Studies on prevalence of gastrointestinal parasites in captive Spotted Deer in and around Bhubaneswar

Transcription

1 Nayak T, M.V.Sc. (Veterinary Parasitology) Thesis, Studies on prevalence of gastrointestinal parasites in captive Spotted Deer in and around Bhubaneswar Studies on prevalence of gastrointestinal parasites in captive Spotted Deer in and around Bhubaneswar Tapaswini Nayak 04PARA/14 DEPARTMENT OF VETERINARY PARASITOLOGY COLLEGE OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY ORISSA UNIVERSITY OF AGRICULTURE AND TECHNOLOGY BHUBANESWAR

2 Studies on prevalence of gastrointestinal parasites in captive Spotted Deer in and around Bhubaneswar THESIS SUBMITTED TO ORISSA UNIVERSITY OF AGRICULTURE AND TECHNOLOGY IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE DEGREE OF MASTER OF VETERINARY SCIENCE IN VETERINARY PARASITOLOGY By Tapaswini Nayak 04PARA/14 DEPARTMENT OF VETERINARY PARASITOLOGY COLLEGE OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY ORISSA UNIVERSITY OF AGRICULTURE AND TECHNOLOGY BHUBANESWAR

3 ORISSA UNIVERSITY OF AGRICULTURE AND TECHNOLOGY DEPARTMENT OF VETERINARY PARASITOLOGY COLLEGE OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY Dr. Mitra Ranjan Panda, Ph.D. Professor and Head Department of Veterinary Parasitology College of Veterinary Science and Animal Husbandry Orissa University of Agriculture and Technology Bhubaneswar , Odisha. Bhubaneswar Date: CERTIFICATE-I This is to certify that the thesis entitled Studies on prevalence of gastrointestinal parasites in captive Spotted Deer in and around Bhubaneswar submitted in partial fulfilment of the requirements for the award of the degree of Master of Veterinary Science (Veterinary Parasitology) to the Orissa University of Agriculture and Technology is faithful record of bonafide and original research work carried out by Tapaswini Nayak under my guidance and supervision. No part of the thesis has been submitted for any other degree or diploma. It is further certified that the assistance and help received by her from various sources during the course of investigation has been fully acknowledged. CHAIRMAN ADVISORY COMMITTEE

4 CERTIFICATE-II This is to certify that the thesis entitled Studies on prevalence of gastrointestinal parasites in captive Spotted Deer in and around Bhubaneswar submitted by Tapaswini Nayak to the Orissa University of Agriculture and Technology, Bhubaneswar in partial fulfilment of the requirements for the degree of Master of Veterinary Science (Veterinary Parasitology) has been approved/disapproved by the students advisory committee and the external examiner. Advisory Committee Chairman Dr. Mitra Ranjan Panda, Ph.D. Professor and Head Department of Veterinary Parasitology C.V.Sc. and A.H, O.U.A.T., Bhubaneswar Members 1. Dr. S. Panda Professor and Head Department of Veterinary Pathology C.V.Sc. and A.H, O.U.A.T., Bhubaneswar 2. Dr. B.N. Mohanty Associate Professor Department of Veterinary Parasitology C.V.Sc. and A.H, O.U.A.T., Bhubaneswar 3. Dr. D.K. Karna Associate Professor Department of Animal Breeding and Genetics C.V.Sc. and A.H, O.U.A.T., Bhubaneswar External Examiner (Name & Designation)

5 C O N T E N T S SL. PAGE CHAPTER NO. NO. 1 INTRODUCTION REVIEW OF LITERATURES MATERIALS AND METHODS RESULTS DISCUSSION SUMMARY AND CONCLUSION REFERENCES i-x

6 LIST OF TABLES TABLE NO PARTICULARS Prevalence of gastro-intestinal parasites in captive spotted deer in and around Bhubaneswar Prevalence of mixed infection in captive spotted deer in and around Bhubaneswar Chi-square test showing Prevalence gastro-intestinal parasites in captive spotted deer at three different selected locations Prevalence (%) of gastro-intestinal parasites identified in fecal samples of captive spotted deer (A.axis) in and around Bhubaneswar Analysis of variance showing comparison of parasitic load of Strongyle spp. among localities in captive spotted deer (A.axis) PAGE NO Analysis of variance showing comparison of parasitic load of Paramphistomum sp. among localities in captive spotted deer (A.axis) 7 Locations wise Mean ± SE of gastro-intestinal parasitic load in terms of EPG (egg per gram) in captive spotted deer 33 8 Seasonal status related prevalence of gastro-intestinal parasites in captive spotted deer (A.axis) at three captive locations (N=292) Mean ± SE (EPG) of different gastro-intestinal parasites in different seasons across locations in captive spotted deer 36

7 LIST OF FIGURES FIGURES NO. PARTICULARS PAGE NO. 1 Flow diagram showing research out line 21 2 Prevalence of gastro-intestinal parasites in captive spotted deer in and 28 around Bhubaneswar 3 Overall prevalence of gastro-intestinal parasites in captive spotted deer at three different selected locations in and around Bhubaneswar 4 Seasonal prevalence of gastro-intestinal parasites in captive spotted deer in and around Bhubaneswar 5 Sex related overall prevalence of gastro-intestinal parasites on captive spotted deer in and around Bhubaneswar 6 A flock of Spotted Deer at Raj Bhawan Deer Park 39 7 Collected faecal samples 39 8 Concentration method 40 9 Floatation method Faecal Culture by petridish method A charged Mc Master slide A dead Spotted Deer Post-mortem of the dead Deer Collection of amphistomes from rumen wall Collected amphistomes Flattening of amphistomes Bottle under running tap water Borax carmine stain Dehydration in Ascending order of alcohol Mounting of slide Ova of Paramthistomum sp. x Ova of Strongyle spp. x Ova of Fasciola sp. x Ova of Strongyloides sp. x single infection of Paramphistomum sp. x single infection of strongyle spp. x Cotylophoron cotylophorum recover from post-mortem of dead 46 spotted deer 28 Dictyocaulus larvae in the concerntrated faecal specimen by floatation 47 technique x Dictyocaulus larvae after staining x Anterior end of Dictyocaulus larva x Posterior end of Dictyocaulus larva x

8 ABBREVIATIONS USED ISFR : India state Forest Report NZP RBDP : Nandankanan Zoological Park : Raj Bhawan Deer Park TDP : Tulasipur Deer Park EPG : Egg per Gram CPG : Cyst per Gram OPG : Oocyst per Gram % : Percent rpm : Revolution Per Minute Viz. : Suchas Fig. : Figure ml : Milliliter 0 C : Degree Centigrade i.e : That is mm : Millimeter SE : Standard Error SPSS : Statistical Package for Social Science C.V.Sc & A.H : College of Veterinary Science and Animal Husbandry O.U.A.T : Orissa University of Agriculture and Technology

9 ACKNOWLEDGEMENTS Words fail to express my deepest sense of gratitude and indebtedness to my esteemed guide and Major Advisor of the Advisory Committee Dr. Mitra Ranjan Panda, Prof. & Head, Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Bhubaneswar for his comprehensive planning, keen interest, scientific supervision, benevolent guidance, support and patience, wholehearted co- operation, meticulous correction and unstinted encouragement with a blend of affection as my esteemed guide major adviser during the entire period of research work to complete this project. I am extremely grateful to Dr. R. C. Patra, the Dean, Faculty of Veterinary Science and Animal Husbandry for providing necessary facilities during the course of my experiment. I am greatly beholden beyond words to express my deep sense of gratitude and obligation to Dr. Baijayendranath Mohanty, Associate Professor, Dept. of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Bhubaneswar for his ever enthusiasm in imparting knowledge, timely help, uncovering encouragement, moral motivation, valuable and generous suggestion, provided during my study as well as carrying forward this research work. It gives me immense pleasure to record deep sense of gratitude and respect to Dr.S.Panda, Professor and Head, Department of Veterinary Pathology, for his constant inspiration generous encouragement and guidance throughout the course of my research work. I am extremely grateful to Dr. D. K. Karna, Associate professor, Department of Animal Breeding and Genetics, for his valuable guidance and technical instructions together in carrying out the statistical analysis related to my research work. I am thankful to Dr. (Mrs.) Manaswini Dehuri, Assistant Professor and Dr. Anant Hembram, Assistant Professor, Department of Veterinary Parasitology, for their moral support and guidance during the whole period of my work. I am thankful to all the staffs of Central Library, Orissa University of Agriculture and Technology for their co-operation and assistance during my research work. I owe a heartful debt of gratitude to Dr. A.K. Das, Senior Veterinary Officer, Dr. D. Mohapatra, Specialist, Dr. S.K. Sahu, Veterinary Assistant Surgeon, Sri. Chandramani, Tempoary Status, Deer enclosure and all other staff members of

10 Nandankanan Zoological Park for their co-operation, constant inspiration and valuable advice rendered during my research work. I am extremely thankful to Dr. S. Panda, I.F.S, Director, Nandankanan Zoological Park, for giving me permission to carry out the research work and providing all necessary facilities from the zoo. I am grateful to Sri. P. R. K. Nair, Forester, Raj Bhawan Deer Park and Sri. S. P. Mishra, Animal keeper, Tulasipur Deer Park, Cuttack for their kind cooperation and help in collection of faecal samples during the course of my research work. I am thankful from the deepest core of my heart to my beloved friends and colleagues Dr. Arabinda Kumar Sethy, Dr. Trilochan Mohapatra, Dr.(Miss) Subhasmita Das, Dr. Samir Nanda, Dr. Manoj Kumar Behera, Dr(Miss). Rachita Sethy for their Valuable and effective suggestions, timely help, encouragement, best wishes and noble involvement in my research work. I have immense pleasure to acknowledge the help and co-operation of all nonteaching staff members of Parasitology Department for their co- operation and kind help in carrying out the laboratory work. I extend my thanks to The Creation s of Siripur market for his sincere effort in preparing this manuscript. I would like to express thanks to my beloved parents for their unequivocal support, stimulating inspirations in every step of my life to excel. My loving thanks to my younger brother Bapu and sister subratini, whose precious help and best wishes led to the completion of my research work. I express my heartful thanks to my beloved husband and in-laws for their support and particularly my cute angel Divyansh who made me feel enchanting and rejuvenate by his graceful presence in my life. At last, I bow my head to my beloved Almighty Sibababa, attributing all my achievements to his kind grace and holy blessings. Bhubaneswar Date: (Tapaswini Nayak)

11 ABSTRACT Aim and objectives: To study the prevalence of gastro-intestinal parasites in captive Spotted deer in zoos and parks with respect to season and sex and to assess the intensity of gastro-intestinal parasites of Spotted deer in captivity,for the adaptation of the best possible prevention and control measures to minimize the spread of infectious diseases of zoonotic importance. Materials and methods: A total of 292 faecal samples from three captive locations viz: Nandankanan Zoological Park, Raj Bhawan Deer Park and Tulasipur Deer Park examined for the gastro-intestinal parasitic prevalence following standard parasitological techniques for their identification based on their morphology and morphometry. Identification of gastrointestinal parasites also carried out on the basis of necropsy findings following the standard procedure of collection of parasites, fixation, preservation of slides, staining and mounting of parasites. Observations and Results: Present studies revealed an overall prevalence of 75.3%. The overall prevalence rate of gastro-intestinal parasites in captive Spotted deer was highest at Nandankanan Zoological Park (82.2%) followed by Raj Bhawan Deer Park (79.0%) and Tulasipur Deer Park (60.7%). The rate of variation in the prevalence of gastro-intestinal parasites in between Nandankanan Zoological Park and Tulasipur Deer Park; Raj Bhawan Deer Park and Tulasipur Deer Park were statistically significant (P<0.05) while a non-significant variation was observed between Nandankanan Zoological Park and Raj Bhawan Deer Park. During the study, an overall prevalence of two types of trematode (25.3%) and two types of nematode (23.2%) were observed and a high rate of prevalence was recorded in Paramphistomum sp. (51.8%) followed by Strongyle spp. (48.3%). Strongyloides sp. (1.4%) and Fasciola sp.(0.6%).no cestode or protozoan cyst/oocyst or coccidian infection recorded in this investigation. Mixed infection was found more common than single infection with a prevalence of 26.8% and ahigh rate was observed in case of Paramphistomum sp. and Strongyle spp. (24.7%). Seasonal investigation revealed a higher prevalence rate of gastro-intestinal parasites in winter (77%) than in summer (73.5%). The gastro-intestinal parasitic infection was recorded higher in male (83.6%) than in female deer (80%) which showed a non-significant difference in prevalence rate between male and female. Lungworm (Dictyocaulus sp.) infection was recorded by finding the larvae of Dictyocaulus sp. in the faecal specimen. The adult parasites of Cotylophoron cotylophorum species of amphistomes were recovered from the rumen wall of Spotted deer through post-mortem. Conclusion: Spotted deer of either sex irrespective of seasons were found infected with varieties of gastro-intestinal parasites viz; Strongyle spp., Paramphistomum sp., Strongyloides sp. and Fasciola sp. in captivity in and around Bhubaneswar which could be prevented by adopting the best preventive and control measures against the identified parasites.

