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Kerala Veterinary and Animal Sciences University, Wayanad

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2021 - 20223
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Subject: Clinical Veterinary Medicine, Ethics and Jurisprudence × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    CLINICO-THERAPEUTIC AND ULTRASTRUCTURAL STUDIES ON CANINE ATOPIC DERMATITIS WITH SPECIAL REFERENCE TO FILAGGRIN GENE POLYMORPHISM
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-11-10) AMBILY V. R.; Dr. Usha Narayana Pillai
    In the present study conducted on canine atopic dermatitis, among the total 2032 dermatological cases in dogs presented to the clinics from different parts of Kerala, 245 cases were diagnosed as various allergies over a period of three years. Out of these, 63 cases were recorded as canine atopic dermatitis based on Favrot’s criteria with the exclusion of other differentials. All the 63 cases diagnosed as CAD were utilized for epidemiological as well as clinical lesion distribution pattern studies using CADESI-04 scoring system. A total of hundred animals, 50 each of atopic and healthy controls were subjected to polymorphism studies. Sixteen atopic animals, which satisfied atleast six out of eight Favrot’s criteria with alesional pruritus at onset and pruritus score above two with clinical lesional scores above 20 were selected for detailed study to reduce variations due to extent and severity of clinical signs. In the present study, occurrence rate of allergic dermatitis recorded was 13 per cent, atopic dermatitis being second to flea allergy dermatitis. Male dogs below 3 years of age were found more affected with CAD. Distribution of lesion follows a pattern in atopic dermatitis with both paws and flexural areas were affected followed by axillae and ear pinnae, which can be better utilized for diagnostic purpose. Haemato-biochemical studies of atopic dogs revealed hypoalbuminaemia with reduced AG ratio, anaemia with leukocytosis, neutrophilia and eosinophilia. Absolute eosinophil count with positive correlation with neutrophil to lymphocyte ratio (NLR) in atopic dogs was of diagnostic significance. Serum Ig E and IL-31 were found significantly elevated in atopic dogs. The IL-31which was correlated significantly with clinical scores, indicated the role of IL-31 as key cytokine in atopic dermatitis. Invitro serum allergen testing revealed allergens such as grass mix, weeds, pollen grains of mulberry and acacia tree, house dust, house dust mites and storage mites sensitive to dogs in the region studied. Histopathological examination of atopic skin was characterized by diffuse epidermal hyperplasia with acanthosis, spongiosis and prominent rete ridges, as well as orthokeratotic hyperkeratosis as major findings. Immunohistochemical staining of filaggrin protein in lesional and non lesional skin of atopic animals exhibited faint granular staining, where as normal skin of healthy animals exhibited intense discrete staining in the stratum granulosum. Transmission electron microscopic studies revealed varying changes in the epidermis, dermis and subepidermal areas. The corneal area, epidermal cells belonging to stratum basale, stratum spinosum, stratum granulosum and stratum lucidum could be seen with retrograde changes of varying intensity. Further non keratinocyte cells that inhabit the epidermis such as melanocytes, Langerhan cells also showed changes. Dermal components were seen loosely textured and contained collagen fibrils, fibroblasts, macrophages and blood vessels. Polymorphism studies revealed 12 SNPs in the amplified fragment of exon 3 FLG, out of which 11 were novel. Genotype frequencies revealed that one novel SNP (Exon 3: c.2584G>T) and other reported SNP (c.2337T>G) were associated with CAD. The novel variation was found to have deleterious effects on protein function as analysed by five bioinformatics tools in combination and, it was also identified as a disease causing one, which affects protein stability and function. The treatment response was evaluated by the periodic reduction in clinical lesional score, heamato-biochemical, serum Ig E and IL-31 values. Cyclosporine @ 5mg/kg body weight was an effective and well tolerated drug for treating atopic dermatitis in dogs, with considerable improvement noticed in pruritus and skin lesions during the first month of treatment followed by tapering the dose by reducing the frequency of administration to every other day. Regular bathing with a non￾irritant shampoo helped in reducing the pruritus and this might be due to the restoration of skin barrier function by its moisturizing and cleansing action. Omega-3 fatty acids could lower Ig E production in allergic individuals and could lower the overall drug requirement. Based on the above studies, it was concluded that follow-up evaluation of these parameters could be a relevant approach to find out the therapeutic effectiveness. A therapeutic plan should consist of both calcineurin inhibitors like cyclosporine and clinically supportive treatments with moisturising shampoos and essential fatty acids to improve epidermal barrier function in atopic skin. Further studies are warranted to uncover new therapeutic targets like filaggrin monomers or drug candidates that upregulate FLG in skin, which can provide more treatment options rather than the current symptomatic therapy, in the increasing eve of atopic dermatitis in dogs
