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ThesisItem Open Access Immunodiagnosis of bovine gastrothylacosis using coproantigens(Department of parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2000) Kandasamy, A; KAU; Devada, KA study was conducted on the prevalence of paramphistomatidosis in Thrissur from June 1999 to May 2000, feasibility of coproantigen detection by ELlSA and comparison of sensitivity of ELlSA using coproantigens and ELlSA using serum antibodies in diagnosis of gastrothylacosis, caused by Gastrothy/ax crumenifer, in cattle. It was noted from the registers maintained at the University Veterinary Hospitals at Kokkalai and Mannuthy and that at the Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Mannuthy, that out of a total number of 1534 faecal samples from bovines examined, 253 (16.5 per cent) animals were found to be positive for amphistome eggs with the maximum prevalence (23 per cent) in June and July. Generally the infection was prevalent throughout the year. An indirect ELlSA using rabbit hyperimmune serum against somatic antigens of G. crumenifer was performed to detect coproantigens in faecal samples collected from 100 known G. crumenifer infected cattle. Seventy four samples were found to contain detectable levels of coproantigen indicating a sensitivity of 74 per cent. Serum samples collected from the same infected cattle were tested for antibodies to G. crumenifer by an indirect ELlSA using somatic antigens. Fifty one samples were found positive for antibodies indicating a sensitivity of 51 per cent. It was seen that when 43 cattle were positive for both coproantigens and serum antibodies, 18 cattle were negative for both of them. Although 31 cattle which were negative for serum antibodies were found positive for coproantigens, eight cattle negative for coproantigens were found positive for serum antibodies. The results showed that coproantigen detection, which revealed a higher sensitivity than the detection of serum antibodies by ELlSA, is feasible for the diagnosis of gastrothylacosis in bovines.ThesisItem Open Access Evaluation of different techniques for the diagnosis of bovine babesiosis(Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2003) Renjitha, T S; KAU; Deveda, KDifferent techniques to diagnose bovine babesiosis were evaluated by screening 71 cattle presented at the various Veterinary Hospitals of Thrissur and Ernakulam districts and those maintained at the University Livestock Farm, Mannuthy and the Cattle Breeding Farm, Thumburmuzhi with symptoms suggestive of the disease. The techniques employed were examination ofGiemsa stained blood smears, examination of ticks collected from suspected animals and serological tests like the Indirect Fluorescent Antibody Test (IF AT) and Slide- Enzyme Linked Immunosorbent Assay (Slide-ELISA). Among the 71 cattle, 11 were suffering from an acute illness whereas 60 showed only subclinical symptoms. Examination of Giemsa stained blood smear could detect Babesia bigemina organisms in 9 (12.68 per cent) of the total samples, which were mainly from the acutely infected cattle, except for one, that was obtained from a subclinically infected calf. The principal vector of babesiosis was identified as Boophilus annulatus in the present study. Parasitic masses indicative of the organisms were identified from the various tick tissues dissected and stained with methyl green pyronine. Considering the ticks as positive on detection of developmental stages of the parasite in any of the tissues, ticks from 27 (54 per cent) out of total 50 animals were interpreted as positive. The IFAT detected B. bigemina antibodies in 37 (52.11 per cent) and Slide-ELISA in 39 (54.93 per cent) sera out of 71 samples that were screened. These two serological techniques were found to be equally efficient, with Slide- ELISA proving more suitable for use as a field diagnostic test. Most of the animals that were detected positive for the organisms by all the tests, were above one year of age. As the number of male and female animals was too uneven, a sex related comparison could not be appropriately done. Similarly, a comparison on the performance of the different tests between the breeds was also not feasible owing to the wide variation between the numbers of breeds studied. On evaluation of the different diagnostic techniques, serology and examination. of ticks were found equally effective in detecting both the clinical and subclinical forms of the disease over and above the conventional method i.e., the blood smear examination. However, examination of Giemsa stained blood smears continues to be the method of choice to detect acute infections.ThesisItem Open Access Immunodiagnosis of canine dirofilariosis using microfilarial antigen(Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2003) Priya, M N; KAU; Subramanian, HA detailed study was conducted on the prevalence on canine dirofilariosis in Thrissur from June 2002 to May 2003, feasibility of microfilarial antibody detection by ELISA and comparison of sensitivity of Indirect ELISA and dot- ELISA in the detection of dirofilariosis. Microfilariae encountered were identified as those of Dirofilaria repens. Out of the total number of2435 blood samples from dogs screened