip formation. investigations of typha latifolia growth revealed that the exposure of plaque-bearing roots to cu, ni, zn, pb, and cd addition did not enhance growth in comparison to plaque-free roots that were exposed to the same metals. experiments with the aquatic plant lobelia dortmanna l. showed that the ips located on the root surface did not a�ect the growth of root diameter (møller, sand-jensen, 2008). �e addition of cu and ni tended to decrease root length, especially when coated by ip. despite this, the presence of ip on the roots of seedlings exposed to slightly toxic conditions of cu and ni improved the shoot growth of rice seedlings. in plants under excess cu and zn toxicity, the ip was demonstrated to positively a�ect the dry weight of roots and shoots and the length of leaves and roots (tripathi et al., 2014). additionally, plants without ip had more leaves displaying chlorosis than those bearing ip when treated with excess cu. �us, it may symbolise the role of ip as a barrier to both toxic and essential metals. conclusion anthropogenic activities are the main sources of recent incremental trends in contaminants loading in wetlands. it is a general belief that ip development on the root surface of hydrophytes is an initial output of ferrous iron oxidation under oxic conditions of the rhizosphere in wetlands. amorphous and crystalline iron oxyhydroxides are fundamental factors of ip constitution and thus account for sequestration of those of nutrients and contaminants having a high binding a�nity to iron oxides. �e so-called sequestration may in�uence the uptake of nutrients and contaminants. �e distribution and evenness of such sequestration are dependent on several components including bio-physicochemical properties of the rhizosphere, di�usion capability of hydrophytes roots and the nutrient and contaminants availability in soil solution. adopting careful management of nutrients and contaminants charging can curb their excess loading in wetlands. above all, much research is still lacking regarding the �uctuate and controversial role of ips (as ine�ectual, barrier or facilitator) concerning the plant uptake of emerging contaminants that need to be addressed. con�ict of interest �e authors declare no con�ict of interest related to this article. references amaral, d.c., lopes, g., guilherme, l.r.g., seyferth, a.l. 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(2007). iron plaque outside roots a�ects selenite uptake by rice (oryza sativa l.) grown in solution culture. plant and soil, 290, 17–28. https://doi.org/10.1007/s11104-006-9072-9 poglądy na koncepcję powstawania blaszki żelazowej i jej cechy charakterystyczne u ryżu siewnego (oryza sativa l.) streszczenie trwały i bioakumulacyjny charakter toksycznych metali(oidów) (tm) jest głównym problemem związanym z ich obecnością w środowisku. skażenie tm w glebie i osadach zwiększa potencjalne ryzyko utraty zdrowia człowieka, przez narażenie na skażenie łańcucha pokarmowego. odkładanie płytki tlenku żelaza na korzeniach hydro�towych (np. ryżu) jest wynikiem różnych czynników biotycznych i abiotycznych. promieniowa utrata tlenu (rol) odgrywa kluczową rolę w utlenianiu żelaza w ryzosferze, a następnie wytrącaniu niskolub wysoko krystalicznych i/lub amor�cznych minerałów żelaza na powierzchni korzeni. biorąc pod uwagę, że każdy gatunek rośliny ma unikalną zdolność tworzenia utlenionej ryzosfery w warunkach beztlenowych gleby, obecność żelaza w ryzosferze ma ogromne znaczenie. grupy funkcyjne (-oh) i specy�czne powierzchnie rem ore insight into the concept of iron plaque form ation and its characteristics in rice (o ryza sativa l.) p ei m an z an di , j oa nn a p uł a, x in g x ia , e lk e b lo em , a m in u d ar m a, y ao sh en g w an g, in gr id t ur is ov á, q ia n li , l uu n go c s in h, n a li 250 agujące w blaszkach żelaza mają wysokie powinowactwo do adsorpcji różnych metali śladowych (toksycznych/ nietoksycznych), wpływając na ich wchłanianie i akumulację w roślinach. w akumulacji różnych pierwiastków ważną rolę odgrywają płytki żelaza (ip). gatunki roślin o niskim ip na swoich korzeniach mogą lepiej akumulować metale ciężkie, niezależnie od tego, czy ip jest barierą, czy buforem. rośliny jadalne o wysokim ip są lepszymi �to-remediatorami potencjalnie �totoksycznych metali(oidów) i mogą być bezpieczniejsze do spożywania przez ludzi. niniejszy przegląd podsumowuje obecną wiedze dotyczącą czynników związanych z tworzeniem i funkcjami płytki żelaza w zarządzaniu transportem metali w systemie korzeniowym. key words: iron oxide plaque, paddy �elds, radial oxygen loss, toxic metals immobilisation received: [2021.07.20] accepted: [2021.10.16] annales universitatis paedagogicae cracoviensis studia naturae, 7: xx–xx, 2022 issn 2543-8832 doi: 10.24917/25438832.7.6 ranjit kakati 1 , dipankar borah 2 , p.k. saikia 1 , ajit hazarika 3 1 department of zoology, gauhati university, guwahati, 781014, assam, india. 2 department of botany, goalpara college, goalpara 783101, assam, india. 3 tyagbir hembaruah college, jamugurihat784180, assam, india. *email of corresponding author: dipankar.borah@rgu.ac.in birds of behali wildlife sanctuary – an important bird area of assam, india introduction avifauna is any country’s environmental health indicator (collar, andrew, 1988; gregory, van strien, 2010). the high and low diversity of bird species is directly related to the environmental condition of an area (devi, saikia, 2010). geographically, india is the 7 th largest country in terms of total area i.e., 3,287,263 km 2 , and ranks 10 th in bird diversity of the world (lepage, 2016). globally 11,158 species of bird are so far known of which 159 are extinct, 5 are extinct in the wild, 225 are critically endangered, 461 endangered, 800 are vulnerable, 1017 are near threatened, 8427 are least concern and 53 are data deficient (birdlife international, 2022a). birds also comprise one of the most threatened taxa of the world, i.e. 13 % of the world’s species (1,313 out of 10,064) are categorised under threatened categories (iucn, 2022). the indian subcontinent has rich avian diversity with about 1263 species (praveen et al., 2016), falling under 23 orders, 107 families and 498 genera which is approximately 12.5% of the world’s bird species (praveen et al., 2016; grimmett et al., 2000). to recognise the important habitats of the highest bird diversity and concentration, 554 important bird areas (ibas) are designated globally and they cover an area of about 19,415,798 ha. (islam, rahmani, 2004). assam, the heart of north-eastern india, harbours around 820 bird species, which includes 280 species of migratory birds (choudhury, 2000). the geographical area of assam is 7.84 million ha., which covers 2.39% of the country’s total area and has 55 recognised important bird areas (ibas) (rahmani et al., 2016). however, many of the ibas are very poorly studied to date, without any complete accounts of the diversity which complicates carrying out conservational programs throughout the landscape. behali wildlife sanctuary (earlier behali reserve forest) is recognised as an “important bird area in-as-05 (a1 and a3)” by the bombay natural history society (rahmani et al., 2016). the forest falls under the east himalayan biodiversity hotspot and is a part of the sonitpur-kameng elephant reserve notified in 2003 (islam, rahmani, 2004). so far a single report on the faunal diversity of the area has been published by kafley (2016-17) in a regional journal, but comprehensive checklist of these different faunal groups is yet to be compiled. hence the present study was carried out with a vision to assess the diversity of bird species in behali wildlife sanctuary and to assess the local status of birds. this present work aims to augment the information about the bird diversity of the iba and the data generated in the present work will also serve as a benchmark for devising better conservation strategies for avian diversity in the future. study area behali wildlife sanctuary (bwls), located in the biswanath district of assam (fig. 1) has a total area of about 14,000 hectares, but now less than 8000 hectares of forested area is left intact (choudhury, 2000). the total area of the wildlife sanctuary is 157.25 sq. km and is located between 26.941701 n and 26.937960 n longitudes and 93.188950 e and 93.466136 e latitudes. the elevation ranges between 80 and 250 m. the mean annual rainfall is > 2300 mm and the temperature ranges from 6 o c to 34 o c (rahmani et al., 2016). many annual and perennial tributaries viz. bihmari, deojan, naharjan, bedeti, dikal, dihiri, thandapani, singlijan, etc. drain the water to the buroi and the borgang river, and finally to the brahmaputra. in the east, flows the singlijan river which is connected to the singlijan reserve forest. to the west, the boundary is marked by the borgang river. numerous annual and perennial streams, wetlands, swamps, and mixed grasslands are also present within the reserve that provides shelter to different forms of life. the northern boundary is shared by the papum reserve forest of arunachal pradesh. the southern boundary has many tea plantations and human habitation areas which include rangagorha tea estate, khorang-line, serelia-pathar, hatimara-pathar, sialmari, bihmari, serelia-bongaon, bihmari-bongaon, etc. this forest is a part of the greater sonitpur elephant reserve and was declared as a reserved forest in 1917. it lies between the two famous protected areas, the nameri national park on its west and kaziranga national park on its south (borah et al., 2009). it acts as an important corridor for migration of species between these protected areas. the reserve forest is an important area for mammals like indian elephant (elephas maximus l.), slow loris (nycticebus coucang boddaert), capped langur (trachypithecus pileatus blyth), malayan giant squirrel (ratufa bicolor sparrman), etc. (islam, rahmani, 2004; borah et al., 2021a). this area is also recognised as an important bird area in 1994 and a key biodiversity area in 2004. fig. 1. map of the study area the area is flat with gentle slopes, typical of bhabar and terai. though the area is dominated by plains, however, some hills lie in the extreme north of the forest. the flora of the forest has been partially worked out (borah et al., 2020a) as well as the plant community structure as elucidated by borah et al. (2021a), shows the presence of various fig (ficus sp.) species as well as other fruit trees forming a suitable refuge for many residential and migratory birds. it is characterised as a semi-evergreen formation with lauraceae, euphorbiaceae, and phyllanthaceae being the most diverse families and magnolia hodgsonii (hook.,thom.) h.keng, mesua ferrea l. and dillenia indica l. are the dominant tree species of the reserve (champion, seth, 1968; borah et al., 2021b). it also includes many rare plants or is of interest to the area’s biodiversity (borah et al., 2019a, b; borah et al., 2020b, c; borah et al., 2021a). the soil is mostly composed of new and old alluvium, which is unconsolidated and loose in structure (bhattacharjya, bhagawati, 2009). agriculture is present on almost all sides of the forest and degradation has severely hampered its boundaries. shifting cultivation is seen in the north boundaries, whereas settled agriculture is predominant in the south and east. methods the study was carried out from september 2016 to may 2022 covering all seasons’ i.e. summer (march–june), monsoon (july–october), and winter (november–february) (devi, saikia, 2010). field visits were conducted at intervals in a month and birds were observed between 5:30 to 9:30 am and 3:30 to 6:30 pm. all the surveys were done by line transect and point count methods (bibby et al., 1992). altogether 25 permanent line transects 1 km long and 50 m wide each were laid randomly. five transects each on all the major habitats of the forest eg. closed canopy, grassland, open canopy, water bodies, and degraded areas near the forest edge were covered (saikia, devi, 2011). also, point counts were conducted along the line transects of each habitat on different days. points were of 20m radius, at least 100m apart from each other to avoid overlap and counts were of 5 min durations. in each habitat, 25-point count surveys were carried out (buckland et al., 1993). the bird species were identified by their primary calls, sight, photographs, and relevant literature (ali, ripley 1987; grimmett et al., 2020, 2011). all recorded bird species are tabulated and characterised by the following characteristics: common name, scientific name, and vernacular names of the birds (datta, 2013) systematic affiliation according to praveen et al., (2015), threat status: cr – critically endangered, en – endangered, vu – vulnerable, nt – near threatened, lc – least concern (iucn, 2022; birdlife international, 2022a); residential status: r – widespread resident, r – very local resident, w – widespread winter visitor, w – sparse winter visitor, p – sparse migrant, v – vagrant or irregular visitor, s – local summer breeder (according to grewal, bhatia, 2014); occurrence in behali wildlife sanctuary (bwls): rare – r, common – c. in addition, a general characterisation of 10 selected species found in bwls with cr, en, and vu categories was carried out. results the study documented altogether 283 species of avifauna belonging to 21 orders, 69 families, and 194 genera in the study area (tab. 1 – appendix 1). among the orders, passeriformes recorded the highest number of birds species i.e. 125 followed by charadriiformes (19 species) and accipitriformes (18 species), piciformes and columbiformes with (14 species) each, coraciiformes with (13 species), pelecaniformes (12 species), strigiformes and cuculiformes with (11 species) each, anseriformes (8 species), ciconiiformes, falconiformes (6 species) each, psittaciformes, bucerotiformes and gruiformes with (5 species) each, galliformes (4 species), suliformes (3 species), podicipediformes (2 species), and lowest of (1 species) each from trogoniformes, caprimulgiformes and apodriformes (fig. 2). fig. 2. comparison of the percentage share of species belonging to 21 orders in the avifauna of behali wildlife sanctuary fig. 3. few avian species of behali wildlife sanctuary: a – white winged duck (asarcornis scutulata); b – great hornbill (buceros bicornis); c – lesser adjutant stork (leptoptilos javanicus); d – woolly-necked stork (ciconia episcopus); e – steppe eagle (aquila nipalensis); f – river tern (sterna auranti); g – osprey (pandion