12 INTRODUCTION Introduction India is rich in wildlife resources. Wildlife found in all ecosystems, like rainforests, deserts, plains and urban areas (zoos and parks); plays a significant role in maintaining the ecological equilibrium of nature. Creation of series of National Parks, Sanctuaries aim at Wildlife conservation which ultimately protects various wild flora and fauna species and their habitats including game reserves and biosphere reserves. Above 33% of geographical areas of India is under forest cover (ISFR, 2015).There are more than 97 National Parks and 481 notified wildlife Sanctuaries in India. In Odisha, there are 2 National Parks, 18 wildlife Sanctuaries and 8 Deer Parks (mini zoos) which have been notified for both in-situ and ex-situ conservation and management of wildlife. Nandankanan Zoological Park is one of the most popular wildlife Sanctuaries of Odisha. The zoo gives shelter to a wide variety of wildlife which includes 67 species of mammals, 18 varieties of reptiles,81 species of birds and 36 species of fishes. In recent years, apart from poaching and destruction of habitats, the health related issue has become a significant threat to wildlife. Parasitic diseases represent a major concern in captive wild animals for the high environmental contamination due to maintenance of animals in confined areas, resulting in loss of condition, reduced body weight gain, reproductive disorder and ultimately death. Some studies have revealed that parasites, particularly gastro-intestinal parasites of wild animals in captivity include some zoonotic species and raise public health concerns (Levecke et al., 2007; Adekunle et al., 2008; Akinboye et al., 2010; Aijbade et al., 2010; Otegbade et al., 2014). In Zoos and Zoological Parks, among all herbivores, the Axis or Spotted Deer (Axis axis) is the most beautiful and abundant cervid species in India (Arora, 1982). It is a prolific breeder, widely distributed all over India and forest regions of Southern Asia and Srilanka. But not common in arid regions of Punjab and Rajasthan. Captivity of wildlife creates an unnatural system and disrupts the balance between parasite and host. It makes a stressful environment due to which animals 1

13 Introduction become diseased or can even die from parasitic loads that they would have survived under natural conditions (VanWyk and Boomker, 2011). Zoo animals living under captivity are susceptible to almost all type of diseases, much so that parasitic diseases (Varadharajan et al., 2000). In captivity wild animals may succumb to parasitic infection due to environmental stress such as change in living conditions and space limitations (Rana et al., 2015; Atanaskova et al., 2011). A wide range of parasitic spp. have been recorded in zoo animals, these parasites not only affect the animal health adversely but sometimes cause mortality, morbidity or both (Borghare, 2009). Helminthic infections in particular can frequently be a major problem causing mortality in captive wild animals (Borghare et al., 2009). In spite of being infected with many species of parasites, massive death or epizootics because of them are rarely seen (Banerjee et al., 2005). Usually, captive animals don t show alarming signs of parasitism if regular deworming practices carried out in the zoo (Parsani et al., 2001). Infections with helminthes are a major health issue in captive and wild deer (Gossens et al., 2005) mainly where herds of animals are kept in relatively small enclosures. Deer are hosts to a wide range of endoparasites such as helminthes, insect larvae and certain protozoa (Rehbein et al., 2001; Vengust, 2003), cestodes (Chapman and Chapman, 1997) and other parasites. Outbreaks of parasitic diseases among captive deer in a limited space and intensive management practices mean that they are more heavily infested with parasites than wild deer (Vengust, 2003). A regular programme of gastro-intestinal parasites surveillance and control measures based on correct diagnosis, effective treatment and proper prophylaxis would be helpful to improve and progress the health of captive zoo animals. Application of existing knowledge of disease control and prevention would significantly reduce economic losses due to gastro-intestinal helminthosis (Barmon, 2013). There are zoological gardens, safari parks, eco-parks which possess different species of wild animals and act as an important source for recreation of all aged people in Odisha. Among these Nandankanan Zoological Park, Raj Bhawan Deer Park and Tulasipur Deer Park are the well-known parks where deer are kept for recreational purpose. 2

14 Introduction Unfortunately, inadequate information on diseases and parasites of zoo animals is a major limiting factor in zoological gardens. Investigations on endoparasitic fauna are important for the study of the prevalence and geographical distribution (Zasityle and Grikiencience, 2002). From the economic point of view, Zoo animals are the most valuable and important animals to keep and restore the ecological balance but, this conception has not been established yet in our country. However, a little work has been carried out to understand the epidemiology of gastrointestinal parasitic disease of captive Spotted Deer in Indian Zoos and a systemic investigation has been undertaken earlier by few research workers (Meshram et al.,2008; Kanungo et al.,2010; Barmon et al.,2014; Mir et al.,2016). This study attempts to determine the prevalence and intensity of parasites particularly in spotted deer in captivity in three different locations present in and around Bhubaneswar, the capital city of odisha i.e at Nandankanan Zoological Park, Raj Bhawan Deer Park and Tulasipur Deer Park, Cuttack. The finding of this study shall be helpful for better understanding of the gastro-intestinal parasites of spotted deer and for the prevention of the spread of infectious parasitic diseases among deer or other animals within zoo or to human. Considering the economic significance of gastro-intestinal parasites in spotted deer (Axis axis) kept at the three different locations respectively, the present work was carried out to investigate the prevalence of gastro-intestinal parasites in spotted deer kept in these three selected places with the following objectives: To study the prevalence of gastro-intestinal parasites in Spotted deer/ Chital in captivity To study the intensity of parasitic infection. To study the seasonal and sex wise variation in prevalence of gastro-intestinal parasites in captive spotted deer. To identify the gastro-intestinal parasites from necropsied animals. 3

15 Review of literature REVIEW OF LITERATURE 2.1 Prevalence of gastro intestinal parasites in captivespotted Deer (A.axis) India Ramaswamy et al., (1991) investigated the prevalence of parasitic infections among the Axis deer (Cervus axis) in three National Parks in India.This investigation revealed infections with the lungworm Muellerius capillaris. Clinical sings were not evident in infected animals,so it was suggested that Cervis axis was probably a carrier of lungworm infection. Cross transmission of Muellerius capillaris occur between wild and domesticated animals in India. Modi et al., (1997) surveyed for prevalence of parasites in zoo animals of Bihar. Examined 105 fecal samples from different animals including spotted deer. The common identified parasites were Ascaris sp., Ancyclostoma sp., Oesophagostomum sp., Trichuris sp., Strongyloides sp., Fasciola sp., and Paramphistomum sp. Varadharajan et al.,(1999) investigated the fecal samples of wild animals at the Zoological Garden, Thiruananthapuram, Kerala and revealed 74% of animals were infected with helminthic and 23% infected with protozoan infections.out of 23 fecal samples of spotted deer(axis axis)18 samples found positive for parasitic infections. Those were Strongyle (6), Strongyloides (1), Amphistomes (7), Spirudid (9), Ascarid (4) and Coccidia (3). Borghare et al., (2000) examined 60 samples of deer from Maharajbag Zoo, Nagpur and found 30 were positive for helminthic eggs and larvae. The parasites reported were Haemonchs sp., Dicrocoelium sp., Paramphistomum sp., Oesophagostomum sp., and Bunostomum sp. etc. Varadharajan et al., (2000) surveyed for gastro-intestinal parasites in wild animals in a typical mini zoo, Coimbatore. Total 60 fecal samples were examined and 4

16 Review of literature examination revealed that 58% of captive animals were positive for helminthes and 6% were positive for protozoan infections. Particularly in Spotted deer, out of 7 animals examined, 4 were found infected with parasites and those were Strongyle and Trichuris (1), Strongyle (1), Ascarid (1), Coccidia (1). Parsani et al., (2001) studied the prevalence of gastro-intestinal parasites of captive animals at Rajkot Municipal Corporation Zoo, Rajkot, Gujrat. A total of 28 fecal samples of different zoo animals were examined out of which 16(57.14%) were found positive for parasitic infection in mammals and birds respectively. Out of 28 fecal samples, 2 were of Spotted deer and examination revealed the parasites, Trichostrongylus sp., (2) and Coccidia (1). Kashid et al., (2002) surveyed the incidence of gastro-intestinal helminthes in captive wild animals at six different locations, viz., Kanha National Park in Kanha Kesari, Gemini Circus Deer Park and Maharajabagh Zoo in Nagpur, Siddharth Zoo in Aurangabad and Peshwe Park in Pune. Fecal sample examination revealed 96.13% of parasitic infection in spotted deer. At Deer Park, Nagpur 100% helminthic infection was observed in Spotted Deer. Hussain et al., (2002) investigated the prevalence of helminthic infection in axis deer from Nagpur. A total of 175 fecal samples were examined and an overall prevalence rate of 90.2% was observed. The parasites recovered from infected deer were Strongyle spp., Strongyloides sp., Trichuris sp., Nematodirus sp., Haemonchus sp. and Oesophagostomum sp. Mandal et al., (2002) investigated the prevalence of helminthic infection in freeranging chital (Axis axis) at Mudumalai Wildlife Sanctuary, Tamilnadu between May,2000 to August, Fecal examination reported maximum infection of Strongyle spp. (41.7%) followed by Paramphistomum sp. (15.6%), Fasciola sp. (13.5%), Strongyloides sp. (11.5%) and Ascaris sp. (5.3%). 5

17 Review of literature Banerjee et al., (2005) examined fecal samples of 79 spotted deer, 161 nilgai, 91 wild pigs and 33 asian elephants from the forest of South Uttaranchal and revealed that 17.7%, 41.6%, 73.6% and 63.6% of spotted deer, nilgai, wild pigs and elephant respectively, were positive for either single or mixed parasitic infection. Amphistomes, Strongyle spp. and Coccidian infections were common in deer, nilgai and elephants, while Fasciola sp. were detected in deer and nilgai only. Singh et al.,(2006) studied the parasitic infection in wild herbivores in the Mahendra Choudhury Zoological Park, Chhatabir, Punjab and found overall prevalence 25.71%.The most commonly detected infection was of Strongyle (89%) followed by Trichuris sp., Eimeria sp.and Amiphostomes. In Spotted Deer (50%) only Strongyle spp. eggs were found in feces. Mohan et al., (2007) investigated the endoparasitic infection in Spotted Deer (Axis axis) in Puducherry. Examination of fecal drooping of 18 stags (>1 yr) revealed Cooperia sp. and Capillaria sp. (in 5 stags), Trichostrongylus sp. (in 2 stags) and Trichostrongylus sp. and Cooperia sp. in another one stag. Meshram et al., (2008) studied the parasitic infection in 200 Axis deer (Axis axis) of Scrub forest of Borgaon Manju in Western Vidarbha region of Maharashtra. Fecal sample examination revealed the presence of Strongyloides sp., Strongyle spp., Trichostrongylus sp., and Oesophagostomum sp. Total 89.05% animal were found positive for parasites, out of which incidence of Strongyloides sp. (31.50%), Strongyle spp.(20.00%), Haemonchus sp.(13.80%), Trichostrongylus sp.(11.50%), Trichuris sp.(8.50%) and Bunostomum sp.(4.00%). Occurance of parasitic infection is highest in winter season followed by rainy season. Singh et al., (2009) studied the prevalence of gastro-intestinal parasitic infection of free ranging wild herbivores; chital or Spotted Deer, Sambar (Rusa unicolor) and Nilgai (Bos elaphus tragocamelus) in the Van Vihar National Park, Bihar. The parasitic prevalence was highest (3.45%) in Sambar followed by Chital (38.19%) and Nilgai (36.84%).The highest prevalence was recorded for Strongyles (26.15%) followed by 6

18 Review of literature Strongyloides sp. (7.13%), Coccidia (6.20%), Fasciola sp. (2.64%), Amphistomes (1.98%) and Trichuris sp. (1.84%). The overall mean EPG was maximum for Strongyles (585.19) followed by Trichuris (410), Amphistome (250), Strongyloides (127.78) and Fasciola (111.19). Overall mean OPG for Coccidia was Jadhav et al., (2010) studied the seasonal variation in prevalence of helminthic infection in captive spotted deer at two different places Balodyan and Maharajabag zoo in Nagpur in summer and rainy season. Fecal examination of 200 samples revealed highest helminthic infection of Strongyloides sp. followed by Strongyle spp., Haemonchus sp., Oesophagostomum sp., and Trichuris sp. in both the season. Among all the parasites Strongyloides infection was found predominant in both the seasons. Gupta et al., (2011) surveyed the incidence of gastro-intestinal parasites in wild ruminants around Jabalpur. Collected 50 fecal samples of wild animals including 15 samples of Chital (Axis axis),15 of Nilgai (Bos elaphus tragocamelus),10 of Gour (Bos gaurus) and 10 of Sambar (Rusa unicolor). Coprological examination revealed highest rate of parasitic infection in Sambar (90.0%) followed by Nilgai (86.6%) and Gaur and Chital (80.0%). Mixed infection was recorded 94% while single infection was only 6%.In Chital the prevalence of Strongyloides (60.0%) was highest followed by Trichuris (53.3%) and Fasciola (6.7%). Ravindran et al., (2011) conducted a survey on parasitic infection in wild animals of Pariyar Wildlife Sanctuary of Kerala. 15 numbers of fecal samples of Sambar deer were examined and reported the presence of Eimeria oocyst, Trichuris, Strongyle ova. Bante et al., (2013) studied the prevalence of helminth parasites in wild animals kept at Kamala Nehru Prani Sangrahalaya, Indore. Examination of 347 fecal samples, revealed 149 positive for helminth parasites indicating 42.90% prevalence. Prevalence of nematode eggs (42.36%) was found highest as compared to trematodes (0.28%) and cestode eggs (0.28%).Out of 347 fecal samples, 10 were of spotted deer, of which 2 were found positive for helminthic infection and the parasite detected was Strongyloides sp. 7

