Browsing by Author "PATEL, SANJAYKUMAR B."
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ThesisItem Open Access OVARIAN STEROIOS AND BIOCHEMICAL CONSTITUENTS OF POLLICULAR FLUID IN RELATION TO FOLLICULAR DEVELOPMENT OF INDIGENOUS GOATS(AAU, Anand, 2005) PATEL, SANJAYKUMAR B.; Pathak, M. M.The present investigation was carried out on non-descript adult female goats of Gujarat State with the objective to study the ovarian steroid and biochemical constituents of follicular fluid of different categories of follicle. Follicular fluid of small (2-3 mm), medium (> 3-5 mm) and large (> 5 mm) sized follicle was collected from 60 goats slaughtered at slaughter house and follicular fluid from preovulatory and anovulatory follicles was collected by laparotomy of adult goats of the farm. Collection of follicular fluid from preovulatory follicle of the normally cyclic goats was made by laparotomising the goat after detection of the animal in heat. Follicular fluid from anovulatory follicle was collected after 72 hrs. of heat in superovulated goats. Superovulation was carried out by PMSG (Folligon®) and all treated goats were subjected to laparotomy operation, 72 hours after breeding and the follicular fluid was aspirated from follicles present on the ovaries at the time of operation. Follicular fluid samples collected from such different stages of follicular development were analysed for hormones (estradiol-17β, progesterone and testosterone) by standard RIA technique and biochemical constituents (total protein and cholesterol) by standard analytical procedure. The results revealed that the level of estradiol-17β increased significantly (P < 0.05) at every stages of follicular development. The smallest follicle (2-3 mm) had lowest concentration of estradiol-17β in follicular fluid (8.95 ± 0.89 ng/ml) which rose gradually with the development of follicles. Preovulatory follicle had significantly higher (32.40 ± 0.78 ng/ml) level of estradiol-17β than small, medium and large sized follicle, whereas the anovulatory follicle showed the highest level (53.00 ± 3.01 ng/ml). Estradiol-7β concentration of follicular fluid showed a significant and positive correlation between different stages of follicular development (r= 0.96). Progesterone concentration in follicular fluid of different developmental stages of follicle varied remarkably. The level of progesterone increased as the follicle grew (small: 3.98 ± 0.40 ng/ml; medium: 8.40 ± 0.35 ng/ml and large: 16.15 ± 1.05 ng/ml) but the differences were statistically nonsignificant. The variation in progesterone concentrations in preovulatory (128.50 ± 5.87 ng/ml) and anovulatory (728.00 ± 33.52 ng/ml) follicles were statistically highly significant (P < 0.05) compared to all developing follicles. Follicular fluid progesterone concentration was positively but nonsignificantly correlated with different stages of follicle (r = 0.83). The level of testosterone in follicular fluid decreased significantly (P < 0.05) as the follicle size increased. The highest value of testosterone (5.68 ± 0.50 ng/ml) was observed in small sized follicle and the lowest value in preovulatory follicle (0.55 ± 0.03 ng/ml). The differences were statistically significant for small and medium sized follicle only (P < 0.05). Testosterone values were negatively correlated with different stages of follicular development (r = -0.66). Ratios of different hormones were also analysed statistically. It revealed that there was a significant (P < 0.05) decrease in estradiol-ivp/progesterone ratio in follicular fluid as the follicle increased in size except small and medium size follicle. The highest ratio of estradiol-17β/progesterone (2.66 ± 0.53) observed in small sized follicle whereas the lowest ratio (0.07 ± 0.01) was observed in anovulatory follicle. Estradiol-lyp/progesterone ratio had significant and negative correlation with follicular development (r = -0.99). The differences of progesterone/testosterone ratio for small (0.63 ± 0.01), medium (6.87 ± 0.34) and large (26.08 ± 0.05) sized follicle were statistically nonsignificant but these ratio differed significantly from that of preovulatory (228.07 ± 14.78) and anovulatory (982.43 ± 87.18) follicle. Progesterone/testosterone ratio correlated positively with follicle development (r = 0.85). The results revealed that there was progressive increase in estradiol- 17β/testosterone ratio in follicular fluid as follicle developed. Lowest value of estradiol- 17β/testerone