SUBCLINICAL GASTROINTESTINAL PARASITOSES IN CROSSBRED CATTLE. ladj xksoa k esa mi k;fud tbjku=h; ijthoh lapkj THESIS MASTER OF VETERINARY SCIENCE

Transcription

1 SUBCLINICAL GASTROINTESTINAL PARASITOSES IN CROSSBRED CATTLE ladj xksoa k esa mi k;fud tbjku=h; ijthoh lapkj BHARAT SINGH THESIS MASTER OF VETERINARY SCIENCE (Veterinary Parasitology) 2014 Department of Veterinary Parasitology College of Veterinary and Animal Science Rajasthan University of Veterinary and Animal Sciences, Bikaner SUBCLINICAL GASTROINTESTINAL PARASITOSES IN CROSSBRED CATTLE ladj xksoa k esa mi k;fud tbjku=h; ijthoh lapkj

2 THESIS Submitted to the Rajasthan University of Veterinary and Animal Sciences, Bikaner In partial fulfilment of the requirements for the degree of MASTER OF VETERINARY SCIENCE FACULTY OF VETERINARY & ANIMAL SCIENCE (Veterinary Parasitology) By BHARAT SINGH 2014

3 Rajasthan University of Veterinary and Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE - I Date... This is to certify that Mr. Bharat Singh has successfully completed the comprehensive examination held on as required under the regulations for Master of Veterinary Science degree. (G.S. Manohar) Head Department of Veterinary Parasitology College of Veterinary and Animal Science Bikaner

4 Rajasthan University of Veterinary and Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE - II Date... This is to certify that this thesis entitled SUBCLINICAL GASTROINTESTINAL PARASITOSES IN CROSSBRED CATTLE submitted for the degree of Master of Veterinary Science in the subject of Veterinary Parasitology embodies bonafide research work carried out by Mr. Bharat Singh under my guidance and supervision and that no part of this thesis has been submitted for any other degree. The assistance and help received during the course of investigation have been fully acknowledged. The draft of the thesis was also approved by the advisory committee on... (G.S. Manohar) (G.S. Manohar) Head (Major Advisor) Department of Veterinary Parasitology College of veterinary and Animal Science, Bikaner (B.K. Beniwal) Dean College of Veterinary and Animal Science, Bikaner Rajasthan University of Veterinary and Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner

5 Date.... CERTIFICATE-III This is to certify that the thesis entitled SUBCLINICAL GASTROINTESTINAL PARASITOSES IN CROSSBRED CATTLE submitted by Mr. Bharat Singh to Rajasthan University of Veterinary and Animal Sciences, Bikaner, in partial fulfillment of requirements for the degree of Master of Veterinary Science in the subject of Veterinary Parasitology after recommendation by the external examiner was defended by the candidate before the following members of the examination committee. The performance of the candidate in the oral examination on his thesis has been found satisfactory. We therefore, recommend that the thesis be approved. (G.S. Manohar) (Major advisor) (S.K. Ghorui) (Advisor) (Hemant Dadhich) (Advisor) (S.P. Joshi) (Advisor) (S.K. Kashyap) (Dean, PGS Nominee) (G.S. Manohar) Head Department of Veterinary Parasitology College of Veterinary and Animal Science, Bikaner APPROVED Dean POST GRADUATE STUDIES RAJUVAS, BIKANER Rajasthan University of Veterinary and Animal Sciences, Bikaner College of Veterinary and Animal Science, Bikaner CERTIFICATE IV Date

6 This is to certify that Mr. Bharat Singh of the Department of Veterinary Parasitology, College of Veterinary and Animal Science, Bikaner has made all corrections/modifications in the thesis entitled SUBCLINICAL GASTRO-INTESTINAL PARASITOSES IN CROSSBRED CATTLE which were suggested by the external examiner and the advisory committee in the oral examination held on The final copies of the thesis duly bound and corrected were submitted on..., and are forwarded herewith for approval. (G.S. Manohar) Head Department of Veterinary Parasitology College of Veterinary and Animal Science, Bikaner Enclosed: One original and two copies of bound thesis. Forwarded to the Dean, Post Graduate Studies, RAJUVAS, Bikaner through the Dean. (G.S. Manohar) Major Advisor (B. K. Beniwal) Dean College of Veterinary and Animal Science, Bikaner Approved Dean Post Graduate Studies RAJUVAS, Bikaner 1. INTRODUCTION The Indian economy is primarily based on agriculture but substantial contribution also comes from livestock wealth. The world cattle population is 1372 million out of which 175 million is in India, contributing 12.7 per cent of total world cattle population (FAO, 2008). Rajasthan has around million cattle population (Census, 2007) including crossbred/exotic cattle (0.81 million) and indigenous cattle (11.30 million). In India economy of farmers in rural set up and dairy industry are directly affected by level of production of dairy animals but high need for animal proteins in tropical countries, especially milk and milk products in recent years, oriented the producers to import high milk producing foreign breeds of cattle or crossbred cattle to meet human consumption requirements. In arid and semi arid region of Rajasthan, agriculture is secondary occupation of animal husbandry due to scanty rainfall and limited source of irrigation. Milk production provides supplementary employment and income, especially for poor farmers. So farmers of this area are mainly dependent on milch cattle for their livelihood.