haliaetus); h– kalij pheasant (lophura leucomelanos) (photo a by chirantanu saikia, b-e,g-h by ranjit kakati; f by sahas gjoel) the study found altogether:  one (0.35%) critically endangered species namely red-necked vulture (sacrogyps calvus);  2 (0.71%) endangered species (white winged duck asarcornis scutulata and steppe eagle aquila nipalensis);  7 (2.47%) vulnerable species (lesser adjutant stork leptoptilos javanicus, wreathed hornbill rhyticeros undulatus, rufous-necked hornbill aceros nipalensis, great hornbill buceros bicornis, greater spotted eagle clanga clanga, eastern imperial eagle aquila heliac) and river tern sterna auranti);  13 (4.59%) near threatend species (white-cheeked partridge arborophila atrogularis, black-necked stork ephippiorhynchus asiaticus, woolly-necked stork ciconia episcopus, black-headed ibis threskiornis melanocephalus, oriental darter anhinga melanogaster, red-necked falcon falco chiquera, pallid harrier circus macrourus, grey-headed fish eagle haliaeetus icthyaetus, alexandrine parakeet psittacula eupatria, blossom-headed parakeet p. roseata, red-breasted parakeet p. alexandri), river lapwing (vanellus duvaucelii) and great stone-curlew (esacus recurvirostris), and other 260 (84.8%) species were least concern. some of them are illustrated in figure (3) (tab. 1 – appendix 1). among 283 bird species, 95 (33.56%) species were very local residents (r), 124 (43.82%) species were widespread residents (r), 9 (3.18%) species were sparse winter visitors (w), 15 (5.3%) species were widespread winter visitors (w), 14 (4.95%) species were widespread resident winter visitors (rw), 8 (2.83%) species were residents and widespread winter visitors (rw), 3 (1.06%) species were residents which were also sparse winter visitors (rw) and 3 (1.06%) species were local summer breeders but widespread winter visitors (sw), one (0.35%) species was widespread summer breeder (sw), one (0.35%) species was vagrant or irregular visitor (v), one (0.35%) species was widespread winter visitor (rw), one (0.35%) species was local migrant (rp) and one (0.35%) species was a sparse visitor (p) (tab. 1 – appendix 1). some of the notable resident and migratory bird species of the behali wildlife sanctuary includes grey peacock pheasant (polyplectron bicalcaratum), white-cheeked partridge (arborophila atrogularis), bar-headed goose (anser indicus), white-winged duck (asarcornis scutulata), great crested grebe (podiceps cristatus), common merganser (mergus merganser), glossy ibis (plegadis falcinellus), oriental honey-buzzard (pernis ptilorhynchus), steppe eagle (aquila nipalensis), greater-spotted eagle (clanga clanga), ruddy kingfisher (halcyon coromanda), common sandpiper (actitis hypoleucos), green sandpiper (tringa ochropus), pacific-golden plover (pluvialis fulva), barred-cuckoo dove (macropygia unchall), vernal hanging parrot (loriculus vernalis), wreathed hornbill (rhyticeros undulatus), taiga flycatcher (ficedula albicilla), blue-bearded bee-eater (nyctyornis athertoni), red-headed trogan (harpactes erythrocephalus), hooded pitta (pitta sordida), maroon oriole (oriolus traillii), white-spectacled warbler (phylloscopus intermedius), silver-eared mesia (leiothrix argentauris), scaly thrush (zoothera dauma) and amur falcon (falco amurensis) (tab. 1. – appendix 1). as many as 29% of species are rare in the study area. short notes on some threatened birds of behali wildlife sanctuary below is a short general characterisation of the selected species that in the bwls belong to the group cr – critically endangered, en – endangered, vu – vulnerable. in the characterisation, previous records of those species noticed in the assam were also taken into account. 1) red-necked vulture, sacrogyps calvus, cr: it is a large-sized scavenging bird, which feeds mainly on the carcasses of dead animals, and was once found on every continent except antarctica and oceania (del hoyo et al., 1994). it is a residential species for india and its population in india has declined at the rate of 91% from the year 1999 to 2003 (cuthbert et al., 2006). now the estimated population of this threatened species is about 2500–9999 individuals. (birdlife international, 2022b). this species has suffered an extremely rapid population reduction in the recent past which is likely to continue into the near future, probably largely as a result of feeding on carcasses of animals treated with the veterinary drug diclofenac (birdlife international, 2022b), perhaps in combination with other causes like demise of wild ungulates, unintentional poisoning, the intensification of agriculture, increased sophistication of waste disposal techniques, direct persecution and disease (clements et al., 2013). earlier records from assam: chakrashila wildlife sanctuary, jamjing and sengajan, kaziranga national park (choudhury, 2000; baruah, sharma, 1999), behali reserve forest (choudhury, 2000), dibru-saikhowa national park (choudhury, 2006a), manas national park (choudhury, 2006b), nameri national park (das, deori, 2010), orang national park (talukdar, sharma, 1995). 2) white-winged duck, asarcornis scutulata, en: it is one of the most endangered avian species in the world according to the iucn red list and also listed as a schedule i species under the indian wildlife protection act, 1972. it inhabits dense tropical evergreen forests, near swamps, and rivers or any form of wetland and lay their eggs in tree cavities nearer to the water sources. only 800 individuals are estimated to be left in the wild globally and out of them, only 450 individuals (choudhury, 2007) are known to be present in the eastern himalayan foothills of northeast india, 200 in laos, thailand, vietnam, and cambodia and 150 in sumatra (birdlife international, 2012). due to habitat destruction, forest fragmentation, loss of large trees with nesting holes (birdlife international, 2022b), hunting, and collection of eggs their population has been tremendously decreased (rahmani et al. 2016). it is locally called “deo-hah” in assamese language, due to their ghostly call. earlier records from assam: dibru-saikhowa national park, dangori reserve forest, dumduma reserve forest & kumsong reserve forests (choudhury, 1996, 2006a; yanha, 1994), joypur reserve forest (choudhury, 1996), morioni (mukherjee, 1961), north cachar hills (green, 1992), dibrugarh, lakhimpur, nowgong, silchar and sonitpur districts (mackenzie, 1985; yanha, 1994), tinsukia and dibrugarh districts (hume, marshall, 18791889), upper dihing east and west complex (choudhury, 1996, 1998; rahmani et al., 2016), tirap-burhidihing (choudhury, 1996, 1998), manas national park (choudhury, 2006b), nameri national park (das, 1995, 1998, 1999; saikia, saikia, 2011; das, deori, 2012), behali reserve forest (choudhury, 2000, sonai rupai wildlife sanctuary (choudhury, 2002), barail range, dhansiri reserve forest, dum duma-dangori, east and north karbi anglong wildlife sanctuaries, garampani, nambor and doigrung, hollongapar gibbon sanctuary, inner line (east), katakhal and barak reserve forests, jamjing and sengajan, jatinga, langting-mupa reserve forest, lumding reserve forest, subansiri dulung (rahmani et al., 2016). 3) steppe eagle, aquila nipalensis, en: this species mainly breeds in eastern europe, russia, republic of kalmykia (karyakin et al., 2016), kazakhstan, kyrgyzstan, china, mongolia (meyburg, boesman, 2013), moldova, romania, ukraine, and turkey (ebird, 2020). in winter they migrate to the middle east, arabia, and southeast africa (meyburg, boesman, 2013). the total global population is estimated at more than 37,000 pairs (karyakin et al., 2016). habitat loss (meyburg, boesman, 2013) and high voltage powerline (birdlife international, 2022b) are the major causes of its decline. earlier records from assam: nameri national park (das, deori 2010), orang national park (chakdar et al., 2019), biswanath district (kakati et al., 2021). 4) lesser adjutant stork, leptoptilos javanicus, vu: they are distributed in cambodia, india, malaysia, nepal, indonesia, sri lanka, bangladesh, myanmar, laos, bhutan, brunei, vietnam, and thailand (birdlife international, 2022a). globally, around 6,500–8,000 mature individuals of the lesser adjutant stork are left in the world and a few substantial populations occur primarily in india (about 2,000 birds in assam, west bengal and bihar). cutting down their nesting trees, habitat destruction, urbanization, collection of eggs and chicks, hunting of adults, conversion of wetlands in to agricultural lands, fisheries and the use of fertilizers and pesticides in agricultural land are threatening the survival of this species. practise of poisoning pools to catch fishes also leads to incidental mortality of this species (gyawali, 2004) earlier records from assam: dibru-saikhowa national park (choudhury, 1995; saikia, 1995), orang national park (saikia, 1995), kaziranga national park (saikia, 1995; bhattacharjee et al., 1996; baruah, sharma, 1999), jengdia beel, kamrup district (saikia, 1995), biswanath district (choudhury, 2000, kakati et al., 2021); dum duma-dangori & kumsong reserve forests (saikia, 1995; rahmani et al., 2016), jamjing beel (choudhury, 1992), deepor beel (saikia, bhattacharjee, 1989), nameri national park (choudhury, 1991), majuli, pobitora wildlife sanctuary, dikhoumukh, nalbari, pani dihing, chakrashilla wildlife sanctuary, sareswar beel (saikia, 1995), laokhowa wildlife sanctuary (kahl, 1971; saikia, 1995), bherjan-borajan-podumoni wildlife sanctuary (choudhury, 1995), manas national park (rahmani et al., 1988; saikia, 1995), jhanjimuk-kokilamukh, jorhat (mahanta et al., 2019), behali reserve forest (kakati et al., 2021) ,amchang wildlife sanctuary, barail range, bauwwa beel, bordoibam-bilmukh bird sanctuary, bordoloni-sampora, chakrashila complex, chandubi lake and adjoining areas, deobali jalah, dhansiri reserve forest, east and north karbi anglong wildlife sanctuaries, garampani, nambor and doigrung, hollongapar gibbon sanctuary, inner line (east), katakhal and barak reserve forests, jamjing and sengajan, satgaon, kuarbari-dalani, laokhowa and burhachapori wildlife sanctuary, lumding reserve forest, majuli island, pabho reserve forest, ripu reserve forest, sibsagar tanks, son beel, subansiri dulung, tamaranga-doloni-bhairab complex, tirap-burhidihing, upper dihing (east) complex, upper dihing (west) complex, urpod beel (rahmani et al., 2016), raimona national park (mahanta et al., 2022). 5) greater spotted eagle, clanga clanga, vu: this species is mainly found in estonia, poland, belarus, russia, ukraine, kazakhstan, china, mongolia (meyburg et al., 1999), small numbers in pakistan and north-west india (birdlife international, 2001), finland, and lithuania (birdlife international, 2015). during winter they migrate to eastern europe, north africa, east africa, the indian subcontinent, south asia, and southeast asia (birdlife international, 2015). the total population is about fewer than 3300-8800 mature individuals (birdlife international, 2022a) and is suspected to be declining at a rate of >30% over three generations as a result of extensive habitat loss due to deforestration, felling of nesting trees and destruction of wetlands (maciorowski et al., 2014) and persistent persecution (birdlife international, 2022a). electrocution, collision with wind turbines, hunting and poisoning during times of their migration as well as heavy metals poisoning from consuming waterbirds are another major threats to them (maciorowski et al., 2014; perez-garcia et al., 2020). earlier records from assam: dibru-saikhowa national park (choudhury, 2006a), kaziranga national park (baruah, sharma, 1999), nameri national park (das, deori, 2010), jhanjimuk-kokilamukh, jorhat (mahanta et al., 2019), orang national park (chakdar et al., 2019), dakra beel, chakrashila wls (choudhury, 2000), biswanath district (kakati et al., 2021). 6) eastern imperial eagle, aquila heliaca, vu: this species mainly breeds in austria, azerbaijan, bulgaria, china, czech republic, macedonia, georgia, hungary, kazakhstan, russia, mongolia, serbia, slovakia, turkey, and ukraine (heredia, 1996). a small population is also found in armenia, croatia, cyprus, greece, kosovo, moldova, and romania (birdlife international, 2015). in winter it migrates to the middle east, east africa, tanzania, arabian peninsula, indian subcontinent, south asia, and east asia (birdlife international, 2015). the global population of this species about 2500-9999 individuals (birdlife international, 2022a). the major threats are loss and alteration of feeding and nesting habitats, shortage of small and medium size prey species, illegal trade, poisoning, hunting and electrocution by powerlines. an average of 450 eastern imperial eagles were killed by powerlines in altai region25% of the total population of the region (karyakin et al., 2009b). earlier records from assam: dibru saikhowa national park (menzies, rao, 2019), kaziranga national park, nameri national park (rahmani et al., 2016), biswanath district (kakati et al., 2021). 7) river tern, sterna aurantia, vu: it is distributed throughout southern asia from india, pakistan, sri lanka, and bangladesh to southeast asia. it is a resident breeder of iran and the indian subcontinent and generally prefers river banks (birdlife international, 2022a). the global population is estimated around 20,000–70,000 mature individuals and 29,577 individuals were recorded during the asian waterbird census during 2008-2015. during the census, india scored with 17,776 individuals (mundkur et al., 2017). flooding, predation and disturbance in wetlands are the factors affecting its population. however, conversion of wetlands into agricultural lands and construction of dams, use of pesticides, deposition of heavy metals in waterbodies, mining, cyclones and other some anthropogenic activities like urbanization are the major causes of their decline (debata, 2019; birdlife international, 2022a). earlier records from assam: nameri national park (das, deori, 2010), dibru-saikhowa national park (choudhury, 2006a), kaziranga national park (baruah, sarma, 1999), orang national park (chakdar et al., 2019), jhanjimuk-kokilamukh, jorhat (mahanta et al., 2019), biswanath district (kakati et al., 2021). 