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Kerala Veterinary and Animal Sciences University, Wayanad
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ThesisItem Open Access EVALUATION OF VITAMIN AND MINERAL STATUS IN DERMATOLOGICAL DISORDERS IN DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) G. SRUJANA SAI; Dr. V. BabithaThe present study was undertaken evaluate vitamin and mineral status in dermatological disorders in dogs. The study included 40 dogs of 2-5 years of age with dermatological disorders presented to University Veterinary Hospitals at Mannuthy and Kokkalai under the Kerala Veterinary and Animal Sciences University, and ten apparently healthy dogs brought to the hospital for routine health check-up which served as control.Ultra high performance liquid chromatography (UHPLC) was employed for estimation of serum levels of vitamin A, D, E, C and vitamin B complex, and the serum zinc, copper and selenium levels were estimated using atomic absorption spectrophotometer (AAS). Among the various vitamins estimated, serum vitamin A concentration was found to be significantly lower (p<0.05) in diseased dogs (1.45 ± 0.06 μmol/L) compared to control group (1.61 ± 0.008 μmol/L), while the serum levels of vitamin E, vitamin D3, vitamin B1, B2, B3, B6, and B12 in diseased dogs did not differ significantly from the control group. Serum vitamin C concentration was found to be significantly lower (p<0.01) in diseased group (41.36 ± 0.20 μmol/L) in contrast to the control group (43.32 ± 0.32 μmol/L). Serum mineral levels analysed through AAS revealed that the concentration of zinc was significantly lower (p<0.01) in diseased group (0.53 ± 0.04 mg/L) than the control group (1.11 ± 0.09 mg/L) and the serum copper concentration was also significantly reduced (p<0.05) in diseased dogs (0.91 ± 0.08 mg/L) with respect to the control animals (1.23 ± 0.11 mg/L). The serum selenium concentration of diseased group of animals, however failed to show any significant difference from the control group of animals. It was concluded from the present study that out of the various vitamin and mineral levels analysed, the serum concentrations of vitamin A, vitamin C, zinc and copper were found to be significantly lower in dogs with skin diseases in comparison to normal healthy animals in control group suggesting that the deficiencies of these vitamins and minerals encountered in diseased dogs might be playing significant roles in either causation and/or consequence of skin ailments.ThesisItem Open Access EVALUATION OF OXIDATIVE STRESS IN ANAEMIA ASSOCIATED WITH HAEMOPROTOZOAN INFECTIONS IN DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE, WAYANAD, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-08-10) ROHIL CHOPRA; Dr. V. RamnathIn order to assess the oxidative stress in anaemic dogs with haemoprotozoan infections, a total of 12 blood samples from anaemic dogs (Hb ≤ 12 g/dL) confirmed positive for Babesia gibsoni and B. canis were collected. The samples were subjected to a complete blood count to assess the RBC count, WBC count, haemoglobin, haematocrit, MCV and MCHC. Oxidative stress was assessed by measuring RBC activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and serum total antioxidant capacity (TAC). Amongst the haematological parameters, the RBC count, WBC count, haemoglobin and haematocrit were all significantly reduced in the infected dogs. All the antioxidant parameters viz. RBC SOD and GPx activities, and serum TAC were found to be significantly increased in the infected dogs. All except one of the findings of this study corroborate with the findings of earlier studies, the exception being the RBC GPx activity. The hallmark of this study is the finding of the RBC GPx activity being elevated in the infected dogs. Significantly elevated GPx activity in dogs in relation to being cytologically and clinically positive for haemoprotozoans is a unique finding that has not been reported till date.ThesisItem Open Access EXPRESSION OF GONADOTROPINS AND THEIR RECEPTORS IN ATTAPPADY BLACK AND MALABARI GOATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR,, 2019-12-31) BHUVANA PLAKKOT; K. RajiThe difference in prolificacy between high prolific Malabari and low prolific Attappady Black goat breeds of Kerala has been evaluated by gonadotropins and their receptors gene expression studies, and analysis of steroid hormones. Six numbers of experimental animals, each from Malabari and Attappady Black breeds were selected for the study from the culling list of University Sheep and Goat Farm, Kerala Veterinary and Animal Sciences University, Mannuthy. Ovaries, pituitary and serum samples were collected from animals slaughtered at the proestrus stage of the oestrus cycle. By radioimmunoassay technique, a significant difference was observed in serum oestradiol concentrations, with Malabari having a higher mean (44.22±7.00 pg/mL) when compared to the Attappady Black breed (13.98±4.35pg/mL). The ovarian antral follicles were isolated and classified into small (1-3 mm) and large (>3 