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Incidence of Gastrointestinal Helminthes in Captive Elephants in Wildlife Reserves of Nepal Dr.Kedar Karki, Senior Vet. Officer. Dr.

Poornima Manandhar Senior Vet.Officer Abstract: This study was conducted to determine the prevalence of parasitic infections among captive elephants of Different Wild life Reserve Park of Nepal .Fecal samples of 20 elephants were collected from Suklaphanta, Bardiya Sauraha and Koshi Tappu during 2007June-November 2007, and examined for the presence of helminthes ova. Of the 20 elephants, all were positive for helminthes. The incidence of helminthes infection as recorded in this study was out of 20 samples 6 samples were positive for Fasciola jacksoni,2 for Paramphistome,15 samples revealed the presence of egg and larvae of Oesophagostomum spp, and Chabertia spp 18 samples were positive for Schistosomes spp 1 sample was found positive for Dicrocoelium spp and 1 sample positive for Monezia spp. Location wise samples from Koshi Tappu wildlife reserve were positive for Fasciola and Schistosomes spp while samples from Chitwan National park were positive for Fasciola,Paramphistome and Oesophagostomum spp .The samples from Suklaphanta wildlife reserve and Bardia wildlife reserve were positive for Schistosomes spp and Oesophagostomum and Chabertia spp.As this is the first screening of the prevalence of Helminthes in Elephant and need to asses seasonalpattern . Background: Elephants are mainly raised in wildlife reserve for security purpose patrolling.Many resort in these wildlife area do keep these animal for tour operation.Very little work has been done to document the disease and parasite condition of this animal has been done in past. Material and Methods: Fecal samples were collected from different wild life reserve and Coprological analysis was done by qualitative method. Carpological findings S.N Koshi tappu 1 2

Fasciola Paramphistomum Oesophagostomum Schistosomes Dicrocoelium Mon spp spp/ Chabertia spp + + + + ++ + -

3 4 5 6 7 Sauraha 8 9 10 Suklaphanta 11 12 13 Bardiya 14 15 16 17 18 19 2o Total

++ ++ ++ + + -

++ + -

--+++ + ++ ++ ++ ++ ++ ++ ++ ++ ++ ++

+ + + ++ -++ ++ ++ ++ ++ ++ ++ ++ ++ ++ ++

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Review of literatures Bivitellobilharzia nairi was first recorded in India by Mudaliar Ramanujachar, Dutta and Srivastava, in 1955. A number of adult worm specimens of this schistosome species were recovered by Agatsuma, T., Rajapakse, R. P., Kuruwita, V. Y., Iwagami, M., and Rajapakse, R. C, from a domestic elephant, which died in 1999 in Sri Lanka. This is the first report of this schistosome from Sri Lanka (1). Saseendran, P. C., Rajendran, S., Subramanian, H., Sasikumar, M., Vivek, G., and Anil, K. S, conducted Study to determine the prevalence of parasitic infections among captive elephants in Guruvayoor town in Kerala, India. The incidence of helminthes infection decreased from 22.73% in 2000 to 12.73% in 2002. Among the positive dung samples, 10 (10.10%) had Strongylidae and 7 (7.07%) had Digenea.They concluded the low incidence of helminth infection among captive elephants in the studied area might be due to the regular annual deworming using albendazole (2.5 mg/kg body weight(2). Easwaran, K. R., Reghu, Ravindran., and Pillai, K. M. reported parasitic infection in 4 wild boars (Sus scrofa cristatus), a calf elephant (Elephas maximus), a sambar deer (Cervus unicolor) and a leopard (Felis bengalensis) at the Thekaddy forest area in Kerala, India. Parasites found in the wild boar, except Gastrodiscoides hominis and Gnathostoma hispidum, commonly infect

