3 17 Afkar M. Hadi Bull. Iraq nat. Hist. Mus. (2011) 11 (4): 17-25 ISOLATION AND IDENTIFICATION OF INTESTINAL PARASITES FROM VEGETABLES FROM DIFFERENT MARKETS OF IRAQ. Afkar M. Hadi Iraq Natural History Research Center and Museum. University of Baghdad ABSTRACT This investigation was designed to determine the occurrence of intestinal parasites in fresh vegetables(Apium graveolense, Lepidium aucheri and Allium porrum), from different markets as a primary effort in Iraq. Eight genera and species of intestinal parasites appear in vegetables, they were as follow: Echinococcus sp. 50%,Oxyuris equi 45%,Habronema sp. 45%,Parascaris equroum 31.6%,Strongyloides westrei 30%,Toxocara sp. 18.3%,Ascaris lumbricoides 11.6% and Hymenolepis sp. 8.3% .The scarcity of fresh water has meant that urban gardeners are increasingly irrigating their plots with wastewater. This poses a threat to public health in addition of roaming dogs in open farms. All studied areas showed high rates of eggs .No significant difference noticed between total rates of north and middle of Iraq. There were highly significant differences in the species of parasites among areas. INTRODUCTION Since 1973, WHO considered reuse of wastewater is special health issue (WHO 1973).In investigations on pathogenic organisms in wastewater and sludge, parasites have received the least attention (WHO 2004). In Iraq, a few studies reported in the literature on parasites transmitted through wastewater and sludge. The total contamination rate with parasites in sewage water was 60% at five regions in Baghdad (Al-Dura, Hi-Al- Maalf, Hi-Al-Jehad, Al- Shabab, and Al-Baya'a) ( Hadi . 2008) This investigation (as primary effort) was designed to determine the occurrence of intestinal parasites in some vegetables which are eaten fresh together in Iraq(Apium graveolense, Lepidium aucheri and Allium porrum), from different markets of Iraq. MATERIALS AND METHODS A total of 60 sample of vegetable groups (Apium graveolense, Lepidium aucheri and Allium porrum) each one weigh 500 gm during study months (November 2009 – April 2010). The samples were collected from markets of north region (Arbil & Kirkuk) and middle region (Baghdad, Najif &Diwania). The samples were tested according to Bariden, (1980). Generally, this method use sedimentation to concentrate the eggs on centrifugal force. Photographs were taken for eggs, then they diagnosed with the help of some Professors. RESULTS Echinococcus sp. showed the highest rate 50% (30) then Oxyuris equi and Habronema sp. 45% (27) (Table 1). Total rates of pollution with the eggs of parasites were 67.3%(97)in the middle area of Iraq compared with 32.6%(47) in the north area ( Table 2). Brief description and measurements of each helminthes is given below (Table 3). 18 Isolation and Identification of Intestinal Table 1: Genera &species of parasites isolated from fresh vegetable groups 60/500mg. Final host % of total No. of veg. positive Parasites Dog Horse Horse Horse Horse Dog &cat Man Man 50% 45% 45% 31.3% 30% 18.3% 11.6% 8.3% 30 27 27 19 18 11 7 5 Echinococcus sp Oxyuris equi Habronema sp. Parascaris equorum Strongyloides westeri Toxocara sp. Ascaris lumbricoides Hymenolepis sp Table 2: Distribution of parasites in the north and middle of Iraq. middle 40 specimens North 20 specimens Parasites 26 15 15 11 9 9 7 5 4 12 12 8 9 2 0 0 Echinococcus sp. Oxyuris equi Habronema sp. Parascaris equorum Strongyloides westeri Toxocara sp. Ascaris lumbricoides Hymenolepis sp 97 67.3% 47 32.6% Total Table.3: Measurements and brief description of parasites eggs from vegetables. Parasites eggs measurements Descriptions of eggs contains Echinococcus sp. 30 -36 µ Spherical, thick, smooth shell. Fig.1 Hexacanth embryo. Oxyuris equi 85 -45 µ Ovoid, slightly asymmetrical, dissimilar side- walls. Fig.2 Larva Habronema sp. 45 - 16 µ Cylindrical, strongly elongated, thick shell. Fig.3 Larva Parascaris equorum 95 -90 µ Nearly spherical, brown yellowish. Thick albominous