by wet film examination 172 (7.06 per cent) dogs were found positive for microfilariae. It has been found that canine dirofilariosis was prevalent throughout the year with more incidence during summer. An Indirect ELISA and dot-ELISA were also conducted in three groups of animals using microfilarial antigen prepared from known microfilaria positive serum samples. Out of the total number of 50 samples of known microfilaria positive dogs 49 (98 per cent) and 47 (94 per cent) were found positive by Indirect ELISA and dot-ELISA respectively. The corresponding figures in 36 dogs with other helminths were 15 (41.67 per cent) and 19 (52.77 per cent) and that of32 totally helminth free dogs were 10 (31.25 per cent) and 12 (37.5 per cent) respectively. The results indicate that both Indirect ELISA and dot-ELISA are feasible for the detection of the microfilarial antibodies though they are less specific.ThesisItem Open Access Immunoprophylaxis against common dog tick using gut antigen(Department of Veterinary Parasitology,College of Veterinary and Animal Science,Mannuthy, 2005) Ajithkumar, K G; KAU; Subramanian, HThe occurrence of tick infestation in dogs in the Corporation of Thrissur, Kerala, India, was studied in 1200 dogs during a period of one year from July 2004 to June 2005. Four hard tick species viz Rhipicephalus sanguineus, R. haemaphysaloides, Haemaphysalis bispinosa, H. bispinosa var. intermedia were identified. Significant difference in species wise prevalence was found on 124 tick positive dogs. The most prevalent species of tick infesting dogs was found to be R. sanguineus (8.58 per cent) followed by H. bispinosa (1.33 per cent), R. haemaphysaloides (0.33 per cent) and H. bispinosa var. intermedia (0.08 per cent). Three species of ticks namely R. haemaphysaloides, H. bispinosa, and H. bispinosa var. intermedia have been recorded in dogs for the first time from Kerala. The influence of month, season, age, gender and breed on the prevalence rate were observed. Prevalence rates were calculated by month, season, age, gender and breed. No significant difference (P>0.05) of tick infestation during different months was observed due to temperature, relative humidity, and rainfall. No clear pattern of seasonality was observed for R. sanguineus, which was present throughout the year. The different variables considered in the present study such as age and gender were not significantly associated with the presence of tick infestation in dogs. There was a highly significant (P<0.01) association between breed and the intensity of infestation with maximum intensity of infestation in German shepherd dogs. Sites of attachment of ticks were ear, neck, interdigital space, dorsum of the body, eyelids, perianal region, withers, thorax and hind limbs. Highly significant variation (P<0.01) was observed between attachment sites with maximum on ear (84.68 per cent) followed by neck and interdigital space. Tick-bite naive guinea pigs inoculated with gut extracts and Freund’s adjuvant revealed induced immunity against R. sanguineus expressed by altered feeding and fertility parameters consisting of the prolonged engorgement period, reduced engorged female weight and feeding efficiency index, prolonged pre oviposition period, reduced oviposition period, egg mass weight, egg rate conversion efficiency and larval mass. Except the incubation period all the reproductive and feeding parameters differed significantly compared to the control. Inradermal test done on immunised guinea pigs to assess cell mediated immunity revealed both immediate and delayed hypersensitivity reaction. Humoral immune response assessed by agar gel immunodiffusion (AGID) and passive haemagglutination (PHA) revealed that the former was less sensitive compared to the latter. Passive haemagglutination test detected anti tick antibodies as early as 14th day post immunisation. Peak titre 1:128 reached on the 35th day post immunisation.ThesisItem Open Access Anthelmintic resistance in gastrointestinal nematodes of goats(Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2005) Deepa, C K; KAU; Devada, KInvestigation on the prevalence of gastrointestinal nematodes of goats in Thrissur Corporation area was carried out by detecting faecal eggs and larvae after coprological examination and coproculture respectively. The faecal samples for coprological examination were collected from animals brought to the University Veterinary hospitals, Kokkalai and Mannuthy, University Goat and Sheep farm and from nearby houses around Mannuthy. Screening of faecal samples of 320 goats during the period of study from June 2004 to May 2005 revealed 114 as positive (35.63 per cent). The type of positive infection noted were Strongyle (95.61 per cent) and Strongyloides (4.38 per cent) infection. Monthwise prevalence of gastrointestinal nematodes showed maximum infection in May (48 per cent) while a lower prevalence in August (28 per cent). Seasonwise, the prevalence of gastrointestinal nematodes was found to be 30.7 per cent, 33.33 per cent and 39.13 per cent during cold wet South West monsoon (heavy rainfall), warm wet North East monsoon (low rainfall) and dry season respectively. The prevalence of gastrointestinal nematodes was observed to be higher in young