J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 102 Short Communication Species Composition and Ecological Aspects of Immature Mosquitoes (Diptera: Culicidae) in Bromeliads in Urban Parks in the City of São Paulo, Brazil Walter Ceretti-Junior 1, Rafael de Oliveira Christe 1,2, Marco Rizzo 3, Regina Claudia Strobel 3, Marco Otavio de Matos Junior 4, Maria Helena Silva Homem de Mello 4, Aristides Fernandes 1, Antônio Ralph Medeiros-Sousa 1, Gabriela Cristina de Carvalho 1, *Mauro Toledo Marrelli 1,2 1Department of Epidemiology, School of Public Health, University of São Paulo, São Paulo, Brazil 2Tropical Medicine Institute, University of São Paulo, São Paulo, Brazil 3Office of Parks and Green Areas, Department of the Environment and Green Areas, São Paulo City Hall, São Paulo, SP, Brazil 4Laboratory for Research into and Identification of Synanthropic Fauna, Zoonosis Control Center, Health Surveillance Coordination Unit, Municipal Health Department, São Paulo City Hall, São Paulo, SP, Brazil (Received 26 Apr 2014; accepted 6 Dec 2014) Abstract Background: Bromeliads can be epiphytic, terrestrial or saxicolous and use strategies to allow water to be re- tained in their leaf axils, where various arthropods can be found. These include mosquitoes, whose larvae are the most abundant and commonly found organisms in the leaf axils. The objective of this study was to look for im- mature forms of mosquitoes (the larval and pupal stages) in bromeliads in municipal parks in São Paulo and to discuss the ecological and epidemiological importance of these insects. Methods: From October 2010 to July 2013, immature mosquitoes were collected from bromeliads in 65 munici- pal parks in the city of São Paulo, Brazil, using suction samplers. The immature forms were maintained until adult forms emerged, and these were then identified morphologically. Results: Two thousand forty-two immature-stage specimens belonging to the genera Aedes, Culex, Trichoprosopon, Toxorhynchites, Limatus and Wyeomyia were found in bromeliads in 15 of the 65 parks visited. Aedes albopictus was the most abundant species (660 specimens collected), followed by Culex quinquefasciatus (548 specimens) and Cx. (Microculex) imitator (444). The taxa with the most widespread distribution were Ae. aegypti and Toxorhynchites spp, followed by Ae. albopictus and Cx. quinquefasciatus. Conclusion: Bromeliads in urban parks are refuges for populations of native species of Culicidae and breeding sites for exotic species that are generally of epidemiological interest. Hence, administrators and surveillance and mosquito-control agencies must constantly monitor these microenvironments as the presence of these species endangers the health of park users and employees as well as people living near the parks. Keywords: Mosquitoes, Bromeliads, Municipal Parks, Culicidae, Breeding sites, Immature, Brazil Introduction Members of the family Bromeliaceae can be epiphytic, terrestrial or saxicolous. The taxon is essentially Neotropical, with more than 3,100 described species distributed in 58 genera. In Brazil, where 40 % of these species are found, the highest diversity is in the Atlantic forest (Givnish et al. 2011). Bromeliads use strategies that allow water and organic debris to be retained in their leaf axils, forming rosettes or tanks from which they acquire nutrients. These structures also act as biotopes for various organisms. Central and lateral bro- meliad tanks provide a permanent habitat for many terrestrial arthropods that have aquatic immature stages (Frank et al. 2004), such as insects of the order Diptera, whose larvae are the most abundant and commonly *Corresponding author: Dr Mauro Toledo Marrelli, E-mail: mmarelli@usp.br http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 103 found organisms in these biotopes (Derraik and Heath 2005). Among the most com- mon dipterans, the culicids include species whose immature forms are specialists in colonizing these microhabitats as well as species that are occasional visitors (Frank and Lounibos 2009). The family Culicidae currently has around 3,500 valid described species worldwide (Harbach 2014). Many culicid species are of great importance in public health be- cause the blood-feeding habits of the fe- males are associated with allergies and the transmission of pathogens (Consoli and Lourenço-de-Oliveira 1994, Forattini 2002). The genera Aedes, Anopheles and Culex include important