Genomic and Computational Analysis of Novel SNPs in TNP1 Gene Promoter Region of Bos indicus Breeding Bulls.

Transition nuclear proteins (TNPs), the principal proteins identified in the condensing spermatids chromatin, have been found to play a key role in histone displacement and chromatin condensation during mammalian spermatogenesis. One such gene belonging to the TNP family called TNP1 gene is abundantly expressed in the regulation of spermatogenesis, and its sequence is remarkably well conserved among mammals. Genomic analysis, by sequencing and computational approach, was used to identify the novel polymorphisms and to evaluate the molecular regulation of TNP1 gene expression in Sahiwal cattle breeding bulls. DNA samples were sequenced to identify novel single nucleotide polymorphisms (SNPs) in the TNP1 gene. Modern computational tools were used to predict putative transcription factor binding in the TNP1 promoter and CpG islands in the TNP1 promoter region. In the TNP1 gene, four SNPs, three TATA boxes, and one CAAT box were identified. One CAAT box was discovered at 89 bp upstream of start site ATG. The computational analyses indicated that the polymorphisms inside the promoter sequence results in an added HNF-1 transcription factor binding site. In contrast, the other variations may remove the naturally occurring SRF transcription factor binding site. The CpG islands in the TNP1 promoter region were predicted to be absent by the MethPrimer program before and after SNP site mutations. These findings pave the way for more research into the TNP1 gene's promoter activity and the links between these SNPs and reproductive attributes in the Sahiwal breeding bulls.

Antioxidant effect of Moringa olifera leaves extract in extender improves post-thaw quality, kinematics, lipid peroxidation, total antioxidant capacity and fertility of water buffalo bull semen.

Our objectives were to determine the antioxidant effect of Moringa olifera leave extract (MLE) in tris extender on post-thaw quality, kinematics, lipid peroxidation, total antioxidant capacity and field fertility of water buffalo bull semen. Semen was collected on a weekly basis for 5 weeks (n = 5 bulls & n = 25). After collection, every sample was diluted individually with extender (tris egg yolk) containing MLE concentrations (0%, 5%, 10%, 15% and 20%) at 37°C. After dilution, semen was filled in 0.54 mL straws and frozen in automatic freezer. After thawing, sperm motility (%) (progressive & total), path velocity (µm/s), straight velocity (µm/s), curvilinear velocity (%), beat cross frequency (Hz), linearity (%), straightness (%), length of average path (µm), length of straight-line path (µm), length of curvilinear path (µm), plasma membrane integrity (%), acrosome membrane integrity (%), DNA integrity (%) and morphology (%) were higher (p < 0.05) in spermatozoa preserved in extender containing 15% MLE as compared to control. The fertility rate (60%, vs. 45%) was higher (p < 0.05) with semen doses frozen with 15% MLE in extender than the control. In conclusion, 15% MLE improves semen quality, kinematics and field fertility in water buffaloes.

Addition of pomegranate juice (Punica granatum) in tris-based extender improves post-thaw quality, motion dynamics and in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa.

The aim of the present study was to determine the protective effects of pomegranate juice in tris-based extender on semen parameters, computer-assisted sperm analysis (CASA) motion characteristics and field fertility of post-thawed Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Two consecutive ejaculates/collection from each of the five adult Nili Ravi buffalo bulls were collected with artificial vagina at 42°C for a period of 7 weeks, diluted in extender containing different concentrations of pomegranate juice (0.0%, 2.5%, 5%, 7.5% and 10%). Diluted samples were packed and frozen in 0.54 ml French straws. The addition of 10% pomegranate juice in extender significantly improved post-thaw sperm morphology (%), motilities (CASA total motility, progressive motility (%) as well as VAP, VSL, VCL, STR, DAP, DSL) compared to the control group (p < 0.05). Plasma membrane, acrosome membrane and DNA integrity were significantly higher in extender with 10% pomegranate juice than the control group (p < 0.05). Field fertility rate (60.39% vs. 46.53%) was higher (p < 0.05) in extender with 10% pomegranate juice as compared to the control. It is therefore concluded that the addition of 10% pomegranate juice in tris-based extender improves post-thaw semen parameters, CASA motion dynamics and field fertility in Nili Ravi buffaloes.