7 Any condition which lowers the production of dairy animals affects the economy of farmers in rural set up and dairy industry directly and to reduce morbidity in cattle, monitoring of health of the animal is of paramount importance. Factors like diseases, genetic makeup, poor nutritional and managemental practices, environmental stress etc. are responsible for the low productivity of our livestock (Wadhwa et al., 2011). The high susceptibility of crossbred cattle to internal parasites plays an important role in dissemination of health problems in dairy farms (Mohammed et al., 2011). Parasitic infections are major constraints of livestock and cause great economic loss to dairy industry by way of reduction in food intake, weight loss, lower milk production, retarded growth, condemnation of meat, lowered fertility, low productivity, reduced work capacity, involuntary culling, mortality in heavily parasitized animals, and increased susceptibility of animals to other infections as well as high expenditure on drugs (Lebbie et al., 1994; Mohammed et al., 2011; Yadav et al., 2004). Most of the economical losses in animals are due to subclinical effects of parasites, although not immediately noticed by the owners. The most complicated part of developing an efficient strategic deworming programme for most dairy farmers is being able to understand the natural occurrence of these parasites in dairy animals. Among, parasitic infections, those caused by helminths are mainly responsible for poor performance of our livestock (Wadhwa et al., 2011). Further, helminth parasitism, especially, gastrointestinal parasitism, is one of the major health problems severely limiting the growth and health of dairy animals. In spite of significant production losses due to parasites, which may run into millions of rupees (Shah and Choudhry, 1995; Jithendaran and Bhatt, 1999), the problem is neglected due to its chronic and insidious nature (Sanyal, 1998; Yadav et al., 2004). Helminthic infections cause nearly one third of total losses due to all animal diseases (Anon, 1990) so time to time strategic treatment is necessary and for this, one has to know about important parasites prevalent in the area. Parasitic infections are of major veterinary importance in most of the developed and under-developed countries of the world. Some parasitic infections even cause death when the control measures are neglected (Hayat et al., 1987). So time to time strategic control measures are necessary for which areawise importance and prevalence of parasites has to be established. It is difficult to give an accurate estimation of economic importance of parasitic diseases; it varies, greatly in different countries, regions, climates and intensiveness of farming in the area. Ecological conditions like climate, texture of soil, population density, type and amount of vegetation, sanitation, management system, host species and age of animals play an important role in the prevalence of parasites. Therefore, study of epidemiology of gastro intestinal parasites can be one of the basic tools to take effective control and preventive measures against the helminths and protozoan parasitic infections In the livestock rich state of Rajasthan, a large number of animal markets are held at different places of Rajasthan. The Gayon ki mandi held on every Thursday at Poogal road, Bikaner is one of the growing mandi of Rajasthan in which mainly crossbred cattle are brought in good numbers for sale from Bikaner, Hanumangarh and Sriganganagar districts of Rajasthan. Although the prevalence of subclinical gastrointestinal parasitoses vary from different agro-ecological areas of Rajasthan and it is considered as a prerequisite for planning effective control measures. However, the study of prevalence of subclinical gastrointestinal parasitoses still requires proper investigation in crossbred cattle in Rajasthan.

8 Considering all these points, the present investigation was therefore, undertaken to carry out the faecal examination for recording subclinical gastrointestinal parasitoses of crossbred cattle at Gayon ki mandi, Poogal road, Bikaner with the following objectives: 1. To record the prevalence of subclinical gastrointestinal parasitoses in crossbred cattle by means of faecal examination 2. To record the intensity of gastrointestinal parasites in crossbred cattle 3. To culture and identify the various infective larvae from strongyle type eggs of these crossbred cattle Subclinical gastroiantestinal parasitoses in crossbred cattle M.V.Sc. Thesis DEPARTMENT OF VETERINARY PARASITOLOGY, COLLEGE OF VETERINARY AND ANIMAL SCIENCE, RAJASTHAN UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, BIKANER Submitted by: Bharat singh Major Advisor: Dr. G.S. Manohar ABSTRACT A total of 516 faecal samples were collected from the crossbred cattle at Gayon ki mandi, Bikaner. The faecal samples were examined by direct smear, flotation and sedimentation methods for detection of any of parasitic infection based on the presence of parasitic egg/oocyst by any or all of these methods. Egg counting for intensity of parasites and coproculture for identification of strongyle larvae were also carried out. The overall prevalence of gastrointestinal parasites in crossbred cattle at Gayon ki mandi, Bikaner was 45.34%. Strongyles type infection predominated followed by Strongyloides spp., Trichuris spp. and Toxocara vitulorum. Among protozoa, the overall prevalence of Balantidium coli was higher than Eimeria spp. Gastrointestinal parasites showed higher prevalence in crossbred cattle below 3 years followed by crossbred cattle of above 5 years and 3 to 5 years of age groups. Among helminths, strongyles infection predominated followed by Strongyloides spp. and Trichuris spp. in 3-5 years age groups studied. Toxocara vitulorum was predominated in below 3 years age group. Among protozoa, Balantidium coli infection was more prevalent than Eimeria spp. infection in all the three age groups studied. Gastrointestinal parasites showed comparatively higher prevalence in HF and Jersy crossbred compared to Rathi crossbred cattle.