vectors of diseases such as malaria, dengue and filariasis in various regions of the planet (WHO 2013a, Bhatt et al. 2013, WHO 2013b). In addition, a number of mosquito-borne diseases, such as the West Nile virus, Chikungunya and yellow fever were or are public health prob- lems in many countries (Weaver 2013). Brazil has a great variety of culicids, 490 species having been described to date (WRBU 2014). In Brazil, malaria, which is transmitted by Anopheles, and dengue, which is transmitted by Ae. aegypti, are to- gether responsible for thousands of cases of morbidity. Also worthy of note is lym- phatic filariasis, which is transmitted by Cx. quinquefasciatus in some cities in the northeast of the country, as well as various arboviruses that circulate in natural envi- ronments and cause sporadic cases of epi- zootics or even human infections such as Saint Louis encephalitis, Mayaro encephalitis, equine encephalitis, Rocio encephalitis and wild yellow fever (Forattini 2002, Oliveira- Ferreira et al. 2010, Viana and Ignotti 2013). The city of São Paulo, located in south- eastern Brazil, is the largest city in South America and has more than 11 million in- habitants. Mosquito-borne diseases in the city include dengue, outbreaks of which occur every year and which was responsi- ble for 2,617 confirmed autochthonous cases in 2013 (São Paulo 2014a). Malaria outbreaks due to Plasmodium vivax and P. malariae in the Atlantic Forest have been detected on the edge of the city of São Pau- lo (Couto et al. 2010, Duarte et al. 2013). The Ilhéus virus was isolated in birds in the Tietê Ecological Park, in the munici- pality of São Paulo (Pereira et al. 2001). In regions of the Atlantic Forest close to the city of São Paulo, antibodies for various arboviruses that cause different types of encephalitis were detected in birds (Fer- reira et al. 1994) and there have been spo- radic cases of wild yellow fever in humans and primates, the virus responsible for the disease having been isolated in mosquitoes from the species Haemagogus leucocelaenus (Souza et al. 2009). As urbanization continues apace, many cities around the world strive to preserve their green areas (Koh and Sodhi 2004). The construction, preservation and reha- bilitation of green areas in parks or plazas is intended to provide leisure facilities for the population, preserve remnants of flora and fauna and reduce the environmental and social stress caused by urbanization (McIntyre 2000, McKinney 2008). The city of São Paulo currently has a hundred pub- lic parks, many of which contain fragments of the Atlantic Forest and representatives of native or exotic flora and fauna (São Paulo 2014b). The diversity of culicids found in some of these green fragments has been investigated in a variety of studies (Urbinatti et al. 2001, Taipe-Lagos and Na- tal 2003, Montes 2005, Ribeiro et al. 2012). A recent preliminary survey in fifty-nine urban parks in the city of São Paulo found that some medically important species of culicids, such as Ae. albopictus, Ae. fluviatilis, Ae. scapularis, Cx. nigripalpus, and Cx. quinquefasciatus, are quite common in these environments (Medeiros-Sousa et al. 2013). These and other species maintain their pop- ulations using a variety of natural or artifi- cial containers found in these green frag- ments that allow their immature forms to develop. Bromeliads can act as breeding grounds for many species of epidemiologic http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 104 interest in the city as they are very common in parks, where they occur naturally and in some cases are planted as part of the land- scape. The aim of this study was to investigate the species composition and ecological aspects of mosquitoes collected in brome- liads in urban parks in São Paulo to deter- mine how important these plants are for the maintenance of populations of native or exotic culicid species, with a particular em- phasis on species of epidemiological interest. Materials and Methods The city of São Paulo (23.54° W 46.63° S) is located in the Atlantic Forest in the southeast of Brazil at an altitude of 750 m above sea level and with a mean relative humidity of 78 % (Fig. 1). The climate is considered temperate tropical, and there is reduced rainfall in the winter. Mean annual temperature is 20.7 °C, winters are mild (15 to 21 °C) and summer temperatures are moderately high (22 to 27 °C). Mean annual rainfall is 1,376 mm, and