19 Review of literature Solanki et al., (2013) surveyed the incidence of parasitic infection in captive wild animals of Rajkot Municipal Corporation Zoo, Rajkot. Examined 2980fecal samples of different zoo animals and revealed 462(15.50%) were positive for parasitic infection. 176(38.10%), 197(42.67%) and 89(19.26%) were found to be positive for nematode, cestode and protozoan infections respectively. Cestode infection of Spirometra sp. was 42.64% and intestinal protozoan infection of Eimeria spp. was 19.26% while the nematode infections were Capillaria spp. (9.52%), Ascaris spp. (2.60%), Trichostrongylus spp. (19.05%), Ancyclostoma spp. (5.19%), Trichuris spp. (1.08%) and Strongyloides spp. (0.65%). Jaiswal et al., (2014) surveyed the prevalence of endoparasitic infection in wild cervids of Army Golf Course, Mathura. Coproscopical analysis of 46 fecal samples revealed 30% single and 12% mixed parasitic infection. Prevalence of Haemonchus sp.(32%) was highest followed by Hookworm(21%), Strongyloides sp.(13%) and Oesophagostomum sp.(5%).coccidian infections with Eimerian sp. found in(28%) and Isospora sp. in (8%) cases. Thawait et al., (2014) investigated the prevalence of gastro-intestinal parasites in captive wild animals of Nandan Van Zoo, Raipur, Chattisgarh. Examination of 210 fresh fecal samples including spotted deer revealed 46.2% (97) prevalence. In Spotted deer, Out of 100 fecal samples 38 found positive and reveal only single parasitic infection with Ascaris sp. The intensity of infection in term of EPG ranged from in spotted deer. Khan et al., (2014) surveyed the captive animals in Siddartha garden at Aurangabad city (Maharasthra) for the prevalence of gastro-intestinal parasites. Examination of 202 fecal samples including spotted deer revealed overall 15.84% of parasitic prevalence. Out of 202, 15 were deer samples, of which 3 found positive with 20.00% of overall parasitic prevalence. Strongylus spp., Balantidium coli and Trichuris sp. were observed in spotted deer. 8

20 Review of literature Thawait et al., (2015) studied the gastro-intestinal parasitic prevalence in captive wild animals of Kanan Pandari Zoo, Bilaspur, Chattisgarh. A total of 145 fecal samples examined and found 37.24% overall prevalence of gastro-intestinal parasites. In Spotted deer, Ascaris sp. of nematodes were found predominant and 35.71% overall prevalence of gastro-intestinal parasites recorded. EPG count varied from Mir et al., (2016) conducted a survey in captive wild animals in Bir Moti Bagh Mini Zoo (Deer Park), Patiala, Punjab for the prevalence of parasitic infection. Examination 0f 31 fecal samples from eight species of captive animals including spotted deer (Axis axis) revealed 20 positive for parasitic ova/oocysts with a prevalence of 68.0%. Out of 8 samples of spotted deer, 4 were found positive indicating an overall prevalence of 50% gastro-intestinal parasitic infection. Parasites recorded were Strongyle (3), Trichuris (1) and Coccidia (1). A mixed infection of (Trichuris and Coccidia) also observed in one sample of spotted deer during this investigation Abroad Mckenzie et al., (1989) surveyed the prevalence of helminthic infection of Axis deer (Cervus axis) among the intermingling herds on the puu-o-hoku Ranch, Molokai, Hawaii and recorded Capillaria sp.(20%), Cooperia sp.(60%), Gongylonema sp.(60%), Hemonchus sp.(10%), Trichostrongylus sp.(30%), Trichuris sp.(30%) and Moniezia sp.(10%). Paul mason (1994) studied on New Zealand farmed deer (Cervus elaphus), European red deer, North American wapiti and their hybrids in New Zealand and recorded lungworm (Dictyocaulus viviparous) and tissue worm (Elaphostrongylus cervi) from them. Pacon (1994) conducted a study on parasites of mouflons, stags and roe deer from lower Silesia forest region from May, 1988 to June, 1992 and a prevalence of gastrointestinal nematodiasis from 50% to 81%. The most frequent identified parasites were Oestertagia sp. (60%) and Nematodirus sp. (30%). 9

21 Review of literature Maia et al., (1999) investigated the parasitic fauna of wild pigs (Sus scrofa), red deer (Cervus elaphus), fallow deer (Dama dama), Mouflon (Ovis musimon), rabbits (Oryctolagus cuiculus) and foxes (Vulpes vulpes silacea) in various national parks in Portugal. The parasites Fasiola hepatica, Haemonchs contortus, Oesophagostomum venulosum and Oesophagostomum radiatum were recovered from red deer for the first time in Portugal. Maia (2001) conducted a survey on parasitic prevalence on 12 red deer and 13 fallow deer over a period of seven months from September, 1992 to March, 1993 in Tapada nacional de Mafra. The study revealed Fasciola hepatica 33.4% and 15.4%; Cysicercus tenuicolis 8% and 7%; Oesophagostomum venulosum 25% and 15%; Haemonchus contortus 8% and 0% in red deer and fallow deer respectively, while a very low EPG count was observed in Trichuris sp., Capillaria sp.and Strongyloides sp. Mukhopadhyay, (2003) carried out a survey on prevalence and aetiopathological aspect of common diseases in spotted deer at 3 National parks, 6 Wildlife Sanctuaries and 5 deer Parks in West Bengal from a period of January 1997 to December death reports were investigated. Coprological examination of dead animals revealed eggs of amphistomes. Three species of amphistomes were identified as Gastrothylax cruminifer, Gigantocotyle explanatum and Cotylophoron cotylophorum. Vengust, (2003) conducted a comparative study of parasitic fauna of fallow deer (Dama dama) from two enclosures in Solvenia. In location A, Fasciola sp. (100%), Capillaria sp. (63%), Oesophagostomum sp. (16%), Trichuris sp. (11%) and location B, Oesophagostomum sp. (67%) and Trichuris sp. (11%) were recovered from the infected fallow deer. Cisek et al., (2003) investigated the prevalence of gastro-intestinal parasites in roe deer, fallow deer and red deer in western Pomerania with in March, 1999 to July, A total of 199 animals were examined and an overall prevalence rate of 73.5%, 96.5% and 92.0% were recorded in red deer, red deer and fallow deer respectively. The individual prevalence of Haemonchus sp. were 9.3%,10.3% and 4%; Oesophagostomum sp. were 10

22 Review of literature 10.5%,10.3% and 44%; Capillaria sp, were 6.98%, 1.5% and 4%; Trichostrongylus sp, were 1.2%, 0% and 0% recorded in roe deer, red deer and fallow deer respectively. Islam et al., (2003) carried out a survey on prevalence of gastro-intestinal parasites of captive deer in Chittagong zoo. Investigation revealed Paramphistomum sp. were 87.5% and 100% in spotted deer and barking deer respectively as well as Trichostrongylus sp. (8.3%), Strongyloides sp. (8.3%), Cooperia sp. (8.3%) in barking deer. It was also found that female were more prone to gastro-intestinal parasitic infection than male. Shibashi et al., (2003) examined a total of 134 fecal samples of sika deer (Cervus nippon) in three public facilities (area A, B and D) and a farm (area C) in Kanagawa, Saitama and Chiba prefectures, Japan to study the gastro-intestinal parasitic infection in deer. Three genera of intestinal parasites (Eimeria sp., Dicrocoelium chinensis and Trichuris ovis) and some undefined nematodes were detected. The prevalence of D. chinensis were 10% and 37% in areas A and D respectively, and Trichuris ovis were 7%, 5% and 3% in areas A, B and C respectively. Satin-Duran et al., (2004) conducted a survey of abomasal parasites in cervids from three localities across Central and Western Spain. Three polymorphic species of helminthes of the Ostertagiinae: Spiculopteragia asymmetrica/s.qudrispiculata, Ostertagia leptospicularis/o.kolchida and O.drozdzi/O.ryjikovi were found commonly. Trichostrongylusaxei was found in very few cases. Gastro-intestinal parasites prevalence in cervids was high, ranging from 97.5 to 100%. Helminthic burdens were higher in fallow deer then in sympatric red deer. Cisek et al., (2004) investigated the parasitic prevalence in wild deer in western Pomerania, Poland. A total of 440 fecal samples were examined, among them 50 from roe deer, 80 from fallow deer, 49 from red deer were analyzed. The prevalence of gastrointestinal nematodes were 84% in roe deer, 47.5% in fallow deer and 71.4% in red deer detected. Pulmonary nematodes were also detected in roe deer (39.1%), fallow deer (30%) and red deer (82.6%) during the investigation. 11

23 Review of literature Gossens et al., (2005) conducted a 12-month survey of the gastro-intestinal helminthes of antelopes, gazelles and giraffids of two Zoos, Antwerp zoo and Animal Park, Planckendael in Belgium and on necropsy of one red deer and three Nelson s elk recovered Camestrongylus mentulatus from the abomasum and Trichostrongylus sp., Nematodirus fillicolis, Capillaria sp. and Trichruris sp. from the intestine. The total worm counts between 950 and Pilarczyk et al., (2005) carried out a survey of the gastro-intestinal parasites of roe deer and red deer in the Western Pomerania voivodeship and reported oocysts of gens Eimeria in roe deer (52%) and red deer (74.5%). Four species of Coccidia in roe deer (E. capreoli. E.panda, E.rotunda, E. ponderosa) and two species in red deer (E. sordida, E. elaphi) were identified. The overall prevalence of gastro-intestinal nematodes recorded in roe deer and red deer were 100% and 47.8% respectively. The nematode species found in the alimentary tract were Spiculopteragia boehmi, Ostertagia kolchida, O. leptospicularis, Haemonchus contortus, Chabertia ovina, Oesophagoatomum venulosum, Nematodirus sp., Trichocephalus ovis and Capillaria bovis. Masuduzzaman et al., (2005) investigated the incidence and pathological changes in Fascioliasis (Fasciola gigantica) of domesticated deer of Chittagong zoo and some houses of Chittagong city. Out of 44 fecal samples, 34(77.3%) were found positive for Fascioliosis, among which 15 were recorded in Spotted deer. In post-mortem examination, Fascioliosis found in 13 (100%) cases, of which 08(61.5%) were from Spotted deer. Overall incidence of Fasciola infection in deer was 82.5%. Balicka-Ramisz et al., (2005) evaluated the gastro-intestinal and pulmonary helminthes infection in fallow deer in North-West Poland. Examined 52 animals and 98 fecal samples and found 12 gastro-intestinal and 2 lung nematodes. The most common gastro-intestinal species were Oesophagostomum venulosum, Spiculopteragia boehmi, Haemonchus contortus and Nematodirus sp., two lung nematode species were Elaphostrongylus cervi and Varestrongylus sagittatus found with prevalence of 59.6% and 46.1% respectively. 12

24 Review of literature Novobilsy et al., (2007) conducted coprological examination of wild cervids of Czech Republic and reported that the prevalence of giant liver fluke (Fascioloides magna) varied from 4% to 95%. Lim et al., (2008) surveyed the prevalence of intestinal parasites of different group of mammals of a Zoological garden in Malaysia. Examined 197 randomly collected fecal samples from various primates (99), hoofed mammals (70) and feline (28) and found 54.5%, 45.7% and 89.3% are infected with intestinal parasites respectively. Intestinal Parasites found in primates where Balantidium Coli (19.2%), Cryptosporodium sp. at (14.1%), hookworm (10.1%), Trichuris sp. (5.1%), Ascaris (4.0%), Blastocystis sp. (2.0%). For hoofed mammals, hookworm had the highest prevalence (34.3%) followed by Trichuris sp. and cryptosporodium sp. (5.7%). For feline, Toxocara cati was most prevalent (64.3%), followed by Cryptosporidim sp. (14.3%), Spirometra sp. (7.1%) and hookworm (3.6%). Satin-Duran et al., (2008) conducted a study on age distribution and seasonal dynamics of abomasal helminthes in wild red deer in Central Spain. Study reported a higher prevalence of Spiculopteragia assymetrica/s.qudrispiculata and lower intensity of Osertagia leptospicularis/o.kolchida and Ostertagia drozdzi/o.ryjikovi. Both intensity and prevalence of abomasal parasitism were recorded higher in male and older animals. Borkovcova et al., (2008) carried out a survey on endoparasitic infection of fallow deer (Dama dama) over a period of in South Moravia (Czech Republic).Fecal examination revealed a maximal prevalence of 60% in the herd in 2004, which reduced to 20% in 2008 and detected parasites were only of species of order Strongylida. Annual dynamics of parasites was found always highest in autumn season. Corden et al., (2008) surveyed the occurrence of gastro-intestinal parasites in the animals of the zoological garden Pena Escrita (almunecar, Spain) between June 2006 and May Collected 432 samples and found 72.5% of animals were infected with pathogenic endoparasites with prevalence of Eimeria sp. (4.5%), Trichuris sp. (5.1%), Strongyloides sp. (4.5%), Cyclospora sp. (4.5%), Cryptosporidium sp., Dicrocoelim 13

