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ThesisItem Open Access COMPARATIVE GENETIC CHARACTERIZATION OF PROLACTIN GENE BETWEEN GIRIRAJA AND INDIGENOUS CHICKEN(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) AJEET SINGH; NAVEEN KUMAR, S.The present study was undertaken to determine and compare the genetic variations in prolactin gene among Giriraja and Indigenous chicken using PCR-RFLP technique. Blood samples were collected from 80 each of Giriraja and Indigenous chicken from Department of Poultry Science, Veterinary College, Bengaluru. The genomic DNA was isolated by following the conventional Phenol: Chloroform: Iso-amyl alcohol (25:24:1) method as described by Khosravinia et al. (2007). A 130/ 154 bp and 439 bp sequences of promoter regions 1 (24 bp indel at -358) and 2 (C-2402T and C-2161G) of prolactin gene were successfully amplified employing published primers. At promoter region 1 (24 bp indel at -358) of prolactin gene, the proportions of DD and ID genotypes were higher in Giriraja (0.45) and Indigenous (0.44) chicken, respectively. However, the frequency of D allele was higher in both the studied populations. The restriction enzymes, AluI and Csp6I were utilized for genotyping polymorphisms of C -2402T and C -2161G, respectively. At C -2402T, TT and CT genotypes were predominant in Giriraja and Indigenous chicken with frequencies of 0.61 and 0.56, respectively. The frequency of T allele was higher in both the studied populations. Whereas at C-2161G, GG and CG genotypes were predominant in Giriraja (0.49) and Indigenous (0.65) chicken, respectively. In both the studied populations the frequency of G allele was higher. Further, three SNPs viz., C>T transition at 161 bp, T>C transition at 371 bp and C>G transversion at 402 bp were detected at promoter region 2 (C-2402T and C-2161G) in T/ G allele compared to C allele.ThesisItem Open Access SPERM DNA FRAGMENTATION AND SIRE CONCEPTION RATE IN HOLSTEIN FRIESIAN BULLS(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) SANTHOSH, G. R.; NAVEEN KUMAR, G. S.The present study was undertaken to know the association of sire conception rate with bull age, sperm phenotypic characters, sperm DNA fragmentation (SDF), protamine polymorphism and PRM1:PRM2 expression ratio in selected HF bulls maintained at State Semen Collection Center, Bengaluru, Karnataka. The overall mean sire conception rate in the study was 50.61 ± 1.17%. Group G-I (Bulls aged < 3 years) had significantly lower sire conception rate (45.0 ± 0.01%), than G-II (3-5.5 years) and G-III (> 5.5 years) with sire conception rates of 50.61 ± 1.17% and 53.0 ± 0.017% respectively. The Single Nucleotide Polymorphism (SNP) analysis of Protamine1 gene revealed presence of six different SNPs in the 5’ promoter region and no SNPs in the exonic and intronic region of Protamine 1 gene. The SNP in TATA box TT > TG (TATATAA), 30 bp upstream from exon 1 was found to be related to the low expression of Protamine1 and hence a lower sire conception rate. No differences in semen and sperm phenotypic traits were observed among bulls of different age group. The average PRM1 to PRM2 ratio and DNA Fragmentation Index% (DFI) observed in present study were 1:0.06 and 8.88 ± 0.22% respectively. Significant lower expression levels of PRM1 was observed in younger (GI) bulls. DFI% was higher in younger aged bulls (9.2 ± 0.39) than aged bulls (8.6 ± 0.36) but not significantly different. DFI% was negatively correlated with PRM1 (r = -0.660) and PRM2 (r = -0.518) expression levels. Sire conception rate was negatively correlated with DFI% also and positively correlated with PRM1 expression (0.564) and Protamine ratio (0.581). However, PRM2 was found to be not significantly correlated with sire conception rate. Present study revealed that lower protamination is a probable reason for lower sire conception rate in younger bulls.