8) wreathed hornbill, aceros undulatus, vu: it inhabits in dense primary rainforest, in the lowlands (poonswad et al., 2013) of southern bhutan, bangladesh, and northeast india, east to laos, cambodia, and vietnam and south through malaysia, thailand, indonesia, and brunei. this species is found in the primary forests of the northeast india especially in arunachal pradesh, assam, nagaland etc. the global population is not estimated yet (del hoyo et al., 2001, birdlife international, 2022a) but is suspected to be declining due to habitat destruction, forest fragmentation, felling of nesting trees and hunting (birdlife international, 2022a). earlier records from assam: kaziranga national park (sarma, barua, 1999), dibru-saikhowa national park (choudhury, 2006), nameri national park (das, deori, 2010), raimona national park (mahanta et al., 2022). 9) rufous necked hornbill, aceros nipalensis, vu: it is a highly threatened hornbill species known from bhutan, northeast india, myanmar, southern yunnan, and southeast tibet, china, thailand, vietnam, and laos. it is locally extinct from many historic ranges. in india, the largest population is found in arunachal pradesh and the northern part of west bengal. relatively high densities (<6 birds per km 2 ) have been recorded from both namdapha national park and eaglenest wildlife sanctuary (naniwadekar et al., 2013). it is also found in eastern nagaland and cherrapunji in meghalaya, lushai hills in mizoram, near jatinga in assam and manipur. from the assessment based on the population of densities from sites in northeast india (arunachal pradesh) and western thailand, of the suitable habitat remaining within the occupied range of the species, it is estimated that the global population is 12,000–15,000 individuals (birdlife international, 2022a). deforestration, shifting cultivation, hunting, forest fragmentation, illegal trade and felling of their nesting trees are the major causes of decline (datta, 2009; naniwadekar et al., 2015a). earlier records from assam: nameri national park (das, deori, 2010), behali reserve forest, dhansiri reserve forest, east and north karbi anglong wildlife sanctuaries, jatinga, langting-mupa reserve forest, manas national park, sonai-rupai wildlife sanctuary, subansiri dulung, upper dihing (east) complex, upper dihing (west) complex (rahmani et al., 2016) , raimona national park (mahanta et al., 2022). 10) great hornbill, buceros bicornis, vu: it is a widely distributed species occurring in china, india, nepal, bhutan, bangladesh, myanmar, thailand, laos, vietnam, cambodia, malaysia and indonesia. in india, it is restricted to the himalayan foothills, hill forests of northeast india, and wet evergreen forest of northeast and west india. mostly it can be seen in protected forests, however is now lost from five out of 16 sites from two protected areas (rahmani, 2012). the estimated population of great hornbill is between 13000-27000 individuals. deforestration, shifting cultivation, hunting, forest fragmentation, illegal trade and felling of their nesting trees are the major causes of decline (birdlife international, 2022a). earlier records from assam: nameri national park (das, deori, 2010), dibru-saikhowa national park (choudhury, 2006a), kaziranga national park (barua, sarma, 1999), behali reserve forest, chakrashila complex, dhansiri reserve forest, dibru-saikhowa complex, inner line (east), katakhal and barak reserve forests, langting-mupa reserve forest, lumding reserve forest, tirap-burhidihing , upper dihing (east) complex, upper dihing (west) complex (rahmani et al., 2016), raimona national park (mahanta et al., 2022). fig. 4. porównanie liczby gatunków awifauny w wybranych obszarach chronionych indii j-k iba – jhanjimukh-kokilamukh iba complex, gwls – gibbon wildlife sanctuary, bwls – behali wildlife sanctuary, rgnp – rajib gandhi orang national park, nnp – nameri national park, knp – kaziranga national park short discussion behali wildlife sanctuary (bwls) holds around 22.41% of the total avian species diversity of india (praveen et al., 2015) and 34.51% of the total avian species diversity of assam (choudhury, 2000). the number of species reported in this study is almost similar to the report of chakdar et al., (2019), reporting 284 bird species from rajib gandhi orang national park (rgnp), assam. devi and saikia (2010) reported 232 species from gibbon wildlife sanctuary and mahanta et al. (2019) reported 205 species from jhanjimukh-kokilamukh iba complex, jorhat. it is less than baruah and sarmah (1999) reporting 479 species from kaziranga national park (knp) and 374 species from nameri national park (nnp) – fig. 4 (barua, sharma, 2005; das, deori 2010), which is due to the greater proportion of protection in both the parks. the habitat heterogeneity in the sanctuary is almost similar to nnp, with almost similar vegetation types and native flora. considering this fact, with increasing protection the reserve will probably host a greater species diversity than here presently recorded. the lower number of threatened (ten) and near threatened species (thirteen) – tab. 3 – appendix 1 – is because despite preferable habitats, the forest was under high human disturbance for several decades losing several of the large trees which might be possible nesting and roosting sites and also these elusive species avoid being seen or disturbed. moreover, bwls lies in between the highlands of arunachal pradesh connecting pakke tiger reserve, nameri national park (nnp), and the lowlands of kaziranga national park (knp), acting as occasional roosting grounds within their migratory routes across the landscapes, as already mentioned in the characteristics of this area. this is also a long patch of foothills where the dominant landscape comprises semi-evergreen forests and secondary forests with a small number of water reservoirs. compared to the surrounding parks (fig. 4), in bwls a small number of migratory birds (mostly threatened ones) flock together in the wetlands. however, the number of threatened species is half the number of knp (twenty-five) (baruah, sarmah, 1999) and more than nnp (nine) (das, deori, 2010), which has a larger area and high protection. until 2021, bwls was a reserve forest (an unprotected area) with very less protection and high reports of hunting as well as habitat loss, extirpating about half of its forests. after a decade-long citizen’s movement, it has recently been upgraded to a wildlife sanctuary (a protected area) with hopes of an increase in vigilance, forest cover, and strict actions on any felonious activities. again, the northern side of the forest which was once a connection to the tropical forests of arunachal pradesh, is now severely exploited and turned into a monoculture of rubber and other cash crops. the only solution to the problem of endangered species of avifauna in the area of bwls is their biotope protection. this is one of the best ways to protect species in the world. preservation of feeding, breeding, and living habitats for these species is the only effective way to protect them. of course, the interests of local farmers must also be taken into account when drawing up a biotopes protection plan in such a way as to reconcile these somewhat opposing aspects. appropriate ecological awareness among farmers will also play an important role, without which no type of protection will be sufficiently effective. activation of local societies for biotope protection is a very important and necessary factor of this protection. conclusion the present study on the avifauna of behali wildlife sanctuary with associated threat factors is a first of its kind for this protected area. the agricultural lands and human settlements surrounding the sanctuary are negatively influencing the biodiversity of the reserve. being a habitat for several species of birds along with some threatened ones viz., great hornbill, wreathed hornbill, woolly-necked stork, lesser adjutant stork, etc., efforts are essential to protect the landscape from anthropogenic pressure. the protection of the forest is not adequate due to the understaffed forest department associated with this reserve. providing attention to this forest, considering the high diversity and threatened representatives, would safeguard it shortly. acknowledgements we are thankful to the department of forest, assam for providing necessary facilities for conducting the work. we are also grateful to parixit kafley for his kind cooperation in the field, his insights and extreme knowledge which helped us to understand the biodiversity of the forest, keshab jyoti borah, niku das, nilim kumar saikia and pradip borah for his accompanying in the field, dr. ravi kiran arigela for his help in drafting the manuscript and to chirantanu saikia and sahas gjoel for their better images of the white winged duck and river tern. this is completely a self-funded work. conflict of interest the author declare no conflict of interest related to this article. references ali, s., ripley, s.d. 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(1994). status of white winged wood duck cairina scutulata and its conservation priorities in india. british ecological society bulletin, 25, 17–22. appendix 1 tab. 1. checklist of avifauna in the behali wildlife sanctuary in assam province (india); abbreviations: iucn status: cr = critically endangered, en = endangered, vu = vulnerable, nt = near threatened, lc = least concern; residential status: r = widespread resident, r = very local resident, w = widespread winter visitor, w = sparse winter visitor, p = sparse migrant, v = vagrant or irregular visitor, s = local summer breeder; occurrence in behali wildlife sanctuary (bwls): rare = r, common =c si. no. family common name scientific name vernacular name occurrence in bwls residential status iucn status (2021) 1. phasianidae red junglefowl gallus gallus bon kukura c r lc 2. kalij pheasant lophura leucomelanos deu dorik c r lc 3. white-cheeked partridge arborophila atrogularis r r nt 4. grey peacock pheasant polyplectron bicalcaratum krishna dorik r r lc 5. anatidae ruddy shelduck tadorna ferruginea sakoi-sokuwa c r, w lc 6. lesser whistling duck dendrocygna javanica soru-sorali c r lc 7. bar-headed goose anser indicus dhritoraj c r, w lc 8. cotton pygmy goose nettapus coromandelianus kiki hah c r lc 9. indian spot-billed duck anas poecilorhyncha futuki hah c r lc 10. white-winged duck asarcornis scutulata deu hah r r en 11. common merganser mergus merganser kokila doria hah r w lc 12. common teal anas crecca ghila hah c w lc 13. podicipedidae great crested grebe podiceps cristatus bor dubi r r, w lc 14. little grebe tachybaptus ruficollis pani dubi c r lc 15. ciconiidae black stork ciconia nigra kalsor r w lc 16. woolly-necked stork ciconia episcopus konuwa c r nt 17. black-necked stork ephippiorhynchus asiaticus telia sareng r r nt 18. lesser adjutant stork leptoptilos javanicus bortukula c r vu 19. asian openbill anastomus oscitans samuk-vonga c r lc 20. ardeidae grey heron ardea cinerea halkheda c r, w lc 21. purple heron ardea purpurea azan c r lc 22. intermediate egret ardea intermedia maju bog c r lc 23. great egret ardea alba bor bog c r, w lc 24. cinnamon bittern ixobrychus cinnamomeus itaguria sorai c r lc 25. indian pond heron ardeola grayii konamusuri c r lc 26. black-crowned night heron nycticorax nycticorax wak sorai c r lc 27. striated heron butorides striata soru-musori c r lc 28. cattle egret bubulcus ibis gubog c r lc 29. little egret egretta garzetta teteri-bog c r lc 30. threskiornithidae black-headed ibis threskiornis melanocephalus kola akuhi bog r r nt 31. glossy ibis plegadis falcinellus itaguria akuhi bog r r, w lc 32. phalacrocoracidae little cormorant microcarbo niger pani kawri c r lc 33. great cormorant phalacrocorax carbo doikola c r, w lc 34. anhingidae oriental darter anhinga melanogaster moniyori c r nt 35. falconidae common kestrel falco tinnunculus bakuhi sorai c r, w lc 36. eurasian hobby falco subbuteo sen r r, p lc 37. red-necked falcon falco chicquera ronga dingi sen r r nt 38. pied falconet microhierax melanoleucos pokhora sen r r lc 39. amur falcon falco amurensis sen r p lc 40. peregrine falcon falco peregrinus pahari sen c r, w lc 41. accipitridae black baza aviceda leuphotes kola tikoni sen r r lc 42. crested serpent eagle spilornis cheela sap-kota c r lc 43. oriental honey buzzard pernis ptilorhynchus sen c r, w lc 44. pallid harrier circus macrourus sen r w nt 45. hen harrier circus cyaneus sen c w lc 46. western marsh harrier circus aeruginosus masuwa sen c w lc 47. black-winged kite elanus caeruleus kola pakhi siloni c r lc 48. black kite milvus migrans siloni c r, w lc 49. grey-headed fish eagle haliaeetus ichthyaetus ukoh c r nt 50. red-headed vulture sarcogyps calvus roja sagun r r cr 51. griffon vulture gyps fulvus pahari sogun c r lc 52. shikra accipiter badius sikari sorai c r lc 53. long-legged buzzard buteo rufinus dighol thengia sen r r, w lc 54. greater spotted eagle clanga clanga futuki eagle r w vu 55. changeable hawk eagle nisaetus cirrhatus eagle r r lc 56. steppe eagle aquila nipalensis eagle r w en 57. eastern imperial eagle aquila heliaca r w vu 58. osprey pandion haliaetus kuruwa c r, w lc 59. burhinidae great stone-curlew esacus recurvirostris dangor silkotora r r nt 60. indian stone-curlew burhinus indicus soru silkotora c r lc 61. turnicidae barred buttonquail turnix suscitator bota sorai c r lc 62. charadriidae river lapwing vanellus duvaucelii balighura c r nt 63. red wattled lapwing vanellus indicus hotitiya c r lc 64. grey-headed lapwing vanellus cinereus dolghura c w lc 65. northern lapwing vanellus vanellus silghura r w lc 66. little ringed plover charadrius dubius loriyoli c r, w lc 67. jacanidae bronze-winged jacana metopidius indicus dolpunga c r lc 68. scolopacidae common snipe gallinago gallinago balituka c r, w lc 69. greater painted-snipe rostratula benghalensis rongsongia bali r r lc 70. common greenshank tringa nebularia pat thengi c w lc 71. green sandpiper tringa ochropus bali boguwa c w lc 72. wood sandpiper tringa glareola murlori bali bagua r w lc 73. common sandpiper actitis hypoleucos bali khusora c s,w lc 74. pacific golden plover pluvialis fulva sunali lorioli c w lc 75. little stint calidris minuta chereka sorai c w lc 76. laridae river tern sterna aurantia gonga siloni r r vu 77. black-headed gull chroicocephalus ridibundus kalfut sagor siloni r r lc 78. rallidae ruddy-breasted crake zapornia fusca jikor c r lc 79. slaty-breasted rail lewinia striata c r lc 80. white-breasted waterhen amaurornis phoenicurus daok c r lc 81. common moorhen gallinula chloropus desi kura dhekor c r lc 82. grey-headed swamphen porphyrio poliocephalus kam sorai r r lc 83. columbidae yellow-footed green pigeon treron phoenicopterus halodhiya thengor haitha c r lc 84. spotted dove streptopelia