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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. Ramnath
In 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.
Open Access
ADAPTABILITY PROFILE OF MALE CATTLE AND BUFFALO CALVES TO VARYING TEMPERATURE HUMIDITY INDEX (THI) IN KERALA
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2016-12-30) AZIZ ZARINA; G. Girish Varma
The present study was conducted to assess the adaptation capacity of castrated buffalo and crossbred male calves to the varying temperature humidity prevalent in central midlands of Kerala using physiological, haematological, growth, endocrine and genetic parameters. Ambient temperature, humidity, rectal temperature, pulse rate and respiratory rate were recorded daily. Relative growth rate and plasma volume were recorded at fortnightly interval. Haematological parameters and endocrine parameters were determined at biweekly interval. The temperature humidity index recorded throughout the year varied from 76.5 to 82.0. Rectal temperature remained invariable throughout the experimental period but respiratory rate and pulse rate exhibited a significant positive correlation (P<0.01) with temperature humidity index. A negative correlation was observed between temperature humidity index and relative growth rate, total erythrocyte count, haemoglobin concentration, volume of packed red blood cells and thyroid hormones of the experimental animals at P<0.01 level. While a positive correlation was observed between plasma volume and temperature humidity index in all the experimental animals. Cortisol levels remained consistent throughout the study period in buffalo and crossbred cattle calves. The rate of increase of plasma volume, respiratory rate and pulse rate were higher in buffalo calves compared to crossbred cattle calves. HSP90 expression showed significant (P<0.05) up regulation at a THI of 82.0 in all the experimental animals, however buffaloes showed a higher fold expression of HSP90 compared to that of crossbred cattle calves.
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. Raji
The 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.
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. Pratheesh
Present 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.
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., Karthiayini
The 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.
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. Babitha
The 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.
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. Karthiayini
The 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.
Open Access
DIETARY SUPPLEMENTATION OF SPIRULINA PLATENSIS ON GROWTH AND IMMUNE RESPONSE OF BROILER CHICKEN
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2008) K. PREETHA; G. GIRISH VARMA
The present experiment was aimed at investigating growth promoting and immunomodulating effects of Spirulina platensis in broiler chicken. Thirty two numbers of day-old broiler (Vencob strain) chicks were procured and reared in battery cages under standard management conditions up to sixth week of age. The birds were randomly divided into four groups, viz. GI, GII, GUI and GIV, having eight birds in each group. Birds in groups GI (control) were fed with standard broiler starter ration (from first to fourth week) and standard finisher ration (from fifth to sixth week). The experimental groups GII, GUI and GIV were fed with standard rations supplemented with Spirulina platensis at the rate of 0.50 per cent, 1.50 per cent and 3.00 per cent, respectively. Both control and experimental rations were made isocaloric and isonitrogenous. Birds were provided with ad libitum feed and clean drinking water throughout the experiment. Bodyweight of individual birds were recorded on day one, eight, 15, 22, 29, 36 and 43. Blood was collected at fourth, fifth and sixth week of age and whole blood/plasma was used for further estimations. Estimation of total erythrocyte count (TEC), blood haemoglobin concentration (Hb), volume of packed red cells (VPRC), total leukocyte count (TEC), differential leukocyte count (DEC), and blood glucose was made using whole blood samples. Blood plasma was utilised for estimation of concentrations of total protein, albumin, globulin, total lipid, total cholesterol, triglyceride, HDL cholesterol, LDL cholesterol, and VLDL cholesterol. Erythrocytic indices (MCV, MCH and MCHC), heterophil: lymphocyte (H:L) ratio and albumin; globulin (A:G) ratio were calculated from the respective values. The data was analysed by Split plot analysis (Split on time) as suggested by Rangaswamy (1995). Bodyweight of Spirulina-fed birds (GII, GUI and GIV) were significantly (P<0.01) higher than that of control group throughout the experiment in an age and dose dependent way. Significant (P<0.01) age and dose dependent increase in TEC, Hb, VPRC, MCH and MCHC and decrease