9 Hanumangarh district showed the higher prevalence of gastrointestinal parasites compared to Bikaner and Sriganganagar districts. Flotation method detected more strongyle, Strongyloides spp. and Toxocara vitulorum infections than sedimentation and direct smear methods. Among protozoa, sedimentation method detected more Eimeria spp. than direct smear and floatation methods. The maximum overall mean counts of epg was recorded for strongyles and followed by epg for Trichuris spp., Strongyloides spp. and Toxocara vitulorum. The crossbred cattle of below 3 years of age showed a significantly higher intencity of gastrointestinal parasites than 3 to 5 and above 5 years of age group. The crossbred cattle of below 3 years of age showed a significantly higher intensity of Toxocara vitulorum infection than 3 to 5 and above 5 years of age group. There was no significant effect of breed and district on intensity. The proportion (per cent) of different third stage strongyle larvae in coprocultures were determined and results indicated that Haemonchus spp. was the major contributor to the strongyle worm population, followed by Oesophagostomum spp., Cooperia spp., Trichostrongylus spp. and Mecistocirrus spp Xkk;kas dh eamh, chdkusj ds ladj xksoa k esa mi k;fud tbjku=h; ijthoh lapkj

10 LukrdksÙkj 'kks/k xzafk i'kq ijthoh fokku fohkkx] i'kq fpfdrlk,ao i'kq fokku egkfo ky;] jktlfkku i'kq fpfdrlk,oa i'kq fokku fo ofo ky;] chdkusj& izlrqrdrkz & Hkjr flag eq[; mikns"vk & MkW- th-,l- euksgj vuq{ksi.k x,a xk;ksa dh eamh chdkusj ds ladj xksoa k ls dqy 516 ey ds uewus,df=r fd;s x, ftuesa ls 88 uewus 3 o"kz ls de] 173 uewus 3 ls 5 o"kz,oa 255 uewus 5 o"kz ls vf/kd vk;q oxksza ls fy;s ey dh tkap ijthoh ladze.k dk irk yxkus gsrq mles flfkr ijthfo;ksa ds vamksa@iqfv;ksa ds vk/kkj ij dze k% vo;ofgr vkysi] Iyou rfkk rynv dk teko fof/k;ksa }kjk dh xbz A v.m x.kuk ijthoh ladze.k dh rhozrk ekius ds fy, o ey lao/kzu dze k% LVªksaxkby ykokz dh igpku ds fy, fd;s x, A xk;ksa dh eamh chdkusj ds xksoa k esa tbjku=h; lw+=dfez;ksa dk loznknh izk;kshkkzo izfr kr Fkk A LVªaksaxkby dk izk;kshkkzo lokzf/kd izfr kr] rri pkr~ LVªksabxkbyksbfMl] VªkbP;wfjl rfkk VksDlksdkjk okbvwyksje dk dzzekuqlkj ik;k x;k A rhuksa vk;qoxksza ¼3 o"kz ls de] 3 ls 5 o"kz rd,oa 5 o"kz ls vf/kd vk;qoxz½ esa LVªksaxkby ladze.k lokzf/kd jgk] rri pkr~ LVªksaxkbyksbfMl] ik;k x;k A izksvkstksvk esa] rhuksa vk;qoxksz esa csysfuvfm;e dksykbz dk izk;kshkkzo lokzf/kd jgk] rri pkr~ vkbesfj;k ladze.k ik;k x;k A tbjku=h; ijthfo;kas dk izk;kshkkzo rqyukred :i ls,p-,q-,oa tlhz ladj es jkbh ladj ls vf/kd jgka guqekux< ftys es tbjku=h; ijthfo;ksa dk izk;kshkkzo rqyukred :i ls chdkusj,oa Jhxaxkuxj ftys ls vf/kd jgka x, A ey ijh{k.k dh Iyou fof/k ls T;knk la[;k esa tbjku=h; ijthfo;kssa ¼LVªksabxkby] LVªksabxkbyksbfMl] VksDlksdkjk okbvqyksje½ dk irk yxk;k x;k tcfd rynv teko fof/k }kjk vkbesfj;k vf/kd ik, dqy rhozrk LVªksaxkby ds fy, lokzf/kd jgh] rri pkr~ VªkbP;wfjl] LVªksabxkbyksbfMl rfkk VksDlksdkjk okbvqyksje dh ikbz xbz A 3 ls 5 o"kzz vksj 5 o"kzz ls vf/kd vk;q oxksza dh rqyuk esa 3 o"kz ls de vk;q oxz ds ladj xksoa k esa fpfugd`r :i ls tbjku=h; ijthfo;kas dh rhozrk vf/kd ik;h xbza 3 o"kz ls de vk;q oxz ds ladj xksoa k esa 3 ls 5 o"kz ls vf/kd vk;q oxksz dh rqyuk esa VksDlksdkjk okbvqyksje dh rhozrk fpfugd`r :i ls vf/kd ik;h xbz A