chinensis kopou c r lc 85. green imperial pigeon ducula aenea porghuma c r lc 86. rock pigeon columba livia bonoria paro c r lc 87. red collared dove streptopelia tranquebarica haruwa kopou c r lc 88. eurasian collared dove streptopelia decaocto seto kopou c r lc 89. pin-tailed green pigeon treron apicauda dighol nejia haitha c r lc 90. thick-billed green pigeon treron curvirostra bor haitha r r lc 91. oriental turtle dove streptopelia orientalis son kopou c r, w lc 92. mountain imperial pigeon ducula badia pahari kopou r r lc 93. ashy-headed green pigeon treron phayrei itaguria haitha r r lc 94. wedge-tailed green pigeon treron sphenurus r r lc 95. asian emerald dove chalcophaps indica sil kopou c r lc 96. barred cuckoo dove macropygia unchall deu kopou c r lc 97. psittacidae vernal hanging parrot loriculus vernalis vatou c r lc 98. rose-ringed parakeet psittacula krameri golmonika vatou c r lc 99. red-breasted parakeet psittacula alexandri golpura vatou c r nt 100. blossom-headed parakeet psittacula roseata ronga moni vatou r r nt 101. alexandrine parakeet psittacula eupatria raj vatou c r nt 102. cuculidae common hawk cuckoo hierococcyx varius sen kuli c r lc 103. asian koel eudynamys scolopaceus kuli c r lc 104. green-billed malkoha phaenicophaeus tristis bomura r r lc 105. lesser coucal centropus bengalensis ulu kukuha c r lc 106. greater coucal centropus sinensis bor kukuha c r lc 107. chestnut-winged cuckoo clamator coromandus badamipakhia kuli r r lc 108. banded bay cuckoo cacomantis sonneratii r r lc 109. plaintive cuckoo cacomantis merulinus mamor petia c r lc 110. fork-tailed drongo cuckoo surniculus dicruroides fesu kuli r r lc 111. asian emerald cuckoo chrysococcyx maculatus sil kuli r r lc 112. indian cuckoo cuculus micropterus keteki c r lc 113. strigidae spotted owlet athene brama futuki fesa c r lc 114. short-eared owl asio flammeus hudo r w lc 115. brown hawk owl ninox scutulata kuo fesa c r lc 116. oriental scops owl otus sunia niu fesa c r lc 117. mountain scops owl otus spilocephalus pahari niu fesa r r lc 118. collared scops owl otus lettia niu fesa c r lc 119. brown fish owl ketupa zeylonensis masuwa hudu r r lc 120. brown wood owl strix leptogrammica kath hudo r r lc 121. asian barred owlet glaucidium cuculoides uruli fesa c r lc 122. collared owlet glaucidium brodiei fesa c r lc 123. tytonidae common barn owl tyto alba lakhi fesa r r lc 124. caprimulgidae large-tailed nightjar caprimulgus macrurus bor dinkona c r lc 125. upupidae common hoopoe upupa epops kakoi sira c r, w lc 126. bucerotidae oriental pied hornbill anthracoceros albirostris pakoidhura, dhekdheki c r lc 127. wreathed hornbill rhyticeros undulatus munathoka dhonesh r r vu 128. rufous-necked hornbill aceros nipalensis ronga dhonesh r r vu 129. great hornbill buceros bicornis raj dhanesh r r vu 130. hemiprocnidae asian palm swift cypsiurus balasiensis tal botahi c r lc 131. coraciidae indian roller coracias benghalensis kao sorai c r lc 132. dollar bird eurystomus orientalis nila kao sorai c r lc 133. alcedinidae white-throated kingfisher halcyon smyrnensis boga bukuwa masruka c r lc 134. pied kingfisher ceryle rudis pokhora masruka c r lc 135. stork-billed kingfisher pelargopsis capensis borthutia masruka c r lc 136. blue-eared kingfisher alcedo meninting r r lc 137. common kingfisher alcedo atthis masruka c r lc 138. ruddy kingfisher halcyon coromanda rongosuwa masruka r r lc 139. oriental dwarf kingfisher ceyx erithaca r r lc 140. meropidae green bee-eater merops orientalis moukhuwa c r lc 141. chestnut-headed bee-eater merops leschenaulti badamimuria moukhuwa c r lc 142. blue-bearded bee-eater nyctyornis athertoni nilapakhi moukhuwa c r lc 143. blue-tailed bee-eater merops philippinus nila nejia moukhuwa c r lc 144. trogonidae red-headed trogon harpactes erythrocephalus kathoni buwari r r lc 145. pramphastidae blue-throated barbet psilopogon asiaticus nilokontho hetuluka c r lc 146. blue-eared barbet psilopogon duvaucelii nilapakhi hetuluka r r lc 147. lineated barbet psilopogon lineatus ghungkulung c r lc 148. great barbet psilopogon virens jomdakini c r lc 149. coppersmith barbet psilopogon haemacephalus hetuluka c r lc 150. picidae black-rumped flameback dinopium benghalense soru sun barhoituka c r lc 151. white-browed piculet sasia ochracea ghila kathruka r r lc 152. rufous woodpecker micropternus brachyurus muga kathruka r r lc 153. grey-capped pygmy woodpecker yungipicus canicapillus soru barhoituka c r lc 154. greater yellownape chrysophlegma flavinucha bor halodhiya barhoituka c r lc 155. lesser yellownape picus chlorolophus soru halodhiya barhoituka c r lc 156. streak-throated woodpecker picus xanthopygaeus c r lc 157. greater flameback chrysocolaptes guttacristatus bor sun barhoituka c r lc 158. fulvous-breasted woodpecker dendrocopos macei pokhora kathruka c r lc 159. pittidae hooded pitta pitta sordida r r lc 160. artamidae ashy woodswallow artamus fuscus botahi sorai c r lc 161. aegithinidae common iora aegithina tiphia krishna sorai c r lc 162. campephagidae large cuckooshrike coracina macei kuli era-khati c r lc 163. scarlet minivet pericrocotus speciosus rupohi sorai c r lc 164. long-tailed minivet pericrocotus ethologus dighol nejia rupohi sorai c r lc 165. short-billed minivet pericrocotus brevirostris rupohi sorai c r lc 166. brown shrike lanius cristatus muga erakhati c w lc 167. grey-backed shrike lanius tephronotus kosai sorai c r, w lc 168. long-tailed shrike lanius schach dighol erakhati c r lc 169. dicruridae hair-crested drongo dicrurus hottentottus kesraj c r lc 170. lesser racket-tailed drongo dicrurus remifer vimraj c r lc 171. greater racket-tailed drongo dicrurus paradiseus bor vimraj c r lc 172. crow-billed drongo dicrurus annectens kawri thutia fesu r r lc 173. black drongo dicrurus macrocercus fesu c r lc 174. ashy drongo dicrurus leucophaeus kojola fesu c r lc 175. bronzed drongo dicrurus aeneus matia fesu c r lc 176. oriolidae black-hooded oriole oriolus xanthornus sokhiyoti c r lc 177. maroon oriole oriolus traillii c r lc 178. rhipiduridae white-throated fantail rhipidura albicollis nasoni sorai c r lc 179. eurylaimidae long-tailed broadbill psarisomus dalhousiae r r lc 180. corvidae large-billed crow corvus macrorhynchos dhura kawri c r lc 181. rufous treepie dendrocitta vagabunda kuklunga c r lc 182. house crow corvus splendens pati kawri c r lc 183. monarchidae black-naped monarch hypothymis azurea kolataluwa nasoni c r lc 184. paridae cinereous tit parus cinereus vodorkoli c r lc 185. sultan tit melanochlora sultanea mukut pindha c r lc 186. hirundinidae barn swallow hirundo rustica teltupi c r, w lc 187. plain martin riperia paludicola teltupi c r lc 188. alaudidae sand lark alaudala raytal c r lc 189. pycnonotidae red-vented bulbul pycnonotus cafer bulbuli c r lc 190. black-crested bulbul rubigula flaviventris halodhiya bulbuli c r lc 191. red-whiskered bulbul pycnonotus jocosus tikoni bulbuli c r lc 192. ashy bulbul hemixos flavala kojola fesluka c r lc 193. black bulbul hypsipetes leucocephalus rongathuti bulbuli c r lc 194. white-throated bulbul alophoixus flaveolus golboga bulbuli c r lc 195. locustellidae striated grassbird megalurus palustris kathi sorai c r lc 196. cisticolidae common tailorbird orthotomus sutorius patsia c r lc 197. plain prinia prinia inornata nolsupi c r lc 198. zitting cisticola cisticola juncidis nasoni sorai c r lc 199. cettiidae brownish-flanked bush warbler horornis fortipes tiposi r r lc 200. yellow-bellied warbler abroscopus superciliaris tiposi c r lc 201. slaty-bellied tesia tesia olivea c r lc 202. acrocephalidae blunt-winged warbler acrocephalus concinens tiposi r r lc 203. phylloscopidae greenish warbler phylloscopus trochiloides tiposi c r, w lc 204. tickell's leaf warbler phylloscopus affinis tiposi c s, w lc 205. chestnut-crowned warbler phylloscopus castaniceps tiposi r r lc 206. whistler's warbler phylloscopus whistleri tiposi r r lc 207. grey-hooded warbler phylloscopus xanthoschistos tiposi c r lc 208. smoky warbler phylloscopus fuligiventer tiposi r s, w lc 209. white-spectacled warbler phylloscopus intermedius tiposi r r lc 210. yellow-bellied warbler abroscopus superciliaris tiposi r r lc 211. whistler's warbler phylloscopus whistleri tiposi c r lc 212. dusky warbler phylloscopus fuscatus tiposi c w lc 213. grey-hooded warbler phylloscopus xanthoschistos tiposi r r lc 214. timaliidae pin-striped tit babbler mixornis gularis r r lc 215. pellorneidae abbott's babbler malacocincla abbotti r r lc 216. puff-throated babbler pellorneum ruficeps khupoti r r lc 217. zosteropidae whiskered yuhina yuhina flavicollis r r lc 218. indian white-eye zosterops palpebrosus boga sakuwa tiposi c r lc 219. leiothrichidae silver-eared mesia leiothrix argentauris c r lc 220. white-crested laughingthrush garrulax leucolophus hahiyoti r r lc 221. greater necklaced laughingthrush pterorhinus pectoralis bairagi hahiyoti r r lc 222. striated babbler argya earlei r r lc 223. vireonida white-bellied erpornis erpornis zantholeuca r r lc 224. sittidae chestnut-bellied nuthatch sitta cinnamoventris jethi sorai r r lc 225. velvet-fronted nuthatch sitta frontalis nila jethi sorai r r lc 226. sturnidae common myna acridotheres tristis ghor salika c r lc 227. asian pied starling gracupica contra kan-kurika c r lc 228. great myna acridotheres grandis kola salika c v lc 229. chestnut-tailed starling sturnia malabarica kath salika c r lc 230. common hill myna gracula religiosa moina c r lc 231. jungle myna acridotheres fuscus sutiya salika c r lc 232. turdidae orange-headed thrush geokichla citrina komolamuria matikotora c r lc 233. scaly thrush zoothera dauma c r lc 234. grey-winged blackbird turdus boulboul r r lc 235. muscicapidae blue whistling thrush myophonus caeruleus nilomoti c r lc 236. little pied flycatcher ficedula westermanni c r lc 237. white-gorgeted flycatcher anthipes monileger r r lc 238. snowy-browed flycatcher ficedula hyperythra c r lc 239. oriental magpie robin copsychus saularis dohikotora c r lc 240. white-rumped shama copsychus malabaricus c r lc 241. slaty-backed forktail enicurus schistaceus c r lc 242. black-backed forktail enicurus immaculatus ketepa tip c r lc 243. bluethroat luscinia svecica c s, w lc 244. siberian rubythroat calliope calliope tez tip c w lc 245. grey bushchat saxicola ferreus c r lc 246. siberian stonechat saxicola maurus c r lc 247. hodgson's redstart phoenicurus hodgsoni c w lc 248. black redstart phoenicurus ochruros c r, w lc 249. white-capped water redstart phoenicurus leucocephalus boga c r lc 250. plumbeous water redstart phoenicurus fuliginosus r r lc 251. blue rock thrush monticola solitarius nila bonjuri r r, w lc 252. slaty-backed flycatcher ficedula erithacus c r lc 253. taiga flycatcher ficedula albicilla r w lc 254. little pied flycatcher ficedula westermanni c r lc 255. red-breasted flycatcher ficedula parva c w lc 256. pale-chinned flycatcher cyornis poliogenys makhiyoti c r lc 257. verditer flycatcher eumyias thalassinus c r lc 258. pale blue flycatcher cyornis unicolor c r lc 259. small niltava niltava macgrigoriae soru nilmoni r r lc 260. stenostiridae grey-headed canary-flycatcher culicicapa ceylonensis bari nasoni r r lc 261. irenidae asian fairy-bluebird irena puella c r lc 262. chloropseidae golden-fronted leafbird chloropsis aurifrons sun kopali pat sorai c r lc 263. orange-bellied leafbird chloropsis hardwickii r r lc 264. dicaeidae fire-breasted flowerpecker dicaeum ignipectus ronga bukua fulthukura c r lc 265. plain flowerpecker dicaeum minullum c r lc 266. scarlet-backed flowerpecker dicaeum cruentatum ronga pithia fulthukura c r lc 267. nectariniidae purple sunbird cinnyris asiaticus moupia c r lc 268. little spiderhunter arachnothera longirostra soru mokora khuwa c r lc 269. streaked spiderhunter arachnothera magna mokora khuwa c r lc 270. crimson sunbird aethopyga siparaja ronga moupia c r lc 271. passeridae house sparrow passer domesticus ghor sirika c r lc 272. eurasian tree sparrow passer montanus c r lc 273. ploceidae baya weaver ploceus philippinus tukura c r lc 274. estrildidae scaly-breasted munia lonchura punctulata tuni c r lc 275. red munia amandava amandava r r lc 276. white-rumped munia lonchura striata tuni c r lc 277. motacillidae white wagtail motacilla alba boga balimahi c r, w lc 278. grey wagtail motacilla cinerea c w lc 279. western yellow wagtail motacilla flava halodhiya balimahi c w lc 280. citrine wagtail motacilla citreola c r, w lc 281. olive-backed pipit anthus hodgsoni matimahi c r, w lc 282. paddyfield pipit anthus rufulus matimahi c r lc 283. rosy pipit anthus roseatus c r lc abstract the avian diversity of behali wildlife sanctuary (india), an important bird area in-as-05 (a1 and a3) is documented here. a total of 283 species of birds belonging to 21 orders, 69 families, and 194 genera were recorded. the study recorded one critically endangered species, 2 endangered species, 7 vulnerable species, 13 near threatened species and 260 species were least concern. distribution of birds within the reserve forest and iucn red list categories of them are also provided here. key words: behali reserve forest, bird sanctuary, threat status, residential status received: [2022.08.09] accepted: [2022.11.20] ostoja behali wildlife – ważny obszar dla ptaków w prowincji assam (indie) streszczenie w artykule udokumentowano różnorodność ptaków w ostoji behali wildlife, ważnego dla ptaków in-as-05 (a1 i a3) obszaru indii. zarejestrowano łącznie 283 gatunki ptaków należących do 21 rzędów, 69 rodzin i 194 rodzajów. wśród gatunków stwierdzono jeden krytycznie zagrożony, 2 zagrożone, 7 wrażliwych, 13 potencjalnie zagrożonych i 260 innych gatunków. udokumentowano również rozmieszczenie ptaków w rezerwacie oraz ich kategorie zagrożeń wg czerwonej listy iucn. słowa kluczowe: behali reserve forest, rezerwat ptaków, stan zagrożenia, status zamieszkania information on the authors ranjit kakati https://orcid.org/0000-0001-5938-2945 he is interested in biodiversity assessments, influence of contaminants on water bodies affecting reproductive health, avifaunal assemblage of protected and non-protected areas, rescue and rehabilitation, etc. dipankar borah https://orcid.org/0000-0002-3016-1070 he is interested in understanding floral assemblage of last remnant forests of northeastern india, taxonomy of endemic plants of ne india, traditional knowledge and plant-animal relationships. p.k. saikia https://orcid.org/0000-0003-4220-9411 he is interested in stork and ibises and animal ecology. ajit hazarika https://orcid.org/0000-0002-6668-9205 he is interested in animal physiology and reproductive biology. 