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ThesisItem Open Access IMMUNE RESPONSE TO COMBINED FOOT AND MOUTH DISEASE, HAEMORRHAGIC SEPTICAEMIA AND BLACK QUARTER VACCINE AND THEIR RESPECTIVE MONOVALENT VACCINES IN CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2001) RAJKUMAR, K.Immune response to combined Foot and Mouth Disease, Haemorrhagic Septicaemia and Black Quarter vaccine and to their respective individual vaccine were studied. Group I animals were vaccinated with combined FMD, HS and BQ oil adjuvant vaccine. Group II animals were vaccinated with FMD polyvalent oil adjuvant vaccine and combined HS, BQ gel vaccine. Group III animals were vaccinated with polyvalent FMD oil adjuvant vaccine, HS gel vaccine and BQ gel vaccine. Vaccinations were done in unvaccinated calves above four month of age. Booster vaccination were carried out after nine month after primary vaccination for oil adjuvant vaccine and six month after primary vaccination for gel vaccine in all group animals. Monthly assessment of antibody titres against O, A, C and Asia-1 antigens were made by liquid phase blocking ELISA and Pasteurella multocida and Clostridium chauvoei by Indirect ELISA. All the three groups maintained protective antibody titre for FMD virus type 'O', 'A', 'C^ 'Asia-1', Pasteurella multocida and Clostridium chauvoei antibody litres throughout the study period. All the three groups did not show any significant variation in antibody litres against FMDV type, 'O', 'A', 'C and 'Asia-l'and Pasteurella multocida and Clostridium chauvoei though they are well above protective litres. From this observation it is concluded that 1. All the three schedule of vaccination provides sufficient protective litre for FMDV type, 'O', 'A', 'C and 'Asia l'and Pasteurella multocida and Clostridium chauvoei though they are well above protective litres. 2. Combined vaccine performance is equally good as that of individual vaccine. 3. Combined vaccine reduces labour cost for injection, number of visit and stress to the animals to a very great extent. There fore combined vaccine may be preferred when compared to individual vaccine.ThesisItem Open Access EPIDEMIOLOGY OF CERTAIN BACTERIAL AND VIRAL DISEASES CAUSING NEONATAL MORTALITYIN PUPS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2004) INDU., S.NAMPUTHIRIThe present study was conducted to assess the role of bacterial and certain viral etiology, to conduct antibiogram and to estimate the nonspecific factors associated with neonatal mortality in pups.The bacterial isolates from milk of bitches were identified as Staphylococcus aureus (29 per cent) Staphylococcus intermedius (six per cent), Klebsiella pneumoniae (47 per cent), Proteus vulgaris (12 per cent) and Pseudomonas aeruginosa (six per cent). Antibiogram of the isolates showed that majority of the isolates from milk were sensitive to ciprofloxacin (71 per cent), followed by amoxycillin (65per cent), gentamicin (41per cent), enrofloxacin (59 per cent), norfloxacin (18 per cent) chloramphenicol (12 per cent) and amoxy-clav (6 per cent).Blood samples taken from two bitches yielded bacterial growth, which were identified as Staphylococcus aureus (50 per cent) and Klebsiella pneumoniae (50 per cent). The bacterial isolates from blood were lOOper cent sensitive to ciprofloxacin and chloramphenicol, 50 per cent to gentamicin, 50 per cent to amoxycillin and resistant to enrofloxacin and norfloxacin. The bacteria isolated from the vaginal samples were identified as Hafnia alvei (Enterobacter a/ve/)(six per cent), Klebsiella pneumoniae (35 per cent), Staphylococcus aureus (12 per cent), Escherichia coli (12 per cent), Proteus vulgaris (23 per cent), Pseudomonas aeruginosa (six per cent) and Staphylococcus intermedius (six per cent). The vaginal isolates were sensitive to ciprofloxacin (88 per cent), gentamicin (53 per cent), amoxycillin (59 per cent), enrofloxacin (71 per cent), chloramphenicol (59 per cent) and norfloxacin (35 per cent). None of the milk, blood or vaginal swabs taken from the control group yielded any bacterial growth. Isolates were obtained from heart blood and from liver and lung of pups. These included Proteus vulgaris{20 per cent),Pseudomonas aeruginosa{40 per cent) and Klebsiella pneumoniae{40 per cent). Fourty per cent of the bacterial isolates from pups were sensitive to gentamicin and chloramphenicol, 80 per cent to ciprofloxacin, 60 per cent to amoxycillin and 20 per cent to norfloxacin and enrofloxacin. The role of parvovirus as a causative agent for neonatal mortality was studied by subjecting the liver, lung,intestine spleen and kidney tissue of dead pups to AGID and HA for detection of parvoviral antigen and one sample was positive by HA. This was confirmed by haemagglutination inhibition test. None of the serum from the bitches gave positive result for antibodies against parvoviral antigen in agar gel immunodiffusion test. The titre of antibodies of bitches were assessed by HI. HI titre of the vaccinated bitches ranged from eight to 64.Only one bitch was having the protective titre. All the unvaccinated bitches