25 Review of literature dendriticum, Metastrongylus sp. and Cylicospirura sp. in Artiodactyla. 70% of animals shown multiple parasitic infection. Ruta et al., (2009) collected fecal samples if 487 red deer, 213 fallow deer, 24 mouflon, 180 wild boars, 19 pheasant and examined for the presence of gastro-intestinal and pulmonary parasites. The prevalence of gastro-intestinal parasites recorded were 73.3%, 63.0%, 87.5%, 26.3% and 100% in red deer, fallow deer, mouflon, wild boar and pheasant respectively. The parasites identified were Protostrongylus sp., Trichostongylus sp., Strongyloides sp., Paramphistomum sp., Dictyocaulus sp. and Coccidia. Strongyloides sp. were the most common gastro-intestinal parasite found in red deer, fallow deer and moufflon. Darabus et al., (2009) investigated the prevalence of parasites from the digestive tract of six species of herbivores namely common deer (Cervus elaphus), fallow deer (Capreolus capreolus), reindeer (Rangifer tarandus), guanaco (Lama guanacue), dwarft goat (Capra aegagrusf. hircus) and the Shetland pony (Equus canallus) from Timisoara zoo by conducting the post-mortem examination of the dead animals. The overall prevalence of gastro-intestinal parasites recorded in this study was 67.8%. Eimeria sp., Namatodirus sp. and Trichostrongylus sp. were identified parasites from red deer, fallow deer, reindeer, guanaco and pygmy goat. Medne et al., (2009) conducted a survey on prevalence of gastro-intestinal parasites in red deer and fallow deer of the deer gardens in Latvia. The identified parasites were Strongyloides sp., Paramphistomum sp., Trichostrongylus sp., Protostrongylus sp., Dictyocaulus sp. and Coccidia. Strongyloides sp. were the most commonly found parasite of red deer and fallow deer. Fangiolini et al., (2010) surveyed the prevalence of gastro-intestinal parasites in primates, carnivores, perissodactyls, artiodactyls and proboscideans at the two Italian Zoos; the zoo safari of Fasano and the Giardino Zoologica of Pistoia between January 2008 to April % of samples revealed parasitic infection with the prevalence of Cryptosporidium sp. (10%), Toxocara cati, Strongyloides stercoralis, Toxascaris leolina 14

26 Review of literature and Hookworms (43.3%) in carnivores, Cryptosporidim sp. (66.7%), Trichuris sp. and Strongyloides sp. (100%) in primates; Eimeria sp. and Cryptosporidim sp. (25%) and Trichuris sp., Toxocara vitulorum, Strongyle and Paramphistomum sp. (57.1%) in artiodactyls for protozoa and helminthes respectively. Kuzmina et al., (2010) surveyed the helminth parasites of roe deer (Capreolus capreolus) from 9 different regions of Ukaraine and recorded 92.4% prevalence of helminthic infection. Examination of 92 roe deer by partial helminthological dissection revealed Setaria cervi (10.9%) in visceral cavity, Dictyocaulus eckerti (6.9%) and D. capreolus (2.3%) in lungs, Taenia saginata larvae (2.3%) in mesentry, Paramphistomum cervi (10.%), Haemonchus contortus (57.6%), Ashworthius sidemi (40.2%), Marshallagia marshalli (15.2%), Nematodirus oiratinus (1.1%), Trichostrongylus axei (3.3%) in stomach, Moniezia expansa (1.1%), Bunostomum phlobotomum (10.90%) in small intestine, Trichocephals ovis (18.5%), Oesophagostomum venulosum (7.6%) and O.dentatum (1.1%) in caecum and Chabertia ovina (28.3%) in large intestine. Gurler et al., (2010) conducted a coprological study to access the prevalence of helminthes infection at the Samsun zoo, Turkey. Examination of 338 numbers of fecal samples from 23 ruminants,3 equines,21 carnivores,10 rabbits,2 primates, 2 kangaroos and 123 birds in four seasons revealed overall 36.4% of helminthic infection with Dicrocoelium sp. (0.91%), Moneizia sp. (1.1%), Toxascaris leolina (7.1%), Toxocara sp.(8.6%), Uncinaria stenocephala (1.4%), Dictyocaulus filarial (1.9%), Mulleria sp.(14.1%), Strongyle (20.7%), Strongylus adentatus (8.3%), Cyathostomum sp. (66.7%), Trichuris sp. (3.9%), Capillarid (9.5%), Ascaridia sp.(1.9%) and Heterakis sp. (12%). Vazquez et al., (2010) conducted a long term study on internal parasitic infections in free ranging roe deer (Capreolus capr eolus) from NW Spain over a period of last 12 years. Survey reported a significant increment in prevalence of infection by endoparasites in the last 12 years. Coprological analysis revealed Eimeria sp., Moniezia sp., Strongylids, Nematodirus sp., Trichuris pp., Capillaria sp., Dictyocaulus sp.and Varestrongylus sp. in roe deer. 15

27 Review of literature Kanungo et al., (2010) studied the prevalence of gastro-intestinal helminthiosis in captive deer of Bangladesh in 3 captive locations; Dhaka zoo, Chittagong zoo and Dulahazara Safari Park, Chokoria. The study revealed 76.2% deer affected with gastrointestinal helminthes and Paramphistomum sp. and Haemonchus sp. were found more prevalent. The prevalence of helminthic infection in Hog deer (90.91%) followed by Barking deer (78.79%), Spotted deer (75%) and Sambar deer (70%). Mixed infection was noted in majority of the deer including spotted deer. Atanaskova et al., (2011) studied the prevalence of endoparasites in the wild animals in the Zoological garden in Skopje, Macedonia over a period of three years from 28 different species of animals including deer (Capreolus capreolus). Fecal examination of deer revealed the presence of gastro-intestinal parasites Strongyloides sp., Trichostrongylus sp., and Trichuris sp. Aukstikainiene et al., (2013) conducted coprological analysis of 10 fecal samples from 30 roe deer (Capreolus capreolus), 60 red deer (Cervus elaphus) and 20 elk (Alces alces) in Lithuania.100% of the samples fond positive for nematode and/or trematode eggs. Paramphistomum cervi (10%), Dicrocoelium dendriticum (45%), Strongyloides papillosus (60%), Trichuris ovis (5%), Capillaria sp. (19%), Nematodirus sp. (36%) and Strongylidae (25%) were commonly found. EPG count was found maximum in farmed red deer fecal samples. Radhy et al., (2013) investigated the gastro-intestinal protozoa of animals of Al- Zawraa Zoo in Bagdad. A total of 160 fecal samples from 19 species of captive animals including deer during November 2012 to May 2013 were examined. Out of 160, 16 were deer samples and100% gastro-intestinal protozoan infection reported with cysts of Cryptosporidium sp. Rahman et al., (2014) investigated the prevalence of gastro-intestinal parasites of herbivores at Dhaka National Zoological Garden of Bangladesh. Examination of 52 fecal samples from 21 herbivores during January to May 2012 revealed 76.9% overall parasitic prevalence, of which 61.5% for helmithic and 55.8% were positive for protozoan 16

28 Review of literature infection. Identified parasites were Balantidim coli (55.8%), Paramphistomum sp. (36.5%), Fasciola sp. (1.2%), Stomach worm (3.9%), Ascaris sp. (1.9%), and Strongyloides sp. (1.9%). Out of 21 herbivores, 15 showed mixed parasitic infection including Spotted deer. Mixed infection revealed Balantidium coli (50%), Paramphistomum sp. (33.33%) and Fasciola sp. (50%). The intensity of infection in terms of EPG/CPG ranged from in Spotted deer. Hidalgo et al., (2014) carried out an investigation in 110 red deer (Cervus elaphus) in the Riano Regional Hunting Reserve, in the province of NW Spain and revealed the presence of Onchocerca flexuosa in the dorsal and flank region of deer. A total of 85.45% of red deer were found parasitized with Onchocerca infection with a mean intensity of 9.53 ± nodules/host, ranging between 1 and 74 nodules/deer. Khatun et al., (2014) conducted a coprological study of gastro-intestinal parasites of captive animals at Rangpur Recreational Garden and Zoo in Bangladesh. 45 fecal samples of different animals including Spotted deer were examined and reported 60% (27/45) overall parasitic prevalence, of which 35.6% (16/45) of helminthic and 24% (11/45) of protozoan infections. In Spotted deer, Out of 36 fecal samples 25 found positive for parasites with the prevalence of Fasciola sp. (13%), Moniezia benedeni (4.4%), Strongyloides sp. (3.7%), Dictyocaulus sp.(4.4%), Coccidia sp.(13%) and Stomach worm(4.4%). Mixed infection of Strongyloides sp. and Coccidia was also observed in Spotted deer. Keidane et al., (2014) carried out an investigation on parasitosis in the red deer (Cervus elaphus), fallow deer (Dama dama), mouflon (Ovis musimon) and wild boar (Sus scrofa) in Latvia over a period of year 2009 to Coprological examination of 487 deer, 213 fallow deer, 24 mouflons and 315 wild boars revealed a highest infestation of digestive system comprised Strongylida in red deer (48.7%), fallow deer (62.5%) and moflons (47.1%). Fasciolosis was found only in red deer with an extensive infestation of 5% while Paramphistomum sp. was found in both red deer and fallow deer with a prevalence rate of 8.6% and 2.5% respectively. 17

29 Review of literature Barmon et al., (2014) studied the prevalence of gastro-intestinal parasites of deer at Char Kukri Mukri Upzilla of Bhola district of Bangladesh. Total 127 fecal samples were examined by Stoll s ova dilution technique and 88 samples were fond positive for gastrointestinal parasites with an overall prevalence rate of 69.29%. The identified parasites were Fasciola sp. (8.66%), Paramphistomum sp. (20.47%), Stomach worm (18.11%), Hookworm (20.47%), Strongyloides sp. (1.57%), Oesophagostomum sp. (1.57%), Eimeria sp. (6.30%) and Balantidium coli (1.57%). The overall prevalence of nematode, trematode, protozoan and mixed infection were 29.13%, 25.19%, 5.51% and 9.45% respectively. The seasonal prevalence of gastro-intestinal parasites in summer and winter were 70.59% and 66.67% respectively. The range of EPG/CPG/OPG varied from 100 to 300 and highest count was found in Paramphistomum sp. (300), Hookworm (300) and Stomach worm (300). Davidson et al., (2014) surveyed the prevalence of helminthic fauna among an isolated Norwegian population of wild red deer (Cervus elaphus).13 red deer culled from the isolated population at the Mongstad Oil Refinery, Norway and investigated for gastro-intestinal helminthes. Osteragia leptospicularis/o.kolchida detected in 83%, Spiculopteragia spiculoptera/s.mathevossiani in 92% and Trichostrongylus axei in 42% of abomasum examined. Capillaria bovis, Cooperia oncophora, Oesophagostomum venulosum, Trichuris globulosa and tapeworm fragments (anoplocephalids) recovered from intestine of red deer. Rana et al., (2015) carried out a comparative study on prevalence of endoparasitic infection in 86 Hog deer (Axis porcinus) kept at Jallo Wildlife Park, Lahore Safari Park and Lahore zoo, Pakisthan. Overall infestation of endoparasites in Hog deer at three Wildlife Parks was found 86.04%. Mixed infection was observed in majority of the Hog deer. Paramphistimum cervi (31.39%), Moniezia expansa (39.53%), Moniezia benedeni (6.98%), Strongyloides papillosus (82.55%), Trichuris globulosa (74.41%), Trichostrongylus sp. (81.39%) and Haemonchus contortus (32.55%) were recorded in these locations. Investigation revealed Lahore zoo (90.00%) mostly susceptible to endoparasites followed by Lahore safari park (87.5%) and Jallo Wildlife Park (83.92%). 18

30 Review of literature Pilaczyk et al., (2015) studied the prevalence and intensity of gastro-intestinal parasitic infection in farmed fallow deer and recorded 57.33% average prevalence in an annual cycle and intensity of 529 EPG. Trichostrongylus (13.33%) and Chabertia (10%) species were found more frequently and Strongyloides sp. (3.33%) had the lowest prevalence. Rana et al., (2015) conducted a comparative study of prevalence of endoparasites in Sambar (Rusa unicolor) and Goral (Naemorhedus goral) kept at Jallo Wildlife Park and Lahore Zoo, Punjab, Pakisthan. Fecal examination revealed, Sambar of Jallo Wildlife Park (81.81%) were mostly susceptible to gastro-intestinal helminthiosis compared to Lahore Zoo (75.0%) and reported an overall prevalence of 77.5% of helminthic infection in Sambar with Trichistongylus sp. and Gaigeria pachyscelis in large numbers. 19

31 Materials and Methods MATERIALS AND METHODS The present investigation was carried out for a period of eight months from October 2015 to May 2016 in different seasons viz; Winter (Oct. to Feb.) and Summer (March to May) during the year at three captive wildlife and deer parks in and around Bhubaneswar. The three selected captive locations were Nandankanan Zoological Park, Raj Bhawan Deer Park, Bhubaneswar and Tulasipur Deer Park, Cuttack under Cuttack Municipal Corporation. 3.1 Prevalence of gastrointestinal parasites Selection of animals The study covered all the ages and both sexes of Spotted Deer, found in the three selected locations. Two hundred ninety two (292) Spotted deer were selected randomly and examined for the prevalence of gastro-intestinal parasites. For sex wise variation in prevalence of gastro-intestinal parasites study, faecal samples from male and female Spotted deer were collected separately from their respective enclosures of Nandankanan Zoological Park, only where male and female deer are kept at separate enclosures. 20