goats below one year (42.10 per cent) and in females (85.96 per cent ) than males (14.03 per cent). There was no significant variation between breeds and management . The species of nematodes encountered were Haemonchus contortus (55.26 per cent), Oesophagostomum columbianum (13.15per cent), Trichostrongylus colubriformis (21.05 per cent), Bunostomum trigonocephalum (6.14 per cent) and Strongyloides papillosus (4.38 per cent). The comparative biometry of various infective larvae of the commonly found nematodes were noted. Resistance to various anthelmintics by the nematodes in goats of the University Goat and Sheep farm, Mannuthy were detected by the methods namely Faecal Egg Count Reduction Test (FECRT) and Egg Hatch Test (EHT). Forty kids aged between three to six months were used for this purpose. Faecal Egg Count Reduction Test revealed resistance to albendazole, ivermectin and morantel citrate by the gastrointestinal nematodes. The drugs namely albendazole, ivermectin and morantel citrate showed a per cent worm reductions of 30, 53, 45 with 52, 34 and 62 as lower 95 per cent confidence limits. Specific resistance to benzimidazole group by EHT showed the ED50 value of albendazole (µg per ml) in EHT to be 0.211556 which further established resistance to albendazole by the gastrointestinal nematodes of goats. Anthelmintic resistance is best controlled in the ground level by the use of correct type of anthelmintics at the correct dose against nematodes in goats. Breeding for disease resistance, development and use of vaccines and biological control using nematophagous fungi are the prospective methods for the control of anthelmintic resistance.ThesisItem Open Access Prevalence, immune response and control of Stomoxys spp. of flies(Department of Veterinary Parasitology, College of Veterinary and Animal Science, Mannuthy, 2000) Basker, N; KAU; Subramanian, HA detailed study on the prevalence, biology, immune response and control of Stomoxys spp. of flies was conducted. The prevalence study on Stomoxys fly infestation on cattle of Thrissur, during June 1999 to May 2000 showed a higher incidence (>90 per cent) during the months of June, July, August and September than in other months. The prevalence increased as temperature decreased and humidity and rainfall increased. The fly infesting cattle was identified. as Stomoxys calcitrans. The biology of the fly was studied in laboratory and were fed with citrated bovine blood. The use of horse dung as oviposition and larval development medium was found to be satisfactory. The life cycle parameters of the fly were also noted. The thoracic muscle antigen prepared from pharate adult fly was used for immunization. Agar gel precipitation test and counter immuno electrophoresis revealed a single clear precipitation line. The passive haemagglutination assay revealed an antibody titre of 1024, on days 21 st and 28th of immunization. Feeding the normal flies with blood from immunized rabbit produced a cumulative mortality of 100 per cent on day 20 against 66.7 per cent mortality in flies fed with control rabbit blood. Abnormal effects like wing paralysis was noted in test group. The insecticidal property of Deltamethrin, Diazinon, Carbaryl and Ethofenprox were evaluated against stable fly infestation and was found that Deltamethrin (0.0025 per cent) was highly effective in controlling flies. At 0.005 per cent concentration Deltamethrin gave better protection. Diazinon (0.3 per cent) and Carbaryl (0.1 per cent) failed to provide good protection after one day post treatment. Ethofenprox (0.002 per cent) was found to be least effective.ThesisItem Open Access Prevalence, pathology and treatment of coccidiosis in rabbits(Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2002) Shameem, H; KAU; Devada, KA detailed study on the prevalence, pathology and treatment of coccidial infections in rabbits belonging to all age groups, breeds and sex was undertaken at the Rabbit Research Station, Mannuthy and at six local rabbi tries in and around Thrissur during the period from October 2001 to September 2002. The overall prevalence was observed as 18.54 per cent. The prevalence of , coccidiosis was found to be high in young rabbits below three months and in female rabbits ofNew Zealand Whiteduring the months of heavy rainfall. The infection was diagnosed by the microscopical examination of faecal samples and clinical signs which included diarrhoea, soiled fur, anorexia and loss of condition. Subclinically affected rabbits showed no apparent signs except growth retardation and reduced feed intake. Six eimerian species causing coccidial infections in rabbits were I identified. Mixed infections were common and the commonly encountered species were E. magna, E. perforans and E. media. They were found as Eimeria media (80 per cent), E. magna (86 per cent), E. perforans (86 per cent), E. coecicola (50 per cent), E. jlavescens (15 per cent) and E. piriformis (8 per cent). Gross lesions found in the dead rabbits brought for postmortem at the Centre of Excellence in Pathology, College of Veterinary and Animal Sciences, Mannuthy comprised of oedematous, thickened intestine with circumscribed greyish white foci on the mucosa. Histopathological examination of the intestine revealed