shell covered with fine dots. One or two cells Strongyloides westeri 45 -30 µ Ovoid, side walls are symmetrical. Similar, wide poles.fig.5 Short thick larva Toxocara sp. 75 -80 µ Nearly spherical, thick rough, pitted shell.fig.6 brown to black granular contents 19 Afkar M. Hadi Ascaris lumbricoides 55 -45 µ Ellipse to round , golden brown thick, rough albuminous outer wall.fig.7 thin yolk membrane Hymenolepis sp. 38 µ Round, grayish, transparent. Smooth, thin membranous shell. Fig.8 oncosphere is 24 µ by 16 µ DISCUSSION Factors affect the occurrence and concentrations of helminthes eggs and protozoan cysts observed in raw wastewater, include the endemicity of disease within the indigenous animal and human population ( Grimason 1995). In current study, a large number of parasites 8 genera and species from fresh vegetable groups this result conjugated with level of incidence of parasitic infection in the community and concentrations of parasitic organisms, such as intestinal nematodes eggs, in the wastewater of such a community ( Dixo 1993). Study shows high rates of pollution with Echinococcus sp and Toxocara sp. eggs of dogs and cats which means high risk to community since Holly (2008) indicated that they may not only be most pathogenic to their specific host, they may also be the major causes of zoonosis. In Iraq, the agriculture of study vegetable groups in open farms where dogs and cats roaming there, which they really risk for human. The presence of many species of horse helminthes (Oxyuris equi, Habronema sp., Parascaris equorum and Strongyloides westeri) in vegetable groups came probably as a result of reuse of feces of equine for manuring. This is accordance with Ram (2009) who found that the reuse of Equine feces in farms lead to re infection with these parasites for horses and unknown results when human ingested them! In England nearly all horses are infected with nematodes. (Thienpont et. al.1986), while in Iraq Faraj & Shabban (2007) found that the total equine infection rate 50.45%. The presence of round worms (Ascaris lumbricoides ) and tapeworm (Hymenolepis sp.) in vegetable groups mean that, they are all readily transmitted by the agricultural use of raw or insufficiently treated excreta and wastewater, indeed, they are the excreted pathogens of greatest public health concern in agricultural reuse schemes. There are highly significant differences P<0.01 among the species of parasites between the two studied areas , Echinococus sp. and Toxocara sp.of dogs were recorded high rates in middle, compared with north area , this result is conjugated with number of roaming dogs in farms. This is similar to Sultan (1997) who showed that infection rate with Toxocara canis 46% of dogs in Najif area. Moreover, in Basrha Al-Emara and Yakub (1999)found that total dogs infection 35.2% with three nematodes: Toxocara canis, Toxocara leonine and Ancylostoma caninum, thus contaminated soil 25% and contaminated gardens grasses 10% from funfair in Basrha. Aside from, infected rate in Mousel area (in north) 25.7% with same parasites (Al-Kalidi, 1983). 20 Isolation and Identification of Intestinal The study shows that infection with eggs of equine worms are high in the north of Iraq, this may be due to prevalence of Equines (horse and mules) in villages and mountains of north area. On the other hand, absence of human parasites (Ascaris lumbricoides and Hymenolepis sp.) in the north area may indicate decreased level of wastewater reuse. Finally, according to present results and in response to claim, WHO (2004) wastewater must use in irrigation of crops to be eaten cooked, sport field, public Parks; cereal crops; industrial crops; fodder and trees; not for crops eaten raw. ACKNOWLEDGEMENTS The author wishes to thank Prof. Mohammad K. Mohammad (Iraqi Natural History Museum and researches center) and wish to thank Prof. Joop Boomker (Department of Veterinary Tropical Diseases, University of Pretoria) for diagnosis the eggs. I also thank Dr. Mike Kinsella. Missoula. MT. America for supporting and helping in diagnosis the eggs of parasites. REFERENCES Al-Emara, G. Yakub. 