11 uly,oa ftys dk rhozrk ij dksbz izhkko ugh ik;k x;k A ey lao/kzu ls 5 izdkj ds LVªksabxkby dze k% gheksudl] blksqsxkslvksee] dwisfj;k] VªkbdksLVªksaxkbyl rfkk esfllvksfljl iztkfr izkir gq,] ftlesa gheksudl iztkfr dh izeq[krk jgh A ACKNOWLEDGEMENT I feel honoured to express my great sense of gratitude to my major advisor, Dr. G.S. Manohar, Professor and Head, Department of Veterinary Parasitology, College of Veterinary & Animal Science, Bikaner and Dean, Post graduate studies, Rajasthan University of Veterinary and Animal sciences,bikaner for his valuable and constant guidance, encouragement and diligent attitude during the course of this study. I am extremely thankful to the members of my advisory committee, Dr. S.K. Ghorui, Principal Scientist, National Research Center on Camel, Jorbeer, Bikaner, Dr. Hemant Dadhich, Professor and Head, Department of Veterinary Pathology, Dr. S.K. Kashyap, Professor and Head, Department of Veterinary Microbiology and Biotechnology, Bikaner and Dr. S.P. Joshi, Veterinary Officer on deputation, Department of Veterinary Parasitology, College of Veterinary & Animal Science, Bikaner for their constructive suggestions throughout the course of study. I have great pleasure to feel obliged to Dr. A.K. Gahlot, Hon ble Vice-Chancellor, Rajasthan University of Veterinary and Animal sciences, Bikaner and Dr. B. K. Beniwal, Dean, College of Veterinary and Animal Sciences, Bikaner, for providing all the necessary facilities throughout the course of study. The financial assistance provided by the RAJUVAS, Bikaner in the form of stipend during the period of studies is thankfully acknowledged. I am very much thankful to Ph.D. scholars of Department of Veterinary Parasitology, Dr. Sanjay Kumar, Scientist, National Research Center on Camel, Jorbeer, Bikaner, Dr. Nitin Kumar Shinde and Dr. Hakim Manzar Alam for their valuable suggestions, timely help and guidance. I feel pleasure in thanking to Dr. Praveen Pilania, Dr. Vivek Khandait, Dr. Shashank Dudhe, Dr. Mukesh Dial and Dr. Mahipal Kajla, College of Veterinary and Animal Science, Bikaner, for their suggestions and required help from time to time. I am thankful to my junior M.V.Sc. student of Department of Veterinary Parasitology, Dr. Rajesh Kumar for his support and help during research phase.

12 Sincere thanks are due to Shri R.P. Sharma, Shri S. Ali, Shri S.K. Vyas, Shri Hanuman, Shri Mukhmal and Smt. Gavra Devi, Department of Veterinary Parasitology for their sincere cooperation in execution of work with a congenial environment and coordinated support during the course of study. It is my pious duty to express fathomless sense of gratitude and indebtedness to my worthy parents, Shri Veer Singh and Smt. Rameshwari Kanwar, my elder brothers, Vijay Singh and Dashrath Singh and sister in-laws Mrs. Manju Kanwar and Kiran Kanwar for their invaluable moral support and encouragement in the accomplishment of the project It gives me immense pleasure to express my gratitude to Miss Dimple Kanwar my fiancée for her co-operation, understanding and encouragement during the study and in preparation of this manuscript. Date: Place: Bikaner (Bharat Singh) 5. DISCUSSION