most of the rain- fall is concentrated in the summer months (IBGE 2014). The species composition of immature cu- licids in fifteen parks with bromeliads in the city of São Paulo was investigated over thirty-four months. Data collected by Medei- ros-Sousa et al. (2013) between October 2010 and February 2011 were used, to- gether with data from collections made be- tween March 2011 and July 2013. Bro- meliads planted in the ground (terrestrial bromeliads) and bromeliads on tree trunks (epiphytic bromeliads) at heights ranging from 1.0 to 5.0 m were sampled. Immature specimens were actively sought out in the central and lateral leaf axils of bromeliads using suction samplers (Fig. 2). The ex- tracted contents were then transferred to small recipients (400ml) and analyzed for the presence of mosquito larvae and pupae. The larvae and pupae collected were taken to the Public Health Entomology Labora- tory at the Faculty of Public Health, Uni- versity of São Paulo, where they were main- tained until the emergence of adult forms, which were identified morphologically based on Lane (1953), Consoli and Lourenço-de- Oliveira (1994) and Forattini (2002), as well as by comparing them with standard spec- imens in the collection at the Faculty of Public Health, University of São Paulo. The abbreviations for genera and subgene- ra used here follow the standardization proposed by Reinert (2009). After all the individuals collected had been identified morphologically, the num- ber of individuals in terrestrial and epi- phytic bromeliads in each park broken down by species was recorded in tabular form (Table 1). The frequency with which each species was found in terrestrial and epi- phytic bromeliads was calculated to deter- mine whether there was an association be- tween culicid species and type of brome- liad (Table 2). Results Overall, 2,042 immature-stage speci- mens belonging to 6 genera (Aedes, Culex, Trichoprosopon, Toxorhynchites, Limatus and Wyeomyia) and 17 species were found in 15 of the 65 parks visited (Fig. 1, Table 1). Table 1 shows the number of specimens of each species collected and the type of bromeliad (epiphytic or terrestrial) in which collections were made. Sixty-six of the specimens collected were classified in the taxon Tx. spp and five in Wy. spp either because they died before the adult form emerged or because of limitations in the keys for morphological identification, which were out of date. The only clear association between culicid species and type of brome- liad (Table 2) was for Cx. quinquefasciatus, one of the most abundant species, which was found exclusively in terrestrial brome- liads. The bromeliads found in the parks be- longed to the genera Aechmea, Alcantarea, Edmundoa, Neoregelia, Nidularium, Quesnelia and Vriesea, all of which are http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 105 common in the Atlantic Forest. Aedes albopictus was the most abundant mos- quito species collected (660 specimens or 32.35% of the total), followed by Cx. quinquefasciatus (548 specimens or 26.86%) and Culex (Microculex) imitator (444 or 21.76%). Another epidemiologically im- portant species collected was Ae. aegypti (122 or 5.98%). The other species together accounted for only 13.05% of the total number collected (Table 1). The taxa with the widest distributions were Ae. aegypti and Tx. spp., found in eight parks, followed by Ae. albopictus (seven parks) and Cx. quinquefasciatus (five parks). The parks with the highest species rich- ness were Anhanguera Park, Ibirapuera Park and Santo Dias Park, each of which had eight species, followed by Previdência Park with six species, Piqueri Park with five and Burle Marx Park with four. No more than three species were recorded for any of the other parks (Table 1). The largest numbers of specimens inde- pendently of species were collected in Piqueri Park (1,019) and Santo Dias Park (632), which together accounted for almost three quarters of all the specimens col- lected and had the largest number of bro- meliads. Ibirapuera Park and Park Mario Covas, where 136 and 102 individuals were collected, respectively, also accounted for a significant number of the sample popu- lation. The remaining parks together ac- counted for just 151 specimens. Fig. 1. Location of the fifteen urban parks in the city of São Paulo, Brazil, where immature stages of culicids were found in bromeliads between October 2010 and July 2013 Fig. 2. Collection