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Kerala Veterinary and Animal Sciences University, Wayanad
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ThesisItem Open Access MINIMAL INVASIVE PERCUTANEOUS CYSTOSTOMY USING PAEDIATRIC SUPRAPUBIC CATHETERS FOR MANAGEMENT OF URETHRAL OBSTRUCTIONS IN TOM CATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) TAMARAISELVI P; Dr. (Maj.) Sudheesh S. NairThe study was conducted with the objective of standardising the technique of minimal invasive percutaneous cystostomy using paediatric suprapubic catheters for management of urethral obstructions in tom cats. Twelve clinical cases of urethral obstruction in tom cats presented to University Veterinary Hospitals of Kerala Veterinary and Animal Sciences University at Kokkalai and Mannuthy during a period of twelve months from October 2021 to September 2022 were selected for the study. The prevalence of urethral obstruction was found to be more in uncastrated Persian breed of cats with age of one to two years. All cats had history of being fed with dry commercial diet, lowered water intake and living in indoor. The stress factors identified were inter-cat conflict, fear, relocation, presence of guests and change in owner’s routine. Cases that had unsuccessful transurethral catheterisation were subjected to minimal invasive percutaneous cystostomy after initial stabilisation. The technique of minimal invasive percutaneous cystostomy demanded distended tensed palpable bladder that was secured firmly in mid ventral abdominal region for trocarisation after careful muscle separation. Controlled trocarisation, introduction of catheter, quick filling of catheter balloon and removal of peel away sheath ensured immediate relief in blocked cats. The complications in the technique were bleeding, chances of through and through puncture of bladder wall and iatrogenic injury to catheterwhile applying stay sutures. The study also identified minor post-operative complications like seepage of urine from incision site, stay sutures loosening and dislodgement of catheter. Average days taken for regaining urethral patency was 17 days and catheters were removed after gaining urethral patency. Out of 12 cases under study, nine animals regained urethral patency. Urinalysis revealed haematuria, proteinuria and leukocyturia and microscopic examination revealed presence of struvite cast in 50 per cent of animals. This study revealed that minimal invasive percutaneous cystostomy with paediatric suprapubic catheter was effective for management of urethral obstruction in tom cats with intact distended bladder.ThesisItem Open Access CHARACTERISATION OF ACID SOLUBLE COLLAGEN FROM DAGGERTOOTH PIKE CONGER (MURAENESOX CINEREUS) AND ITS APPLICATION IN WOUND HEALING(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) NIMNA AJAY; Dr. Varuna P. PanickerCollagen, the abundant structural protein in the extracellular matrix has wide application in medical, tissue engineering and pharmaceutical industry. The present study was conducted to characterize and evaluate the wound healing potential of isolated ASC from Daggertooth pike conger (Muraenesox cinereus).Conger eels were collected from Kochi harbour and pre-treatment for removal of fats and non-collagenous proteins. ASC was isolated by treating with 0.5 M acetic acid for 3 days and supernatants were collected. Protein precipitation was carried out by adding NaCl to final concentration of 2.3 M. Precipitated samples were dissolved in 0.5 M acetic acid and dialysed against 0.1 M acetic acid for one day and in distilled water until neutral pH was obtained. Yield of ASC was 12.78 and 31.95 per cent on wet and drymatter basis respectively. ASC on Lowry’s test showed a concentration of 8.8µg/ml. On electrophoretic separation revealed the presence of characteristic α (2 α1 and α2), β, γ subunits similar to that of type I calf collagen. An absorption peak of 221 nm characteristic to triple helix was observed on UV-visible spectroscopy. FTIR spectrum of ASC showed Amide bands similar to that of collagen. Analysis for secondary structure, by CD spectroscopy revealed a CD spectra with positive absorption maximum at 222 nm, and negative at 214 nm. Denaturation temperature of the extract was obtained at 38.2℃ which in turn confirm that collagen can withstand very high temperature. Solubility assay revealed a maximum solubility at low salt concentration and acidic pH. Hydroxyproline concentration in the extract was 9.9g. Invivo wound healing study showed that group treated with paste has increased wound healing compared to gel and control group. Results were confirmed by histopathology, immunohistochemistry and relative quantification of selected growth factors and cytokines. On histopathological evaluation increased fibroblast proliferation, collagen