were not having any titre of antibodies against parvo by HI testThesisItem Open Access EPIDEMIOLOGY AND CLINICO-THERAPEUTIC STUDIES ON DERMATOMYCOSIS IN DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2004) DEVI., T.; vijayakumar, KThe present study was mainly focused on epidemiological, diagnostic and clinico-therapeutic aspects of dermatomycosis in dogs. A total of 94 animals showed positive results for the presence of arthrospores on direct microscopic examination, out of the total 11016 animals brought to University Veterinary Hospitals Kokkalai and Mannuthy from April 2003 to March 2004, were included in the infected group. No age, sex, breed predilection and seasonal variation for dermatophyte infection were observed. Clinical signs of the infected group are non-specific and quite variable, hence the diagnosis mainly relied on the examination of skin scrapings by direct microscopy and cultural method. Direct microscopic examination is considered to be an ideal method of diagnosis for detecting dermatophytes, since cultural method requires minimum of three weeks time and delays the treatment, even though, it is conclusive. Isolates of Microsporum spp and Trichophyton spp were identified by cultural method using Sabouraud's dextrose agar with chloramphenicol and cycloheximide supplement. Haematological abnormalities recorded were low haemoglobin content and leucopaenia and serum biochemistry revealed increased levels of total protein, albumin, globulin and A/G ratio in infected group compared to that of control group. No significant variations were observed on serum minerals such as copper, iron and zinc contents in infected group compared to control group. But mean values of copper and iron in infected and control groups were lower than the respective normal range in canine, where as zinc levels in both the groups were within the normal range. Three different therapeutic trials were conducted in three different experimental groups. Group 1 animals treated with ketoconazole at the dose rate of 5 mg per kg bodyweight daily orally for two weeks, showed a dramatic clinical response. Clinical examination of dogs at the end of treatment, revealed complete remission of lesions and signs of regrowth of hairs in almost all affected areas in all the animals. Terbinafme at the dose rate of 4 mg per kg body weight once daily orally given in-group 2 animals was proved to be very effective in treatment of dermatophyte infected dogs. Combination of intramuscular injection of "Biocan M" vaccine adsorbed with aluminium hydroxide and topical application of ketoconazole shampoo in-group 3 animals was not found to be effective. Hence the results of present study confirms the importance of the diagnosis of dermatophyte infections by direct microscopy, applicability of the cultural method of isolation of dermatophytes and proved the efficacy of ketoconazole and terbinafme as the good choice of drugs for the effective treatment of dermatomycosis.ThesisItem Open Access SEROSURVEILLANCE OF FOOT AND MOUTH DISEASE IN CATTLE BASED ON STRUCTURAL AND NON-STRUCTURAL PROTEIN ELISA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2015) MINI, K.V.; Tresamol, P.V.Post-vaccination sero-monitoring is a vital component of any intensive vaccination strategy. The main aim of the study was to assess the sero-conversion following FMD vaccination in pre-sensitized adult cattle using Sd-LPB ELISA. In addition, immune response in primi-vaccinated calves against oil adjuvanted trivalent FMD vaccine was also assessed. Differentiation of immune response due to vaccination and infection was studied using 3ABC NSP ELISA. Dynamics of maternally derived antibody in new born calves was also monitored. Adult animals responded well to FMD booster vaccination and maintained protective immunity up to six months. Sero-positivity to NSP 3ABC in apparently healthy animals corroborated persistence of subclinical infection within vaccinated herd. Maternally derived antibody found to persist in the system of colostred calves for up to the age of five months and its interference toward induction of primary immune response to vaccination was further documented. The efficacy of a booster dosage in calf-hood vaccination was evaluated and found to drastically improve the immune response. Carrier status could be detected among FMD convalescent cattle even six months post-outbreak, with higher molecular prevalence of serotype O in their oropharyngeal secretion. Presence of previous exposure to infection, subclinical infection or carrier state did not significantly influence the sero-conversion to subsequent vaccination.ThesisItem Open Access MOLECULAR CHARACTERIZATION AND SEROMONITORING OF CANINE PARVOVIRUS FOLLOWING MODIFIED LIVE CPV-2b VACCINATION IN PUPS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2017) LOBO ANDREA, ROBIN; Vinodkumar, K.This study focused on the influence of maternally derived antibodies on the development of serum antibody titres among pups subsequent to vaccination with a commercially available multicomponent vaccine containing high-titre low-passage