of immature mosquitoes in leaf axils of bromeliads using suction samplers in urban parks in the city of São Paulo, Brazil, between October 2010 and July 2013. (A) Terrestrial bromeliad, (B) Epiphytic bromeliad http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 106 Table 1. Number of immature mosquitoes collected by species for each urban park and type of bromeliad (terrestrial or epiphytic) collected between October 2010 and July 2013 in the city of São Paulo, Brazil Taxon A n h an gu er a B u rl e M ar x C ar m o C ol in as d e S ão F ra n ci sc o C h ic o M en d es G u ar ap ir an ga Ib ir ap u er a L aj ea d o M ár io C ov as N ab u co P iq u er i P re vi d ên ci a R od ri go d e G as p er i S an to D ia s S ev er o G om es T ot al F re q u en cy ( % ) T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic T er re st ri al E p ip h yt ic Aedes (Stegomyia) aegypti (Linnaeus, 1762) 3 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 2 0 0 0 72 19 2 2 0 7 0 13 0 0 122 5.97 Aedes (Stegomyia) albopictus (Skuse, 1895) 16 0 7 0 0 0 0 0 0 5 0 0 28 14 0 0 0 0 0 0 90 467 2 7 0 0 3 21 0 0 660 32.3 Aedes (Ochlerotatus) fluviatilis (Lutz, 1904) 0 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 6 0.3 Culex (Culex) chidesteri Dyar, 1921 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0.05 Culex (Culex) quinquefasciatus Say, 1823 15 0 0 0 0 0 0 0 0 1 0 0 70 0 0 0 100 0 0 0 362 0 0 0 0 0 0 0 0 0 548 26.8 Culex (Microculex) imitator imitator Theobald, 1903 0 0 0 0 0 0 1 0 0 0 0 0 3 0 0 0 0 0 0 0 0 0 0 0 0 0 78 362 0 0 444 21.7 Culex (Microculex.) Group Imitator (Theobald, 1903) 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 13 23 0 0 40 1.9 Limatus durhami Theobald, 1901 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7 0 0 1 0 0 0 0 0 0 8 0.4 Trichoprosopon (Trichoprosopon) pallidiventer (Lutz, 1905) 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 3 0.15 Toxorhynchites bambusicola (Lutz , Neiva, 1913) 1 0 0 0 0 0 0 0 0 0 0 0 3 2 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 2 9 0.4 Toxorhynchites solstitialis (Lutz, 1904) 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0.1 Toxorhynchites spp. Theobald, 1901 9 0 29 0 0 0 0 2 0 0 5 0 8 0 0 0 0 0 0 1 0 0 0 8 0 0 1 3 0 0 66 3.2 Wyeomyia (Phoniomyia) davisi (Lutz, 1904) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7 52 0 0 59 2.9 Wyeomyia (Miamyia) oblita (Lutz, 1904) 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0.15 Wyeomyia serratoria (Dyar , Nunez Tovar, 1927) 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 2 0.1 Wyeomyia (Phoniomyia) cf galvaoi (Correa, Ramalho, 1956) 0 0 0 0 0 0 0 0 0 0 9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 51 0 0 64 3.1 Wyeomyia. spp Theobald, 1901 0 0 4 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 0.2 Total no. of individuals 49 0 41 0 0 3 1 2 0 7 14 0 119 17 1 0 102 0 0 1 533 486 6 19 0 7 106 526 0 2 2,042 100 Total no. of species 8 4 3 2 3 2 8 2 2 1 5 6 1 8 1 http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 107 Table 2. Number and frequency of culicids in decreasing order of abundance in terrestrial and epiphytic bromeliads collected between October 2010 and July 2013 in urban parks in the city of São Paulo, Brazil Species N. in terrestrial bromeliads Frequency in terrestrial bromeliads N. in epiphytic bromeliads Frequency in epiphytic bromeliads Total no. of specimens Aedes (Stegomyia) albopictus (Skuse, 1895) 146 0.22 514 0.77 660 Culex (Culex) quinquefasciatus Say, 1823 547 0.99 1 0.002 548 Culex (Microculex) imitator imitator Theobald, 1903 82 0.18 362 0.81 444 Aedes (Stegomyia) aegypti (Linnaeus, 1762) 80 0.65 42 0.34 122 Toxorhynchites spp Theobald, 1901 52 0.78 14 0.21 66 Wyeomyia (Phoniomyia) cf. galvaoi (Correa, Ramalho, 1956) 13 0.20 51 0.79 64 Wyeomyia (Phoniomyia) davisi (Lutz, 1904) 7 0.11 52 0.88 59 Culex (Microculex.) Group Imitator (Theobald, 1903) 15 0.37 25 0.62 40 Toxorhynchites bambusicola (Lutz , Neiva, 1913) 4 0.44 5 0.55 9 Limatus durhami Theobald, 1901 7 0.87 1 0.12 8 Aedes (Ochlerotatus) fluviatilis (Lutz, 1904) 6 1.0 0 0.0 6 Wyeomyia spp Theobald, 1901 5 1.0 0 0.0 5 Trichoprosopon (Trichoprosopon) pallidiventer (Lutz, 1905) 3 1.0 0 0.0 3 Wyeomyia (Miamyia) oblita (Lutz, 1904) 3 1.0 0 0.0 3 Toxorhynchites solstitialis (Lutz, 1904) 1 0.5 1 0.5 2 Wyeomyia serratoria (Dyar , Nunez Tovar, 1927) 1 0.5 1 0.5 2 Culex (Culex) chidesteri Dyar, 1921 0 0.0 1 1.0 1 Total no. of individuals per type of