157 annales universitatis paedagogicae cracoviensis studia naturae, 6: 157–184, 2021, issn 2543-8832 doi: 10.24917/25438832.6.10 barbara kubik1,2, alina stachurska-swakoń3* 1institute of biology, pedagogical university of krakow, podchorążych 2 st., 30-084 kraków, poland 2centermed hospital św. łazarza, św. łazarza 14 st., 31-530 kraków, poland 3jagiellonian university, gronostajowa 3 st., 30-387 kraków, poland; *alina.stachurska-swakon@uj.edu.pl plants supporting the treatment of cardiovascular diseases introduction cardiovascular diseases have always accompanied human, however nowadays they occur much more o�en due to an intense lifestyle, daily dose of stress, excessive alcohol consumption, smoking, sedentary work combined with physical inactivity, environmental pollution, poor diet dominated by high-fat products and many other external factors. �at is why it is so important not to ignore the �rst symptoms of cardiovascular disorders, such as: palpitations, excessive fatigue or breathlessness. symptoms of this type should stimulate specialist examinations and undertake appropriate treatment or prophylaxis (beaglehole et al., 2001; noskowicz-bieroniowa, 2006; górnicka, 2012; wojtyniak et al., 2013; senderski, 2016; liperoti et al., 2017). ignoring the �rst symptoms of abnormal heart function and vascular insu�ciency leads to a gradual intensi�cation of these ailments. it can be directly life-threatening in the later stages of the disease (sroka, 1988; palmieri et al., 2018). despite the ongoing preventive measures, diseases from this group are recognised as the leading cause of death globally. in 2019, an estimated 17.9 million people died from cardiovascular diseases, which was responsible for 32% of all global deaths (www.who. int). �e number of deaths unfortunately is increasing across the years: in 2002, 16.7 million people died of this cause, including 6.9 million from coronary heart disease and 5.1 million from stroke (matyjaszczyk et al., 2011). statistics show that across europe more women than men die from heart-related diseases. cardiovascular diseases are classi�ed as civilisation diseases, which are currently one of the biggest health problems in highly developed and developing countries. among the civilisation diseases of the circulatory system, the most frequently mentioned are: hypertension, low blood pressure, coronary heart disease, atherosclerosis and myocardial infarction (matyjaszczyk et al., 2011; lewkowicz-mosiej, 2017). for example, inadequately treated or b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 158 untreated high blood pressure can disrupt a variety of organs including the kidneys, brain, heart, and arteries, which can lead to haemorrhage, paralysis, and even death (noskowicz-bieroniowa, 2006; górnicka, 2012). with atherosclerotic lesions, fatty and calcium deposits are deposited in the lumen of the blood vessels, which in turn results in narrowing, thickening or loss of elasticity of the vessels. �e consequence of this type of changes are anal varices (haemorrhoids), shins or purpura (ożarowski, jaroniewski, 1987; noskowicz-bieroniowa, 2006). �e causes of cardiovascular diseases can be di�erent, therefore many groups of medicines are used to treat them. �e treatment strategy is based on the use of medicines that lower blood pressure, regulate the heart rhythm or have an antiarrhythmic e�ect. in recent years, herbal preparations are being used more and more o�en because their side e�ects are usually less burdensome for the patient. at the same time, the use of plant-based medicines, especially those containing cardiac glycosides, cannot take place without medical supervision, because these raw materials are not indi�erent to human health and life (capasso et al., 2000; nowak, 2009; kohlmünzer, 2010; liperoti et al., 2017; länger et al., 2018; bejček et al., 2021). �e aim of this study was to analyse the composition of commercially available medications, herbal preparations and dietary supplements on the polish market, and on this basis to create a systematic list of plants most o�en used in the treatment and prevention of cardiovascular diseases. methods �e compositions of medicines and preparations of plant origin (dietary supplements, mixtures), available without a prescription in pharmacies and herbal stores in southern poland were analysed in order to recognise plant taxa used. a total of 100 preparations, randomly chosen, were taken into consideration. most of which were sold as capsules, infusing herbs and pills (fig. 1; tab. 1 – appendix 1). in few cases, the medications were attainable in various forms. in overall, 61% of analysed medications have a solid form (capsules, dragée, �lm-coated tablets, pills, etc.). on this base, a list of species used in the treatment of cardiovascular diseases was compiled. �e properties of plants were characterised in terms of this group of diseases, based on the available bibliography, including: ożarowski and jaroniewski (1987), świejkowski (1990), broda and mowszowicz (2001), jeziorski (2009), kohlmünzer (2010), błach-olszewska et al. (2014), senderski (2016), and others. when characterising plants, their systematic position, geographic range, type of plant substance (raw material), content of active compounds, therapeutic e�ect in the analysed group of diseases and available preparations (trade names) were taken into account. systematic position of taxa was assumed on the basis of stevens (2001), authors of family p lants supporting the treatm ent of cardiovascular diseases 159 names were given according to reveal (2007). latin nomenclature of native plants followed mirek et al. (2020), and alien plant species according to podbielkowski and sudnik-wójcikowska (2003), or other available sources (e.g. powo, 2021). results �e compiled systematic list of plants used in analysed medicaments includes 65 taxa, two of which are given in the rank of genus – rubus and salix, and the rest in the rank of species. �e most common species used in the analysed group of preparations are seed plants (63 taxa) – the other two are: a species representing algae from the phaeophyta divisio (fucus vesiculosus l.) and a species from the tracheophyta, equisetopsida classes (equisetum arvense l.). in the case of genera allium, cola, crataegus and panax, double species are given as their raw materials as they are o�en used together in a mixture (tab. 2 – appendix 1). �e taxa recorded in this group of herbal plants belong to 40 families. �e most species represent the rosaceae, asteraceae and lamiaceae families. a large group consists of single species representatives of as many as 30 families (fig. 2). among the noticed taxa, 23 are native to central europe, while 20 are alien species for the european �ora, and their raw materials are imported from di�erent regions of the world (fig. 3; table 2 – appendix 1). fig. 1. �e forms of medications, dietary supplements and plant substances supporting the treatment of cardiovascular diseases available on the polish market (blue columns represent solid form, orange are for liquid) b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 160 fig. 2. taxonomical representation of seed plant species used in the medications of the cardiovascular diseases fig. 3. �e origin of raw plant materials from medications available in polish markets used in cardiovascular diseases p lants supporting the treatm ent of cardiovascular diseases 161 fig. 4. �e frequency of plant species used as ingredients of medicaments of cardiovascular diseases (found at least in �ve percentage of preparations) some species are cultivated due to the better quality of the raw material – this applies to both native and alien plants. �e most common raw material used in preparations available on the market is the genus crataegus and species such as: aesculus hippocastanum or ruscus aculeatus (fig. 4). most plants show vasoprotective activity (lat. vasotonicum); its mechanism is to seal the walls of blood vessels, thus preventing them from breaking. �is group of species contains 23 taxa e.g. aesculus hippocastanum, arnica montana, gingko biloba, malpigia glabra (tab. 3 – appendix 1). in addition, a large group of plants lowers blood pressure (lat. hypotonicum), (e.g. achillea millefolium, crataegus laevigata, fucus vesiculosus, kalmia latifolia), has a diastolic e�ect (lat. spasmolyticum) (e.g. ammi visnaga, leonurus cardiaca, ruta graveolens) and strengthens the heart (e.g. adonis vernalis, ophiopogon japonicus, passi�ora incarnata). discussion on the streets of many cities, herbal shops and bio-supermarkets specialising in the sale of herbs and other ecological products are more and more o�en found, proving a  great interest in natural methods of treatment. some of the clients see them as an opportunity to improve their health or want to stop using synthetic medicines for othb ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 162 er reasons. for patients struggling with numerous side e�ects of medicines obtained through a synthetic route, herbal therapy may be an alternative method of treatment (brzeziński, 2014; senderski, 2016; liperoti et al., 2017; easley, horne, 2019). for medicinal purposes, plant raw materials are o�en obtained from wild specimens and then purchased by herbal or pharmaceutical companies. trade preparations of this type of companies have become the basis for compiling the list of medicinal plant species most o�en used in the treatment of cardiovascular diseases. �e systematic list prepared for this study includes 65 taxa of herbal plants. �ese taxa belong to 40 families (tab. 2 – appendix 1). most of the plants used in this group of diseases belong to native to central europe �oristic components (fig. 3). however, there is also a large group of plants origin e.g. from the mediterranean area; they are o�en cultivated for the needs of the pharmaceutical industry. �ese include, for example, toothpick-plant ammi visnaga l., pot marigold calendula o�cinalis l., maritime squill urginea maritima bak and others (senderski, 2016). some species alien to central europe with proven medicinal properties (e.g. bayan et al., 2014; romero et al., 2016; marques et al., 2017) belong to plants cultivate for consumption, e.g. garlic allium sativum l. or globe artichoke cynara scolymus l., olive olea europaea l. or are ornamental plants, e.g. ginkgo gingko biloba l., witch-hazel hamamelis virginiana l., horse chestnut aesculus hippocastanum l. (e.g. isah, 2015; pizzorno et al., 2015; zampieron, 2017; abbas et al., 2020). �e plants whose herbal substrates are imported from distant regions of the world include: thorny bamboo bambusa arundinacea (retz.) willd. (e.g. rathod et al., 2011), barbados cherry malpighia glabra l. (e.g. johnson, 2003), maypop passi�ora incarnata ker.-gawl. (e.g. krenn, 2002), night-blooming cereus cactus selenicereus grandi�orus (l.) brit. & rose (haque et al., 2015), indian snakeroot rauvol�a serpentina (l.) benth. (e.g. mittal et al., 2012), ginseng panax sp. (e.g. pan et al., 2012) and others. �ese herbal substances are generally expensive due to indirect costs (transport, storage, etc.), therefore they are used much less frequently in the discussed group of preparations. however, their exoticism o�en makes them more attractive to the client who is looking for new products in supplements and drugs supporting synthetic therapies. each herbal plant has a di�erent spectrum of activity. it is possible due to the various accumulation of active substances, such as: cardiac glycosides, �avonoids, alkaloids, anthocyanins and others (kohlmünzer, 2010; pizzorno et al., 2015; fecowicz et al. 2020). preparations with the content of cardiac glycosides, mainly digoxin, obtained from woolly foxglove digitalis lanata erth., are of signi�cant importance in severe, chronic diseases. preparations with pheasant’s eye adonis vernalis l., may bells convallaria majalis l. and strophanthus strophantus gratus l. are also of great importance (sroka, 1988; nowak, 2009). in addition to these plants with strong e�ects, herbs with mild antihypertensive properties are also helpful in treatment (de souza et al., 2011; al p lants supporting the treatm ent of cardiovascular diseases 163 disi et al., 2016; anwar et al., 2016), toning, sealing the vessels, and relaxing (tab. 3) (górnicka, 2012). in the lamiaceae family, there are many plants containing essential oils, which are widely used not only in pharmacy or herbal medicine, but also in cosmetology and in the kitchen (podbielkowski, sudnik-wójcikowska, 2003; senderski, 2016; kliszcz et al., 2021). examples of this type of aromatic plant include rosemary rosmarinus o�cinalis l. and lemon balm melissa o�cinalis l. �ey are also species that are used in supporting the treatment of cardiovascular diseases. however, not all species with known healing properties in cardiovascular diseases are currently used in medicine, because they are very strong, with high content of glycosides, dangerous to health (broda, mowszowicz, 2001; noskowicz-bieroniowa, 2006; nowak, 2009). some of them have been withdrawn from circulation due to their toxic e�ects and high accumulation properties in the organism, e.g. common foxglove (digitalis purpurea l.), oleander (nerium oleander l.) or white waterlily (nymphaea alba l.), however, some medicaments are still available. dioscorides has already written about the poisonous properties of oleandrin, an active substance isolated from common oleander. its operation was also known during the times of alexander the great (bohne, dietze, 2008). in the history of herbal medicine, however, this plant is recorded not as highly toxic, but because of its therapeutic e�ects. �e group of plants most o�en used in the treatment and support of cardiovascular therapies includes such taxa as: crataegus sp., aesculus hippocastanum, or ruscus aculeatus (fig. 4). many studies describe their therapeutic e�ect in the treatment of cardiovascular diseases. a. hippocastanum and r. aculeatus show primarily vasoprotective properties, while crataegus additionally strengthens and improves the work of the heart (tab. 2). however, there is still extensive research into other bene�cial e�ects of these plants on the cardiovascular system (shatoor, 2013; wang et al., 2013). �is especially applies to species used in asian medicine based on traditional herbal folk medicine (xiong et al., 2013; khan et al., 2016; liu, huang, 2016). �e mere use of synthetic or herbal medicines may be insu�cient in cardiac therapies, therefore it is important to change the diet, increase physical activity, and even change the environment or workplace (sroka, 1988). centuries of observations of taking various herbal remedies allow us to state that it is more e�ective and reliable to use many herbs in appropriately selected mixtures than individually (senderski, 2016). �anks to this, they can support each other and complement each other’s action (ożarowski, jaroniewski, 1987; matyjaszczyk et al., 2011; górnicka, 2012). however, it is also important that in the case of serious diseases of the heart or the circulatory system, it should always take place under the strict supervision of a specialist doctor – cardiologist (rogowska, giermazaik, 2018). b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 164 conclusions �e paper presents an analysis of the group of medicines and herbal substances commonly available on the pharmaceutical market in poland and used in the treatment of cardiovascular diseases. its aim was to create a systematic list of plants used in overthe-counter medicinal preparations. �e compiled systematic list includes 65 plant taxa. among them, the most species belong to the families: rosaceae (7), asteraceae (6), lamiaceae (4). �e most common raw materials used in over-the-counter preparations are crataegus sp., aesculus hippocastanum and ruscus aculeatus. most plants show vasoprotective properties, lower blood pressure and diastolic. �e systematic list includes 23 species of plants native to central europe, while plant substances from 20 are imported from more or less distant areas of the world. some of the analysed species, rich in cardiac glycosides, were withdrawn from sale due to their toxicity or strong accumulation of active substances. dosing of medicines with the content of cardiac glycosides must be under the constant supervision of a doctor, because they can easily be overdosed. con�ict of interest �e authors declare no con�ict of interest related to this article. references abbas, t., abbas, m., jarad, a. 