domestic pigs. The elephant calf was heavily infected with stongyle worms and maggots of Cobboldia elephantis. The sambar was infected with Linguatula serrata larvae, whereas the leopard was found to be infected with Echinococcus granulosus.(3). Kashid, K. P., Shrikhande, G. B., and Bhojne, G. R. isolated worms included Amphistoma, strongyles, Trichuris spp., Moniezia spp., Ascaridia galli, Toxascaris leonina, Raillietina tetragona, Paragonimus westermani, Filaroides osleri [Oslerus osleri], F. hirthi, Ascaridia galli + R. tetragona, Taenia taeniaeformis, Trichuris + P. westermani and Ascaridia galli + Subulura spp.(4). Carreno, R. A., Neimanis, A. S., Lindsjo, J., Thongnoppakun, P., Barta, J. R., and Peregrine, A. S. reported faecal examinations were negative for nematode eggs in all elephants and negative for fluke eggs in two of the animals. However, adult parasites were recovered from faeces from each of the animals 36-72 hours after deworming. These included Murshidia falcifera, M. neveulemairei, a Quilonia species, and the amphistome Pfenderius papillatus, 1910. The finding of P. papillatus constitutes the first record of this species in Thailand. Specimens of P. papillatus were examined by scanning electron microscopy. In contrast to earlier descriptions of this species, no prominent papillae were found at the anterior end. Structures on the acetabulum that had previously been described as papillae were actually elevated pores that were spread over the acetabulum. These pores differ from all previous descriptions of P. papillatus and indicate a highly modified acetabulum in P. papillatus relative to other Pfenderius species.(5). Raman, M., Jayathangaraj, M. G., and Malik, P. K. reported under laboratory conditions, larvae of Murshidia sp. revealed high motility until 4 months and maintained moderate motility until 7 months. In comparison, larvae of Quilonia sp. and Decrusia sp. displayed a high degree of motility during the first 2 months, but this subsequently declined. These observations suggest that, under in vitro conditions, strongylid larvae from captive Asiatic elephants of the Chennai region may survive for 4 to 7 months.(6). Suresh, K., Choudhuri, P. C., Kumari, K. N., Hafeez, M., and Hamza, P. A conducted faecal egg counts (EPG) by Stoll's dilution method. Analysis of old records revealed that in NZP strongylosis was predominant in summer (52.63%) and the prevalence was lower in animals below 15 years of age. Seven animals (63.64 and 87.5%) each tested positive for ova of strongyles in NZP and SVD, respectively. On treatment with albendazole (Kalbend, 5 mg/kg BW, PO), the animals completely recovered on the seventh day. Therapy resulted in decreases in the pretherapeutic mean EPGs of 700±128.89 (SVD) and 671.4±123.20 (NZP) to 78.57±30.53 and 50±21.79, respectively. The animals were monitored up to four weeks after therapy.(7). Raman, M., Jayathagaraj, M. G., Rajavelu, G., and John, M. C.reported Strongylosis in captive elephants (8).Saidul, Islam, Abdul, Mukit, Manoranjan, Das, Islam, S., Mukit, A., and Das, M. studied the pathology of both immature and mature Gastrodiscus secundus and Pseudodiscus collinsi were recovered from the caecum of 2 captive Asian elephants (Elephas maximus) at Kaziranga National Park, Assam, India. Oedema, pin head size haemorrhages and ulcerative patches in the caecal mucosa were prominent. Mild lymphocytic infiltration in the mucosa with focal necrosis at the tip of the villi were