CPV strain. The pups were divided into two groups based upon the estimation of CPV antibody titres on day 0 by HI and ELISA. Estimation of the titres by HI test revealed that by day 15, 16 out of 23 pups from group One and 24 out of 27 pups from group Two had protective titres. All 50 pups attained protective titres by day 30 but only 33 (66 per cent) retained it by day 180. Results of the ELISA showed 19 out of 20 puppies in group One and 19 out of 30 in group Two were having protective titres on day 15. All but one of the 50 pups became protected by day 30, but only 37 including the pup that was non-protected on day 30 remained with protective serum titres. These results indicate that vaccination with a high-tire low-passage CPV strain can induce protective immunity by day 30 post vaccination irrespective of the MDA status of the vaccinates. Extracts of faecal DNA followed by PCR and sequencing coupled with comparison against existing sequences revealed presence of New CPV-2a in 19 samples while the other two were CPV-2a indicating that New CPV-2a was the prevailing strain in Thrissur districtThesisItem Open Access ACUTE PHASE PROTEINS IN SERUM AND MILK AS DIAGNOSTIC TOOLS IN BOVINE SUBCLINICAL MASTITIS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2017) AARSHA, RAJ; .Vinodkumar, .KThis study aimed to evaluate the utility of acute phase proteins (APP) levels in the serum and milk as diagnostic markers for subclinical mastitis in dairy cattle. The concentrations of various acute phase proteins (APP) like Haptoglobin (Hp), C- reactive protein (CRP), α- 1 acid glycoprotein (AGP) and albumin in milk as well as Hp,serum amyloid A (SAA), CRP and albumin in serum of 40 normal cows as well as 40 subclinically infected cows were compared in the study. The milking animals were screened by California mastitis test (CMT), electrical conductivity, somatic cell count (SCC) and bacterial culture and isolation. Comparison was also made between the variations of APP levels based on the CMT, EC and SCC scores as well as the correlation with lactoferrin (Lf) levels in milk. The pathogen specific variations in the APP values were also investigated. The values of Hp, SAA and AGP showed significant difference between the groups of normal and subclinically affected cows. On comparing the means based on CMT scores, Hp and AGP values in milk were significantly different between groups. Hp, CRP and AGP in milk showed significant difference between groups when compared based on EC values. The cows with different SCC ranges showed significant changes in case of Hp and AGP in milk. Hp in milk and SAA showed positive correlation with Lf. There was no significant difference noticed in the APP values of cows with single and mixed infections. The study showed that the use of APP like Hp and AGP in milk very effective for the early diagnosis of subclinical mastitis in bovines. The APP in serum can be altered by any other factors making it non specific whereas in milk the APP values will increase only in case of mammary gland inflammation and hence is of high diagnostic significance.ThesisItem Open Access MOLECULAR DETECTION AND THERAPEUTIC MANAGEMENT OF CANINE BABESIOSIS(College of Veterinary and animal Science,Mannuthy, 2017) Arthi, A; Tresamol, P VCanine piroplasmosis caused by Babesia canis and B. gibsoni is increasingly being detected among dogs in Kerala. The present work was carried out to study the epidemiology, protozoal etiology, haemato-biochemical alterations and ultrasonographic changes in liver and spleen, in canine babesiosis, to diagnose babesiosis using blood smear examination and PCR and to compare the efficacy of various treatment protocols. Diagnosis of babesiosis was performed by direct microscopic observation of intra-erythrocytic piroplasms in stained blood smears and by PCR. A total of 200 blood samples from dogs suspected for canine babesiosis were collected from different breeds, gender and age group of animals. Blood smear examination revealed B. canis piroplasms in 38 dogs and B. gibsoni piroplasms in 112 dogs. Polymerase chain reaction using genus specific PCR yielded amplicons specific for Babesia spp. in 156 samples. Species specific PCR for B. canis yielded positive results in 38 samples and that of B. gibsoni in 118 samples. The sub-species specific primers confirmed the B. canis organism as B. canis vogeli, in all 38 positive samples. Overall prevalence of canine babesiosis was estimated to be 78 per cent and 75 per cent by conventional staining technique and PCR respectively. Higher incidence of B. canis infection was seen in the age group of six months to one year in breed Rottweilers and in female dogs. Higher incidence of B. gibsoni infection was seen in the age group of one to five years in breed Labrador retrievers and in female dogs. The most frequent clinical signs recorded in B. canis infection include pyrexia, anorexia, vomiting, pallor or congestion of conjunctival mucous membrane and lymphadenopathy whereas, in B. gibsoni infection signs include pyrexia, anorexia, lethargy, pallor of mucous