in MCV values were observed in Spirulina-fed groups, suggesting strong erythropoietic effect of Spirulina. Significant (P<0.01) age and dose dependent increase in total leukocyte and lymphocyte counts and decrease in heterophil, eosinophil, and monocyte counts were noticed in Spirulina-fed groups. Total leukocyte count was higher in Spirulina-fed birds, but without increase in heterophil count, ruling out the possibility of inflammatory responses. Additionally, reduced stress and more vigilant humoral immunity could be the implication. Also, Spirulina induced significantly (P< 0.01) more plasma albumin and plasma globulin, and thus, total plasma protein output in the recipients. Since the albumin: globulin (A:G) ratio did not change significantly, both these classes of proteins were equally promoted. This would mean a general boost up in plasma protein output by the liver as well as more immunoglobulin production by the lymphocytes. Increased number of lymphocytes in Spirulina-fed birds could be attributed to the higher plane of globulins in this experiment. Plasma lipid profile showed age and dose dependent lowering of total lipid, cholesterol, triglyceride, LDL and VLDL cholesterol in Spirulina-fed groups compared to control group, indicating that Spirulina would reduce lipid level, especially the 'undesirable' fractions, signifying the hypolipidaemic / lipolytic activity of the alga. The significant (P<0.01) hypoglycaemic effect of Spirulina supplementation in the present study may indicate an anti-diabetic effect, which may indicate, in turn, propensity to have more lean meat than fatty meat in the subjects. The growth-promoting and erythropoietic effects of Spirulina platensis in broiler chicken may indicate its effectiveness as a nutritional supplement, while the hypolipidaemic and hypoglycaemic effects may imply better consumer safety and desirability to the broiler meat, which intum, can potentially fetch premium price to the produce in the market.
Open Access
INFLUENCE OF COCONUT OIL AND SUNFLOWER OIL ON PLASMA AND LIVER LIPID PROFILE AND PRODUCTION PERFORMANCE IN JAPANESE QUAIL (Coturnix coturnix japonica)
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 1997) MINI. K. P.; P.T. Philomina
The role of dietary factors in the genesis and development of atherosclerosis has been associated wich elevated serum cholesterol levels, both in man and animals. Coconut oil has been a dietary component for Keralites from time immemmorial . There are several reports implicating coconut oil (a saturated fat) as one of the major factors involved in the production of increased blood cholesterol level leading to the increased incidence of cardio vascular disease (CVD) in human beings and animals. No systematic investigation has been carried out so far to study how increasing unsaturation of the oil affects lipid metabolism in warding off atherosclerosis. In addition to the chain length of fatty acids their relative position in the triglyceride molecule appears to affect their potency for atherogenicity. Gingelley oil is commonly used as one of the energy source in poultry feed and sunflower oil is also gaining popularity now a days, since many reports indicated that feeding of unsaturated fat decreased the cholesterol content in the oodv Hence it was thought worthwhile to study the influence cT these three oils on the levels of total lipid, triglyceride, total cholesterol and phospholipid in plasma and liver, the concentration of HDL-cholesterol and (VLDL+LDL)-cholesteroJ m plasma, weight of the liver and the level of total lipid and total cholesterol in egg yolk. The production performance under these dietary oils was also assessed in Japanese qua;' Is by recording the egg production, egg weight and egg mass, feed consumption, body weight and feed efficiency. A total number of 72 (36 males and 36 females), four-week old Japanese quails of the same strain (egg type) and hatch were procured from the Kerala Agricultural University Poultry Farm, Mannuthy and divided into three main groups (12 males and 12 females in each main group viz. GI, Gil, GUI) and then subdivided to 12 males and 12 females as M-I, M-II and M-III (males) and F-I, F-II and F-III (females) . The birds were provided grower ration upto sixth week of age and then adult ration, from the sixth to the 10th week of age in males and 16th week of age in females. The standard ration was incorporated with the different dietary oils at 2 per cent level viz. GI (MI and F-I) with gingelley oil, GII (Mil and F-II) with coconut oil and GUI (Mill and F-III) with sunflower oil. Feed consumption, egg production and egg weight were recorded daily and body weight recorded weekly. The eggs from the three groups (F-I, F-II and F-III) were collected on the last day of 14th, 15th and 16th week of age, weighed and stored at 4°C for biochemical analyses. The male birds were sacrificed at the 10th week of age and females