characteristic lesions of intestinal coccidiosis with schizonts and gametogonic stages in the / epithelium of the intestine. Active inflammatory reaction with mononuclear infiltration was noted. The tips of villi showed necrotic changes and desquamation of epithelial cells resulted in villous atrophy. Haematological studies of infected animals revealed reduction in the values of PCV, Hb and erythrocyte count and a slight increase in leucocyte count when compared to the apparently healthy animals. Therapeutic trials against rabbit coccidiosis usmg drugs like sulphadimidine sodium (200 mg per kg body weight for three days), furazolidone (10 mg per kg body weight for five days) and metronidazole (40 mg per kg body weight for three days) orally, showed sulphadimidine sodium as the drug of choice. The other two drugs did not give satisfactory results. The mean clearance values of the three drugs were 99.68 per cent, 85.48 per cent and 69.92 per cent respectively.ThesisItem Open Access Development of a precocious strain of Eimeria tenella(Department of Veterinary Parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2001) Sangeetha, R; KAU; Devada, KStudies on the prevalence of caecal coccidiosis in chicken, by referring to the postmortem registers maintained at the centre of Excellence in Pathology, College of Veterinary and Animal Sciences~ Mannuthy revealed that 48.4 per cent of 550 coccidia positive viscera was due to Eimeria tenella. A pure line of E. tenella was established in the laboratory by single oocyst isolation technique from a virulent field case of caecal coccidiosis in chicken. A stabilised strain of E. tenella was developed by administering a dose of 1.5x103 sporulated pure line E. tenella oocysts per bird orally to six three to five day old coccidia free White Leghorn chicks and harvesting the caecal contents at 168 hr pi for 15 cyclic~l passages. For the development of a precocious strain of E. tenella a dose of 1.5x103 sporulated E. tenella oocysts per bird was administered orally to six, three to five day old White Leghorn chicks. The caecal contents were harvested at 136 hr pi and the oocysts were allowed to sporulate and subsequently repassaged by applying selection pressure in increments of one hour reduction at each passage. At the end of 18 cyclical passages, oocysts obtained at 125 hr pi were designated E. tenella. as the precocious strain of. A comparative study between the precocious and stabilised strain of E. tenella revealed the following observations~ The prepatent period of the precocious strain was 125 hr while that of the stabilised strain was 135 hr. The oocyst morphology did not reveal any significant difference between the two strains. The precocious strain was less pathogenic and produced milder clinical signs than the stabilised strain. The gross lesions of caeca were less morbid in the precocious strain infected group. Weight gain was more in the precocious strain infected group than in the stabilised strain infected group. The precocious strain produced less mortality and showed a drop in oocyst production. Endogenous study revealed an accelerated growth in the early stages of the asexual life cycle in the precocious strain infected group.ThesisItem Open Access Prevalence and treatment of bovine coccidiosis(Department of Parasitology, College of Veterinary and Animal Sciences, Mannuthy, 2000) Sreevidya Nambiar, K; KAU; Devada, KA detailed study on the prevalence, clinical pathology and treatment of coccidial infections in cattle belonging to all age groups was undertaken at the Veterinary Hospitals, University Livestock Farm, Buffalo Breeding Station, Cattle Farm, Pattambi, Kerala Agricultural University and other localities in Thrissur during the period from June 1999 to May 2000. The prevalence of the infection was found to be 2.2 per cent. On the whole, the incidence of coccidiosis was found to be high in animals below one year, and female animals, during the rainy and humid months. Clinical signs; predominantly blood tinged or foul smelling diarrhoea and the microscopical examination of the faecal samples from suspected cases were made use of for diagnosis. Eight Eimerian species causing coccidial infections in cattle were identified and they were Eimeria bovis (35 per cent), E. zuernii (55 per cent), E. subspherica (10 per cent), E. ellipsoidalis(60 per cent), E. cylindri ca (fi ve per cent), E. bareillyi (five per cent), E. brasiliensis (five per cent) and E. wyomingensis (10 per cent). The most commonly encountered species were E. ellipsoidalis, E. bovi s, and E. zuernii. Haematological studies of clinically infected animalsrevealed reductio in the values of haemoglobin, packed cell volume, total leukocyte and lymphocyte counts. A reduction in serum total protein and globulin fractions was also observed during the assay of serum protein fractions in these animals. Treatment trials against bovine coccidiosis using drugs, Sulphadimidine sodium (125 mg per kg body weight for three days), Amprolium hydrochloride (20 mg per kg body weight for five days) and Salinomycin (2 mg per kg body weight for 21 days) orally resulted in nearly cent per cent efficacy in the clinically affected animals. All the three drugs were found to be equally effective.