mm) follicles. RNA isolated from the follicles and pituitaries were used for expression studies. By performing quantitative real-time PCR technique, it was observed that between the breeds, there was no significant difference in the level of FSHβ expression in the pituitary, FSHR expression and LHR expression in follicles. However, there was a significant difference in the pituitary LHβ expression between breeds, with a significantly (P<0.01) high level in Attappady Black compared to Malabari breed. A non-significant decrease in (fold-change of 0.64) FSHR expression and a non-significant increase in (fold-change of 1.4) LHR expression were observed from small to large follicles in goats. Irrespective of breeds, a non-significant negative correlation (-0.49) was observed between oestradiol and FSHβ expression. Also, a non-significant negative correlation (-0.15) was observed between LHβ expression and progesterone concentration. A positive correlation although not very significant was found between FSHβ expression and FSHR expression in goats irrespective of breeds. Similarly, between FSHβ expression and LHR expression in goat antral follicles, a significant positive (P<0.05) correlation was observed. The antral follicle FSHR expression irrespective of breeds, showed a significant (P<0.05) positive correlation (0.46) with the oestradiol concentration. No correlation was found between pituitary overall LHβ and follicle LHR expression in antral follicles irrespective of the breed. A significant (P<0.05) positive correlation was also observed between oestradiol and oestrus signs and Malabari breed exhibited prominent oestrus signs compared to Attappady Black breed.ThesisItem Open Access EFFECT OF CELL DENSITY AND SERUM STARVATION ON VASCULAR ENDOTHELIAL GROWTH FACTOR GENE EXPRESSION IN CANINE ADIPOSE TISSUE DERIVED CELLS(Kerala Veterinary and Animal Sciences University,Pookode, Wayanad, 2021-09-20) HARSH JAMDA; M. D. PratheeshPresent experiment was conducted to study the effect of cell density and serum starvation on Vascular Endothelial Growth Factor (VEGF) gene expression in canine adipose tissue derived mesenchymal stromal cells (MSCs). Subcutaneous fat tissue sample was collected from adult dog presented at the Department of Veterinary Surgery and Radiology, CV & AS, Pookode. The tissue pieces were washed and digested using 0.2 % collagenase type 3 and filtered using 40μm filter to isolate the adipose tissue derived stromal cells which were seeded in DMEM (Dulbecco's Modified Eagle's Medium) supplemented with 15 % FBS (Fetal Bovine Serum) for the primary cell culture. As primary culture attained 70-80 % confluency, subculture was done till fourth passage. Growth curve and population doubling time of the in vitro expanded canine adipose tissue derived MSCs of cells were calculated by using 0.4 % trypan blue exclusion test. Fourth passage cells were seeded in different seeding densities (10 4 /cm 2 ; 3x10 4 /cm 2 ; 6x10 4 /cm 2 ) within six well cell culture plates and were grown for 72 hrs (pre- incubation) in normal serum rich (+) growth media. After pre-incubation, cells were further incubated for 36 hrs replacing with either serum rich (+) or serum free (-) media at respective time intervals (Zero h; 12 h; 24 h; 36 h). At the end of incubation period, treated/adherent cells were harvested from each well and total RNA was isolated and cDNA was synthesized. Expression of VEGF in different groups was analyzed by real time PCR using specific primers designed for VEGF keeping the expression of GAPDH as reference. Among the different seeding density and serum starvation combination experimented, we could not find any significant impact on the expression of VEGF, but we observed a significant increase in VEGF expression in canine adipose tissue when used at a higher cell seeding density (6x10 4 /cm 2 ). These findings suggest that cell therapy using stromal cells derived from subcutaneous canine ASCs, regulated with high seeding density may be a novel therapeutic option to enhance angiogenesis thereby provides therapeutic benefits in pathological vascular conditions including wound repair, ischemic damage, microvascular permeability and diabetes.ThesisItem Open Access EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) AND ITS BIOLOGICAL RECEPTOR, VEGFR2 IN GRANULOSA CELLS OF MALABARI GOAT(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2016-07-05) D., SUPRIYARAJ; K., KarthiayiniThe aim of the present study was to quantify the relative gene expression levels of Vascular Endothelial Growth Factor (VEGF) and its biological receptor, Vascular Endothelial Growth Factor Receptor-2 (VEGFR-2) in Granulosa cells (GCs) of medium (Gp-I; 3-5mm diameter) and large (Gp-II; >5mm diameter) goat ovarian follicles, by real-time PCR. Quantification of Progesterone (P4) and Estradiol-17β (E2) was also performed in the follicular fluid (FF) of two different follicular groups by enzyme-linked immunosorbent assay (ELISA). Pooled samples (n=7) of FF from two groups of follicles were used for the study. The