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[in polish/latin/english/french/german/russian] powo, (2021). plants of the world online, facilitated by the royal botanic gardens, kew. http://www. plantso�heworldonline.org/retrieved 14 october 2021. rathod, j.d., pathak, n.l., patel, r.g., jivani, n.p., bhatt, n.m. (2011). phytopharmacological properties of bambusa arundinacea as a potential medicinal tree: an overview. journal of applied pharmaceutical science, 1(10), 27–31. reveal, j.l. (2007). classi�cation of extant vascular plant families – an expanded family scheme. http:// www.plantsystematics.org/reveal/pbio/fam/vascplfam.html. rogowska, m, giermazaik, w. (2018). wpływ roślin leczniczych na farmakokinetykę i metabolizm leków syntetycznych (e�ect of medicinal plants on the pharmacokinetics and metabolism of synthetic medicines). postępy fitoterapii, 19(4), 274–282. [in polish] http://dx.doi.org/10.25121/pf.2018.19.4.274 romero, m., toral, m., gómez-guzmán, m., jiménez, r., galindo, p., sánchez, m., olivares, m., gálvez, j., duarte, j. (2016). antihypertensive e�ects of oleuropein-enriched olive leaf extract in spontaneously hypertensive rats. food and function, 7(1), 584–593. http://dx.doi.org/10.1039/c5fo01101a senderski, m.e. (2016). prawie wszystko o ziołach. seria mądrość natury (almost everything about herbs. �e wisdom of nature series), 3 ed. podkowa leśna: wydawnictwo mateusz e. senderski. [in polish] shatoor, a.s. (2013). in vivo hemodynamic and electrocardiographic changes following crataegus aronia syn. azarolus l administration to normotensive wistar rats. saudi medical journal, 34(2), 123–134. sroka, o.g.f. (1988). poradnik ziołowy (herbal tutorial). warszawa: wydawnictwo iwzz. [in polish] stevens, p.f. (2001). angiosperm phylogeny website, version 14, july 2017 [and more or less continuously updated since]. http://www.mobot.org/mobot/research/apweb/. p lants supporting the treatm ent of cardiovascular diseases 167 świejkowski, l. (1990). rośliny lecznicze i  przemysłowe: klucz do oznaczania (medicinal and industrial plants: the key to marking). warszawa: wydawnictwo libra. [in polish] szot-radziszewska, e. (2005). sekrety ziół. wiedza ludowa, magia, obrzędy, leczenie (secrets of herbs. folk knowledge, magic, rituals, treatment). warszawa: wydawnictwo trio. [in polish] szweykowska, a., szweykowski j. (2003). słownik botaniczny (botanical dictionary). warszawa: państwowe wydawnictwo wiedza powszechna. [in polish] wang, j., xiong, x., feng, b. (2013). e�ect of crataegus usage in cardiovascular disease prevention: an evidence-based approach. evidence-based complementary and alternative medicine, 2013, 149363. http://dx.doi.org/10.1155/2013/149363 1-16 wojtyniak, k., szymański, m., matławska, i. (2013). leonurus cardiaca l. (motherwort): a review of its phytochemistry and pharmacology. phytotherapy research, 27(8), 1115–1120. http://dx.doi.org/10.1002/ ptr.4850 xiong, x., yang, x., liu, y., zhang, y., wang, p., wang, j. (2013). chinese herbal formulas for treating hypertension in traditional chinese medicine: perspective of modern science. hypertension research, 36(7), 570–579. http://dx.doi.org/10.1038/hr.2013.18 zampieron, e. (2017). horse chestnut (aesculus hippocastanum) for venous insu�ciency. international journal of complementary and alternative medicine, 5(3), 00153. https://doi.org/10.15406/ijcam.2017.05.00153 b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 168 appendix 1 tab. 1. �e herbal medicines sold in the polish market used in cardiovascular diseases form of the herbal medicine trade name of the herbal medicine capsules alitol, allio�l forte, alliogal, alliomax, bilberin, bilobil, bodymax plus, diosmina plus, extraspasmina, garlicin, ginkgo caps, ginkogins, kapsułki aronia, krążenie i love herbs, liść oliwny, na ciśnienie kapsułki, rosavit c, ruscoven plus, rutina c max, sapoven t, veno 33 complex, venocorector, venox, żeń-szeń vita complex, dragée convafort, venescin, drops cardiol c krople, heel cralonin krople, kelicardina, neocardina, ne-ospasmina, ribes nigrum – macerat glicerynowy, venol, �lm-coated tablets ginkofar, ginselin, proscillaridin, sylicynar, varixinal, venosol bon-ifraters, gel aescin żel, arnical żel, esceven, essaven gel, latan żel, neo-aesculan żel, preparation h żel, venescin, venoczar żel, herbs to brew barwinek pospolity (vina minor), bio hibiscus, bratek �x, cardiosan, ciśnienie norma – herbatka ziołowa, herba asperulae, herba betonicae, herbatka �x żylaczek, krwiściąg lekarski ziele, nervosan �x, neurosin tea, owoc jarzębiny herbatka ziołowa, rektosan, sklerosan, viola – �x, ziele krwiściągu, juice aronia sok 100%, ointment aesculan, arcalen, esceven, maść nagietkowa, ruscogenin, ruscolan, venescin, oral �uid cravisol, doppel herz vital, intractum hippocastani, intractum visci, nervosol, passispasmina, rutisol, venoforton pills bratek, colladen, cynarex, diostrin, esceven, głóg zioła w tabletkach, hemorigen femina, neospasmina, rutinosal c, serpina, strophanthus comp., tabletki tonizujące, tabletki z czosnku, suppositories hemorol, ruskorex, syrup syrop malina z kwiatem bzu czarnego z wit. c, tincture cardiactiv – nalewka, cardio farm – nalewka, cardiotonic – nalewka, tinctura adonidis vernalis, tinctura ammi visnagae, tinctura arnicae, tinctura ginkgo bilobae, tinctura sophorae japonicae tonic plusz gold vital p lants supporting the treatm ent of cardiovascular diseases 169 ta b. 2 . s ys te m at ic li st ; s ho rt c ha ra ct er is tic s o f h er bs u se d in su pp le m en ta tio n an d tr ea tm en t o f c ar di ov as cu la r d is ea se s sp ec ie s g eo gr ap hi c ra ng e pl an t m at er ia l a ct iv e co m po un ds e� ec t o cc ur re nc e in m ed ic in es or s up pl em en ts (p ol is h tr ad e na m es ) fu ca ce ae a da ns on 1) f uc us ve sic ul os us l . n or th s ea a nd n or th a tla nt ic th al lu s or ga ni ca lly b ou nd io di ne , po ly sa cc ha ri de s (a lg in ic ac id , f uc oi di n, la m in ar in ), m an ni to l, fu co xa nt hi n, br om in e, b v ita m in s, m in er al s al ts in th e pr ev en tio n of hy pe rt en si on , d ila te s bl oo d ve ss el s, pr ev en ts at he ro sc le ro si s sk le ro sa n eq ui se ta ce ae m ic hx . e x d c . 2) e qu is et um ar ve ns e l. co sm op ol ita n he rb (e qu is et i he rb a) �a vo no id s ( e. g. is oq ue rc et in , ap ig en in ), si lic on co m po un ds , p he no lic a ci ds (e .g . c a� ei c ac id ), sa po ni ns , vi ta m in s, m in er al s, po ly en ic ac id s a nd p yr id in e de ri va tiv es in cr ea se s t he a m ou nt of e ry th ro cy te s a nd ha em og lo bi n in th e bl oo d, an tiat he ro sc le ro tic , a nt iha em or rh ag ic c iś ni en ie n or m a – he rb at ka zi oł ow a, h er ba tk a �x ży la cz ek g in kg oa ce ae e ng l. 3) g in kg o bi lo ba l. ja pa n, c hi na ; cu lti va te d in a m er ic a an d eu ro pe le af (g in kg on is fo liu m ) se sq ui te rp en es (b ilo ba lid ), di te rp en es (g in ko go lid es a , b , c , j , m ), �a vo no id s, st er ol s, fa tty a ci ds im pr ov es c ir cu la tio n in th e br ai n, m em or y an d co nc en tr at io n, ha s a nt ic oa gu la nt a nd va so pr ot ec tiv e pr op er tie s bi lo bi l, g in kg o ca ps , g in ko fa r, g in ko gi ns , k rą że ni e i l ov e h er bs -c ap s, ti nc tu ra g in kg o bi lo ba e, ve no fo rt on , v en ol a lli ac ea e bo rk h. 4) a lli um sa tiv um l, a .u rs in um l . c en tr al a si a; cu lti va te d bu lb (a lli i s at iv i bu lb us , a lli i s at iv i pu lv is) , h er b/ le af (a lli i u rs in i h er ba / fo liu m ) al lic in a nd it s d er iv at iv es hy po te ns iv e, a nt iat he ro sc le ro tic , a nt ico ag ul an t a lit ol , a lli o� l f or te , a lli og al , a lli om ax , g ar lic in , t ab le tk i z cz os nk u b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 170 h ya ci nt ha ce ae b at sc h ex b or kh . 5) u rg in ea m ar iti m a ba k (= sc ill a m ar iti m a l. ) m ed ite rr an ea n ba si n bu lb (b ul bu s sc ill ae ) am on g ot he rs : s ci lla re n a , pr os ci lla ri di n a ca rd io to ni c: a nt iar rh yt hm ic , st re ng th en in g th e ac tiv ity o f th e he ar t m us cl e pr os ci lla ri di n c on va lla ri ac ea e h or an 6) c on va lla ri a m aj al is l. eu ro pe , n or th a m er ic a, w es t a si a he rb (h er ba c on va lla ri ae ) co nv al la to xi n, c on va llo si de , co nv al la to xo l ca rd io to ni c: a nt iar rh yt hm ic , st re ng th en in g th e ac tiv ity o f th e he ar t m us cl e c ar di ol c , c on va fo rt , k el ic ar di na ru sc ac ea e sp re ng . 7) o ph io po go n ja po ni cu s ( l. f. ) k er .-g aw l. so ut hea st a si a rh iz om e, ro ot (r ad ix o ph io po go ni s) op hi og en in , r us co ge ni ns , sp ir os ta ns , o ph io si de a , m on ot er pe ne g lic os id e, op hi op og on ol an tisw el lin g, v as op ro te ct iv e, in th e tr ea tm en t o f v ar ic os e ve in s a nd h ae m or rh oi ds ; st re ng th en s t he h ea rt a nd im pr ov es b lo od c ir cu la tio n; an tic oa gu la nt ; u se d as a n au xi lia ry a �e r a st ro ke ru sc og en in 8) r us cu s ac ul ea tu s l . m ed ite rr an ea n ba si n rh iz om e (r us ci rh iz om a) st er oi d sa po ni ns (r us co ge ni n, ru sc in , r us co si de ), es se nt ia l oi l an tisw el lin g, v as op ro te ct iv e ag en ts , u se d in th e tr ea tm en t of v ar ic os e ve in s i n th e le gs an d ha em or rh oi ds d io sm in a pl us , d io st ri n, k rą że ni e i l ov e h er bs -c ap s, ru sc og en in , r us co la n, ru sc ov en p lu s, ru sk or ex , va ri xi na l, ve no 3 3 co m pl ex , ve no c or ec to r, ve no x po ac ea e ba rn ha rt (= g ra m in ea e) ju ss . 9) b am bu sa ar un di na ce a (r et z. ) w ill d. (= b. b am bo s ( l. ) vo ss ) so ut h c hi na , in di an su bc on tin en t le af , s ee d, ro ot am in o ac id s, si lic a, po ly sa cc ha ri de s, m in er al s, vi ta m in s ( ri bo �a vi n, th ia m in e, v ita m in c , n ia ci n an d βca ro te ne ), re si ns , w ax es an tiat he ro sc le ro tic , va so pr ot ec tiv e, a nt iha em or rh ag ic ve no c or ec to r p lants supporting the treatm ent of cardiovascular diseases 171 ra nu nc ul ac ea e ju ss . 10 ) a do ni s ve rn al is l. eu ro pe , a si a �o w er in g he rb (h er ba a do ni di s ve rn al is) ca rd en ol id e gl yc os id es (e .g . ad on ito xi n, a do ni to xo l), �a vo no id s ( e. g. v ite xi n, lu te ol in ), ph yt os te ro ls , ch ol in e, a do ni to l ca rd io to ni c: a nt iar rh yt hm ic – st re ng th en s a nd im pr ov es th e ac tiv ity o f t he h ea rt m us cl e, c al m in g k el ic ar di na , t in ct ur a a do ni di s v er na lis (c ur re nt ly on p re sc ri pt io n) h am am el id ac ea e r . b r. 11 ) h am am el is vi rg in ia na l . n or th a m er ic a; cu lti va te d in eu ro pe le af (h am am el id is fo liu m ) ta nn in s ( ga la to ni ns , g al lic ac id ), �a vo no id s, sa po ni ns as tr in ge nt , a nt iha em or rh ag ic la ta n że l, pr ep ar at io n h ż el , ru sc ov en p lu s, ve no cz ar ż el , ve no so l b on ifr at er s g ro ss ul ar ia ce ae d c . 12 ) r ib es n ig ru m l. eu ro pe , a si a le af (f ol iu m ri bi s n ig ri ), fr ui t (f ru ct us r ib is ni gr i ) �a vo no id s ( ru to si de ), ta nn in s, an th oc ya ni ns , vi ta m in c as tr in ge nt , v as op ro te ct iv e; ha s a p os iti ve e �e ct o n ci rc ul at io n an d he ar t fu nc tio n r ib es n ig ru m – m ac er at gl ic er yn ow y v ita ce ae ju ss . 13 ) v iti s v in ife ra l. m ed ite rr an ea n ba si n an d so ut hw es t a si a; c ul tiv at ed fr ui t ( v iti s v in ife ra f ru ct us ex tr ac tu m ), se ed (v iti s v in ife ra s em en ex tr ac tu m ), le af (v iti s v in ife ra f ol iu m ex tr ac tu m ) ta nn in s, a nt ho cy an in s, �a vo no id s, w ax es , v ita m in s, pr oc ya ni di ns , p ec tin s, po ly sa cc ha ri de s, a ro m at ic s, ca ro te no id s; s til be ne de ri va tiv es : r es ve ra tr ol , tr an sre sv er at ro l a nd vi ni fe ri ne in c ar di ov as cu la r d is or de rs , i t pr ev en ts a th er os cl er os is a nd is ch em ic h ea rt d is ea se c ol la de n, d op pe l h er z v ita l, k rą że ni e i l ov e h er bs -c ap s, ru sc ov en p lu s b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 172 m al pi gh ia ce ae ju ss . 