deposit and re-epithelisation were observed on collagen treated group. Difference in the collagen deposition in response to treatment was studied using staining like Picrosirus and Masson’s trichome. Angiogenesis was studied by immunohistochemistry using CD31 antibody. Observations on histopathology and immunohistochemistry could correlate with the gene expression study of TGFβ-1, VEGF, IL-10 and TNF-α. Upregulation of TGFβ-1, VEGF and IL-10 and downregulation of TNF-α was observed in collagen treated groups which might be due to the wound healing potential of collagen.ThesisItem Open Access EFFECT OF MORINGA (MORINGA OLEIFERA) LEAF EXTRACT AND NISIN ON QUALITY AND SHELF LIFE OF JAPANESE QUAIL MEAT(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) ANJITHA JOSE K; Dr. Irshad AThe effective utilisation of Moringa oleiferia leaves aqueous extract and nisin as natural antioxidant and antimicrobial ingredients in quail carcasses during refrigerated storage was investigated, and its efficiency was evaluated. During the 12 days storage period at 4 ± 1ºC, the quality of the carcass was assessed via physico-chemical, sensory attributes and microbiological quality characteristics at three days intervals. The mean value of total phenolics and flavonoid content of prepared moringa leaves extract (MLE) was 72.16 ± 1.8mg GAE/ml and 12.65 ± 0.83 mg QE/ml, respectively. The antioxidant activity of MLE was analysed and expressed in terms of IC50 (half maximal inhibitory concentration) value of 4.60 ± 0.20 per cent of MLE. The best four concentration was selected, such as 5, 10, 15 and 20 per cent, having mean values of 52.22 ± 1.09, 71.50 ± 2.23, 82.53 ± 1.84 and 87.30 ± 1.46 of antioxidant activity, respectively and used as dipping solutions and conducted raw and cooked quail carcasses sensory analysis. No antimicrobial activity was observed for prepared moringa leaf aqueous extract against E. coli and Staphylococcus aureus susp. aureus. All the results were statistically analysed. There was no significant difference (p>0.05) between treatments and controls in the sensory parameters of raw quail carcasses. But there was a significant (p<0.05) difference in sensory attributes of cooked quail carcasses, and carcasses treated with 20 per cent MLE had lower flavour values and overall acceptability scores compared to other treatments and controls. Quail carcass treated with MLE concentration up to 15 per cent having higher scores compared to other treatments and controls. Hence, 15 per cent concentration of MLE was selected for the third phase of the study. In the third experiment, various studies were conducted on Quail carcass treated with 15 per cent moringa leaf extract along with 500ppm and 1000ppm nisin against positive and negative control. All the control and treated Japanese quail meat were packed aerobically in polyethylene bags and stored at 4±1 ⁰C. Samples were evaluated for pH, TBARS, tyrosine value, colour values, microbiological quality and sensory attributes for 12 days of storage. Findings showed that TBARS value, tyrosine value, b* colour values and microbiological parameters were significantly (p<0.05) lower in treatments as compared to the negative control. There was no significant difference between treatments and controls and during storage days for pH, L* values. There was significant (p<0.05) difference in sensory attributes of raw and cooked quail meat throughout the storage period. From the studies, it can be concluded that MLE alone or in combination with nisin can be utilised as a natural preservative in quail carcass to extend shelf-life and quality during refrigerated storage upto 12 days.ThesisItem Open Access IMMUNOHISTOCHEMICAL EVALUATION OF THE EXPRESSION OF GLUTAMATE AMMONIA LIGASE IN THE BRAIN OF RABIES AFFECTED DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) PULLAGURA NIKHITHASREE; Dr. Divya CThe current research work was designed with the objective of studying immunohistochemical localisation of glutamate ammonia ligase (GLUL) protein in different anatomical sites of brain in canine rabies as well as the comparison of immunohistochemical signals in furious and dumb forms of the disease along with gross and histopathological changes in rabid dog brains. The carcasses of 219 rabies suspected dogs that were brought to the Department of Veterinary Pathology, CVAS, Mannuthy, for necropsy between January 2021 and August 2022 formed the source of the samples in this study. One hundred and thirty-threebrain samples tested positive for dFAT were further confirmed by employing polymerase chain reaction targeting N gene of rabies which yielded amplicon size of 533bp. Gross, cytological and histopathological alterations in brain of rabies were recorded. Immunohistochemical staining revealed localisation of GLUL in glial cells