1999. Epidemiology study of Nematode in digestive Tract of Dogs in Basrha area. MSc. Thesis. Coll. Vet. Med. Uni. Baghdad. Al-Kalidi, N. W.; Daoud, M.S.; Shubber, A. H. and Al-Alousi; T. I. 1983. A survey for internal and external parasites in dogs in Mosul(Iraq). Iraq. J. Vet. Sc., (1-2): 9-14. Bariden, K. (1980). Factors modifying the prevalence of bovine ostertagiasis with special reference to infective larvae in soil. M. tyySc. Thesis, University of Glasgow, U. K. Dixo, N. G., Gambrill, M. P., Catunda, P. F. and Van Haandel, A. C. (1993). Removal of pathogenic organisms from the effluent of an up flow anaerobic digester using waste stabilization ponds. Proc. 2nd IAWQ Special Conference on waste stabilization Ponds and the Ruse of effluents. Brazil. Faraj, A. A.; Shabban F. (2007). Prevalence of internal parasites (stomach &intestine) in horses in Iraq. Iraq. J. Vet, Sc., Vol. 31 No.2. Grimason, A.M., Smith, H. V., Young, G. and Thitai, W. N. (1995). Occurrence and removal of Ascaris spp. ova by waste stabilization ponds in Kenya. 3rd IAWQ International Specialist Conference and Workshop "waste stabilization ponds Technology and Application". Brazil, preprint volume. Hadi, A. M.; Faraj, A. A. (2008). Distribution of intestinal parasites in drinking water in some regions in Baghdad. Al-Qadisiya Journal of Vet. Med. Sci. vol.17 no.12 2008. Holly N., (2008). Roundworms (Toxascaris leonine, Toxocara cati). Veterinary services department, Drs. Foster and Smith , Inc. Pet Education.com Ram Chandra S. (2009). A Report on Prevalence of Helminthes Parasites in Mules of Brick Kiln of Lalitpur District. www.animalnepal.org/documents/donkey/ Sultan, B. E. (1997). Epidemiology study of Toxocara canis in Najef area. PHD. Thesis. Coll. Vet. Med. Uni. Baghdad. http://www.animalnepal.org/documents/donkey/ 21 Afkar M. Hadi Thienpont E., Rochette F., Vanparijs O. F. J.(1986). Diagnosing helminthiasis by coprological examination. Turnhoutsebaan 30, 2340 Beerse, Belgium). Available from: Vet lab Services, UNIT 11. Station Road, South water, Sussex RH 13 7HQ. World Health Organization (1973). Reuse of Effluents: Methods of Wastewater Treatment and Health Safeguards. Technical Report Series No.517. World Health Organization (2004). Integrated Guide to Sanitary Parasitology. Regional office for the Eastern Mediterranean. Regional Center for Environmental Health Activities Amman- Jorden 2004. 22 Isolation and Identification of Intestinal Fig.1: Eggs of Echinococcus sp Fig.2: Egg of Oxyuris equi. Fig.3: Egg of Habronema sp. 23 Afkar M. Hadi Fig.4: Egg of Parascaris equorum. Fig.6: Egg of Toxocara sp. Fig. 5:Strongyloides westeri. 24 Isolation and Identification of Intestinal Fig.7: Egg of Ascaris lumbrecoides. Fig.8: Egg of Hymenlepois sp. 25 Afkar M. Hadi Bull. Iraq nat. Hist. Mus. (2011) 11 (4): 17-25 ض من بع م وية ت ال ط يليا ص ال خي ل وت ن عز م ختلفة و ق ن أس ت ضروا ال ق .العرا ي هاد كا مسل أف ي ف التار خ ال بيع ح ح ث ومت ز ب جامعة بغداد -مرك ة ص الخال ل ؤك وا ال ت ضر خل ع ا ت ن ب ط يليا ص ال خي ش ع ل ت س ب درا ذ ال ه م اهت ق را يف ل عا ث(م كرا د ال شا س الر كرف ث ) ال راق ي ع يف ا ول م ة خم ل ة و ق وا س ن م ة وية ا ختلف ت ملع ع م الطف ليا سة انية أجنا و نوا را هرت ا د مايلي أظ :وك Echinococcus sp. 50%, Oxyuris equi 45%, Habronema sp. 45%, Parascaris equroum 31.6%, Strongyloides westrei 30%, Toxocara sp. 18.3%, Ascaris lumbricoides 11.6% and Hymenolepis sp. 8.3% . وت س ن وبن ب مت ا ب واخليو واإلن ص ب الك يت ت وهذا بسبب الزراعة املفتوحة و جتوال ةال مل يف قي ا ة ت ال ائل ف عما ا خل ست دة ع إعا ضال ة في ا ف ب ل ائب جة ال ال عا نتي و زر ة ذب مليا لع ص ا زارع النق وأي ا تس يد ا ة عا ة ل صح ى ل عل سي ا ج ط ا ل خ ك ذا ي وه تب ت احلي انا ال ض ق .ف ق لعرا ضر مناط خل ث ا ت ل يف عدال س رتفا ا ت لدرا ظهر ة (أ مالي ش ال ى ط س ض الخ الفا) والو ع ت تعلى حد واء مع ب طفيليا و ع ل ني أن صائية ح إل .ا