results showed that the mean relative expression levels of VEGF and VEGFR-2 mRNA in GCs of Gp-II follicles was higher when compared to Gp-I follicles but it was non-significant (p>0.05). The mean (± S.E) increase in relative expression values of VEGF and VEGFR-2 was 1.47 ± 0.78 and 3.84 ± 2.51 respectively. The follicular fluid P4 and E2 concentration was significantly (p≤0.05) higher in the Gp-II follicles than the Gp-I follicles. The mean (± S.E) P4 concentration (ng/mL) in FF of Gp-I and Gp-II follicles were 10.56 ± 2.3 and 20.53 ± 3.82 respectively. The mean (± S.E) E2 concentration (ng/ml) in FF of Gp-I and Gp-II were 12.08±2.78 and 22.08±1.97 respectively. The study revealed the GCs VEGF do not play a significant role in transition of medium (3-5mm) to large (>5mm) follicle in caprines. Increase in concentration of E2 with the follicle size might be due to increase in number of GCs with increase in the size of follicle because GCs are the principal site of aromatization activity which enhanced the production of E2. Similarly, increase in P4 concentration in Gp-II follicles might be due to the production of P4 in the synthetic pathway of follicular production of E2.ThesisItem Open Access EXPRESSION OF ANGIOPOIETIN-1 (ANPT-1) AND AROMATASE (CYP19) IN OVARIAN ANTRAL FOLLICLES OF MALABARI AND ATTAPPADY BLACK BREEDS OF GOATS.(College of Veterinary and animal Science,Mannuthy, 2019) SHIJI S. SAJU; V. BabithaThe expressions of ANPT-1 and CYP19 genes in small and large follicles were studied in Malabari and Attappady Black breeds of goat. Animals (n=6) were selected from each breed and given PGF2α, after confirming oestrous by detecting visible external heat signs and open cervix using vaginal speculum, and ultra sound scanning was done in the animals for one entire cycle and observed the ovarian activities. We noticed a significant difference in the exhibition of heat signs between two breeds with a greater mean [(± S.E) 12±1.88] in Malabari than Attappady black breed [with a mean (± S.E.) 5.67 ±1.12]. On detecting the next heat, scanning was done from day 15 onwards and the animals were slaughtered on reaching pro-oestrous stage and the ovaries and blood samples were collected. Studied the expression pattern of ANPT-1 and CYP19 in different categories of ovarian antral follicles and estimated the serum oestradiol and progesterone concentration by RIA. Significant difference was found in serum oestradiol concentration between Malabari [mean (± S.E) 44.23 ± 7.00 pg/mL] and Attappady black breed [mean (± S.E) 13.95 ± 4.35 ng/mL], with no significant difference detected in serum progesterone concentration between breeds. The expression of ANPT-1 and CYP19 in follicles at different developmental stages within and between breeds failed to show any significant difference. The expression of ANPT1 was found to be decreased in large follicles of both breeds compared to small follicles whereas the expression of CYP19 was found to be increased in the large follicles of both breeds compared to small follicles. Hence it is concluded that changing pattern of ANPT-1 and CYP19 expression have a definite role in controlling the follicular development in these two breeds.ThesisItem Open Access MYOSTATIN AND INSULIN LIKE GROWTH FACTOR-1 GENE EXPRESSION IN BROILER CHICKEN EXPOSED TO MONOCHROMATIC LIGHT(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2018) QUEENIE CANDICE FERNANDES; K. KarthiayiniThe study was conducted to evaluate the effect of different LED monochromatic light (green, λ= 560 nm; blue, λ= 480 nm; red, λ= 660 nm) on growth of broiler chicken in comparison with white LED light and incandescent bulb light. Relative expression of IGF-1 and MSTN gene in breast muscle, muscle fibre diameter of breast and thigh muscle of 17, 24 and 31 day old broiler chicks as well as weekly body weight and body weight gain was used to assess the growth. The study showed that on day 17 the birds under green light had more IGF-1 and MSTN expression, followed by those under blue light. A better breast and thigh muscle fibre diameter was also observed in birds under green and blue light. On day 24 and 31 birds reared under blue light showed a better breast and thigh muscle fibre diameter, IGF-1 and MSTN gene expression than others. However, on day 31, they had MSTN expression similar to other birds except the birds under red light. Though changes in IGF-1 expression were noticed at different ages in birds reared under blue, green, red, white LED and incandescent light; there were corresponding changes in MSTN expression which followed the same pattern. Light colour effected the growth and expression of MSTN and IGF-1 genes, breast and thigh muscle fibre diameter, body weight and body weight gain. In the early period up to day 17, green light was most stimulatory to growth, breast and thigh muscle fibre diameter and the expression of MSTN and IGF-1 genes under study. After day 17 up to day 31, blue light resulted in maximum expression of MSTN and IGF-1 genes, breast and thigh muscle fibre diameter and growth. Chicken