13 The present investigation revealed that 45.34% of crossbred cattle at Gayon ki mandi, Bikaner suffered from subclinical gastrointestinal parasitism. The prevalence of gastrointestinal parasites in cattle has been reported earlier by many workers in different parts of world viz., 64.83% in Mandya district, Karnataka (D Souza et al., 1988), 38.86% in Kheda district, Gujarat (Hirani et al., 1999), 74.5% in Charsaddh district, Pakistan (Shah and Rehman, 2001), 50.27% in Orissa (Sahoo et al., 2003), 18.22% in Southern taluks of central dry zone, Karnataka (Muraleedharan, 2005), 59.68% in R.S. Pura, Jammu (Yadav et al., 2005), 51% in Punjab, Pakistan (Raza et al., 2007), 32.18% in Nagpur, India (Chavhan et al. 2008), 82.35% in Patiala (Kaur and Kaur, 2008), 30% in Namakal, Tamilnadu (Saravaran et al., 2009), 33.68% in Toba Tak Singh district, Punjab, Pakistan (Khan et al., 2010), 37.97%% in Punjab, Pakistan (Haque et al., 2011), 61.40% in Panjab, India (Jyoti et al., 2011) 11% in Bikaner, Rajasthan (Wadhwa et al., 2011) and 40% in Karimanagar, Andhra Pradesh (Murthy and Rao, 2012). There was slightly higher prevalence of gastrointestinal parasites found in the crossbred cattle of below 3 years of age group than above 5 years and 3-5 years of age groups which might be due to higher susceptibility of these animals than older animals to parasitic infections. The prevalence of Toxocara vitulorum was higher in cattle below 3 years of age compared to 3 to 5 years and above 5 years age groups. Ishlam et al. (2005) observed that ascariasis was significantly prevalent in calves upto 3 months of age followed by 3 to 6 months in comparison to > 6 to 12 months of age. Raza et al. (2007) observed that prevalence of helminths was higher in young animals compared with adults and also found that the prevalence of Toxocara vitulorum was higher in calves than adults. Manya et al. (2008) observed high infection of gastrointestinal nematodes in young bovine calves (4-12 months) than adults. Biu et al. (2009) found that the younger ruminants were more infected than older ruminants. Bilal et al. (2009) also observed that infections were higher among bovine calves aged between 1 to 6 months than in 7 to 12 months age group. Degefu et al. (2011) found a significantly higher prevalence of Toxocara vitulorum in young than adult animals. Haque et al. (2011) recorded higher prevalence of gastrointestinal parasites in cow calves than adults. Alim et al. (2012) found that Toxocara vitulorum was predominant in calf. In the present study the prevalence of gastrointestinal parasites was higher in HF in comparison to Jersy and Rathi crossbred cattle. Sardar et al. (2006) recorded the maximum rate of infection of gastrointestinal parasites in crossbred cattle compared to native cattle. Azam et al. (2011) found higher prevalence of gastrointestinal parasites in crossbred cattle compared to indigenous cattle. Mahmuda et al. (2012) while studying of seasonal prevalence of gastrointestinal nematods in calves in nigeria reported that the prevalence was low in indigenous breed, sokotu gudali compared to crossbreeds and exotic, Friesian breeds probably because of more resistence to helminthes in indigenous breed. Maximum prevalence of gastrointestinal parasites was recorded in the crossbred cattle of Hanumangarh district than Bikaner and Sriganganagar districts. Kaur and Kaur (2008) detected a high prevalence of gastrointestinal parasites in Patiala and its adjoining areas. Manya et al. (2008) recorded high incidence of bovine coccidiosis at Patna (Bihar) and its surrounding areas. Godara and Sharma (2010) also reported higher prevalence of gastrointestinal parasites in and around semi arid Jaipur. Wadhwa et al. (2011) reported a comparatively low prevalence of gastrointestinal helminth in Bikaner. Gastrointestinal parasitic prevalence in the cattle has been reported from India and abroad earlier by many workers (Costa, 1978; Surez et al., 1990; Aumount et al., 1991; Deore and Narsapur, 1995; Raman et al., 1995; Kashyap et al., 1997; Funmayer et al., 1999; Jithendran and Bhatt, 1999; Yadav and Sadna, 1999; Sardar et al., 2006; Sarvaran et al., 2009; Azam et al., 2011 and Alim et al., 2012). However, such study on crossbred cattle at Gayon ki mandi, Bikaner could not be detected in the literature. In the prevalence of parasites, season and climatic conditions like humidity, temperature, availability of vectors for parasites having indirect life cycle may also play an important role (Gundach et al., 2004). The free living stages of parasites survive for sufficient length of time and continuous cycle of infection is maintained between the host and pasture due to the high