32 Materials and Methods Study design Selection of animals Collection of fresh faeces from spotted deer Preservation of faeces in 10% formalin Qualitative and quantitative examination of faeces Identification of ova/larva/oocyst/cyst of different gastro-intestinal parasites Compilation and tabulation of data Statistical analysis Interpretation Figure-1: Flow diagram showing research out line 21

33 Materials and Methods Collection of samples Fresh faecal samples were collected from the enclosure ground after defecation with the help of animal care takers in the early morning. During collection of faecal samples proper care was taken to prevent any type of extraneous contamination Preservation of samples After collection of faecal samples each sample of about grams were kept in the separate clean interlocked polythene bags containing 10% formalin. The sample were marked according to collection place, sex of animal and date of collection of faecal matter with a marker. The properly labelled and correctly numbered polythene bags containing fresh faecal samples were brought to the laboratory of the Department of Parasitology, College of Veterinary Science and Animal Husbandry, Bhubaneswar for parasitological examination Coprological examination The collected samples were subjected to detail routine parasitological analysis for the presence of parasitic eggs/oocysts by direct smear examination, standard sedimentation and flotation techniques. The ova/larvae/oocyst/cyst of different parasites were identified as per the morphology and morphometry and quantitative estimation done by using Modified Stoll s ova dilution technique and Mc Master technique to determine eggs per gram (EPG), oocyst per gram (oocyst) and cyst per gram (CPG) of faeces as described by Soulsby (1982) Direct Method To prepare the direct faecal smear, a drop of water was placed on a clean grease free slide. A small quantity of faeces was placed on the drop, stirred until the entire sample was suspended. Any fibres or particles were removed and the slide was covered with a clean cover slip avoiding any air bubble. Glass slide was then examined under low power microscope (10x). 22

34 Materials and Methods Sedimentation Method About 1 gram of faeces was taken in a glass pestle and a little quantity of water was added to it and mixed well. Suspension was strained to remove the debris and poured into a centrifuge tube up to an inch below the brim. Centrifuged at revolutions per minute (rpm) for 2-3 minutes. The supernatant was discarded and from the sediment, a drop was taken and examined under low power objective (10x) by covering with a cover slip. The presence of eggs were identified through their morphological characteristics (Bowman, 1999) Flotation Method About 2 grams of faeces was taken in a glass pestle and little quantity of saturated solution of flotation fluid (magnesium sulphate/zinc sulphate/sugar solution) was added and mixed well. Suspension was strained to remove the debris. The suspension was centrifuged at 1500 revolutions per minute (rpm) for 2-3 minutes. The surface layer was examined under low power microscope by covering with a cover slip. The presence of eggs was identified through their morphological characteristics (Bowman, 1999) Modified Stoll s Ova Dilution Technique The faecal sample was mixed well and 3 grams of faeces were weighed with the help of a balance and taken in a 100 ml graduated beaker. Beaker was filled with water up to 42 ml mark. The faeces was thoroughly mixed with water with the help of a stirrer. Faecal mixture was strained with a tea strainer. The strained mixture was again shaken and 0.15 ml of the mixture was taken using a 1 ml clean pipette and put on a clean grease free glass slide. Glass slide was covered with a cover slip. Special care was taken to avoid any bubble formation under the cover slip. Slide was placed under a microscope and parasitic eggs/cyst/oocyst were identified and counted. Total number of eggs/cyst/oocyst of parasites found was multiplied by 100 to get the egg/cyst/oocysts per gram of faeces (EPG/CPG/OPG). 23

35 Materials and Methods Mc Master Method The Mc Master technique depends upon the examination of a precise volume of a suspension of faeces on flotation solution and counting the eggs that are floated up in the counting chambers of Mc Master slide. A known volume of faeces (5 to 10 gm) was thoroughly suspended in a known volume (50 to 100 ml) of saturated salt solution (sp.gr, 1.12). The suspension was strained through a 150 mm mesh sieve to remove the coarse particles. A portion of the suspension was withdrawn with the help of a plastic pipette and allowed to run into the chambers of the Mc Master slide. Slide was allowed to stand for 3 to 5 minutes to allow the eggs to float. Eggs in the two chambers were counted under the lower power objective (10x) of the microscope. Number in onegram Number in twochambers dilution factor* 0.3 Totalvolume of suspensionin ml Dilution factor * Totalweight of faeces 3.2 Identification of gastro-intestinal parasites Identification of egg and larva The identification of eggs was based on morphology and micrometric studies (Bowman, 1999). The eggs recovered from the faecal samples by various methods as described above, were studied for morphological characteristics and identified Culture of faeces eggs The faecal sample highly positive for nematode eggs were selected for culture. The faeces were broken up and mixed with charcoal and placed in a glass jar with a lid and stored at a temperature of 26 0 C for seven days.the faecal culture was examined at interval for not be too dry nor too wet. After seven days of incubation the sample was transferred to Baermann s Apparatus. It consists of a glass funnel in which wire mess is placed and rubber tubing with a pinchcock is attached to the 24

36 Materials and Methods funnel stem. The funnel was filled with warm water up to one fourth to half an inch above the sieve. The cultured faeces was placed in the wire mess and the funnel was slowly filled with water until the faeces immersed.the larvae which migrate to water were collected and the larvae were identified by their morphological features and by micrometry (Bowman,1999). 3.3 Identification of gastro-intestinal parasites by necropsy examination Necropsy examination Necropsy examination of six numbers of necropsied spotted deer was conducted to assess the parasitic burden directly as faecal egg and larva counts did not always give a reliable indication of worm burdens Collection of helminth parasites During post-mortem, efforts were made to collect the adult parasites alive for identification. The worms recovered were killed immediately putting in the 70% alcohol and preserved in a suitable preservative (10% formalin). For both Fasciola and Dicrocoelium, livers were removed and cut into slices of approximately 1 cm thick squeezing the liver slices, grossly visible flukes were removed. For intestinal paramphistomes, first 4 meters of duodenum was tied off, opened, washed and examined for adherent trematodes. For recovery of nematodes of the alimentary tract, the abomasums and small intestine were ligated and removed from the animal and washed separately. Intestinal scrapping was taken and examined. Stomach was cut open over a bowl and content were collected. Stomach wall was washed thoroughly under a stream of water. The mucous membrane of stomach was carefully rubbed with the fingers to remove any worms adhered to it. Contents of the bowl were poured through a wire mesh of 0.15 mm aperture and washed with a stream of water from a rubber tube attached to the tap. The screened material was inverted over a pneumatic trough and worms were collected and preserved. 25

37 Materials and Methods Fixation and preservation of slides The collected worms (amphistomes) were washed 2-3 times in water to make them free from debris. Kept in water for an hour in a refrigerator for relaxation of muscles. They were placed in filter paper to remove the excess water. Transferred to a slide with the ventral sucker in contact with the slide surface. Then another slide was placed over the fluke and pressed gently with even pressure. Two ends of the slides were fixed together with thread or secured rubber rings. The whole material was kept in 10% formalin and left overnight Staining and mounting of amphistomes Flattened amphistomes were removed and washed in running water for several hours to remove excess fixative. This was done by putting them in a wide mouth bottle and securing it by a piece of cheese cloth over the top of the bottle with a rubber band. Submerging the bottle in running water and letting water run slowly through the cheese cloth from the tap. Amphistomes transferred from water to borax carmin stain and left for several hours till they were over stained. Excess stain was washed in 70% alcohol. Then flukes were put in ascending order of alcohols i.e 50%, 70%, 90%, 100% alcohol for one or several hours each. Washed momentarily in Xylol. Placed in a microscopic slide. Mounted with DPX mountant. Slide was kept to dry at room temperature. After complete drying, fluke was kept under a dissecting microscope and identified. 3.4 Statistical analysis The data regarding the effect of sex and season on prevalence of gastrointestinal parasitic infection were subjected to statistical analysis as per the standard methods suggested by Snedecor and Cochran, Statistical analysis were carried out by Statistical Package for Social Science (SPSC) using Chi Square test. 26

38 RESULTS Results 4.1 Prevalence of Gastrointestinal Parasites Prevalence of gastro-intestinal parasites in captive Spotted Deer The overall prevalence of gastro-intestinal parasites in captive Spotted Deer at the three selected locations viz; Nandankanan Zoological Park, Raj Bhawan Deer Park, Bhubaneswar and Tulasipur Deer Park, Cuttack was studied by examination of a total of 292 fecal samples of Spotted deer of three different locations and examination of gastro-intestinal tracts of six Spotted deer that died during the period of experiment for the presence of any parasitic eggs and gastro-intestinal parasites respectively. Out of 292 fecal samples, 72 samples were found negative for any parasitic ova while 220 samples were found positive for parasitic ova indicating an overall prevalence rate of 75.3%. A total of four species of gastro-intestinal parasites were indentified. Out of them, two species were trematode and two species were nematode. No tapeworm or protozoan cyst/oocyst or coccidian infection recorded in this investigation. The overall prevalence of trematode 25.3% (74/292) and nematode 23.2% (68/292) were recorded during the study (Table 1).A mixed infection of four different types of parasites were found in 78 samples indicating an overall prevalence of mixed infection of 26.8% (78/292). (Chart 1). Different type of mixed infection recorded in this study were Paramphistomum and Strongyle spp. (n=72), Paramphistomum and Strongyloides sp. (n=4), Fasciola sp. and Strongyle spp. (n=1), Paramphostomum and Fasciola sp. (n=1) and their prevalence rate were 24.7%, 1.4%, 0.4% and 0.4% respectively. (Table 2). 27

39 Results Table 1. Prevalence of gastro-intestinal parasites in captive Spotted deer in and around Bhubaneswar Serial no Type of infection No.of positive Prevalence % Cases 1. Nematode(single) % 2. Trematode(single) % 3. Mixed infection % Fig. 2: Prevalence of gastro-intestinal parasites in Captive Spotted Deer in and around Bhubaneswar Nematode (23%) 25% 23% 27% 25% Trematode (25%) Mixed infection (27%) Negative (25%) 28

40 Results Table 2. Prevalence of mixed infection in captive Spotted deer in and around Bhubaneswar Serial no Type of mixed infection No. of cases found Prevalence % Paramphistomum sp. and Strongyle spp % Paramphistomum sp.and Srtongyloides sp % Fasciola sp. and Strongyle spp % Paramphistomum sp. and Strongyle spp % Total % Out of 292 samples. 118 numbers of samples from Nandankanan Zoological Park, 95 from Raj Bhawan Deer Park and 79 from Tulasipur Deer Park, Cuttack were collected during the study period in two different seasons. Fecal examination of the collected samples revealed 97, 75 and 48 numbers of samples were positive for gastro-intestinal parasitic infection in the three places respectively indicating a prevalence rate of 82.2% at Nandankanan Zoological Park, 7.90% at Raj Bhawan Deer Park and 60.7% at Tulasipur Deer Park, Cuttack (Chart 2).The perusal of Chart 2 illustrated that the prevalence of gastro-intestinal parasitic infection was highest at Nandankanan Zoological Park followed by Raj Bhawan Deer Park, Bhubaneswar and Tulasipur Deer Park, Cuttack. A non-significant differences in the prevalence rate of gastro-intestinal parasites was observed between Nandankanan Zoological Park and Raj Bhawan Deer Park while a significant was recorded in the prevalence gastro-intestinal parasites in between Raj Bhawan Deer Park and Tulasipur Deer park and between Nandankanan Zoological Park and Tulasipur Deer Park (P<0.05). (Table 3) 29

41 Results Fig. 3. Overall prevalence of gastro-intestinal parasites in captive spotted deer at three different selected locations in and around Bhubaneswar 90.00% 80.00% 82.20% 79.00% 75.30% 70.00% 60.00% 60.70% 50.00% 40.00% 30.00% NZP-82.2% RBDP-79.0% CDP-60.7% 20.00% 10.00% 0.00% Nandankanan Zoological Park Raj Bhawan Deer Park Tulasipur Deer Park Total 30

42 Results Table 3: Chi-square test showing prevalence of gastro-intestinal parasites in captive Spotted Deer in three different selected locations Serial Number of faecal Number of cases Prevalence Locations No samples examined found positive (%) 1. NZP % a 2. RBDP % a 3. TDP % b N:B:- Prevalence (%) bearing different superscripts differs significantly at 5% (P<0.05). *NZP= Nadankanan Zoological Park, RBDP= Raj Bhawan Deer Park, TDP= Tulasipr Deer Park During the investigational study a total of four types of gastro-intestinal parasites were indentified in captive Spotted deer from three different locations. Out of them two were nematode and two were trematode. Examination of 292 fecal samples revealed that 141 number of Spotted deer were infected with Strongyle spp., 151 were infected with Paramphistomum sp. while Strongyloides sp. and Fasciola sp. were reported only in four and two number of Spotted deer (Table 4) indicating the overall prevalence (%) of the helminthic parasites were Strongyle spp. 141/292(48.3%), Paramphistomum sp. 151/292 (51.8%), Strongyloides sp. 4/292(1.4%) and Fasciola sp. 2/292 (0.6%). The perusal of Table 4 illustrated that the prevalence rate of Paramphistomum sp. was highest followed by Strongyle spp. while Strongyloides sp. and Fasciola sp. showed a very low prevalence rate. In this study,epg/cpg/opg (eggs/cysts/oocysts per gram of feces) were also determined. The EPG count ranges from among the different identified parasites. The highest EPG was counted in case of Strongyle spp. (300)followed by Paramphistomum sp. (200), Strongyloides sp. (100) and Fasciola sp. (100). This indicates a very low gastro-intestinal parasitc burden in case of Fasciola sp. and Strongyloides sp. 31