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    ThesisItemOpen Access
    DIAGNOSIS AND MANAGEMENT OF CARDIOVASCULAR RENAL AXIS DISORDER SUBTYPE H IN DOGS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2021-12-31) GHAG RUPALEE SUNIL; N. Madhavan Unny
    Thirty non azotaemic dogs with clinical signs suggestive of cardiovascular dysfunction formed the part of this study. Dilated cardiomyopathy (DCM) and acquired valvular disease were the two types of cardiac disorders diagnosed in these dogs. In dogs with DCM, tall R waves were observed on electrocardiogram (ECG), elevated vertebral heart score on radiography, gross enlargement of all cardiac chambers and increased left atrium to aorta diameter ratio, large E- point septal separation, lowered values of ejection fraction and fractional shortening were documented on echocardiography. In dogs with valvular disease, variability in R-R interval and low amplitude QRS complex on ECG, bronchial and vascular pattern on radiography and varying degrees of valvular regurgitation on echocardiography were recorded. Urine specific gravity varied between 1.000 – 1.025. The values of ionised calcium observed in the present study indicated mild hypocalcemia. A decline in serum inosine values on day 45 from that of day 0 was suggestive of reduced renal functional status. The urine microalbumin values documented were consistent with microalbuminuria. Microalbuminuria was found to be a reliable biomarker of glomerular damage and developing nephropathy suggestive of CvRDH. On day 45, 26 of the 30 cardiac cases were clinically stable and were responding to the treatment. On day 90 of presentation, cough of reduced intensity and frequency was reported in four dogs. Six dogs with acute decompensated heart failure, volume overload and refractory to diuretics were subjected to ultrafiltration. Ultrafiltration rate in all these dogs was determined on the basis of clinical findings and estimate of volume overload. Four out of six dogs had favourable prognosis with stabilisation in condition and resolution in volume overload. Extracorporeal ultrafiltration was found to be a useful and yielding therapy for acute decompensated heart failure dogs which were refractory to diuretics.
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    ThesisItemOpen Access
    CLINICO-BIOCHEMICAL AND MOLECULAR INVESTIGATIONS OF HAEMOLYTIC ANAEMIA IN DOGS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES THRISSUR, KERALA, 2022-01-01) ARUN GEORGE; Usha Narayana Pillai
    Haemolytic anaemia is a cause of concern in all breeds of dogs. Dogs brought to the University Veterinary Hospitals at Mannuthy and Kokkala in Thrissur district were screened for haemolytic anaemia. One hundred and one dogs with haemolytic anaemia were subjected to detailed clinical, biochemical, and molecular investigation. The primary reasons for haemolysis were blood parasites, oxidative injury, and micro-angiopathic haemolytic anaemia and these causative factors occurred as single-etiology or multi-etiology cases. Among the blood parasites, Babesia gibsoni was the major reason for haemolysis followed by Babesia canis vogeli and Ehrlichia canis. Trypanosoma evansi and Anaplasma platys appeared as co-infection in a few cases. Other minor reasons for haemolysis were immune-mediated haemolytic anaemia, stomatocytosis, and an unidentified bacterial organism. Thrombocytopenia was a consistent finding in all haemolytic cases. Eighty-one per cent of dogs with haemolytic anaemia were non-regenerative. Serum phosphorus, total cholesterol and LDL-cholesterol were not significantly different from the healthy controls at p<.05 and did not appear to influence erythrocyte membrane stability as determined by erythrocyte osmotic fragility test and erythrocyte sodium dodecyl stability test. The values of erythrocyte malondialdehyde, reduced glutathione and glutathione peroxidase were not significantly (p<.05) different from the healthy controls. The major erythrocyte morphological abnormalities identified were anisocytosis, microcytosis, hypochromasia, schistocytes, and blister cells. Genotyping of the pre-documented mutations in erythrocyte pyruvate kinase gene, phosphofructokinase gene, α spectrin and β-spectrin genes did not reveal any mutation. To conclude, the major reasons for haemolytic anaemia in dogs were blood parasites, oxidative injury, and microangiopathic haemolytic anaemia, whereas the minor reasons were IMHA, stomatocytosis and bacterial infection. Anti-rabies and multi-component vaccinations, deworming, serum phosphorus, total cholesterol and serum LDL cholesterol did not increase fragility of erythrocytes. The pre-documented gene mutations in erythrocyte pyruvate kinase enzyme, erythrocyte phosphofructokinase enzyme, α-spectrin and β-spectrin proteins were not detected in this study.