14 ) m al pi gh ia gl ab ra l . so ut h pa rt o f n or th a m er ic a, c en tr al a m er ic a, no rt he rn pa rt o f s ou th a m er ic a fr ui t e xt ra ct ( f ru ct us ex tr ac tu m m al pi gh ia g la br a ) so ur ce o f v ita m in c se al s c ap ill ar ie s r os av it c v io la ce ae b at sc h 15 ) v io la tr ic ol or l. s. st . eu ro pe , n or th a m er ic a, n or th a fr ic a, a si a (s ib er ia ) he rb (h er ba v io la e tr ic ol or is) �a vo no id s ( ru to si de , qu er ce tin ), an th oc ya ni ns (v io la ni n) , p he no lic a ci ds , ca ro te no id s ( vi ol ax an th in ), m uc ila ge s a nd ta nn in s va so pr ot ec tiv e, to ni ng br at ek , b ra te k �x , r ek to sa n, v io la – � x pa ss i� or ac ea e ju ss . e x r ou ss el 16 ) p as si� or a in ca rn at a k er .g aw l. so ut hea st u sa ; c ul tiv at ed in c en tr al a nd so ut h a m er ic a he rb (h er ba pa ss i� or ae ) in do le a lk al oi ds , � av on oi ds , cy an og en ic g ly co si de , ph yt os te ro ls , m in er al sa lts , pa si �o ri n lo w er s b lo od p re ss ur e, st re ng th en s t he h ea rt , r ed uc es sp as m s o f t he c or on ar y ve ss el s pa ss is pa sm in a sa lic ac ea e m ir b. 17 ) s al ix sp . co sm op ol ita n ba rk (c or te x sa lic is) sa lic in , � av on oi ds , c ha lc on es , ph en ol ic g ly co si de s, st er ol s, te rp en es a nd o th er s an tico ag ul an t, se al in g on bl oo d ve ss el s ru tin os al c fa ba ce ae l in dl . ( le gu m in os ae ju ss . = p ap ili on ac ea e g is ek e) 18 ) m el ilo tu s o� ci na lis (l .) pa ll. eu ro pe , c en tr al a si a; in tr od uc ed in a m er ic a he rb (m el ilo ti he rb a) co um ar in , d ic ou m ar ol , co um ar ic a ci d, m el yl to si de , al la nt oi n, � av on oi ds , t an ni ns an tico ag ul an ts , a nt isw el lin g, in th e tr ea tm en t o f po or ly h ea lin g w ou nd s a nd ha em or rh oi ds ta bl et ki to ni zu ją ce p lants supporting the treatm ent of cardiovascular diseases 173 19 ) s ar ot ha m nu s sc op ar iu s ( l. ) w im m . e x w .d .j. k oc h c en tr al a nd so ut h eu ro pe he rb (h er ba sa ro th am ni ) qu in ol iz id in e al ka lo id s (s pa rt ei ne , l up an in e, hy dr ox yl up an in e, sa ro ta m in e) , a m in es , �a vo no id s an tiar rh yt hm ic , s tr en gt he ns th e he ar t, in cr ea se s b lo od pr es su re a nd d ila te s b lo od ve ss el s c ar di os an , h em or ol 20 ) s op ho ra ja po ni ca l . ( = st yp hn ol ob iu m ja po ni cu m (l .) sc ho tt) c hi na , k or ea ; cu lti va te d �o w er (s op ho ra e ja po ni ca e �o s) , fr ui t ( so ph or ae ja po ni ca e fr uc tu s) �a vo no id s ( qu er ce tin , r ut in ), ph os ph ol ip id s, al ka lo id s, po ly sa cc ha ri de s, es se nt ia l oi ls , o rg an ic a ci ds , v ita m in s c , p su pp or ts th e w or k of th e he ar t, no rm al is es b lo od pr es su re , s tr en gt he ns b lo od ve ss el s, an tico ag ul an ts k rą że ni e i l ov e h er bs -c ap s, ti nc tu ra s op ho ra e ja po ni ca e ro sa ce ae ju ss . 21 ) a ro ni a m el an oc ar pa (m ic hx .) el lio tt n or th a m er ic a; cu lti va te d in eu ro pe fr ui t ( a ro ni ae fr uc tu s) an th oc ya ni ns , � av on oi ds , ta nn in s, or ga ni c ac id s, pe ct in s hy po te ns iv e, su pp or ts th e fu nc tio ni ng o f t he c ir cu la to ry sy st em , i n hy pe rt en si on , i n ha em or rh oi ds a ro ni a so k 10 0% , h er ba tk a �x ż yl ac ze k, k ap su łk i a ro ni a, n a ci śn ie ni e ka ps uł ki 22 ) c ra ta eg us la ev ig at a (p oi r.) d c . a nd c . m on og yn a ja cq . eu ro pe , a si a, n or th a fr ic a in �o re sc en ce ( i n� or es ce nt ia c ra ta eg i ) , f ru it (c ra ta eg i f ru ct us ) �a vo no id s ( vi ta m in , r ut os id e, ap ig en in ), am in es (c ho lin e, ac et yl ch ol in e) , p he no lic a ci ds (c a� ei c ac id ), co um ar in s (e sc ul in ), ph yt os te ro ls , b vi ta m in s, vi ta m in c ca rd io to ni c, h yp ot en si ve , re la xa nt , v as op ro te ct iv e c ar di ac tiv – n al ew ka , c ar di ol c , c ar di ot on ic , c iś ni en ie n or m a – he rb at ka zi oł ow a, c ra vi so l, d op pe l h er z v ita l, ex tr as pa zm in a, g łó g zi oł a w ta bl et ka ch , k el ic ar di na , n eo ca rd in a, n eo sp as m in a, n eu ro si n te a, p as si sp as m in a, p lu sz go ld v ita l, sk le ro sa n, st ro ph an th us c om p. , ta bl et ki to ni zu ją ce , ve no fo rt on , 23 ) p ot en til la er ec ta (l .) r ae us ch . c en tr al a nd no rt he rn eu ro pe , a si a rh iz om e (t or m en til la e rh iz om a) ta nn in s, el la gi c ac id , q ui no ni c ac id , t or m en to si de an tiha em or rh ag ic , d ia st ol ic , as tr in ge nt h em or ol b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 174 24 ) r os a ca ni na l. no rt he rn he m is ph er e �o ra l c up – hy pa nt hi um (f ru ct us r os ae ) �a vo no id s, an th oc ya ni ns , ca ro te no id s, xa nt ho ph yl ls , ta nn in s, vi ta m in s c , e , k , p an d gr ou p b st re ng th en in g ca pi lla ri es , an tisp as m od ic , t on in g c ar di os an , n eo ca rd in a, pl us z go ld v ita l, r os av it c , 25 ) r ub us sp . eu ro pe , a si a fr ui t ( fr uc tu s r ub i) an th oc ya ni ns , t an ni ns , vi ta m in c su pp or ts th e w or k of th e ci rc ul at or y sy st em : i nc re as e th e el as tic ity o f c ap ill ar ie s an d st re ng th en s a nd de to xi �e s t he h ea rt c ar di os an 26 ) s an gu is or ba o� ci na lis l . eu ro pe , a si a, n or th a m er ic a rh iz om e (r ad ix sa ng ui so rb ae ), he rb (h er ba sa ng ui so rb ae ). ta nn in s, sa po ni ns , � av on oi ds in h ae m or rh oi ds , a nt iha em or rh ag ic z ie le k rw iś ci ąg u 27 ) s or bu s au cu pa ri a l. em en d. h ed l. eu ro pe , a si a �o w er (s or bi � os ), fr ui t ( so rb i f ru ct us ) or ga ni c ac id s, ca ro te no id s, ta nn in s, bi tte rn es s, so rb ito l, vi ta m in c an tiat he ro sc le ro tic , a nt iin �a m m at or y, as tr in ge nt o w oc ja rz ęb in y he rb at ka zi oł ow a r ek to sa n, s kl er os an u rt ic ac ea e ju ss . 28 ) u rt ic a di oi ca l. co sm op ol ita n le af (f ol iu m u rt ic ae ), ro ot (r ad ix u rt ic ae ), se ed (s em en u tr ic ae d io ic ae ) ta nn in s, ca ro te no id s, �a vo no id s, m ic ro el em en ts , am in es , v ita m in s b , c , k hy po te ns iv e, to ni c, an tiha em or rh ag ic , a nt isp as m od ic , a nt ian ae m ic c ar di o fa rm h ip po ca st an ac ea e d c . 29 ) a es cu lu s hi pp oc as ta nu m l . ba lk an pe ni ns ul a; cu lti va te d in eu ro pe ko ra (c or te x h ip po ca st an i) , �o w er (f lo s h ip po ca st an i) , fr ui t ( fr uc tu s h ip po ca st an i im m at ur us ), se ed (s em en h ip po ca st an i) sa po ni ns (e sc in ), co um ar in s (e sc ul in , e sc ul et in ), �a vo no id s, �a vo ne s va so pr ot ec tiv e, a nt ied em a, an tiha em or rh ag ic , i n th e tr ea tm en t o f p er ip he ra l sy st em d is or de rs , v ar ic os e ve in s o r h ae m or rh oi ds a es ci n że l, a es cu la n, es ce ve n, h em or ol , h er ba tk a �x ż yl ac ze k, in tr ac tu m h ip po ca st an i, k rą że ni e i l ov e h er bs , l at an ż el , n eo -a es cu la n że l, r ek to sa n, sa po ve n t, v ar ix in al , ve ne sc in , v en of or to n, v en ol p lants supporting the treatm ent of cardiovascular diseases 175 ru ta ce ae ju ss . 30 ) c itr us au ra nt iu m l . so ut hea st a si a pe ri ca rp (a ur an tii am ar i e pi ca rp iu m et m es oc ar pi um ), �o w er (a ur an tii am ar i � os ) es se nt ia l o ils , r ut in , n ar in gi n, he sp er id in , v ita m in p se al s b lo od v es se ls ; i nc re as es bl oo d pr es su re k rą że ni e i l ov e h er bs , ru tin a c m ax , v en o 33 co m pl ex , v en oc or ec to r, ve no x 31 ) c itr us li m on bu rm . f . so ut hea st a si a; c ul tiv at ed in m an y co un tr ie s pe ri ca rp (c itr i pe ri ca rp iu m ) ru tin , v ita m in s c a nd b 1 a nd ci tr ic a ci d se al s b lo od v es se ls d io sp ir in 32 ) r ut a gr av eo le ns l . eu ro pe , n or th a fr ic a he rb (h er ba ru ta e) , l ea f ( fo liu m ru ta e) �a vo no id s ( ru to si de , qu er ce tin ), fu ra no co um ar in s, qu in ol in e, a cr id in e an d fu ra nc in ol in e al ka lo id s va so pr ot ec tiv e, d ia st ol ic , hy po te ns iv e, a nt ied em a; im pr ov es p er ip he ra l ci rc ul at io n ve no c or ec to r st er cu lia ce ae v en t. 33 ) c ol a ni tid a (v en t.) s ch ot t & e nd l. an d c . ac um in at a (p . be au v. ) s ch ot t & en dl . w es t a fr ic a em br yo s f ro m se ed s ( c ol ae se m en ) al ka lo id s, am on g ot he rs ca �e in e, th eo br om in e ca �e in e di la te s t he c er eb ra l an d co ro na ry v es se ls o f th e he ar t a nd n ar ro w s t he ab do m in al v es se ls ; i nc re as es bl oo d pr es su re c ar di ol c m al va ce ae ju ss . 34 ) h ib is cu s sa bd ar i� a l. tr op ic al a fr ic a; cu lti va te d in a ra b co un tr ie s an d in di a �o w er (h ib is ci sa bd ar i� ae � os ) an th oc ya ni ns , p ol yp he no ls , �a vo no id s, or ga ni c ac id s (c itr ic , m al ic , o xa lic , hi bi sc us ), so ur ce o f v ita m in c lo w er s b lo od p re ss ur e an d ch ol es te ro l – im pr ov es h ea rt fu nc tio n bi o hi bi sc us , c ar di os an b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 176 br as sic ac ea e bu rn et t 35 ) c ap se lla bu rs apa st or is (l .) m ed ik . ea st er n pa rt o f m ed ite rr an ea n ba si n; c ur re nt ly co sm op ol ita n he rb (h er ba b ur sa e pa st or is) �a vo no id s ( e. g. ru tin , di os m os is , h es pe ri di n) , am in o ac id s ( e. g. or ni th in e, α -a m in ob ut yr ic ac id ), m on ot er pe no id s, gl uc os in ol at es (s yn ig ri n) , e tc . as tr in ge nt , a nt iha em or rh ag ic ; i n ha em or rh oi ds h em or ig en fe m in a lo ra nt ha ce ae ju ss . 36 ) v is cu m al bu m l . eu ro pe , a si a he rb (h er ba v is ci ) �a vo no id s ( qu er ce tin ), am in es (h is ta m in e, c ho lin e, ac et yl ch ol in e) , v is co to xi ns , ph en ol ic a ci ds (c a� ei c ac id , fe ru lic a ci d) se da tiv e, h yp ot en si ve ; re gu la te s t he w or k of th e he ar t a nd d ila te s t he v es se ls c ra vi so l, in tr ac tu m v is ci , sk le ro sa n, v en of or to n po ly go na ce ae ju ss . 37 ) f ag op yr um es cu le nt um m oe nc h eu ro pe , c en tr al a si a. he rb (f ag op yr i he rb a) �a vo no id s ( qu er ce tin de ri va tiv e, ru to si de ), ph en ol ic ac id s, vi ta m in c va so pr ot ec tiv e; g en tly lo w er s bl oo d pr es su re es sa ve n ge l, h em or ig en fe m in a, k el ic ar di na , r ut is ol , ve no c or ec to r 38 ) p ol yg on um av ic ul ar e l. eu ro pe he rb (h er ba po ly go ni a vi cu la ri s) �a vo no id s ( hy pe ro si de , qu er ce tin ), ph en ol ic a ci ds (c a� ei c, c ho lo ro ge ni c) , ta nn in s an tiha em or rh ag ic , t on in g; cl ea ns in g th e bl oo d h er ba tk a �x ż yl ac ze k, sk le ro sa n c ac ta ce ae ju ss . 39 ) s el en ic er eu s gr an di �o ru s ( l. ) br it. & r os e c en tr al a nd so ut h a m er ic a �o w er in g st em s fa vo no id s (c ac ci tin , r ut in , qu er ce tin -3 -r ut os id e) , hy pe ro si de (h yp er in or k em pp er ry th ri n) , bi og en ic a m in es (t yr am in e, n -m et hy lty ra m in e) , di m et hy l t yr am in e (h or de ni ne ), m uc us , r es in w ax es , g ly co si de s in th e he ar t f ai lu re , he ar t w ea kn es s an d at he ro sc le ro si s st ro ph an th us c om p. p lants supporting the treatm ent of cardiovascular diseases 177 � ea ce ae m ir b. e x k er g aw l. 40 ) c am el lia sin en sis (l .) o . k un tz e c hi na , i nd ia , bu rm a; cu lti va te d le af (f ol iu m � ea e ) al ka lo id s (c a� ei ne , th eo br om in e, th eo ph yl lin e) , ta nn in s, m in er al s al ts , co um ar in s, � av on oi ds (q ue rc et in , k ae m pf er ol ), sa po ni ns a nd e ss en tia l o ils re gu la te s th e co nt en t o f su bs ta nc es th at c on st ri ct an d ex pa nd b lo od v es se ls – pr ev en tio n of a rt er ia l hy pe rt en si on ; i n la rg e do se s it in cr ea se s bl oo d pr es su re k rą że ni e i l ov e h er bs er ic ac ea e ju ss . 41 ) k al m ia la tif ol ia l . ( = c ha m ae da ph ne la tif ol ia (l .) k un tz e) ea st p ar t o f n or th a m er ic a le af (f ol iu m k al m ia e) di te rp en es (e .g . gr ay an ot ox in s) , � av on oi ds , ta nn in s, ph yt os te ro ls (s ito st er ol ), te rp en es , ph en ol ic g ly co si de s ( ar bu tin ) hy po te ns iv e; in h yp er tr op hy an d pa lp ita tio ns o f h ea rt st ro ph an th us c om p. 42 ) o xy co cc us pa lu st ri s p er s. n or th a nd c en tr al eu ro pe , n or th a si a, n or th a m er ic a fr ui t ( o xy co cc i fr uc tu s) , l ea f (o xy co cc i f ol iu m ) ta nn in s, an th oc ya ni ns , or ga ni c ac id s ( ci tr ic , q ui ni c, ur so lic , b en zo ic ); so ur ce o f vi ta m in c lo w er in g bl oo d pr es su re , as tr in ge nt , v as op ro te ct iv e c ol la de n 43 ) v ac ci nu m m yr til lu s l . eu ro pe , n or th a m er ic a, so ut he rn a si a fr ui t ( fr uc tu s m yr til li) , l ea f (f ol iu m m yr til li) fr ui t – ta nn in s, an th oc ya ni ns , pr os cy an id in , p he no lic ac id s, ur so lic a ci d; le af – ta nn in s, gl yc os id es (a rb ut in ), �a vo no id s, qu in ol iz id in e al ka lo id s, ph en ol ic a ci ds , tr ite rp en es as tr in ge nt , v as op ro te ct iv e bi lb er in , c ol la de n, h er ba tk a �x ż yl ac ze k, v ar ix in al ru bi ac ea e ju ss . 44 ) g al iu m od or at um (l .) sc op . eu ro pe , s ou th a fr ic a he rb (h er ba a sp er ul ae ) co um ar in s, �a vo no id s, or ga ni c ac id s, ta nn in s, vi ta m in c se da tiv e, a nt iin �a m m at or y, re la xa nt ; i m pr ov es b lo od ci rc ul at io n, lo w er s b lo od cl ot tin g h er ba a sp er ul ae 178 b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń lo ga ni ac ea e r . b r. ex m ar t. 45 ) s pi ge lia an th el m ia l . a nt ill es a nd so ut h a m er ic a he rb al ka lo id s: is oq ui no lin e an d ac tin id in ety pe a lk al oi d ca rd io to ni c pr op er tie s; ac ce le ra te s t he h ea rt ra te , re du ce s h ea rt rh yt hm di st ur ba nc es st ro ph an th us c om p. , h ee l cr al on in k ro pl e a po cy na ce ae ju ss . 46 ) r au vo l� a se rp en tin a (l .) be nt h. so ut he rn a nd so ut hea st er n a si a ro ot (r ad ix r au w ol �a e ) in do le a lk al oi ds (a jm al in e, re se rp in e) , p hy to st er ol s se da tiv e, h yp ot en si ve , an tiar rh yt hm ic : i n ca rd ia c ar rh yt hm ia s, pa lp ita tio ns a nd at ri al � br ill at io n, d ia st ol ic se rp in a 47 ) s tr op ha nt hu s gr at us (w al l. & h oo k. ) b ai ll. w es te rn a fr ic a se ed (s em en st ro ph an th i g ra ti ) st ro ph an th in g (u ab ai n) a nd k (c om be tin ) ca rd ia c: a nt iar rh yt hm ic , st re ng th en s a nd im pr ov es th e ac tiv ity o f t he h ea rt m us cl e st ro ph an th us c om p. 48 ) v in ca m in or l. eu ro pe , a si a he rb ( h er ba v in ca e m in or is ) in do le a lk al oi ds (v in ca m in e, vi nc in in e, v in ca m in or in ), �a vo no id s, an th oc ya ni ns , tr ite rp en es , t an ni ns , v ita m in c an tiha em or rh ag ic , hy po te ns iv e, d ia st ol ic : di la te s p er ip he ra l v es se ls an d st re ng th en s t he h ea rt fu nc tio n ba rw in ek p os po lit y (v in ca m in or ) so la na ce ae ju ss . 