14 humidity in wet areas. Summer and autumn seasons are suitable for the development of infections in animals in dry areas. Season, sex, age and agro-ecology show an association with prevalence. (Kuchai et al., 2011) The rate of infection varies from place to place in a district and season to season with in the same region. It was found that crossbreed cattle were also suffering from mixed infection of different gastrointestinal parasites. The mixed infections of gastrointestinal parasites in cattle have been reported earlier also (D Souza et al., 1988; Sahoo et al., 2003; Yadav et al. 2005; Chavhan et al., 2008; Kaur and Kaur, 2008; Saravanan et al., 2009; Singh et al., 2009; Rahman and Samad, 2010 ; Raffiullah et al., 2011 and Raza et al., 2013). In the present investigation, among helminths, subclinical infection of strongyles was maximum followed by Strongyloides spp., Trichuris spp. and Toxocara vitulorum. Almost similar observations for helminths have been recorded earlier by many workers (Kashyap et al., 1997; Waruiru et al., 2000; Chavhan et al., 2008). Kashyap et al. (1997) also observed a higher prevalence of strongyles while conducting a study in Malva region of Madhya Pradesh. A very high prevalence of strongyles infection was recorded by Godara and Sharma (2010) while conducting a study on parasitic infections of ruminants and pigs in Jaipur, In this study, next to strongyles, Strongyloides infection was observed to be very common in crossbred cattle. The Strongyloides infection has been reported to be predominant in crossbred cattle in similar studies conducted by many workers (Raman et al., 1995; Shah and Rehman, 2001; Yadav et al. 2005; Khan et al., 2010). Among the strongyles infection, Haemonchus spp. was the main contributor followed by Oesophagostomum spp., Cooperia spp., Trichostrongylus spp. and Mecistocirrus spp. Among strongyles, Haemonchus spp. has been reported to be the main contributor by other workers also (Raman et al., 1995; Yadav and sadana, 1999; Shah and Rehman, 2001; Khan et al., 2010). However, Oesophagostomum spp. (Anosike et al., 2005) and Trichostrongylus spp. (Indre et al., 2009) have also been reported to be predominant among strongyles. Raza et al. (2013) while studying the prevalence of gastrointestinal helminths in 500 faecal in cattle at the Vicinities of tehsil jatoi, punjab, Pakistan and found infections of Trichostrongylus spp., Haemonchus placei, Oesophagostomum radiatum, Bunostomum phlebotomum and Cooperia spp. In the present study average total length and tail sheath length of recovered infective third stage larvae of Haemonchus spp. were ±02.58 and 64.22±00.37 µm, of Oesophagostmum spp ±06.73 and ±02.48 µm, of Cooperia spp ±03.58 and 63.08±01.30 µm and 53.02±01.69 µm, of Trichostrongylus spp ±05.78 and 25.83±00.96 µm and of Mecistocirrus spp ±05.44 and 59.43±00.99 µm. Keith (1953), Soulsby (1965) and Levine (1968) also gave the measurement of total length and tail sheath length of the infective third stage larvae. According to Keith (1953), total length and extension of sheath tail for Haemonchus spp., Oesophagostmum spp., Bunostomum spp. and Trichostrongylus spp. were and , and , and 59-83, and and µm, repectively. In present study, overall prevalence of Balantidium coli and Eimeria spp. was reported 15.69% and 2.09%, respectively among protozoa. Shah and Rehman (2001) reported 51.0% prevalence of Eimeria spp. infection in village Bahlola, District Charsaddah (Pakistan). Kaur and Kaur reported 50% prevalence of Eimeria spp. infection in Patiala and its adjoining areas. Sarvaran et al. (2009) also recorded 4.30% incidence of Eimeria spp. infection in local village of Namakal. Rahman and Samad (2010) recorded 25.29% prevalence of Balantidium coli infection in Bangladesh.

15 It was revealed in the present study that flotation and sedimentation methods were better as compared to direct smear method as these could detect more parasitic infections than direct smear method. Flotation and sedimentation methods have been found better than direct smear method by earlier studies also for parasite detection (Rehbein et al., 2002 and Cristopher and Bernadette, 2008). It was also observed that direct method usually failed to detect the mild infection having epg below 150 (Kayum et al., 1992). These results also showed that flotation method was better to detect strongyles, Strongyloides spp., Trichuris spp. and T. vitulorum whereas the sedimentation method was found better for detection of Eimeria spp. infections. It was revealed in investigation that the overall intensity of gastrointestinal parasites in the crossbred cattle of Gayon ki mandi, Bikaner was ±39.87 epg. Cox and Lemisky (1989) recorded mean count of gastrointestinal parasites as 16 epg in Western Canada with minimum and maximum individual counts of 0.7 and 78.1 epg, respectively. Jitendran and Bhat (1999) recorded the average epg of nematodes ranging from 85 to 1720 epg in crossbred cattle in Himachal Pradesh. It was reported that the highest overall intensity was of strongyles (506.16±34.97) followed by Trichuris spp. (259.37±30.96), Strongyloides spp. (167.06±26.77) and Toxocara vitulorum (123.83±14.94). Nwosu et al. (2007) found that mean faecal egg counts were generally moderate in both sheep (1052 ± 922 strongyle, 1000 ± 590 Strongyloides and 380 ± 110 Trichuris eggs, respectively, per g of faeces) and goats (2092 ± 3475 strongyle, 958 ± 854 Strongyloides and 683 ± 512 Trichuris eggs, respectively, per g of faeces) in semiarid zone of Nigeria. Degefu et al. (2011) recorded that mean egg counts of faeces was moderate (319.40±62.30 epg) in small holder dairy farms of Ethiopia. Wadhwa et al. (2011) observed that the egg count for Strongyle was range between with an average of ± No significant difference in intensity of gastrointestinal parasites due to difference of breeds was observed in the present study. Degefu et al. (2011) also recorded that the intensity of parasites showed no variation when breeds were compared. No significant difference of district on the intensity was observed. However, such study on cattle could not be detected in the literature. Significantly higher intensity (356.89±43.34) of gastrointestinal parasites was found in the crossbred cattle up to 3 years of age than 3-5 years (297.67±23.56) and 5 to above years (213.45±34.78) age group. Waruiru et al. (2000) recorded that age of the animals had a significant influence on intensity of infection. The prevalence of gastrointestinal parasites present and their intensities depended upon hosts, natural resistance or immunity, grazing habit, climate, character of the soil, nature and amount of the vegetation, stocking rate, species and number of other domestic and wild ruminants grazing on the same land (Lodha, 1977). On the basis of present investigation it could be concluded that crossbred cattle of Gayon ki mandi, Bikaner harboured subclinical infection of gastrointestinal parasites. The prevalence and intensity of infection varied slightly from age to age and district to district and among breed. 7. LITERATURE CITED Adama, J.Y., Alemede, I.C., Ogunbanjo, S. and Isah, S Assessment of gastrointestinal helminths from faecal samples of cattle at federal university of technology farm, Minna. J. Vet. Sci., 3: 1-6.