can be reared under green light up to day 17 and then afterwards under blue light for more growth during same period. Further studies can be conducted to evaluate the meat quality parameters and different cuts of meat, feathering, lighting management practices, welfare, blood physiological and biochemical variables and the functional development of the eye and brain. At fourth week of age birds under blue light had a better body weight gain followed by birds under green light. At fifth week of age birds under blue light had a significantly high body weight gain when compared to all other groups. The study revealed that blue and green monochromatic LED light is better for growth of broiler chicken than other lights used.ThesisItem Open Access DIFFERENTIAL FOLLICULAR MORPHOLOGY AND BMPR1B GENE EXPRESSION IN MALABARI AND ATTAPPADY BLACK BREEDS OF GOATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2015) ABHINA M; Raji KProlificacy difference between two native goat breeds of Kerala, Malabari and Attappady black has been evaluated by morphometric, histological, hormonal and gene expression studies. Six numbers each of Malabari and Attappady black goat breeds were used for the study. Even though there was no significant difference, the morphometric parameters (weight, length and breadth) recorded were more in Malabari than Attappady black breed of goats. The follicular development was studied by classifying the follicles based on size into very small (<1mm), small (1-3mm) and large (3-5mm). The total number of follicles was more in Malabari when compared to Attappady black. However, follicles classified under small size were more in Attappady breed of goats whereas, follicles under very small size and large size were more in Malabari. Haematoxylin and eosin stained sections were examined for histological studies and the different categories of follicles namely primordial, primary, secondary and antral were compared between breeds. The diameter of secondary follicle and the oocyte were significantly different with a higher mean value for Attappady breed. The ratio of granulosa cell to secondary follicle diameter and the ratio of oocyte to secondary follicle diameter were found to be significantly different between the breeds with higher mean in Malabari breed. The mean diameter of the antrum in Attappady breed was higher than Malabari breed. The ratio of granulosa cell to the antral follicle diameter was not significantly different between breeds. The serum oestrogen concentration was found to be significantly different between the breeds with Malabari having a higher mean (± S.E) 41.69 ± 7.50 pg/mL when compared to the Attappady breed with a mean (± S.E) 20.20 ± 4.40 pg/mL. There was no significant difference in the BMPR1B gene expression level between breeds. However, there was significant down regulation of BMPR1B expression in large follicles compared to small ones.ThesisItem Open Access ASSESSMENT OF STRESS RESPONSE AND SUPPORTIVE ROLE OF VITAMIN E IN CROSSBRED FEMALE CALVES(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2015) JANIPALLI NIKHIL KUMAR TEJ; Girish Varma GA study was conducted to assess the heat stress responses and supportive role of vitamin E in crossbred female calves using physiological, haematological, biochemical and hormonal parameters during pre-monsoon, monsoon and post monsoon seasons in Kerala. Fourteen calves were randomly selected, divided in to two groups while treatment group was additionally supplemented with vitamin E @ 5 mg/kg of body weight orally for 30 days in all the three seasons. The highest value of ambient temperature (AT) and temperature-humidity index (THI) was recorded in pre-monsoon while highest relative humidity (RH) was recorded in monsoon. Respiratory rate (RR) was significantly (p<0.05) higher in pre monsoon compared with monsoon and post-monsoon. Heart rate (HR) and pulse rate was significantly (p<0.05) higher in pre-monsoon and monsoon than in post monsoon. Rectal temperature (RT) did not differ significantly (p>0.05) between seasons. Total erythrocyte count (TEC) was significantly lower in monsoon than other seasons, while total leucocyte count (TLC), haemoglobin (Hb) and volume of packed red cells (VPRC) showed no significant (p>0.05) difference between seasons. Season had no significant (P>0.05) effect on cortisol, total protein, albumin, globulin and albumin globulin ratio in contrary glucose and cholesterol levels were significantly (p<0.05) higher in post-monsoon compared with other seasons. Glutathione peroxidase (GSH-Px) was significantly (p<0.05) higher in pre-monsoon compared with other seasons. Vitamin E had no effect on physiological, haematological, biochemical and hormonal parameters studied whereas significantly lower glutathione peroxidase (GSH-Px) was noticed in supplemented group. From the study it may be concluded that the crossbred calves have the adaptive capability to alter their physiological responses to maintain their body temperature and vitamin E may be helpful to ameliorate heat induced oxidative stress.