membrane, emaciation, jaundice, voiding of dark yellow or coffee coloured urine. Atypical signs like syncope, abortion, ascites, vasculitis, limb oedema and scrotal oedema were also observed. Tick infestation was observed in both infections. Ultrasonographic evaluation in 20 B. canis affected dogs and 33ThesisItem Open Access MOLECULAR DETECTION AND OCCURRENCE OF EHRLICHIOSIS AND ANAPLASMOSIS IN DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2017) SUKANYA SUKUMARAN, V.; Tresamol, P.V.Ehrlichiosis and anaplasmosis comprises a major part among the vector borne infectious diseases affecting the canine population in Kerala. The present study aims to evolve a better diagnostic tool for the diseases at the molecular level along with assessment of various clinicopathological and epidemiological factors. Hundred dogs presented to the Teaching Veterinary Clinical Complex, Mannuthy and University Veterinary Hospital, Kokkalai having fever, lymphadenopathy and thrombopenia were screened for the presence of E. canis and A. phagocytophilum. Iniatially screening was done by peripheral blood/buffy coat smear examination. Out of hundred animals screened monocytic inclusions were noticed in sixty per cent of cases and neutrophilic inclusions in three per cent. Buffy coat smears were more sensitive to detect the organism than blood smear examination. Polymerase chain reaction was performed using DNA isolated from hundred dogs with clinical signs suggestive of ehrlichiosis and anaplasmosis. Conventional PCR was employed for E. canis and A. phagocytophilum using species specific primer targeting 16S rRNA, which yielded positive results in four and eleven samples respectively. Nested PCR was employed for amplification of Ehrlichial DNA which yielded fifty five positive samples using genus specific primers and twenty positive samples using species specific primer. Nested PCR amplification for Ehrichia ewingi gave positive results in four samples. Labrador Retrievers were the breed found to be predominantly affected with ehrlichiosis and anaplasmosis. More incidence of ehrilichiosis was noticed in males and anaplasmosis in females. Most prominenet clinical signs noticed were pyrexia, lymphadenopathy, pallor or congestion of mucus membrane, opthalmic lesions, epileptic seizures and bleeding episodes in ehrlichiosis and in anaplasmosis, pyrexia, lymphadenopathy, change in colour of mucus membrane, splenomegaly and lameness. Haematological abnormalities noticed were microcytic hypochromic anaemia, thrombocytopenia and monocytosis in ehrlichiosis and anaemia and granulocytosis in anaplasmosis. Serum biochemical profile revealed hyperprotinaemia with hypergammaglobulinaemia in ehrlichiosis and elevated serum alkaline phosphatase level and creatinine in anaplasmosis. The result of present study revealed a higher prevalence of ehrlichiosis and anaplasmosis among dogs in Kerala and it also proved PCR as a sensitive diagnostic tool for detecting these diseases.ThesisItem Open Access OCCURRENCE AND CLINICO-THERAPEUTIC STUDIES ON FELINE INFECTIOUS ANAEMIA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2017) AMEL DEV, P.; Tresamol, P.V.The present study was designed to investigate the clinical, haemato-biochemical profile and risk factors associated with feline mycoplasma infection and its molecular diagnosis. Hundred anaemic cats presented at UVH, Kokkalai and Mannuthy during January to December 2016, were formed the subjects for study. Peripheral blood smears, were collected from each animal and subjected to Romanowsky staining for initial screening and further confirmed by acridine orange staining. The presence of small coccoid organism in the periphery of red blood cells were identified in blood smears of eleven out of hundred anaemic cats, suggestive of haemotropic Mycoplasmas. Most frequent clinical signs noticed in affected cats were pale mucous membranes, flea infestation, reduced appetite, pyrexia, splenomegaly and lymphadenopathy. Haematological studies revealed leucocytosis with granulocytosis, reduction in RBC count, haemoglobin and platelet count. Biochemical alterations such as Hyperglobulinaemia, elevated creatinine and ALT were noticed in cats with haemotropic mycoplasmosis. More number of cases were observed in summer season and among adult male non - pedigree cats with outdoor access. Whole blood samples were subjected to DNA isolation followed by polymerase chain reaction targeting 16s rRNA gene. Twenty four out of hundred animals were found to be positive for genus specific PCR assay and twenty three animals were found to be positive for Candidatus Mycoplasma haemominutum and only one cat was positive for Mycoplasma haemofelis. Additionally a multiplex PCR assay was also standardised for the simultaneous detection of both mycoplasma species. The infected animals were treated successfully with parenteral administration of oxytetracycline followed by oral doxycycline or doxycycline alone depending upon condition of animals. Response to the treatment was assessed by resolution of clinical signs and absence of organism in the blood smear.