bromeliad 972 0.48 1,070 0.52 2,042 http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 108 Discussion This study, the first to investigate im- mature mosquitoes in bromeliads in mu- nicipal parks in the city of São Paulo, Bra- zil, found seventeen species or groups from the genera Aedes, Culex, Trichoprosopon, Toxorhynchites, Limatus and Wyeomyia in these plants. The findings were similar to those of previous studies except that we failed to find species from the subgenus Kerteszia of Anopheles, a group of mos- quitoes found almost exclusively in bro- meliads, some species of which are consid- ered vectors of “bromeliad-malaria” in the Atlantic Forest of South and Southeast Brazil (Consoli and Lourenço-de-Oliveira 1994, Marques et al. 2012, Ribeiro et al. 2012). This absence could be due to the urban or peri-urban nature of the parks visited, as this subgenus is considered a bioindicator of preserved environments (Dorvillé 1996). Of the Sabethini tribe, which includes mosquito species with diurnal habits usu- ally associated with less anthropic environ- ments, only species of the genera Limatus, Trichoprosopon and Wyeomyia were found. Of these species, Limatus durhami tends to be adapted to anthropic environments (Consoli and Lourenço de Oliveira 1994), and immature forms of this species have been reported in artificial breeding sites such as waste tires near houses (Zequi et al. 2005). Like most other Culicidae, the Sabethini are potential vectors of several arboviruses and have been found carrying the Pixuna, Bussuquara, Wyeomyia, Ilhéus and yellow fever viruses (Forattini 2002). In our survey, we recorded Ae. aegypti and Ae. albopictus in eight and seven parks, respectively. These species are the main vehicles of dengue and Chikungunya virus in various parts of the world but are con- sidered exotic species in Brazil (Bhatt et al. 2013). They use the same breeding sites in the urban environment, although Ae. albopictus is also often found inhabiting rural and natural environments. The finding of Ae. albopictus and Ae. aegypti in bromeliads demonstrates the high ecological valence of both species. Ac- cording to Crovello and Hacker (1972), the use of bromeliads by Ae. aegypti may be considered a remnant of primitive behavior of the species, as outside its original envi- ronment (Northwest of Africa) this species is found strictly only in synanthropic envi- ronments, is highly dependent on clean water in containers associated with intradomicile environments and exhibits anthropophilic behavior (Forattini 2002). Aedes albopictus is considered the wilder and more exophilic of the two species, with the potential to disperse through various habitats, and is often found breeding in phytotelmata, such as those in bromeliads and bamboos (Natal et al. 1997). Our results indicate that in ur- ban parks Ae. albopictus is better able to colonize bromeliads, a finding that may be related to the fact that these environments possess similar characteristics to those of rural and wild environments despite being embedded in an urban landscape. Another Aedeini registered was Ae. fluviatilis, a suspected vector in the trans- mission of arboviruses to humans that is found in wild or modified environments and is associated with small, usually tem- porary rupiculous breeding sites with little shade (Forattini 2002). In the present study, only six specimens were found (in Ibirapuera Park), showing that, although not their pre- ferred breeding sites, bromeliad phytotelmata may be used by this species for the devel- opment of its immature form. Culex quinquefasciatus is a recognized vector of the worm that causes Bancroftian filariasis infection, Wuchereria bancrofti. This cosmopolitan mosquito species is also associated with the transmission of West Nile virus in the United States, Canada and Mexico (Murray et al. 2010). The species is adapted to environments that have been changed by anthropogenic activities, main- ly in areas where the disorderly use of ur- ban spaces is associated with a lack of in- http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 109 frastructure and failures in sewage and drainage systems, resulting in the accu- mulation of dirty still waters, which favor the proliferation of this culicid (Morais et al. 2006). Although the species is characteristic of large breeding sites, it has high ecologi- cal valence and may occupy small natural and artificial containers, such