43 Results Table 4: Prevalence (%) of gastro-intestinal parasites identified in the faecal samples of Spotted deer (Axis axis) in and around Bhubaneswar Serial no. Type of gastrointestinal parasites Total No. of samples examined No. of positive cases Prevalence% 1. Strongyle spp % 2. Paramphistomum sp % 3. Strongyloides sp % 4. Fasciola sp % A significant difference(p < 0.05) in the load of Strongyle sp. in the fecal samples of Spotted Deer (A. axis) at all the three captive locations was recorded (Table 5)whilea non-significant effect of locations was observed forparamhistomum sp. in the fecal samples of Spotted deer at all the three captive locations investigated (Table 6). Table 5: Analysis of variance showing comparison of parasitic loadof Strongyle sp.among localities in captive Spotted deer (A. axis). Source Degree of Sum of Maen F Value Pr > F freedom squares squares Location * Error Corrected total *=Significant (P < 0.05). 32

44 Results Table 6: Analysis of variance showing comparison of parasitic loadof Paramphistomum sp. among localities in captive Spotted deer (A. axis). Source Degree of Sum of Mean F Value Pr > F freedom squares sqares Location NS Error Corrected Total NS= Non-significant (P > 0.05). A non-significant variation in the load of Paramphistomum sp.(3.60 ± 0.65 a ) and Strongyle spp.(5.50 ±1.35 a ) at Nandankanan Zoological Park was observed, while a significant difference(p < 0.05) was recorded in the load of Paramphistomum sp. (3.90 ± 0.75 a ) and Strongyle spp. (4.25 ± 0.88 ab ) at Raj Bhawan Deer Park and of Paramphistomum sp. (3.90 ± 0.71 a ) and Strongyle spp. (2.1 ± 0.43 b ) of Tulasipur deer park, Cuttack, where as there is no significant difference was observed in the parasitic load of Paramphistomum sp. at all the three captive locations (Table 7). Table 7: Location wise Mean ± SE of gastro-intestinal parasitic load in terms of EPG (egg per gram) in captive spotted deer Serial no. Locations Paramphistomum sp. Strongyle spp. 1. NZP* 3.60± ±1.35 a 2. RBDP* 3.90± ±0.88 ab 3. TDP* 3.90± ±0.43 b Means bearing different superscripts in a column are statistically significant (P<0.05). *NZP= Nadankanan Zoological Park, RBDP= Raj Bhawan Deer Park, TDP= Tulasipr Deer Park 33

45 4.2 Seasonal prevalence of gastro-intestinal parasites in captive spotted deer Results This investigation revealed a profound effect of season on the prevalence of gastro-intestinal parasites in Spotted deer. Out of 292 fecal samples, 152 sample were examined in winter season and 140 samples examined in summer season. Examination revealed that 117 Spotted deer were found infected with gastro-intestinal parasites in winter while 103 were found infected in summer, indicating a prevalence rate of 77% (117/152) in winter and 73.5% (103/140) in summer(chart 3). In winter season, prevalence was relatively higher in case of Strongyle spp. (50.0%) followed by Paramphistomum sp. (49.3%), Strongyloides sp. (2.7%) and Fasciola sp. (1.4%), while in summer season, prevalence was higher in case of Paramphistomum sp. (54.3%) followed by Strongyle spp. (45.7%)(Table 8). There was a significant difference (P < 0.05) in the mean parasitic load of Strongyle sp. and Paramphisomum sp. with respect to both the two seasons viz; winter and summer at all the three captive locations was observed but interestingly, the mean parasitic load of Strongyle sp. was recorded relatively more in winter season than summer, where as the mean parasitic load of Paramphistomum sp. was recorded relatively more in summer season than winter at all the captive locations (Table 9) 34

46 Results Fig. 4:Seasonal prevalence of gastro-intestinl parasites in captive spotted deerin and around Bhubaneswar 60.00% 50.00% 50.00% 49.30% 45.70% 54.30% 40.00% 30.00% winter summer 20.00% 10.00% 2.70% 1.40% 0.00% Strongyle sp. Paramphistomum sp. Strongyloides sp. Fasciola sp. Table 8: Seasonal status related prevalence of gastro-intestinal parasites in captive Spotted deer (A. axis) at three captive locations. (N=292) Summer (140) Winter (152) Serial no. Name of parasites No. infected Prevalence % No. infected Prevalence % 1. Strongyle spp % % 2. Paramphistomum sp % % 3. Strongyloides sp % 4. Fasciola sp % 35

47 Table 9: Mean ± SE (EPG) of different gastro-intestinal parsites in different seasons across locations in captive spotted deer Results Serial no. Season Paramphistomum sp. Strongyle sp. 1. Winter 2.83±0.48 b 5.70±0.98 a 2. Summer 4.76±0.60 a 2.20±0.42 b Means bearing different superscripts in a column are statistically significant (P<0.05). 4.3 Prevalence of gastro-intestinal parasites in captive spotted deer in relation to sex Present study alo revealed, sex had an important effect on prevalence of gastro-intestinal parasites in captive Spotted deer. Out of 118 samples of Nandankanan Zoological Park, 55 fecal samples from male Spotted deer and 63 fecal samples from female Spotted deer were examined for the study of sex effect on theprevalence of gastro-intestinal parasites, as only at Nandanankanan Zoological Park, the male and female Spotted deer kept at separate enclosure.fecal examination revealed 46 male and 51 female Spotted deer were between male and female in the fecal samples of Spotted deer of Nandankanan infected with helminth parasites indicating an overall prevalence rate of 83.6% (46/55) in male deer and 80% (51/63) in female deer. (Chart 4). The persual of Chart 4 illustrated that the prevalence of gastro-intestinal parasitic infection was higher in male than in female. The prevalence (%) comparison Zoological Park showed a non-significant variation in the presence of gastro-intestinal parasites. 36

48 Results Chart 4. Sex related overall prevalence of gastro-intestinal parasites in captive spotted deer in and around Bhubaneswar 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% 83.60% 80.00% Male Female female male 4.3 Identification of parasites Identification of egg and larva The eggs collected from sedimentation and flotation method were subjected to morphological examination and larval culture for identification. Four type of eggs were recovered from the fecal samples; Paramphistomum sp., Strongyle spp., Fasciola sp. and Strongyloides sp., were identified (Figure 21, 22, 23 & 24) as per the characters described in Soulsby. The larvae detected from the faecal samples were sbjected to faecal cultre and after 7 days of incubation, the sample was transferred to Baermann s apparatus. Larvae were recovered from the Baermann s apparatus and identified as Dictyocaulus sp.(fig. 28)(Lungworm) by their morphological features as described by Van Wyk (Van Wyk et al., 2013). 37

49 Results Identification of adult parasites Adult parasites were recovered from the rumen wall conducting post-mortem of Spotted deer died during the reaseach study. Recovered parasites were subjected to serial steps of preservation, washing, staining, dehydration, clearing and fixation (Figure 16, 17, 18, 19 & 20) for the study of morphological charcteristics of the parasites. Adult flukes were oval in shape. Oral sucker situated at the anterior extremity and vental sucker situated at the posterior extremity. Pharynx was absent. Testes were two in number and tandem in position. Ovary is post-testicular in position. Vitellaria were well developed laterally. A genital sucker enclosed the genital opening just behind the intestinal bifurcation (Figure 27). All these characteristic features led to the parasite to be Cotylophoron cotylophorum. No cestode or nematode parasites or coccidia were recovered from post-mortem of Spotted deer during the study period. 38

50 DISCUSSION Discussion The prevalence of gastro-intestinal parasites in zoo herbivores, especially in spotted deer kept in captivity is mostly influenced by various factors like size of the enclosure viz; small or large enclosure, management of enclosures, feeding and watering pattern of the animals, hygiene and sanitation of the zoo keepers or attendants. 5.1 Prevalence of gastro-intestinal parasites in captive spotted deer A total of 292 spotted deer were examined and 202 (75.3%) were found to be affected with gastro-intestinal helminthosis, which was in conformity with the findings of (Kanungo et al., 2010; Barmon et al., 2014 and Mir et al., 2016). It was revealed that deer was highly susceptible to gastro-intestinal nematodiasis which supports the earlier findings reported by (Varadharajan et al.,1999; Pilarczyk et al., 2005; Satin-Duran et al., 2008; Singh et al., 2009 and Thawait et al., 2015). During the study, it was observed that the overall incidence rate of mixed infection was 26.8%. Mixed infection of Paramphistomum sp. and Strongyle spp. was found more common in captive spotted deer which was in conformity with the findings of (Rahman et al., 2014 and Barmon et al., 2014) at Dhaka National Zoological garden of Bangladesh and at Char Kukri Mukri in Bhola district, Bangladesh respectively. Similarly Kanungo et al., (2010) who reported that a higher rate of mixed infection was noted in majority of the deer including spotted deer at three captive locations; Dhaka zoo, Chittagong zoo and Dulahazara safari park, Chokoria of Bangladesh. Single infection of Paramphistomum sp. was found common in captive spotted deer of Nandankanan zoo and at the deer parks of Raj Bhawan and Tulasipur, Cuttack and the prevalence recorded was 51.8%. The high prevalence of Paramphistomum sp. was also reported previously by Rahman et al., (2014) and Barmon et al., (2014) at Dhaka National Zoological garden and from Char Kukri Mukri in Bhola district, Bangladesh respectively. Varadharajan et al., (1999); Borghare et al., (2000); Banerjee et al., (2005) and Singh et al., (2006) also reported 49

51 Discussion the presence of Paramphistomum ova in spotted deer of Thiruvananthpuram zoological garden, Kerala, Maharajbag zoo, Nagpur, South Uttaranchal and Mahendra Choudhury zoological park, Chattabir, Punjab respectively. This indicates that Paramphistomum sp. infection are common in spotted deer in zoological gardens. Our observation also encountered that, single infection of Strongyle spp. were common in captive spotted deer which is supported by record of Strongyle ova in spotted deer at Mahendra Choudhury zoological park, Punjab and at Siddhartha garden, Maharashtra by Singh et al., (2006) and Khan et al., (2014) respectively. Meshram et al., (2008) and Singh et al., (2009) reported presence of Strongyle eggs in free-ranging spotted deer at Western Vidharbha region of Maharashtra and at Van Vihar National Park, Bihar respectively. The Fasciola sp. was recorded 0.60% which was much less than reported by Gupta et al., (2011) and Barmon et al., (2014). They reported the prevalence of Fasciola sp. was only 6.7% and 8.66% in deer around Jabalpur and at Char Kukri Mukri in Bhola district, Bangladesh. These variations may be due to climatic condition and density of intermediate host. Overall Prevalence of Strongyloides sp. recorded in the study was 1.4% in spotted deer which was similar as reported by Barmon et al., (2014) previously. More or less similar prevalence rate of Strongyloides sp. have been reported earlier by Varadhrajan et al., (1999); Jaiswal et al., (2014); Khatun et al., (2014) and Pilaczyk et al., (2015). However, some variations in the prevalence rate of gastro-intestinal parasites existed which might be due to topography of study area, environment, duration of study and season. In this study, the prevalence of gastro-intestinal helminth infection was recorded as (75.3%) which was more or less similar with the report of Varadharajan et al., (1999) who revealed that 74% of animals were infected with gastro-intestinal helminthes at the zoological garden, Thiruananthapurum, Kerala. The present study also confirms the report of Varadharajan and Kandasamy (2000), who recorded that 58% of animals were positive for helminthic infection and only 6% were positive for protozoan infections in wild animals in a mini zoo of Coimbatore. Khatun et al., (2014) reported 35.6% of animals were positive for helminths infection and 24% 50

52 Discussion positive for protozoan infections which is much lower than the present study, while Khasid et al., (2002) 96.13% of parasitic infection in wild animals and a 100% helminthic infection was observed in spotted deer at Gemini Circus Deer Park, Nagpur. Such variations in the prevalence of gastro-intestinal helminthic infection may be due to geographical conditions, method of sample collection and use of anthelmintic in captive animals. During the present study, no cestode or protozoan parasites were recorded in captive spotted deer which is similar with the finding of Borghare et al., (2009) who reported the absence of cestode and protozoan infection in captive deer at Maharajbag zoo, Nagpur. Rana et al., (2015) had also reported absence of protozoan infection in captive deer at Jallo wildlife park, Lahore safari park and Lahore zoo in Pakistan. The intensity of different parasites in deer was calculated and measured in this study. The intensity of infection in terms of EPG ranged from in spotted deer which was more or less similar with the findings of Thawait et al., (2014); Barmon et al., (2014); Rahman et al., (2014) and Thawait et al., (2015). 5.2 Seasonal prevalence of gastro-intestinal parasites in captive spotted deer Seasonal fluctuation of the year had a significant (P < 0.05) effect on the prevalence of gastro-intestinal parasitic infection in spotted deer. A relatively higher infection with gastro-intestinal parasites were observed in winter season (77%) than in summer (73.5%). From the result of present study, it is noted that the incidence of helminth parasites in Axis deer touched its peak during winter followed by summer season. This can be attributed to the fact that after rains during winter (Oct., Nov. and Dec.) the atmosphere is conducive for increased pasture contamination and development of infective larvae and also increased infestation of parasites. Reports of this study was in conformity with the findings of Meshram et al., (2008); who reported that the incidence of gastro-intestinal parasites in Axis deer was highest in winter season followed by rainy season in the scrub forest of Borgaon Manju in Western Vidarbha region of Maharashtra. Our observations also corroborates well with the finding made by Kafil Hussain et al., (2002); who reported 51