at the 16th week of age. The weight of the liver noted and plasma and liver stored at -20°C for analyses. Total lipid, triglyceride, total cholesterol , HDL-cholesterol (VLDL+LDL)-cholesterol and phospholipid in plasma of male and female Japanese quails were not significantly different among the groups, since the normal level (2%) of oils used in the present study was not able to exert any significant influence on the lipid metabolism in quails. The total lipid content in the liver in male quails was not significantly different among the groups. In the female quails the total lipid content in liver of gingelley oil fed group (F-I) was significantly higher than that of coconut oil fed (F-II) and sunflower oil fed (F-III) groups. Lower triglyceride lipase activity in coconut oil and sunflower oil fed groups, which causes decreased break down of triglyceride in adipose tissue and lower transportation of fatty acids to liver may be the reason for the lower total lipid content in liver in these two groups. Irrespective of sex the triglyceride and total cholesterol content in liver were not significantly different among the groups. The liver phospholipid content in male quails of gingelley oil fed group (MI) was significantly higher than that of sunflower oil fed group (Mill) . Sunflower ocj (unsaturated fatty acids) causes enhanced faecal excretion of free fatty acids. The total lipid, triglyceride, (VLDL+LDL)-cholesterol and phospholipid in plasma were significantly higher in adult female quails compared to males in all the groups. In the laying bird lipids are synthesised in the liver and transported to the ovary in the form of lipoproteins . This is the reason for higher total lipid, triglyceride and phospholipid in plasma of laying hen (VLDL + LDL)- cholesterol content was also higher in the female quails since they are the transport form of cholesterol from liver to the ovary. The total cholesterol and HDL-cholesterol were higher in male quails compared to females, since the cholesterol is not eliminated through the egg yolk and are mainly found along with the HDL fraction in males unlike females. The total lipid, triglyceride and total cholesterol content in liver of female quails were significantly higher than that of male quails. There was higher lipid synthesis in the liver of female quails under the influence of oestrogen. There was no significant difference in the liver phospholipid content between male and female quails. No significant difference in the weight of the liver among the groups in both male and female Japanese quails could be noticed. However, weight of the liver in females was ® fi^sntly higher than the males in each group. There was no significant difference in the total lipid and total cholesterol content in egg yolk among the groups, at the 14th, 15th and 16th week of age. This was because the total lipid and total cholesterol content in the egg yolk has to be maintained at a constant level in order to create a favourable environment for the development of the embryo. Female quails attained sexual maturity earlier at the end f fifth week (38 to 41 days of age). Egg production started the 38th day in groups F-II and F-III while it was on the 41st day in group F-I. The egg production was lower in gingelley oil fed group (F-I) than coconut oil fed group (F-II) may be due to lower mineral absorption in gingelley oil fed group. The egg weight was higher in sunflower oil fed group (F-III) than coconut oil fed group (F-II) since sunflower oil in the diet causes higher protein retention. The egg mass was not significantly different among the groups. However, the egg weight and egg mass significantly increased with age. There was no significant difference in feed intake among the groups in both male and female Japanese quails as tiie caloric value of the feeds were the same. Female quails had higher feed intake than males due to their higher growth rat:e. The body weight of the male quails was not significantly among the groups. The body weight in gingelley oil fed group was the lowest among the female quails during most part of the experimental period, since there is lower mineralisation of bones in that group. The body weight of female quails in the three groups was significantly higher than that of males. Feed efficiency was not different among the groups in male Japanese quails. Among the female Japanese quails coconut oil (F-II) and sunflower oil (F-III) fed groups had better feed efficiency than gingelley oil fed group (F-I), lower rate of absorption of minerals in gingelley oil fed group (F-I) may the reason for the lowest feed efficiency anu body weight. In order to arrive at a conclusion as to which of the particular oil is ideal for health and for better production performance in Japanese quails, higher levels of oils are to be incorporated and a more detailed study is required.

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