as bromeli- ads, tree holes, bamboo and tires (Consoli and Lourenço de Oliveira 1994, Forattini 2002). Our results suggest that this species is found more abundantly in terrestrial bro- meliads in parks in the city of São Paulo. The subgenus Microculex Theobald, 1907 of Culex displays a strong preference for breeding in natural containers, especially bromeliads and tree hollows, the former typically used by species Cx. (Mcx) imita- tor (Muller and Marcondes 2006). In the present study, Cx. imitator was recorded in five parks, but in high abundance only in Santo Dias Park, where it was found in terrestrial and epiphytic bromeliads. The genus Toxorhynchites includes spe- cies that typically develop in bromeliads, tree holes and bamboo internodes and rare- ly in artificial breeding sites such as tin cans, barrels and tires (Lopes 1997). The adults are exclusively phytophagous and are considered to be the only culicid that is harmless to man and other vertebrates. Their larvae are equipped with powerful jaws adapted to predation of smaller im- mature aquatic arthropods, including other culicids. For this reason, species of this genus have been suggested as tools for bi- ological control in the context of integrated pest management (Nyamah et al. 2011). Bromeliads are considered highly selective breeding sites, and some species of culicids, such as those of the genera Wyeomiya and Trichoprosopon, are closely associated with these plants (Mocellin et al. 2009). The spatial distribution of these culicids is therefore quite restricted. The bromeliads found in parks may represent ecological refuges for these species, which may have been introduced in these green areas unintentionally when these plants were used for ornamentation. Previous studies have shown that vari- ous species of culicids are present in con- servation areas and public parks in the city of São Paulo (Urbinatti et al. 2001, Taipe- Lagos and Natal 2003, Montes 2005, Ribeiro et al. 2012, Medeiros-Sousa et al. 2013). Contributing to this body of knowledge, our study has shown that bromeliads in urban parks in the city of São Paulo not only act as refuges for populations of na- tive species of culicids, but are also used for breeding by exotic and epidemiologi- cally important species such as Ae. aegypti, Ae. albopictus and Cx. quinquefasciatus, which have a high ecological valence, are synanthropic and play an important role as vectors of human diseases. These green ar- eas constitute highly favorable niches, as they increase the supply of resources (food and shelter), provide protection against nat- ural enemies and physical changes in the environment (such as changes in tempera- ture and rainfall) and may improve the sur- vival of species that can exploit small and isolated remnants of native habitats and therefore tolerate a greater degree of urban development than species that are more sensitive to the same transformations in en- vironments occupied by humans (McIntyre 2000). Conclusion The bromeliads present in urban parks of the city of São Paulo not only act as refuges for populations of native species of culicids but also allow exotic and epidemi- ological important species such as Ae. aegypti, Ae. albopictus and Cx. quinque- fasciatus, to use these habitats for prolifer- ation of their populations. It is important for administrators and mosquito control and surveillance agencies to constantly monitor this microenvironment, since the presence of these species endangers the health of park users and employees as well as people living near the parks. http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 110 Acknowledgements We would like to thank the Fundação de Amparo à Pesquisa do Estado de São Paulo (BIOTA Program, Project 2010/51230-8) for providing financial support. We are also grateful to the field team from the De- partment of Epidemiology in the Faculty of Public Health, University of São Paulo; to the staff at the Office of Parks and Green Areas, Department of the Environment and Green Areas, São Paulo, and to the staff at the Zoonosis Control Center, São Paulo City Hall, who helped collect and identify the specimens. We extend our special thanks to M Bicudo de Paula for assistance with the identification of the specimens and to Dr PR Urbinatti for assistance with the fieldwork and for the pictures. The authors declare that there is no conflict of interests. References Bhatt S, Gething PW, Brady OJ, Messina JP, Farlow AW, Moyes CL, Drake JM, Brownstein JS, Hoen AG, Sankoh O, Myers MF, George DB, Jaenisch T, Wint GR, Simmons CP, Scott TW, Farrar JJ, Hay SI (2013) The global distribution and burden of dengue. Nature. 