ratio was observed in small (1.67 ± 0.19) sized follicle, while highest value of ratio was observed in anovulatory follicle (69.51 ± 4.88). Ratio of estradiol- 17βand testosterone was correlated significantly (P < 0.05) and positively with the different categories of follicles (r = 0.93). The studies on biochemical constituents indicated that the differences observed in cholesterol concentrations in follicular fluid of different category of follicle were statistically significant (P < 0.05). Follicular fluid cholesterol levels showed significant and positive correlation with various stages of follicular development (r = 0.90). The variation of total protein content of follicular fluid of different categories of follicle was found statistically significant (P < 0.05). Average total protein content ranged from 3.13 ± 0.20 gm/dl in small sized follicle to 12.59 ± 0.31 gm/dl in anovulatory follicle. Follicular fluid protein concentration correlated positively but nonsignificantly with different stages of follicle development (r = 0.80). Overall, it was found that follicular fluid concentration of steroid hormones and biochemical parameter has important relative association with the physiology of follicle and oocyte development, their maturation and ovulation.ThesisItem Unknown STUDY ON BLOOD BIOCHEMISTRY IN RELATION TO MILK PROTEIN WITH SPECIAL REFERENCE TO KAPPA CASEIN IN INDEGENOUS GOAT(AAU, Anand, 2010) PATEL, SANJAYKUMAR B.; Pande, A. M.Caprine casein genes exiiibit an unusual and extensive polymorphism that affects milk quality and composition. The Kappa casein gene includes 5 exons, 4 of them carrj'ing more than 90 per cent of the information to encode for the mature protein. Caseins comprise about 80 per cent of the total protein content of milk and present polymorphism with changes in the amino acid sequence. Within this abundance of proteins. Kappa casein is noteworthy, since it has been associated with differences in milk yield, composition and processing. The objective of this study was to observe the existence of polymorphism in the Kappa casein gene and to compare the levels of plasma and milk biochemical parameters in lactation stage of Zalawadi goats. For this purpose, blood samples from 50 lactating goats, collected with vacutainer by needle puncture of the jugular vein. Plasma separated from blood and cellular parts (WBCs) was used for genomic DNA extraction. Milk samples were collected in glass test tube (50 ml capacity) v^ith boric acid preservative and milk samples were preserved at -20 °C till laboratory analyses. The milk biochemical profiles were estimated by using the ECOMILK analyzer and milk urea nitrogen and total casein were estimated by Auto method and kjeidahl method, respectively. The blood biochemical profiles were estimated by using the BS 120 CHEMISTRY analyzer. Plasma sodium and potassium were measured by Flame photometer. DNA was extracted from cellular part of blood. Goat Kappa Casein (CSN3) gene (exon 4) was amplified from genomic DNA samples by PCR. The PCRs were carried for the amplification of Kappa casein {CSN3) gene with specific primers (F: - 5' TCC CAA TGT TOT ACT TTC TTA ACA TC 3') and (R: - 5' GCG TTG TCC TCT TTG ATG TCT CCT TAG 3') gene loci. Amplified products were visualized as a single compact band of expected size of 645bp under UV light by gel documentation system. PCR products and restriction enzymes were incubated overnight in water bath at 37 °C for digestion with Haelll and Alw441 RE and at 65 °C for digestion with BseNI RE. Digested products were visualized as per restriction patterns with compact band size under UV light by gel documentation system. There were monomorphic restriction patterns with HaeIII-RFLP digestion, whereas polymorphic restriction patterns shown with BseeNI-RFLP and Alw44I-RFLP. In this study, genotypes AA, AG, GG ware observed with BseNI and CC, CT, TT with Alw44I in these animals except HaeIII RFLP. The frequency of allele A and allele G was 0.87 and 0.13, respectively and the frequency of allele C and allele T was 0.88 and 0.12, respectively. There were non-significant and positive correlations between blood glucose with milk fat, plasma total proteins with milk protein and milk casein and plasma total cholesterol with milk fat. There was non-significant and negative correlation between plasma triglycerides with milk fat. There was highly significant (P<0.01) and positive correlation between blood urea nitrogen and milk urea nitrogen.