observed. (9). Bhat, M. N. and Manickam, R. conducted coproculture and demonstration of third stage larvae of Murshidia sp. in elephants (Elephas maximus).(10). On Postmortem examination Matsuo, K., Hayashi, S., and Kamiya, M. found that the gastrointestinal tracts of all 3 animals were also infected with Murshidia falcifera (Nematoda), Hawkesius hawkesi and Pfenderius papillatus (Digenea) and Cobboldia elephantis (Diptera). The elephant louse, Haematomyzus elephantis, was a common cause of dermatopathy in elephants kept in the national park.(11). Islam, S. conducted Study on some aspects of fascioliasis in Asian elephants (Elephas maximus). Wild elephants had an overall prevalence rate of 33.78%. Captive elephants showed prevalence rates of 42.50, 62.28 and 18.18% according to locality. The egg, miracidium and adult stages of F. jacksoni were studied by light and scanning electron microscopy, and their morphology is described. A diurnal fluctuation in faecal egg count was recorded, with average counts of 4.89, 2.47 and 2.76 during the morning, noon and evening, respectively. Young animals were most affected by the parasite and showed anorexia, constipation, diarrhea, anaemia and icterus, with death occurring in severe cases. Some old adults survived the disease with no apparent clinical manifestations. The adult parasites caused massive liver damage. Treatment with triclabendazole (9 mg/kg, not exceeding 7200 mg/animal) and oxyclozanide (7.5 mg/kg, not exceeding 6.8 g/animal) were 100 and 72.16% effective, respectively.(12). Matsuo, K. and Suprahman, H reported three Sumatran elephants (Elephas maximus sumatranus) in Way Kambas National Park, Indonesia, that died of clostridiosis were infected with 1 species of nematode (Murshida falcifera), 2 trematodes (Hawkesius hawkesi and Pfenderius papillatus), and 1 larval botfly species (Cobboldia elephantis) in the gastrointestinal tract. This is the first report of H. hawkesi, P. papillatus and C. elephantis infection in Sumatran elephants in Indonesia(13). Watve, M. G. conducted study on Helminthes Parasites of Elephants: Ecological Aspects. The helminthes parasites of free ranging as well as captive elephants of the Mudumalai Wildlife Sanctuary were studied quantitatively by analysing over 200 dung samples and 7 autopsy examinations. The prevalence and intensities of infection were high in both captive as well as wild elephants. The helminth communities of both were, however, species poor as compared to other mammalian host species. The high prevalence and intensities are thought to be related to the absence of predation and the low species diversity may be a result of absence of other closely related host species. The age and sex of individuals sampled did not affect their parasite loads significantly. The faecal propagule densities were significantly greater during the dry season as compared to the wet season. Stronglid nematodes of the genus Quilonia dominated the helminth communities. Tapeworm infection was significantly greater in captive elephants than the wild ones. The possible reasons for this difference are discussed.(14). Singh, K. P., Srivastava, V. K., Prasad, A., and Pandey, A. P conducted study of Pathology due to Fasciola jacksoni in Indian elephants (Elephas indicus). F. jacksoni recovered from infected liver and lungs were almost round, pearshaped measuring 10-16 mm in length and 8.5-14 mm in width with ill-defined

cephalic end between indistinct shoulders. The intestine was extensively branched. The yellowish tinged ova were oval with an operculum at one end and measured 0.112-0.160 (mean 0.13) mm in length and 0.054-0.096 (mean 0.07) mm in width. Infected liver showed haemorrhagic tracts, thickening of bile ductules, cirrhotic changes and pseudolobulations. In the lungs, the bronchial lumen contained desquamated cells admixed with fibrinohaemorrhagic exudate.(15). Li, C. X., Rong, Y. M., and Xie, Q. P.conducted a study of helminth parasites of elephants (Elephas maximus).In between 1985 and 1990, the following parasites were recovered from Elephas maximus in China: Choniangium epistomum, Murshidia falcifera, M. murshida, M. neveulemairei, Quilonia travancra, Chabertia erschovi, Gasterophilus pecorum, and G. intestinalis.(16) Chakraborthy, A. and Chaudhury, B. conducted study on Pathology of Fasciola jacksoni infestation in elephants. Fasciola jacksoni infection was discovered in 2 out of 3 elephants autopsied at Assam State Zoo, India, during 1985 to 1989. The parasites were attached to biliary epithelium. Microscopy demonstrated that the biliary epithelium was distorted by necrotic tissue which contained erythrocytes and ova of F. jacksoni. The epithelium was analyzed by X-ray microanalysis, which showed that the infected epithelium contained aluminum, silicon, calcium and iron, while non-infected, normal biliary epithelium contained only phosphorus and sulfur. Scanning electron microscopy demonstrated that both the dorsal and ventral surfaces of the parasite possessed spines.(17). Maske, D. K., Sardey, M. R., and Bhilegaonkar, N. G, Studied the Helminth parasites in zoo animals of Maharaj Bag, Nagpur, Maharashtra State on faecal samples obtained from 28 animals during 1979 to 1981 were examined by direct smear, flotation and dilution count techniques. 17 animals were positive for parasitic infections. Toxascarids and Ancylostoma were found in lions and tigers in rainy and winter seasons. Isospora felis, Paragonimus westermani and Taenia pisiformis were also found in winter. Ancylostoma sp. was found in leopard cubs, and ascarids in lion cubs. More than 1 helminth species were not usually found together. Strongyles were found in elephants and a cestode in a python. A slothbear, Himalayan bear, Russian bear and white and black monkeys were negative for helminth parasites.(18). Li, C. X., Rong, Y. M., and Lan, J. G. conducted a study on antihelmintic efficacy of albendazole against parasites in Indian elephants 4 Indian elephants treated orally with albendazole at 20, 30 or 35 mg/kg were negative for nematode and trematode eggs after one month.(19). Eric R. Morgan1,2,6, Blok Shaikenov3, Paul R. Torgerson2,4, Graham F. Medley1 and E. J. Milner-Gulland5 reported Saiga antelope (Saiga tatarica) graze extensively on livestock pasture, potentially enabling transmission of a wide range of parasitic helminths between saigas and domestic ruminants. Thirty-six of the 38 species of helminth that have been found in saigas in Kazakhstan in the past have been found also in domestic livestock. We examined 133 saigas culled for meat in autumn 1997, and found three species of cestode and 12 nematodes (nine in the abomasum), but no trematodes or lungworms. The most abundant species were Marshallagia marshalli, Marshallagia mongolica, and Nematodirus gazellae in the abomasum, Nematodirus gazellae in the small intestine, and