in different anatomical sites of brain with different staining intensity. Diffuse strong immunopositivity was noticed in the microglial cells, astrocytic end feet process and axonal fibres in cerebellum, hippocampus, brainstem and cerebrum in furious form. Whereas weak immunopositivity was noticed in all the four parts of brain in dumb form. Comparison of immunofluorescence with clinical forms of rabies revealed that furious form exhibited higher grade of fluorescence where as lower grade of fluorescence was noticed in dumb form. Further, in different clinical forms of rabies, immunohistochemical localisation of GLUL was compared with immunofluorescence of the viral antigen. In furious form of rabies, immunofluorescence was higher and immunopositivity for GLUL was stronger. The dumb form, in contrast, demonstrated low immunofluorescence and weak immunopositivity. Immunofluorescence and immunopositivity were positively correlated in both furious and dumb form. However, the significant assosciation and correlation between immunofluorescence and immunopositivity were greater in hippocampus and cerebellum (p< 0.01) compared to brainstem and cerebrum (p>0.05).Mechanism by which GLUL is increased and decreased in furious and dumb form respectively was substantiated from the present study. It is postulated that increased levels of GLUL leads to excitotoxic conditions manifested as furious form and decreased levels of GLUL leads to neurodegenerative changes manifested as dumb form.ThesisItem Open Access FREEZABILITY AND IN VITRO FERTILITY OF MALABARI BUCK SEMEN CRYOPRESERVED IN LIPOSOME-BASED EXTENDER(COLLEGE OF VETERINARY AND ANIMAL SCIENCS, MANNUTHY, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) URMILA S.; Dr. Lekshmi Bhai K.The present study evaluated the freezability and in vitro fertility of Malabari buck semen cryopreserved in liposome based extender. Fresh semen ejaculates collected on six different days from three adult, healthy Malabari bucks were used for the study. Initially, they were evaluated for volume of the ejaculate (mL), progressive motility (%), concentration of spermatozoa (x106/mL), viability of spermatozoa (%), morphological abnormalities of spermatozoa (%), functional membrane integrity of spermatozoa (%), acrosome integrity of spermatozoa (%), malondialdehyde concentration (nmol/mL) and cervical mucus penetration distance by spermatozoa (mm). After evaluation, fresh semen ejaculates were pooled and divided into two groups (n=6) as Group I (semen extended with tris-egg yolk based extender and Group II (semen extended with liposome based extender) and subjected to cryopreservation. Post thaw evaluation of Group I and Group II showed that progressive motility of spermatozoa (%), viability of spermatozoa (%), functional membrane integrity of spermatozoa (%), acrosome integrity of spermatozoa (%) and cervical mucus penetration distance by spermatozoa (mm) were significantly higher in Group II (p<0.05) compared to Group I. The morphological abnormalities of spermatozoa (%) and malondialdehyde concentration (nmol/mL) were significantly higher in Group I (p<0.05) compared to Group II. In vitro fertilisation rate of oocytes fertilised by semen extended with liposome based extender (Group II) was significantly higher (p<0.05) compared to those fertilised by semen extended with tris-egg yolk based extender (Group I). Therefore, it can be concluded that liposome based extender is more efficient than tris egg yolk based extender for cryopreservation of Malabari buck semen.ThesisItem Open Access SUB-ACUTE TOXICITY OF INDOXACARB AND ITS REVERSAL WITH HESPERIDIN AND ASCORBIC ACID IN RATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCS, MANNUTHY, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) ARCHANA RAJ; Dr. Suresh N. NairThe present study was conducted to evaluate the sub-acute toxicity of indoxacarb and its reversal with hesperidin and ascorbic acid in rats. Female Wistar albino rats were divided into five groups of six animals each. Group I received 1 per cent carboxy methyl cellulose (CMC) as vehicle control and group II received indoxacarb (IND) at the dose of 30 mg/kg body weight in 1 per cent CMC. Group III and IV received hesperidin (HSP) and ascorbic acid (ASC) at 100 and 50 mg/kg body weight dose levels respectively while Group V received hesperidin and ascorbic acid together at the above doses, along with indoxacarb in 1 per cent CMC. The vehicle/test substances were administered orally for 28 days on daily basis. During the treatment period the daily body weight gain, feed and water intake were measured along with the weekly assessment of hepatic and renal function through serum biomarkers. After 28 days of treatment, the animals were euthanized and organs were collected. The mean weekly weight gain and feed intake