496(7446): 504–507. Consoli RAG, Lourenço-de-Oliveira R (1994) Principais mosquitos de importância sanitária no Brasil. FIOCRUZ, Rio de Janeiro, p. 225. Couto RDA, Latorre MDRDD, Di Santi, SM, Natal D (2010) Autochthonous malaria notified in the State of São Paulo: clinical and epidemiological characteristics from 1980 to 2007. Rev Soc Bras Med Trop. 43(1): 52–58. Crovello TJ, Hacker CS (1972) Evolution- ary strategies in life table character- istics among feral and urban strains of Aedes aegypti (L). Evol. 26(2): 185–196. Derraik JGB, Heath ACG (2005) Immature Diptera (excluding Culicidae) inhab- iting phytotelmata in the Auckland and Wellington regions. New Zealand J Marine Freshw Res. 39(4): 981–987. Dorvillé LFM (1996) Mosquitoes as bioindicators of forest degradation in Southeastern Brazil, a statistical eval- uation of published data in the litera- ture. Stud Neotrop Fauna Env. 31(2): 68–78. Duarte AMR, Pereira DM, de Paula MB, Fernandes A, Urbinatti PR, Ribeiro AF, Homem de Mello MHSH, Matos Jr MO, Mucci LF, Fernandes LN, Natal D, Malafronte RS (2013) Natu- ral infection in anopheline species and its implications for autochtho- nous malaria in the Atlantic forest in Brazil. Parasit Vectors. 6(1): 50–58. Ferreira IB, Pereira LE, Rocco IM, Marti AT, de Souza LT, Iversson LB (1994) Surveillance of arbovirus in- fections in the Atlantic Forest Re- gion, State of São Paulo, Brazil. I. Detection of hemagglutination-inhib- iting antibodies in wild birds be- tween 1978 and 1990. Rev Inst Med Trop São Paulo. 36(3): 265–274. Forattini OP (2002) Culicidologia Médica. Vol. 2. Edusp, São Paulo, p. 864. Frank JH, Sreenivasan S, Benshoff PJ, Deyrup MA, Edwards GB, Halbert SE, Hamon AB, Lowman MD, Mockford EL, Scheffrahn RH, Steck GJ, Thomas MC, Walker TJ, Welbourn WC (2004) Invertebrate animals extracted from native Ti- llandsia (Bromeliales: Bromeliaceae) in Sarasota County, Florida. Florida Entomol. 87(2): 176–185. Frank JH, Lounibos LP (2009) Insects and allies associated with bromeliads: a review. Terr Arthropod Rev. 1(2): 125– 153. Givnish TJ, Barffus MHJ, Van EB, Riina R, Schulte K, Horres R, Gonsiska PA, Jabaily RS, Crayn DM, Smith JAC, Inverno K, Brown GK, Evans TM, Holst BK, Luther H, Até W, Zizka http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 111 G, Barry PE, Sytsma KJ (2011) Phy- logeny, adaptative radiation, and his- torical biogeography in Bromeliaceae: insights from an eight-locus plastid phylogeny. Am J Botany. 98(5): 872– 895. Harbach RE (2014) Mosquito Taxonomic Inventory. Available at: http:// mos- quito-taxonomic-inventory.info/. IBGE (2014) Instituto Brasileiro de Geog- rafia e Estatística. Cidade de São Paulo. Available at: http://cidades. ibge.gov.br/xtras/home.php. Koh LP, Sodhil NS (2004) Importance of reserves, fragments, and parks for butterfly conservation in tropical ur- ban landscape. Ecol Appl. 14(6): 1698–1708. Lane J (1953) Neotropical Culicidae. Vol. 2. Universidade de São Paulo, São Paulo, Brazil. pp. 565. Lopes J (1997) Mosquito (Diptera: Culi- cidae) ecology in natural and arti- ficial rural breeding places in north- ern Parana, Brazil. V. Larvae collec- tion in artificial containers installed in ciliary Forest. Rev Saude Publ. 31(4): 370–377. Marques TC, Bourke BP, Laporta GZ, Sallum MAM (2012) Mosquito (Dip- tera: Culicidae) assemblages asso- ciated with Nidularium and Vriesea bromeliads in Serra do Mar, Atlantic Forest, Brazil. Parasit Vectors. 5(1): 1–9. McIntyre NE (2000) Ecology of urban ar- thropods: a review and a call to ac- tion. An Entomol Soc Am. 93(4): 825–835. McKinney ML (2008) Effects of urbaniza- tion on species richness: a review of plants and animals. Urban Ecosyst. 11(2): 161–176. Medeiros-Sousa AR, Ceretti Jr W, Urbinatti PR, Carvalho GC, Bicudo De Paula M, Fernandes A, Matos JR MO, Orico LD, Araujo AB, Nardi MS, Marrelli MT (2013) Mosquito fauna in Municipal parks of São Paulo City, Brazil: A preliminary survey. J Am Mosq Control Assoc. 29(3): 275–279. Mocellin MG, Simões TC, Nascimento TFS, Teixeira MLFT, Lounibos LP, Oliveira RP (2009) Bromeliad-in- habiting mosquitoes in an urban bo- tanical garden of dengue endemic Rio de Janeiro. Are bromeliads pro- ductive habitats for the invasive vec- tors Aedes aegypti and Aedes albopictus? Mem Inst Oswaldo Cruz. 104(8): 1171–1176. Montes J (2005) Culicidae fauna of Serra da Cantareira, Sao Paulo, Brazil. Rev Saúde Pública. 