Skrjabinema ovis in the large intestine. There was no clear relationship between intensities of abomasal nematodes and body condition. Age-intensity patterns differed between species: N. gazellae intensities were highest in saigas around 2–3 yr old, and declined in older animals, whereas the intensity of Marshallagia spp. rose asymptotically with age. Fecal egg density was directly proportional to adult worm intensity across ages for Marshallagia spp., but only in young animals for N. gazellae. There was no evidence that helminths, at the intensities observed, adversely affect saiga populations. The host range of many of the parasites found is broad, and transmission between saigas and livestock in both directions might become important to agriculture and conservation as livestock numbers recover. Simplified sampling techniques used in this study, and statistical analysis based on bootstrapping, could prove useful in other parasitologic surveys of wildlife in remote areas. (20) Result and Discussion: The incidence of helminthes infection as recorded in this study was out of 20 samples 6 samples were positive for Fasciola jacksoni,2 for Paramphistome,15 samples revealed the presence of egg and larvae of Oesophagostomum spp, and Chabertia spp 18 samples were positive for Schistosomes spp 1 sample was found positive for Dicrocoelium spp and 1 sample positive for Monezia spp. Location wise samples from Koshi Tappu wildlife reserve were positive for Fasciola and Schistosomes spp while samples from Chitwan National park were positive for Fasciola,Paramphistome and Oesophagostomum spp .The samples from Suklaphanta wildlife reserve and Bardia wildlife reserve were positive for Schistosomes spp and Oesophagostomum and Chabertia spp.As this is the first screening of the prevalence of Helminthes in Elephant and need to asses seasonal pattern . References: 1.Agatsuma, T., Rajapakse, R. P., Kuruwita, V. Y., Iwagami, M., and Rajapakse, R. C. Molecular taxonomic position of the elephant schistosome, Bivitellobilharzia nairi, newly discovered in Sri Lanka. Parasitology International 53[1], 69-75. 2004. 2:Saseendran, P. C., Rajendran, S., Subramanian, H., Sasikumar, M., Vivek, G., and Anil, K. S. Incidence of helminthic infection among annually dewormed captive elephants. Zoos' Print Journal 19[3], 1422. 2004. 3:Easwaran, K. R., Reghu, Ravindran., and Pillai, K. M. Parasitic infection of some wild animals at Thekkady in Kerala. Zoos' Print Journal 18[2], 1030. 2003. 4:Kashid, K. P., Shrikhande, G. B., and Bhojne, G. R. Incidence of gastrointestinal helminths in captive wild animals at different locations. Zoos' Print Journal 18[3], 1053-1054. 2003. 5;Carreno, R. A., Neimanis, A. S., Lindsjo, J., Thongnoppakun, P., Barta, J. R., and Peregrine, A. S. Parasites found in faeces of Indian elephants (Elephas maximus) in Thailand following treatment with mebendazole, with observations