of the indoxacarb alone treated group was lower than other groups during the entire study period while a significant reduction in mean weekly water intake was observed in this group from weeks 2 to 4. The treatment groups III, IV and V with HSP, ASC and a combination of both significantly rectified the above parameters. The indoxacarb induced anomalous levels of serum biomarkers (ALT, AST, creatinine and BUN) and the reduction in the weight of the liver were significantly restored after treatment with HSP, ASC and their combination. The variation in the antioxidant parameters, catalase and superoxide dismutase, in brain, heart, liver, spleen, kidney, lungs and reproductive organs were also rectified by the treatment with the above agents. The histological examination of the organs revealed hepatotoxicity and nephrotoxicity in indoxacarb alone group, whereas it was reversed in treatment groups.From the present study, it can be concluded that both HSP and ASC can reverse the toxicity of indoxacarb in rats where the combination of these agents offers the most effective management of sub-acute toxicity. However, studies with further synergistic doses of HSP and ASC can be conducted for selecting the appropriate treatment regimenThesisItem Open Access DECELLULARISED PORCINE CORNEA AS AN ACELLULAR SCAFFOLD GRAFT FOR THE MANAGEMENT OF CORNEAL ULCERS IN DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCS, MANNUTHY, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) ASWATHY MOHAN A; Dr. S. AnoopThe study was undertaken to evaluate the efficacy of decellularised porcine cornea as an acellular scaffold graft for the management of corneal ulcers in dogs. All dogs presented with corneal lesions were clinically examined and six dogs were selected randomly irrespective of age, breed and sex. Out of six dogs selected, five were Chinese pug and one was a Cane corso. The age of affected animals ranged from four months to 48 months with a mean value of 25.33 ± 6.44 months and three were male and three were female. Right eye was affected in four animals while left eye was affected in two animals. Thorough ophthalmic examination was carried out in all animals with bilateral evaluation of menace response, palpebral reflex, pupillary light reflex, cotton ball test, Schirmer tear test, Fluorescein dye test, direct ophthalmoscopy and slit lamp biomicroscopy. Decellularised porcine corneas were prepared using 0.1per cent Sodium dodecyl sulphate and efficiency of decellularisation was assessed by histological examinations. Under general anaesthesia, corneal grafting was done, after corneal wound debridement or iridectomy, with decellularised porcine cornea. All the corneas were protected using temporary tarsorrhaphy which was retained up to seven days and observations such as corneal clarity, oedema, neovascularisation, extent of pigmentation and scarring were serially recorded on day 7, 14, 21 and 60 post-grafting. Three corneas showed negative FDT on day 7 post grafting while complete epithelialisation was obtained in three corneas by day 21. Corneal oedema subsided by day 21 in all cases and neovascularisation was at peak on day 7 and 14 post-grafting which regressed completely by day 60 in all the cases. Corneal clarity improved gradually through the observation period. Vision impairing corneal melanosis and scarring was observed only in one out of six corneas. The main complications observed post operatively were development of corneal melanosis, scarring and anterior synechia formation in one cornea. It was concluded that re-epithelialisation and corneal wound healing were improved with minimal pigmentation and scarring by corneal grafting with decellularised porcine cornea as an acellular scaffold graft, in comparison to normal corneal healings reportedThesisItem Open Access CLINICAL AND LABORATORY EVALUATION OF RESPONSE TO CHEMOTHERAPY IN CANINE TRANSMISSIBLE VENEREAL TUMOUR(COLLEGE OF VETERINARY AND ANIMAL SCIENCS, MANNUTHY, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-02) AJA TAJ NAZER; Dr. B. Bibin BechaA retrospective study was conducted for a three-year period at the University Veterinary Hospitals and the overall occurrence of CTVT were recorded as 0.15 percent among total cases, 0.88 per cent among total reproductive cases, 21.42 per cent among total tumour cases and 4.74 per cent among dogs with vaginal bleeding. Highest occurrence was found in Labrador Retrievers (41.30%), intact female dogs (92.22%), dogs aged 2-5 years (73.91%), pluriparous dogs (40.85%) and during south-west monsoon season (33.70%). Thirty-one CTVT confirmed dogs were randomly allocated to three treatment groups. The group I (n=13) dogs were treated with vincristine sulphate @ 0.025 mg/Kg bodyweight intravenously at weekly intervals till tumour regression. The group II (n=9) dogs were treated with ivermectin @ 200μg/Kg subcutaneously, followed by vincristine sulphate @ 0.025 mg/Kg bodyweight intravenously after 24 h and repeated at weekly intervals till tumour regression. The group III (n=9) dogs were treated with vincristine @ 0.0125mg/Kg body weight intravenously, followed by cyclophosphamide @ 5mg/Kg orally for 10 days. Vital parameters during different days of treatment were within normal limits. The body weight of CTVT affected dogs ranged from 7.0– 40.5 Kg with an average body weight of 22.79 ± 1.73 Kg. Body Condition Scores (BCS) were 4–5 in all dogs on all days of observation. Majority of tumours were located in the vulva (51.61%), followed by vagina (19.35%) and vestibulo-vaginal junction (16.13%), and appeared as cauliflower like friable mass that bled easily. The size of the tumour ranged from 0.9–8.5 cm with an average size of 3.90 ± 0.35 cm. A significant decrease in Canine Karnofsky Score was observed in three groups by day 21, which increased gradually. On TNM staging, majority of dogs scored T3N0M0 (54.84%), followed by T2N0M0 (32.26%) and T1N0M0 (6.45%). Two cases of metastasis (6.45%), were also reported (T3N0M1 and T4N0M1). The haemato-biochemical parameters like TEC, Hb, VPRC, TP, BUN, creatinine, albumin, globulin and ALT were within normal range. A significant difference was observed in differential leucocyte count, thrombocyte count and mean corpuscular volume (p<0.05). Significant difference were observed in AST, ALP and bilirubin values (p<0.05). On cytomorphological classification, majority of tumours were plasmacytoid (74.19%), followed by lymphocytoid (22.58%) and mixed type (3.23%). The combined malignancy scores were found to decrease after successive treatments from marked (+++) malignancy sore to absent (0) by day 21 in groups I and II and day 28 in group III. The immunocytochemical scoring with vimentin showed strong immunoreactivity on day of presentation in all dogs, and it was scored absent in group I by day 21, in group II by day 14 and in group III by day 28. The duration of treatment in vincristine, vincristine-ivermectin combination and vincristine-cyclophosphamide combination were 23.10 ± 1.49, 22.00 ± 1.00 and 30.00 ± 2.00 days, respectively. Duration of treatment and side-effects were found to be less in vincristine-ivermectin combination therapy. No recurrence of tumour was observed in three treatment groups. Considering the response to treatment, duration of treatment and intensity of side effects, combination therapy of vincristine with ivermectin gave better response in CTVT treatment. Cytology, mitotic index, apoptotic index and immunocytochemistry signalling with vimentin could be used to predict the proliferation and regression of tumour mass and thus response to therapy.ThesisItem Open Access PROCESS STANDARDIZATION FOR LUTEIN INCORPORATED CALCIUM AND VITAMIN D FORTIFIED SHRIKHAND(VERGHESE KURIEN INSTITUTE OF DAIRY AND FOOD TECHNOLOGY,MANNUTHY, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-07-14) AJMI S SHAMS; Dr. S. N. RajakumarThe present study was conducted to explore the feasibility of utilization of shrikhand as a carrier of lutein, calcium and Vitamin D for added nutritional benefits to the consumer. The levels of ingredients were optimized by using Central Composite Rotatable Design of Response Surface Methodology (RSM). The highest desirability of optimized shrikhand was 94.3 per cent with the levels of addition of tricalcium citrate tetrahydrate, vitamin D3 and lutein at 162 ppm, 558 IU/L and 0.59 mg/100g respectively. The effect of enrichment on the optimized shrikhand was assessed based on its sensory quality, physico-chemical characteristics and microbiological parameters. When compared to control, the mean sensory scores for the attributes colour and appearance body and texture and overall acceptability of the optimized product were higher at one per cent significance level. The optimized shrikhand had significantly higher values for protein (10.84 ± 0.12), ash (0.57 ± 0.02), total solids (61.42 ± 0.31) and pH (4.43 ± 0.03) as compared to control shrikhand. In optimized shrikhand, an estimated content for calcium and vitamin D3 were 166.81 mg/100 g and 22.3 IU/100 g respectively, whereas the recovery (per cent) was 84.8 and 39.8 respectively. The total carotenoid content and antioxidant activity (IC50value) of the optimized shrikhand were 0.49 mg/ 100g and 949.23 mg/kg respectively. The shrikhand (control and optimized) had a shelf life of 28 days when packed and stored in polystyrene cups at 7±1℃. The extra cost incurred for the enrichment was 0.32 paise/ 100g of the optimized product. Based on the results, it can be suggested that incorporation of tricalcium citrate tetrahydrate, vitamin D3 and lutein can improve the functionality of shrikhand.