49 ) a tr op a be lla do nn a l. eu ro pe , a si a m in or , n or th a fr ic a an d a m er ic a he rb (h er ba be lla do nn ae ), le af (b el la do nn ae fo liu m ), ow oc e (f ru ct us be lla do nn ae ), ro ot (r ad ix be lla do nn ae ) tr op an e al ka lo id s ( at ro pi ne , hy os cy am in e, sc op ol am in e) , be lla do ni ne , c ou m ar in s, �a vo no id s, ta nn in s di as to lic ; s pe ed s u p th e he ar t ra te h em or ol p lants supporting the treatm ent of cardiovascular diseases 179 o le ac ea e ju ss . e x r . b r. 50 ) o le a eu ro pa ea l . eu ro pe , a fr ic a, a si a; c ul tiv at ed le af (f ol iu m o le ae ) ph en ol ic s ( ol eu ro pe in , hy dr ox yt yr os ol ), �a vo no id s (lu te ol in , r ut in ), qu in ol in e al ka lo id s an tiat he ro sc le ro tic , a nt ith ro m bo tic ; i m pr ov in g bl oo d �o w in th e co ro na ry v es se ls li ść o liw ny la m ia ce ae m ar tin ov (= l ab ia ta e ju ss .) 51 ) b et on ic a o� ci na lis l . eu ro pe , n or th a si a he rb (h er ba be to ni ca e) , l ea f (f ol iu m b et on ic ae ) ta nn in s, ph en ol ic a ci ds , �a vo no id s, ch ol in e, v ita m in c an tiin �a m m at or y, as tr in ge nt , a nt iha em or rh ag ic , i nc re as in g bl oo d cl ot tin g h er ba b et on ic ae 52 ) l eo nu ru s ca rd ia ca l . eu ro pe , a si a, n or th a m er ic a (t em pe ra te zo ne ) he rb (l eo nu ri ca rd ia ca e he rb a) ca rd ia c gl yc os id es , a lk al oi ds (s ta ch yd ri ne , l eo nu ri n, co nc re te ic in e, fu ri ci n) , ph en yl pr op an oi ds (l eo no si de a , b ), �a vo no id s ( qu er ce tin , ka em pf er ol , a pi ge ni n) , an th oc ya ni ns , t an ni ns , �t os te ro ls , u rs ol ic a ci d, ch ol in e ca rd ia c, h yp ot en si ve , t on ic , se da tiv e, d ia st ol ic , r eg ul at es th e w or k of th e he ar t c ar di os an , t ab le tk i to ni zu ją ce 53 ) m el iss a o� ci na lis l . m ed ite rr an ea n ar ea le af (m el iss ae fo liu m ) te rp en oi ds (c itr al , l in al oo l, ci tr on el la l), ta nn in s, ph en ol ic ac id s ( ca �e ic , r os em ar y, fe ru lic a ci d) , t ri te rp en es , �a vo no id s, vi ta m in c se da tiv e, h yp ot en si ve , a nt isp as m od ic ; i n ar rh yt hm ia s o r ca rd ia c ne ur os is c ar di o fa rm , c ra vi so l, d op pe l h er z v ita l, n er vo sa n �x , n er vo so l, n eu ro si n te a, pl us z go ld v ita l, ta bl et ki to ni zu ją ce b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 180 54 ) r os m ar in us o� ci na lis l . m ed ite rr an ea n ar ea le af (r os m ar in i fo liu m ), ol ej ek (r os m ar in i o le um ) es se nt ia l o il (b or ne ol , lim on en , c am ph or ), �a vo no id s, ta nn in s, ro sm ar in ic a ci d, sa po ni ns , ph yt os te ro ls an tisp as m od ic , t on ic , an tiat he ro sc le ro tic ; d ila te s ca pi lla ri es a nd st im ul at es bl oo d ci rc ul at io n pl us z go ld v ita l, c ar di ac tiv , es sa ve n ge l a st er ac ea e be rc ht . & j. p re sl (= c om po sit ae g is ek e) 55 ) a ch ill ea m ill ef ol iu m l . s .s. eu ro pe , a si a, n or th a m er ic a, a us tr al ia a nd n ew z ea la nd he rb (m ill ef ol ii he rb a) , k w ia to st an (m ill ef ol ii �o s) , l ea f (m ill ef ol ii fo liu m ) es se nt ia l o il (c ha m az ul en e, ac hi lle in , b et ai ne ), se sq ui te rp en es , � av on oi ds , ch ol in e, ta nn in s an tiha em or rh ag ic , a nt isp as m od ic , h yp ot en si ve ; i n th e tr ea tm en t o f v ar ic os e ve in s a nd h ae m or rh oi ds h em or ig en fe m in a, h em or ol , n er vo sa n �x , n er vo so l, r ek to sa n, sk le ro sa n 56 ) a rn ic a m on ta na l . c en tr al eu ro pe , a si a an d n or th a m er ic a in �o re sc en ce (a rn ic ae � os ) se sq ui te rp en es (a rn ic ol id es a , b , c a nd d , h el en al in ), po ly ph en ol s ( cy na ri n, c a� ei c ac id ), �a vo no id s ( as tr ag al in e, is oq ue rc et in ), tr ite rp en es , ca ro te no id s, ph yt os er ol s an tiin �a m m at or y, ca rd ia c, an tied em a, a nt ico ag ul an t, va so pr ot ec tiv e, in cr ea se s bl oo d pr es su re a rc al en , a rn ic al ż el , st ro ph an th us c om p. ti nc tu ra a rn ic ae , v en oc za r że l, ve no fo rt on , v en os ol bo ni fr at er s 57 ) c al en du la o� ci na lis l . m ed ite rr an ea n ar ea ; c ul tiv at ed in �o re sc en ce (c al en du la e �o s) tr ite rp en e sa po ni ns , ca ro te no id s, �a vo no id s, co um ar in s ( sc op ol et in , es cu le tin ) va so pr ot ec tiv e, a nt ied em a, re du ce s t he p er m ea bi lit y an d fr ag ili ty o f c ap ill ar ie s a rc al en , m aś ć na gi et ko w a, ve no cz ar ż el , v en ol , v en os ol bo nf ra te rs 58 ) c ha m om ill a re cu tit a (l .) r au sc he rt eu ro pe , a si a, n or th a m er ic a, a us tr al ia in �o re sc en ce (m at ri ca ri ae � os ) es se nt ia l o il (c ha m az ul en e, αbi sa bo lo l), � av on oi ds (a pi ge ni n, q ue rc et in , pa lu le tin ), co um ar in s (u m be lli fe ro ne ), ch ol in e, vi ta m in c di as to lic ; u se d as a n au xi lia ry in h ae m or rh oi ds a es cu la n, h em or ol , h er ba tk a �x ż yl ac ze k, n eo ae sc ul an ż el , n er vo sa n �x , n eu ro si n te a, v en ol 59 ) c on yz a ca na de ns is (l .) c ro nq ui st co sm op ol ita n he rb (h er ba er ig er on tis ca na de ns is) �a vo no id s ( ru to si de ), ph en ol ic a ci ds (c a� ei c, fe ru lic ), ta nn in s, ph yt os te ro ls , vi ta m in c as tr in ge nt , a nt iha em or rh ag ic h em or ig en fe m in a p lants supporting the treatm ent of cardiovascular diseases 181 60 ) c yn ar a sc ol ym us l . et hi op ia ; cu lti va te d in th e m ed ite rr an ea n, n or th a fr ic a an d a m er ic a he rb (c yn ar ae he rb a) , l ea f (c yn ar ae fo liu m ) cy na ri n, c a� ei c an d ch lo ro ge ni c ac id , cy na ro pi cr in , � av on oi ds , ph yt os te ro ls an tiat he ro sc le ro tic ; l ow er s ch ol es te ro l a nd lo w er s t he co nt en t o f l ip id s i n th e bl oo d c yn ar ex , s yl ic yn ar a ra lia ce ae ju ss . 61 ) p an ax g in se ng c .a . m ey er a nd p. q ui nq ue fo liu s l. cu lti va te d in a si a an d n or th a m er ic a ro ot (g in se ng ra di x, g in se ng ex tr ac tu m si cc um ) tr ite rp en e sa po ni ns (g in se no si de s) , a lc oh ol s (p an ax id ol , p an ax in ol , fa lk ar in ol ), �a vo no id s, am in o ac id s, ch ol in e an tiat he ro sc le ro tic , st re ng th en in g th e he ar t m us cl e; in cr ea se s b lo od pr es su re bo dy m ax p lu s, g in ko gi ns , g in se lin , ż eń -s ze ń vi ta co m pl ex a pi ac ea e li nd l. (= u m be lli fe ra e ju ss .) 62 ) a m m i vi sn ag a (l .) la m . m ed ite rr an ea n ar ea ; in tr od uc ed to c en tr al an d no rt he rn eu ro pe fr ui t ( a m m i vi sn ag ae fr uc tu s) py ra no co um ar in s ( vi sn ad in e, sa m id in e) , f ur an oc hr om on e de ri va tiv es (k el in , v is na gi ne ), �a vo no id s, ph en ol ic a ci ds , st er ol s di as to lic , d ila te s b lo od ve ss el s; us ed in c or on ar y ar te ry d is ea se , i n pr e a nd po st -i nf ar ct io n co nd iti on s k el ic ar di na , t in ct ur a a m m i vi sn ag ae 63 ) c en te lla as ia tic a (l .) u rb . so ut h an d c en tr al a si a an d a fr ic a, a us tr al ia , so ut h an d c en tr al a m er ic a he rb (c en te lla e as ia tic ae h er ba ) tr ite rp en e sa po ni ns (a si at ic os id e, m ad ec as so si de , ce nt el lo si de , s cc ep ho le os id e) an d ac id s ( as ia tic , ce nt el ic ), m on ot er pe ne s, se sq ui te rp en es , p ol ya ce ty le ne co m po un ds , � av on oi ds hy pe rt en si on , p er ip he ra l ci rc ul at io n an d el as tic ity of b lo od v es se ls – in at he ro sc le ro si s ru sc ov en p lu s, va ri xi na l, ve no c or ec to r b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 182 c ap ri fo lia ce ae ju ss . 64 ) s am bu cu s ni gr a l. eu ro pe , w es te rn a nd c en tr al a si a ro ot (r ad ix sa m bu ci ), co rt ex (c or te x sa m bu ci ), le af (f ol iu m sa m bu ci ), �o w er (f lo s s am bu ci ), fr ui t ( fr uc tu s sa m bu ci ) �a vo no id s, ph en ol ic a ci ds (c a� ei c an d ch lo ro ge ni c ac id ), ta nn in s, m uc ila ge s va so pr ot ec tiv e, su pp or ts m ic ro ci rc ul at io n sy ro p m al in a z kw ia te m bz u cz ar ne go z w ita m in ą c ; j ui ce s a nd d ec oc tio ns o f �o w er s a nd d ri ed fr ui t va le ri an ac ea e ba ts ch 65 ) v al er ia na o� ci na lis l . eu ro pe , a si a, n or th a m er ic a. ro ot (r ad ix va le ri an ae ) te rp en e al ka lo id s ( va le ri ne , sc in ta nt in , a ct in id in e) , ir id oi ds , e ss en tia l o il (v al er ic ac id , b or ne ol , i so eu ge no l) re la xi ng , d ia st ol ic ; i n ar rh yt hm ia s o r c ar di ac ne ur os is c ar di ol c , c ar di ot on ic , ex tr as pa zm in a, n eo ca rd in a, n eo sp as m in a, n er vo sa n �x , n er vo so l, pa ss is pa sm in a p lants supporting the treatm ent of cardiovascular diseases 183 tab. 3. list of plants with a therapeutic e�ect in cardiovascular diseases healing action list of species number of species generally cardiac adonis vernalis, arnica montana, convallaria majalis, crataegus laevigata, c. monogyna, hibiscus sabdari�a, kalmia latifolia, leonurus cardiaca, strophanthus gratus, styphnolobium japonicum, urginea maritima, viscum album 12 strengthening the heart muscle adonis vernalis, ammi visnaga, camellia sinensis, convallaria majalis, ophiopogon japonicus, panax ginseng, p. quinquefolius, passi�ora incarnata, rosa canina, rubus sp., sarothamnus scoparius, selenicereus grandi�orus, strophanthus gratus, urginea maritima, vinca minor, vitis vinifera 16 anti-arrhythmic adonis vernalis, convallaria majalis, rauvol�a serpentina, sarothamnus scoparius, strophanthus gratus, spigelia anthelmia, urginea maritima 7 anti-atherosclerotic allium sativum, a. ursinum, bambusa arundinacea, centella asiatica, cynara scolymus, fucus vesiculosus, equisetum arvense, olea europaea, panax ginseng, p. quinquefolius, rosmarinus o�cinalis, selenicereus grandi�orus, sorbus aucuparia, vitis vinifera 14 anti-coagulants allium sativum, a. ursinum, arnica montana, betonica o�cinalis, ginkgo biloba, melilotus o�cinalis, olea europaea, ophiopogon japonicus, salix sp., styphnolobium japonicum 10 anti-edema aesculus hippocastanum, arnica montana, calendula o�cinalis, melilotus o�cinalis, ophiopogon japonicus, ruscus aculeatus, ruta graveolens 7 anti-haemorrhagic achillea millefolium, aesculus hippocastanum, bambusa arundinacea, betonica o�cinalis, capsella bursa-pastoris, conyza canadensis, equisetum arvense, hamamelis virginiana, polygonum aviculare, potentilla erecta, urtica dioica, vinca minor 12 anti-spasmodics melissa o�cinalis, rosa canina, rosmarinus o�cinalis 3 diastolic achillea millefolium, ammi visnaga, atropa bella-donna, chamomilla recutita, cola nitida, c. acuminata, crataegus laevigata, c. monogyna, galium odoratum, leonurus cardiaca, potentilla erecta, rauvol�a serpentina, ruta graveolens, urtica dioica, vinca minor 15 anti-hypertensive (lowering blood pressure) achillea millefolium, allium sativum, a. ursinum, aronia melanocarpa, camellia sinensis (small doses), centella asiatica, crataegus laevigata, c. monogyna, fagopyrum esculentum, fucus vesiculosus, hibiscus sabdari�a, kalmia latifolia, leonurus cardiaca, melissa o�cinalis, oxycoccus palustris, passi�ora incarnata, rauvol�a serpentina, ruta graveolens, urtica dioica, valeriana o�cinalis, vinca minor, viscum album 22 hypertensive (raising blood pressure) arnica montana, camellia sinensis (large doses), citrus aurantium, cola nitida, c. acuminata, panax ginseng, p. quinquefolius, sarothamnus scoparius, 8 vasoprotective (protect the structure of blood vessels) aesculus hippocastanum, arnica montana, bambusa arundinacea, calendula o�cinalis, centella asiatica, citrus aurantium, citrus limon, crataegus laevigata, c. monogyna, fagopyrum esculentum, ginkgo biloba, malpighia glabra, ophiopogon japonicus, oxycoccus palustris, ribens nigrum, rubus sp., ruscus aculeatus, ruta graveolens, salix sp., sambucus nigra, styphnolobium japonicum, vaccinum myrtillus, viola tricolor 23 toning leonurus cardiaca, polygonum aviculare, rosa canina, rosmarinus o�cinalis,urtica dioica, viola tricolor 6 astringent (causing the blood vessels to contract) betonica o�cinalis, capsella bursa-pastoris, conyza canadensis, hamamelis virginiana, oxycoccus palustris, potentilla erecta, ribens nigrum, sorbus aucuparia, vaccinum myrtillus 9 b ar ba ra k ub ik , a lin a s ta ch ur sk as w ak oń 184 rośliny wspomagające leczenie chorób układu krwionośnego streszczenie choroby układu sercowo-naczyniowego należą do grupy chorób cywilizacyjnych odpowiedzialnych za główną część zgonów na świecie. w leczeniu i zapobieganiu tym chorobom stosuje się liczne leki i preparaty, w tym zawierające substancje roślinne, po które szczególnie chętnie sięgają pacjenci we wczesnych stanach chorobowych. na rynku obecnych jest wiele preparatów roślinnych dostępnych bez recepty. celem opracowania było sporządzenie wykazu systematycznego roślin, które są stosowane powszechnie przez pacjentów przy leczeniu i suplementacji różnych schorzeń związanych z układem krwionośnym. na podstawie 100 preparatów dostępnych w aptekach lub sklepach zielarskich bez recepty w polsce sporządzono wykaz gatunków roślin, które scharakteryzowano pod kątem ich pochodzenia, rodzaju surowca zielarskiego, składów chemicznych i właściwości leczniczych wykorzystywanych w chorobach krążenia. sporządzony wykaz systematyczny obejmuje 65 gatunków, głównie roślin naczyniowych. najbardziej powszechnym surowcem stosowanym w preparatach dostępnych na rynku jest crataegus sp., aesculus hippocastanum oraz ruscus aculeatus. większość odnotowanych tu roślin wykazuje działanie wazoprotekcyjne (np. ginkgo biloba, ophiogon japonicus, ruscus aculeatus, ribes nigrum), obniża ciśnienie tętnicze krwi (np. aronia melanocarpa, fucus vesiculosus, passi�ora incarnata), działa rozkurczająco (np. leonurus cardiaca, polygonum aviculare). lista systematyczna obejmuje 23 gatunki roślin rodzimych dla europy środkowej, natomiast substancje roślinne otrzymywane z 20 gatunków sprowadzane są różnych obszarów świata. keywords: blood vessel and heart diseases, dietary supplements, herbal therapies received: [2021.09.10] accepted: [2021.11.17]