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20 Sreedhar S., Mohan M.E. and Babu S.D Prevalence of parasitic infections in cattle and buffaloes of Anantapur district of Andhra Pradesh. Indian J. Anim. Res., 43: Suarez, V.H., Ciminari, O.E., Bedotti, D.O., Busetti, M.R. and Bello, E.M Epidemiology, effect and control of nematodes infections on Zebu crossbred, Hereford and Hereford Brahman calves of Argentina Western Pampas. J. Vet. Parasitol., 35: Swai, E.S., Mtui, P.F., Mbise, A.N., Kaaya, E., Sanka, P. and Loomu, P.M Prevalence of gastro intestinal parasite infections in Maasai cattle in Ngorongoro District, Tanzania. Livestock Research for Rural Development., 18. Thaper, D.S Systematic survey of helminth parasites of domesticated animals in India. Indian J. Vet. Sci., 26: Theodoropoulos, G., Peristeropoulou, P., Kouam, M. K., Kantzoura, V. and Theodoropoulou, H Survey of gastrointestinal parasitic infections of beef cattle in regions under Mediterranean weather in Greece. J. Parasitol. Int., 59: Wadhwa, A., Tanwar, R.K., Singla, L.D., Eda, S., Kumar, N. and Kumar, Y Prevalence of gastrointestinal helminths in Cattle and buffaloes in Bikaner, Rajasthan. J. Vet. Parasitol., 25: Waruiru, R.M., Kyvsgaard, N.C., Thamsborg, S.M., Nansen, P., Bogh, H.O., Munyua, W.K. and Gathum, J.M The Prevalence and Intensity of Helminth and Coccidial Infectios in Dairy Cattle in Central Kenya. Vet. Res. Commu., 24: Whitlock, H.V The recovery and identification of the first stage larvae of sheep nematodes. Aust. Vet. J., 35: Yadav, A., Khajuria, J.K. and Raina, A.K Gastrointestinal parasitic infection profile of bovines at R.S. Pura. J. Vet. Parasitol., 19: Yadav C.L., Kumar, R.R., Vatsya, S., Garg, R. and Baneerjee, P.S Epidemiological studies on gastrointestinal nematodosis in cattle and buffaloes. J. Vet. Parasitol., 22: Yadav, C.L. and Sadna, J.R An outbreak of gastrointestinal nematodiosis in dairy cattle in Hissar, India. J. Vet. Parasitol., 13: Yeneneh, A., Kebede, H., Fentahun, T. and Chanie, M Prevalence of cattle flukes infection at Andassa Livestock Research Centerin north-west of Ethiopia. Vet. Res. Forum., 3: MATERIALS AND METHODS The study was undertaken to know the prevalence of sub clinical gastrointestinal parasitoses and to record the intensity of gastrointestinal parasites in crossbred cattle of different age groups and of different regions brought to the Gayon ki mandi, Bikaner. The coprocultures were also set up to identify the infective strongyle larvae Animals and their management

21 The animals for the study were crossbred cattle, brought at Gayon ki mandi, Bikaner mainly from Bikaner, Hanumangarh and Sriganganagar districts of Rajasthan (Plates 1, 2 and 3). These crossbred cattle were generally reared in open and/or close herd system in groups irrespective of their age and sex. They were offered dry / green fodder and concentrate and provided tube well / well / pond / canal water ad lib Collection of faecal samples A total 516 faecal samples of crossbred cattle were collected randomly from the Gayon ki mandi, Bikaner on Thursday during September 2012 to December The faecal sample of individual animal was collected directly from the rectum and during whole of the study period the samples were collected, as far as possible, during morning hours and kept in individually marked polythene bags. The relevant information regarding age, sex, breed, feed, water, deworming history and region etc. were also recorded Faecal examination To know the prevalence and intensity of parasitic infections of crossbred cattle, the collected faecal samples were processed and examined as follows: Plate 1. Crossbred cattle at Gayon ki Mandi, Bikaner