on Pfenderius papillatus (Cobbold, 1882) Stiles and Goldberger, 1910 by scanning electron microscopy. Helminthologia 38[2], 75-79. 2001. 6:Raman, M., Jayathangaraj, M. G., and Malik, P. K. In vitro survivability of strongylid larvae of elephants. Indian Journal of Animal Sciences 71[11], 10431044. 2001. 7;Suresh, K., Choudhuri, P. C., Kumari, K. N., Hafeez, M., and Hamza, P. A. Epidemiological and clinico-therapeutic studies of strongylosis in elephants. Zoos' Print Journal 16[7], 539-540. 2001. 8:Raman, M., Jayathagaraj, M. G., Rajavelu, G., and John, M. C. Strongylosis in captive elephants -- a report. Indian Journal of Animal Health 39[2], 85-86. 2000. 9;Saidul, Islam, Abdul, Mukit, Manoranjan, Das, Islam, S., Mukit, A., and Das, M. Pathology of concurrent Gastrodiscus secundus and Pseudodiscus collinsi infection in two captive Asian elephants (Elephas maximus). Journal of Veterinary Parasitology 13[2], 151-152. 1999. 10:Bhat, M. N. and Manickam, R. Coproculture and demonstration of third stage larvae of Murshidia sp. in elephants (Elephas maximus). Indian Veterinary Journal 75[12], 1140-1142. 1998. 11:Matsuo, K., Hayashi, S., and Kamiya, M. Parasitic infections of Sumatran elephant in the Way Kambas National Park, Indonesia. Japanese Journal of Zoo and Wildlife Medicine 3[2], 95-100. 1998. 12:Islam, S. Studies on some aspects of fascioliasis in Asian elephants (Elephas maximus). Journal of Veterinary Parasitology 11[1], 109. 1997. 13:Matsuo, K. and Suprahman, H. Some parasites from Sumatran elephants in Indonesia. Journal of the Helminthological Society of Washington 64[2], 298299. 1997. 14;Watve, M. G. Helminth Parasites of Elephants: Ecological Aspects. Daniel, J. C. A Week with Elephants; Proceedings of the International Seminar on Asian Elephants. 289-295. 1995. Bombay, India, Bombay Natural History Society; Oxford University Press. 15:Singh, K. P., Srivastava, V. K., Prasad, A., and Pandey, A. P. Pathology due to Fasciola jacksoni in Indian elephants (Elephas indicus). Indian Journal of Animal Sciences 64[8], 802-804. 1994. 16;Li, C. X., Rong, Y. M., and Xie, Q. P. A study of helminth parasites of elephants (Elephas maximus). Chinese Journal of Zoology 28[5], 43-44. 1993. 17:Chakraborthy, A. and Chaudhury, B. Pathology of Fasciola jacksoni infestation in elephants. Indian Journal of Veterinary Pathology 16[2], 98-101. 1992. 18:Maske, D. K., Sardey, M. R., and Bhilegaonkar, N. G. Helminth parasites in zoo animals of Maharaj Bag, Nagpur, Maharashtra State. Indian Journal of Animal Science 60[8], 952. 1990. 19;Li, C. X., Rong, Y. M., and Lan, J. G. Anthelmintic efficacy of albendazole against parasites in Indian elephants. Chinese Journal of Veterinary Science and Technology 9, 42-43. 1988. 20:Eric R. Morgan1,2,6, Blok Shaikenov3, Paul R. Torgerson2,4, Graham F.

Medley1 and E. J. Milner-Gulland5 Journal of Wildlife Diseases, 41(1), 2005, pp. 149-162 Acknowledgement: We would like to acknowledge our sincere thanks to the officer and technical staffs of different wild life reserve of Nepal for their cooperation in collection of faecal samples as well all officers and technical staffs of Central Veterinary Laboratory Tripureshwor involved in the processing and identification the parasite spp of this study