53 Discussion a higher incidence of helminthic infection in winter and rainy season in Axis deer from Nagpur while Barmon et al., (2014) had recorded the seasonal prevalence of gastro-intestinal parasites in summer and winter were 70.59% and 66.67% respectively. The contrast in between the present and earlier findings can be explained by the fact of variation in the geographical location of the study area also the methods used in this study. However, in this investigation, year was divided into two seasons, but in other parts of the world there were four seasons. So, this difference in classification of seasons had made some over lapping of months and seasons. Therefore, that might have created some dissimilation. 5.3 Sex wise prevalence of gastro-intestinal parasites in captive spotted deer Sex had a significant (P < 0.05) effect on the prevalence of gastro-intestinal parasitic infection in captive spotted deer. A relatively higher infection was observed in male deer (83.6%) than in female (80.0%) which was similar with the report of Satin-Duran et al., (2008); who had also observed a higher intensity and prevalence of parasitism in male animals than in female. A similar finding was also observed by Rana et al., (2015) who reported that male had a higher infection of endo-parasites as compared to females in deer. 5.4 Identification of eggs and larva Based on morphological observations (Bowman, 1999) nematode and trematode eggs recorded in this study were identified as eggs of Strongyle spp., Strongyloides sp. and eggs of amphistomes and Fasciola sp. respectively. Similar morphological observations on ova of deer were also recorded by Barmon et al., (2014) during their study at Char Kukri Mukri in Bhola district, Bangladesh. The lungworm larvae detected in the fecal samples of spotted deer during this study were identified as Dictyocaulus species on the basis of their morphological features which is supported earlier by Nashiruddullah et al., (2005) and Ruta et al., (2009) who had reported the prevalence of Dictyocauls sp. in spotted deer and also recovered the lungworm parasites from the lungs of deer through post-mortem of dead ones. Earlier observations by Puttanaiah et al., (2011) who had reported the presence 52

54 Discussion of Dictyocaulus sp. in deer, causing verminous pneumonia in deer also supports the present finding. 5.5 Identification of adult parasites In this study, the adult parasites recovered from the rumen wall conducting post-mortem of dead spotted deer were identified as Cotylophoron cotylophorum species of amphistomes on the basis of their morphological characteristics which was in conformity with the findings of Mukhopadhayay, (2003) who had also reported the presence of Cotylophoron ctylophorum species of amphistomes in spotted deer from the post-mortem examination of dead ones. 53

55 SUMMARY AND CONCLUSION Summary and Conclusion The present investigation was carried out from October, 2015 to May, During the prevalence study, 292 faecal samples and gastro-intestinal tract of six dead spotted deer were collected from the three different selected captive locations viz; Nandankanan Zoological Park, Raj Bhawan Deer Park, Bhubaneswar and Tulasipur Deer Park, Cuttack. Collected fecal samples were examined by direct, sedimentation and floatation methods. Out of the 292 fecal samples, 220 were found positive for ova of trematode and nematodes indicating an overall prevalence of 75.3%. No cestode or protozoan parasites were recorded in this study. The study revealed that the overall gastro-intestinal parasitic prevalence rate was highest at Nandankanan Zoological Park (82.2%) followed by Raj Bhawan Deer Park (79.0%) and Tulasipur Deer Park (60.7%). Coproscopical examination of captive spotted deer of the three selected locations revealed infection with two types of trematodes (Paramphistomum sp. and Fasciola sp.), two types of nematodes (Strongyle spp. and Strongyloides sp.) and Lungworms. The incidence of Paramphistomum sp. and Strongyle spp. was found most common in captive spotted deer. Overall prevalence of Paramphistomum sp. infection was observed as 51.8%, Strongyle spp. was 48.3%, while a very low prevalence rate was observed in case of Strongyloides sp. (1.4%) and Fasiolasp. (0.6%). The ovaerall incidence of trematode infection was recorded as 25.3%, nematode infection was 23.2% and a mixed infection of 26.8% was reported in captive spotted deer in this study. Mixed infection was found more common than single infection in deer. Paramphistomum sp. and Strongyle spp. was common in all the three captive locations, while Strongyloides sp. and Fasciola sp. was detected only at Raj Bhawan Deer Park and Lungworm infection was reported in spotted deer of Nandankanan Zoological Park only. In the cases of mixed infection, the infection rate of Paramphistomum sp.and Strongyle spp. was found highest as 24.7%, followed by Paramphistomm sp. and Strongyloides sp. (1.4%), Fasciola sp. and Strongyle spp. (0.4%) and Paramphistomum sp. and Fasciols sp. (0.4%). 54

56 Summary and Conclusion The seasonal investigation on prevalence of gastro-intestinal parasites in captive spotted deer at the three captive locations revealed a relatively higher prevalence rate of gastro-intestinal parasites in winter (77%) than in summer (73.5%). In winter season, the prevalence of Strongyle spp. (50.7%) was relatively higher than Paramphistomum sp. (4.3%) while the reverse was recorded in summer season with a prevalence rate of Paramphistomum sp. (54.3%) and Strongyle spp. (45.7%). The coprological examination of male and female spotted deer of considerable sample size kept in separate enclosure at Nandankanan Zoological Park revealed that the overall prevalence of gastro-intestinal parasitic infection was higher in case of male 46/55(83.6%) than in female deer 51/63(80.0%). In this study, post-mortem examination of six numbers of dead spotted deer were carried out, which revealed infection with Paramphistomum sp. Mature amphistomes were recovered from the ruminal papillae of spotted deer. The helminthes eggs were identified on the basis of their morphological characteristics. The larvae detected in the fecal samples of spotted deer were properly recovered by using Baermann s technique and identified as Dictyocaulus species on the basis of their morphological features. The adult parasites recovered from postmortem were processed and the morphological features were observed and identified as Cotylophoron cotylophorum species of amphistomes. In conclusion, it can be said that the prevalence of gastro-intestinal parasites in spotted deer in these three captive locations reported during this study indicates that these zoos and parks are not free from parasitic infections. The high prevalence of gastro-intestinal parasites determined from the current study indicating that it might have some detrimental effect on the health of these captive deer. It means that an undetermined number of deer had the gastro-intestinal infection without showing any outward physiological signs of infection. It is zoonotically very important because these animals may be serving as reservoir of hosts for gastro-intestinal parasites that are pathogenic to man. The incidence of gastro-intestinal parasites like Paramphistomum sp., Strongyle spp., Strongyloides sp. and Fasciola sp. either as single or mixed infection 55

57 Summary and Conclusion recorded in captive deer at a higher rate during the present investigation shows that; it is highly necessary to take efficacious control measures to reduce their environmental contamination. Frequent dung removal and treatment of infected animals are very important strategies for control of gastro-intestinal parasites. Moreover, husbandry practices, the routine monitoring of parasitic disease and the use of selective anthelmintic can represent crucial measures for the control of gastro-intestinal parasitic infections in captivity. The higher incidence of gastro-intestinal nematodiasis in captive spotted deer in this study suggests that the soil of the enclosure of the selected locations may be a possible persistent potential source of gastro-intestinal nematode infection for these captive animals should be examined systematically which may help in effective control and minimize the reoccurrence of such parasites. The prevalence of Dictyocaulus sp. (Lungworm) infection in the captive axis deer, those appeared apparently healthy even when they were harboring the parasites suggested that infections routinely are sub-clinical in these animals and they may serve as carriers or reservoir host for this infections. So, more investigation is also needed to have an idea of the prevalence of vectors and intermediate hosts of the prevailing gastro-intestinal parasites in these captive animals. The present study has generated an important baseline data and provides a clear concept regarding parasitism of these captive spotted deer which is of immense significance from epidemiological point of view. Moreover, It can be suggested that further a long term epidemiological study of gastro-intestinal parasitic infection is highly essential for a better understanding of parasitism in detail and necessity of future research work to ensure better management and to find out the best possible ways to prevent the possible reoccurrence of existing infection in spotted deer in zoos and parks. Ultimately, it may be able to minimize the chances of zoonotic infections among visitors, zoo and deer park workers, also can bring normal appearance of captive animals and can save the valuable lives of these animals as well as the economic losses of the government. 56

58 References REFERENCES Aukstikalniene R. and Bukelskis E A contribution to the knowledge of endoparasitic helminthes: Fauna of wild and farmed cervidae in Lithuania, Society Journal of Biological Science. 2(1): 1-4 Atanaskova E, Kochevski Z, Stefanovska J and Nikolovski G Endoparasites in wild animals at the Zoological garden in Skopje, Macedonia, Journal of Threatened Taxa. 3(7): Adekunle BA and Olayide JA Preliminary investigation zooanthroponosis in a Nigerian Zoological garden, Vet. Res. 2(3-4): Akinboye DO, Ogunfetimi AA, Fawole O, Agbolade O and Ayinde OO Control of parasitic infections among workers and primates in Nigerian zoo, Nig. J. Parasitol. 31: Ajibade WA, Adeyemo OK and Agbede SA Coprological survey and inventory of animals at Obafemi Awolowo University and University of Ibadan Zoological gardens, World J Zool. 5(4): Arora BM Bibliography on microbiological, pathological, parasitological and clinical studies in Indian Wild fauna-mammals, Aves and Reptiles. Divison of Epidemiology, Indian Veterinary Research Institution, Izatnagar, India. 236 pp. Banerjee PS, Rajat G, Yadav CL and Hira R Parasite infections in some wild animals of Uttaranchal, Indian Journal of Animal Science, 75: Balicka RA, Pilarczyk B, Ramisz A and Cisek A Occurance of gastro-intestinal and pulmonary nematodes of fallow deer (Dama dama L.) in North West Poland, Acta Parasitologica, 50: i

59 References Barmon BC, Begum N, Labony SS, Kundu UK, Dey AR and Dey TR Study of gastro-intestinal parasites of deer at Char Kukri Mukri in Bhola District, Bangladessh Journal of Veterinary Medicine, 2(1) Bante S, Bagherwal RK and Agrawal V Prevalence of helminth parasites in wild animals of Zoological Park at Indore, Indian Veterinary Journal, 90(7): Borghare AT, Bagde VP. Jaulkar AD, Katre DD, Jumde PD, Maske DK and Bhangale GN Incidence of gastro-intestinal helminthiasis in captive deer at Nagpur, Veterinary World, 2(9); Bowman DD Georgis Parasitology for Veterinarians, 7 th Edn. W.B. Saunders. Philadelpha. Corden P, Prados GH, Romero A, Sanchez MS, Pontes M, Osna A and Rosales MJ Intestinal parasitism in the animals of the Zoological garden Pen~aEscrita (Almun~ecar, Spain), Veterinary Parasitology. 156: Cisek A, Balicka-Ramisz A, R amisz A and Pilarczyk B Monitoring of parasitic fauna in wild animals in the western Pomeranian region, Folia Universitatis Agriclturae Stetinensis, Zootechnica. 46: Chapman and Chapman N Fallow deer: their history, distribution and biology: Machynllenth Coch-y-bonddu Books, Cisek A, Balicka-Ramisz A and Pilarczyk B Occurance of gastrointestinal nematodein cervids (cervidae) of Western Poland, Electronic Journal of Polish Agricultural Universities, 6: Davidson RK, Kutz SJ, Madslien K, Hoberg E and Handeland K Gastrointestinal parasites in an isolated Norwegian population of wild red deer (Cervus elaphus), Acta Veterinaria Scandinavica, 56(1): 59. ii

60 References Darabus GH. Afrenie M, Llie MS and Indre D The researches regarding digestive parasitism in herbivores from zoological garden. Timisoara, Lucrari Stintifice Veterinara, XLII (1): Fagiolini M, Riccardo PL, Pierol L, Riccardo M, Claudia C, Riccardo F and Stefania P Gastro-intestinal parasites in mammals of two Italian Zoological gardens, Journal of Zoo and Wildlife Medicine, 41(4): Gurler AT, Beyhan YE, Acici M, Bolukbas C and Umar S Helminths of mammals and birds at the Samsun Zoo, Turkey, Journal of Zoo and Wildlife Medicine, 41(20): Gupta A, Dixit AK, Dixit P, Mahajan C and Srivastava AB Incidence of gastrointestinal parasites in wild ruminants around Jabalpur, India, Journal of Threatened Taxa, 3(11): Gossens E, Dorny P, Boomker J, Vercammen F and Vercruysse J A 12-month survey of the gastro-intestinal helminthes of antelopes, gazelles and giraffids kept at two zoos in Belgium, Veterinary Parasitology, 127: Hidalgo MR, Martinez A, Carreno R and Natividad DB Level of infection, pathology and nodule size of Onchocerca flexuosa (Nematode: Oncercidae) in red deer (Cervus elaphus) from northern Spain, Journal of Helminthology, 89 (03): 1-9. Hussain K, Sarode DB, Dakshinkar NP, Rode AM, Kothekar MD and Maske DK Prevalence of helminthic infection in axis deer at Nagpur, Indian Veterinary Journal, 79 (12): Islam SKMA, Ahmed S, Hoque MA, Alim MA and Hassan MM Gastro-intestinal parasites of captive deer and their response to selected anthelmintics, Bangladesh Veterinary Journal, 37: iii