22 Plate 2. Crossbred cattle at Gayon ki Mandi, Bikaner

23 Plate 3. Crossbred cattle at Gayon ki Mandi, Bikaner

24 Qualitative faecal examintion Qualitative examination was conducted to record the sub clinical gastrointestinal parasitoses in crossbred cattle on the basis of presence/absence of eggs/oocyts in the faeces of animals. The samples were qualitatively examined by direct smear, flotation and sedimentation methods Direct smear method A small amount of faeces was placed on a clean microglass slide and emulsified in 3-4 drops of water with the help of an applicator stick, then a micro coverslip was put over it and was examined under low power objective (10X) of the microscope Floatation method 1-5 g of faecal sample was mixed with ml of ordinary water and triturated by pestle and mortar. It was strained and taken in 15 ml centrifuge tube and centrifuged for 1-2 min. at rpm. The supernatant was removed and after emulsifying the decant centrifugation was repeated one or more times till the colour of faeces was cleared. After last washing in water, the faecal decant at the bottom of the tube was mixed with Zinc sulphate solution (32.5%), filling the tube to its brim and centrifuged similarly. The tube was placed in vertical position in the tube stand, covered with a clean cover slip for 2 minutes. The coverslip was then picked up gently and put over a slide for examination under low power objective (10X) Sedimentation method 1-5 g of faecal sample was mixed with ml of ordinary water and triturated by pestle and mortar. It was strained and taken in 15 ml centrifuge tube and centrifuged for 1-2 min. at rpm. The supernatant was removed and after emulsifying the decant centrifugation was repeated one or more times till the colour of faeces was cleared. After last washing in water, the faecal decant at the bottom of the tube was mixed, a drop placed on a clean microglass slide, a micro coverslip was put over it and was examined under low power objective (10X) of the microscope. The presence was recorded if parasitic infection was shown by any or all of these methods i.e., direct, sedimentation and floatation methods Quantitative faecal examination To obtain accurate information with regard to the severity of infection, eggs/oocysts counting (quantitative faecal examination) methods were used to determine the number of eggs/oocysts per gram of faeces. It was done by modified McMaster counting technique as described by Coles et al. (1992). Three grams of faeces were taken in a mortar and soaked in 42 ml water for few minutes and then emulsified by using pestle. The emulsion was poured through a sieve and after stirring, 15 ml of emulsion was centrifuged at 1500 rpm for 5 minutes. The supernatant was gently poured off. The tube was agitated to loosen sediment and saturated salt solution was added to make the final volume up to 15 ml. The tube was inverted five to six times and immediately sample/emulsion was withdrawn and both the chambers of the McMaster slide were charged by using pasture pipette. Eggs/oocysts were counted under low power objectives in two ruled squares containing 0.3 ml of total volume of emulsion. The number of eggs/oocysts counted in both these squares was multiplied by 50 to get eggs/oocysts per gram of faeces in the samples.

25 Coproculture and larval identification Coprocultures were set to identify the various Strongyle larvae on the basis of gut cells number and morphological peculiarities of third stage infective larvae of Strongyles. Faecal cultures provide an environment suitable for the hatching of strongyle eggs and for their development to the infective third stage larvae. Different eggs require different conditions but the general method given below was suitable for the culture of strongyle eggs in crossbred cattle faeces. For this purpose strongyle nematode egg positive faecal samples of crossbred cattle were pooled and cultured for three different samplings following the method of Roberts and Sullivan (1950). About grams of faeces were broken up finely, using a large pestle and mortar along with spatula. If the lump remained harder a small quantity of water was added to make it desirably soft and if the faeces were too soft in consistency animal charcoal was added to get the required consistency. A large lump of faeces of desirable consistency was taken in a small Petri dish and spread evenly and this dish was placed in another larger Petri dish having a small quantity of water in it. The larger Petri dish was then covered with another Petri dish to minimize evaporative losses and incubated at 25-27ºC in BOD incubator for 7 days (Plate 4). The water in larger Petri dish contained larvae which migrated from the faecal mass in small Petri dish after hatching out from the eggs. Many of the larvae reached the infective third stage by that time facilitating specific diagnosis. The infective larvae migrating to water in outer Petri dish were pipetted out and centrifuged at 1500 rpm for 3 minutes. The supernatant was discarded and the sediment was warmed over the

26 Plate 4. Photograph showing coproculture sets in BOD incubator for larval culture