39(4): 578–584. Morais AS, Marrelli MT, Natal D (2006) Aspects of the distribution of Culex (Culex) quinquefasciatus Say (Dip- tera: Culicidae) in the region of the Pinheiros River, in the city of São Paulo, State of São Paulo, Brazil. Rev Bras Entomol. 50(3): 413–418. Muller GA, Marcondes CB (2006) Brome- liad-associated mosquitoes from At- lantic forest in Santa Catarina Island, southern Brazil (Diptera, Culicidae), with new records for the State of Santa Catarina. Iheringia Sér Zool. 96 (3): 315–319. Murray KO, Mertens E, Desprès P (2010) West Nile virus and its emergence in the United States of America. Vet Res. 41(6): 67. Natal D, Urbinatti PR, Taipe-Lagos CB, Ceretti-Jr W, Diederich ATB, Souza RG, Souza RP (1997) Aedes (Stego- myia) albopictus (Skuse) breeding in Bromeliaceae in the outskirts of an urban area of the city of São Paulo, Brazil. Rev Saude Publ. 31(5): 517– 518. Nyamah MA, Sulaiman S, Omar B (2011) Field observation on the efficacy of Toxorhynchites splendens (Wiede- mann) as a biocontrol agent against Aedes albopictus (Skuse) larvae in a cemetery. Trop Biomed. 28(2): 312– 319. http://jad.tums.ac.ir Published Online: June 27, 2015 J Arthropod-Borne Dis, March 2016, 10(1): 102–112 W Ceretti-Junior et al.: Species Composition … 112 Oliveira-Ferreira J, Lacerda MV, Brasil P, Ladislau JL, Tauil PL, Daniel- Ribeiro CT (2010) Malaria in Brazil: an overview. Mal J. 9: 115. Pereira LE, Suzuki A, Coimbra TLM, Souza RD, Chamelet ELB (2001) Ilheus arbovirus in wild birds (Sporo- phila caerulescens and Molothrus bonariensis). Rev Saúde Pública. 35 (2): 119–123. Reinert JF (2009) List of abbreviations for currently valid generic-level taxa in family Culicidae (Diptera). Eur Mos Bull. 27: 68–76. Ribeiro AF, Urbinatti PR, de Castro Duarte AM, de Paula MB, Pereira DM, Mucci LF, Fernandes A, de Mello MH, de Matos Júnior MO, de Oliveira RC, Natal D, dos Santos Malafronte R (2012) Mosquitoes in degraded and preserved areas of the Atlantic Forest and potential for vec- tor-borne disease risk in the mu- nicipality of São Paulo, Brazil. J Vector Ecol. 37(2): 316–324. São Paulo (2014a) Guia dos Parques Municipais de São Paulo. Prefeitura Municipal de São Paulo, SP, Brazil. 4th ed. Secretaria do Verde e Meio Ambiente, São Paulo, p. 193. São Paulo (2014b) Informe sobre Dengue. Secretaria de Saúde do Município de São Paulo, SP, Brazil. Available at: http://www.prefeitura.sp.gov.br/cida de/secretarias/upload/chamadas/infor me. Souza RPD, Petrella S, Coimbra, TLM, Maeda AY, Rocco IM, Bisordi I, Silveira VR, Pereira LE, Suzuki A, Silva SJS, Silva FG, Salvador FS, Tubaki RM, Menezes RT, Pereira M, Bergo ES, Hoffmann RC, Spinola RMF, Tengan CH, Siciliano MM (2011) Isolation of yellow fever virus (YFV) from naturally infected Hae- magogus (Conopostegus) leucoce- laenus (Dipteral: Culicidae) in São Paulo State, Brazil, 2009. Rev Inst Med Trop São Paulo. 53(3): 133–139. Taipe-Lagos CB, Natal D (2003) Abun- dância de culicídeos em área metro- politana preservada e suas implica- ções epidemiológicas. Rev Saúde Pública. 37(3): 275–279. Urbinatti PR, Sendacz S, Natal D (2001) Imaturos de mosquitos (Diptera: Culi- cidae) em parque de área metropo- litana aberto à visitação pública. Rev Saúde Pública. 35(5): 261–266. Viana DV, Ignotti EA (2013) Ocorrência da dengue e variações meteoroló- gicas no Brasil: revisão sistemática. Rev Bras Epidemiol. 16(2): 240–256. WRBU (2014) Walter Reed Bio- systematics Unit. Systematic catalog of Culicidae. Smithsonian Institution, Washington DC, USA. Available at: http://www. mosquitocatalog.org. Weaver SC (2013) Urbanization and geo- graphic expansion of zoonotic arbo- viral diseases: mechanisms potential strategies for prevention. Trends Microbiol. 21(8): 360–363. WHO (2013a) World Health Organization. Malaria entomology and vector con- trol: Guide for participants. Geneva, Switzerland. WHO (2013b) World Health Organization. Lymphatic filariasis: managing mor- bidity and preventing disability: an aide-mémoire for national programme managers. Geneva, Switzerland. Zequi AC, Lopes J, Medri IM (2005) Imaturos de Culicidae (Diptera) en- contrados em recipientes instalados em mata residual no município de Londrina, Paraná, Brasil. Rev Bras Zool. 22(3): 656–661. http://jad.tums.ac.ir Published Online: June 27, 2015