61 References ISFR Available from, survey 2015, Jaiswal AK, Srivastava A, Sudan V, Singh R, Shanker D and Parashar R Prevalence of endoparasitic infections in wild cervids of Army Golf Course, Mathur, Journal of Parasitic Diseases, 38(4): Jadav P, Jadav K, Chavan P, Baviskar B and Maske DK Seasonal variation in prevalence of helminthic infection in captive spotted Deer at Nagpur, Zoo s Print Magazine, XXV(5): Kasid KP, Srikhande GB and Bhojne GR Incidence of gastro-intestinal helminthes in captive wild animals at different locations, Zoo s Print Journal, 18(3): Kanungo S, Das A, Das GM and Shakif-ul-Azam Prevalence of gastro-intestinal helminthiasis in captive deer of Bangladesh, Wayamba Journal of Animal Science, ISSN: Keidane D, Kruklite A and Ganola K Deer, fallow deer, moufflon and wild boars prevalent parasitosis in Deer Parks. Abstract: Proccedings of Conference, Reasearch and Pratice in Veterinary Medicine, th -28 th November, Jelgava, Latvia. Kuzmina TA, Kharchenko VA and Malege AM Helminth fauna of Roe Deer (Caprelous capreolus) in Ukarine: Biodiversity and Parasite Community, Vestnik Zoologii, 44(1): Khatun MM, Begum N, Mamun MAI, Mondal MMH and Azam MSU Coprological study of gastrointestinal parasites of captive animals at Rangpur Recreational Garden and Zoo in Bangladesh, Journal of Threatened Taxa, 6(8): iv

62 References Khan IA, Shaikh JD, Kale MK, Jadhav VD, Dama LB and Ragade Vinod R Prevalence of gastro-intestinal parasites in captive animals of Siddhart Garden Zoo at Aurangabad City, Maharashtra, India, Trends in Parasitology Research, 3(1): ISSN: X (Print); (online). Lim YAL, Ngui R, Shukri J, Rohela M, Mat NHR Intestinal parasites in various animals at a zoo in Malaysia, Veterinary Parasitology, 157: Levecke B, Dorny P, Geuren T, Vercammen F and Vercrusse J Gastro-intestinal protozoa in primates of four zoological gardens in Belgium, Vet. Parasitol., 148: Maia MJ, Vilavicosa MJ and Sioes AJL Endoparasitas da fauna silvastre ramamente ou naoassinalados em Portugal, Acta Parasitologica Portuguesa, 2: Maia MJ The helminth fauna of the red deer (Cervus elaphus) and fallow deer (Dama dama) in Tapada Nacional de Mafra, Revista-Portuguesa-de- Ciencias-Veterinarias, 96: Mohan M and Coumarane K Endoparasitic infection of Spotted Deer (Axis axis) in Puducherry, Zoo s Print Journal, 22(12): Meshram MD, Shirale SY and Khillare KP Incidence of Helmithic infection in Axis deer, Veterinary World, 1(1): 10. Mc Kenzie ME and Davidson WR Helminth parasites of intermingling axis deer, wild swine and domestic cattle from the island of Molokai, Hawaii, Journal of wildlife diseases. 25(2): Mason P Parasites of deer in New Zealand, New Zealand Journal of Zoology, 21: v

63 References Mir AQ, Dua K, Singla LD, Sharma S and Singh MP Prevalence of parasitic infection in captive wild animals in Bir Moti Bagh mini zoo (Deer Park), Patiala, Punjab, Veterinary World, 9(6): Masuduzzaman M, Raman ML and Hossain MA Incidence and pathological changes in Fascioliasis (Fasciola gigantica) of domesticated deer, Bangladesh Journal of Veterinary Medicine, 3(1): Mandal P, Jayathangaraj MG, John L, Latha BR and Raman M Prevalence of helminthic infection in free-ranging chital (Axis axis) at Mudmalai Wildlife Sanctuary, Tamil Nadu, Proceeding of the XIII Nationl Congress of Veterinary Parasitology, (February, 14-16), Kolkata. Modi GS, Prasad BN and Sinha BK Seasonal effect on prevalence of parasitic zoonotic diseases among zoo animals of Bihar, Zoos Print, 12:8-11. Mukhopadhayay SK, Studies on prevalence and aetiopathological aspect of common diseases in spotted deer in West Bengal. Ph.D thesis abstract, West Bengal University of Animal and Fishery Sciences. Novobilsky A, Horackova E, Hirtova L, Modry D and Koudela B The giant liver fluke Fasciolodies magna (Bassi, 1875) in cervids in the Czech Republic and potential of its spreading to Germany, Parasitology Reasearch, 100: Nashiruddullah N, Dazzi MM, Mir MS and Shahardar RA Recovery of Dictyocaulus species from the lungs of a kashmir red deer (Cervus elaphus hanglu), The Veterinary World, 157(19): 591. Otegbade AC and Morenikeji OA Gastro-intestinal parasites of birds in zoological gardens in South-West Nigeria, Trop. Biomed., 31(1): vi

64 References Opara MN, Osuji CT and Ooara JA Gastro-intestinal parasitism in captive animals at the zoological gardens, Nekede Owerri, Southeast Nigeria, Rep. Opin., 2(5): Parsani HR, Momin RR, Maradin MG and Veer S A survey of gastro-intestinal parasites of captive animals at Rajkot Municipal Corporation Zoo, Rajkot, Gujrat, Zoo s Print Journal, 16(10): Puttaniah GB, Seshadri SJ, Mraleedharan K and SrinivasaGowda RN Verminous pneumonia in a Hog deer (Axis porcins) caused by Dictyocalus eckerti skrajabin, 1931 with remarks on the present status of this species, Abstract, April 2011, Researchgate.net. Pilarczyk B, Tomza-MarciniakA, Udala Jan and Kuba J The prevalence and control of gastrointestinal nematodes in farmed fallow deer (Dama dama L.), Veterinariski Archiv, 85(4): Pacon Parasites of mouflons, stags and roe deer from the lower Silesia region, Wild Parasitology, 40: Pilarczyk B, Balicka-Ramisz A, Ramisz A and Lachowska S The occurance of intestinal parasites of roe deer and red deer in the western Pomerania voivodeship, Wiadomosci Parazytogiezne, 51: Ravindran R, Ajith Kumar KG and Abdul Gafoor VM Parasitic infections in wild animals of Kerala, Zoo s Print Journal, XXVI (5): 34. Radhy AM, Khalaf JM and Faraj AA Some gastro-intestinal protozoa of zoonotic importance observed in captive animals of Al-Zawraa zoo in Bagdad, International Journal of Science and Nature, 4(3): vii

65 References Rehbein S, Walburga L and Visser M Winter Beitrage Zur Kenntnis der parsitenfauna deswildes in Nordrhein-Westfalen, Deer Endoparasitenbefall des Dameildes. Z. Jagdwiss, 47:1-16. Rana MA, Jabeen F, Shabnam M, Ahamad I and Hassan MMU Comparative study of endo-parasites in captive hog deer (Axis porcinus), International Journal of Biosciences, 6(1): Rana MA, Ahmad I, Jabeen F, Naureen A and Shabnam M Comparative study of endo-parasites from fecal samples of Sambar (Rusa unicolor) and goral (Naemorhedus goral) in captivity, Journal of Biodiversity and Environmental Science, 6(5): Ramaswamy K and Arora BM Prevalence of Mullerius capillaries in free-ranging Spotted deer (Cervus axis) in India and its experimental cross-transmission to goats, Journal of wildlife diseases, 27(1): Rahman SM, Dey AR, Kundu UK and Begum N Investigation of gastro-intestinal parasites of herbivores at Dhaka National Zoological Garden of Bangladesh, Journal of Bangladesh Agricultural University, 12(1): Ruta M, Anna K, Peteris K, Edgars L, Dace K, Evija E, Daina IMR, Kruklite A, Keidans P, Liepins E, Keidane K, Eihvalde E and Ikauniece D Parasitic infestation of animals in deer garden in Latvia, Acta Biologica Universitatis Daugavpiliensis, 9: Singh P, Gupta MP, Singla LD, Sharma S, Sandhu BS and Sharma DR Parasitic infections in wild herbivores in the Mahendra Choudhury Zoological Park, Chhatbir, Punjab, Zoo s Print Journal, 21 (11): Singh S, Shrivastav AB and Sharma RK The epidemiology of gastro-intestinal parasitism and body condition in free-ranging herbivores, Journal of Threatened Taxa, 1(10): viii

66 References Shibashi T, Shimamura A, Izumo A and Nogami S A survey of parasites in the feces of Sika deer (Cervus Nippon) from Kanagawa, Sitama and Chiba prefectures, Japan, Japanese Journal of Zoo and Wildlife Medicine, 8: Satin-Duran M, Alunda JM, Hoverg EP and Fuente CDL Age distribution and seasonal dynamics of abomasal helminthes in wild red deer from Central Spain, Journal of Parasitology, 94 (5): Satin DM, Alnda JM, Hoberg EP and Fuente COL Abomasal parasites in wild sympatric cervids, red deer (Cervus elaphus)and fallow deer (Dama dama) from three localities across Central and Western Spain: relationship to host density and Park management, Journal of Parasitology, 90(6): Soulsby EJL Helminths, Arthropods and Protozoa of Domesticated animals. Bailliere and Yindall, London. Thawait VK and Maiti SK Prevalence of gastro-intestinal parasites in captive wild animals of Kanan Pandari Zoo, Bilaspur, Journal of Animal Research, 5(1): ThawaitVK, Maiti SK and Dixit AA Prevalence of gastro-intestinal parasites in captive wild animals of Nandan Van Zoo, Raipur, Chhattisgarh, Veterinary World, 7(7): Van Wyk JA and Mayhew E Morphological identification of parasitic nematode infective larvae of small ruminants and cattle: A practical lab guide, Onderstpoort Journal of Veterinary Research, 80 (1): Vardharajan A and Pythal C A preliminary investigation on the parasites of wild animals at the Zoological garden, Thiruvananthapuram, Kerala, Zoo s Print Journal, I-XIV (3-12): ix

67 References Vazquez L, Painceira A, Dacal V, Pato FJ, Panadero R, Lopez C, Diaz P, Arias MS, Francisco I, Diez-Banos P and Morrondo P Long-term study of internal parasitic infection in free-ranging roe deer (Capreolus capreolus) from NW Spain, Rev. Ibero-Latinoam Parasitol, 69 (2): Vengust G Comparison of thr parasitic fauna of fallow deer (Dama dama) from two enclosures in Solvenia, Solvenia Veterinary Research, 40: Varadharajan A and Kandasamy A A survey of gastro-intestinal parasites of wild animals in captivity in the V.O.C Park and Mini Zoo, Coimbatore, Zoo s Print Journal, 15(5): Van Wyk IC and Boomker J The prevalence of helminthes in some common antelopes, warthogs and a bushpig in the Limpopo province, South Africa. Parasites of South African Wildlife. XIX, Onderstepoort Journal of Veterinary Research, 78: Zasityte R and Grikienciene T Some data in endoparasites of common mole in Lithnani, Act Zoological Lituanica, 4:403. x

68 FIGURES Fig.6 A flock of Spotted Deer at Raj Bhawan Deer Park Fig.7 Collected faecal samples 39

69 FIGURES Fig. 8 Concerntration method Fig.9 Floatation method 40

70 FIGURES Fig.10 Faecal Culture by petridish method Fig.11 A charged Mc Master slide 41

71 FIGURES Fig.12 A dead Spotted Deer Fig. 13 Post-mortem of the dead Deer 42

72 FIGURES Fig.14 Collection of amphistomes from ruminal wall Fig.15 Collected amphistomes Fig.16 Flattening of amphistomes 43

73 FIGURES Fig.17 Bottle under running tap water Fig.18 Borax carmine stain Fig.19 Dehydration in Ascending order of alcohol Fig.20 Mounting of slide 44

74 FIGURES Fig.21 Ova of Paramthistomum sp. x 10 Fig.22 Ova of Strongyle sp. x 10 Fig.23 Ova of fasciola sp. x 10 Fig.24 Ova of strongyloides sp. x 10 45

75 FIGURES Fig.25 single infection of Paramphistomum sp. x 10 Fig.26 single infection of strongyle sp. x 10 Fig.27 Cotylophoron Cotylophorum recover from post-mortem of dead spotted deer 46

76 FIGURES Fig.28 Dictyocaulus larvae in the concerntrated feacal specimen by floatation technique x 10 Fig.29 Dictyocaulus larvae after staining x 10 47

77 FIGURES Fig.30 Anterior end of Dictyocaulus larva x 40 Fig.31 Posterior end of Dictyocaulus larva x 40 48