Aim:An experiment was designed to evaluate the role of Vitamin E and glutathione in improving the seminal parameters during hypothermic storage of liquid semen at 4°C for 72 h.Materials and Methods:Thirty-six semen ejaculates were collected by artificial vagina from 6 bucks (Beetal) during the normal reproduction season (September to November) at weekly interval. The samples were centrifuged, and the seminal plasma was removed. The sperm pellet was diluted with Tris-based extender and divided into three groups. Group T1: Control samples without antioxidants, Group T2: Samples supplemented with tocopherol at 3 mM, and Group T3: Samples supplemented with glutathione at 1 mM. The samples were evaluated for progressive motility, percent liveability, percent abnormal spermatozoa, and acrosome integrity after liquid preservation for 0, 24, 48, and 72 h. The level of lipid peroxidation and antioxidant enzymes, namely, glutathione peroxidase (GPx) and superoxide dismutase (SOD) were estimated after liquid preservation for 0 and 72 h.Results:It was observed that, after storage of semen at 4°C up to 72 h, the progressive sperm motility, percent liveability, percent abnormal spermatozoa, and percent intact acrosomes were significantly (p<0.05) higher in group T2 and T3 samples as compared to control. However, the level of lipid peroxidation in T2 and T3 groups was significantly (p<0.05) lower after 72 h of incubation at 4°C. Similarly, GPx and SOD values were significantly (p<0.05) increased in T2 and T3 groups after 72 h of storage at 4°C.Conclusion:Thus, it can be concluded that Vitamin E and glutathione supplementation at 3 mM and 1 mM, respectively, while preserving the semen samples at 4°C helped in maintaining the seminal parameters up to 72 h and protected the spermatozoa from oxidative damage.

Background and Aim:Probiotics are the living microorganism which when administered improves the digestion and health of the animal. Saccharomyces cerevisiae (SC) improves the humoral and innate immunity of the animal. Prilled fat is a hydrogenated palm oil triglyceride which has been reported to promote the release of cytokines from macrophages. The aim of the study was to evaluate the immunomodulatory effect of probiotic and prilled fat during transition stage in Karan Fries (KF) cows.Materials and Methods:A total of 12 KF cows at 21 days prepartum were selected and divided into two groups of six animals each. The control group was fed as per the standard feeding practices and the supplemented group cows were supplemented daily with prilled fat at 100 g/cow, SC at 25 g/cow, and sweetener at 1 g/cow in addition to the standard feeding practices from −30 days of prepartum to 21 days of lactation. The sweetener was added to improve the palatability of the feed. The natural sweetener of an African plant leave had 105 times more sweetness than glucose with good aroma. The dry matter intake of the animal was recorded. Plasma samples were collected weekly from all cows for the analysis of blood metabolite beta-hydroxybutyric acid (BHBA). Lymphocytes were isolated from the blood for studying the expression of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) and for estimating lymphocyte proliferation index (LPI).Results:The upregulated IL-1β and TNF-α around calving might be possibly associated to the metabolic changes occurring during the transition period and suggest a higher degree of inflammation around parturition. High concentrations of BHBA caused increased expression and synthesis of the pro-inflammatory factors such as TNF-α and IL-1β in supplemented group in primary calf hepatocytes. The LPI was higher in supplemented group as compared to control which suggests a stimulatory effect of unsaturated fatty acids on mitogen-stimulated T-cell proliferation.Conclusion:Dietary supplementation of probiotics, prilled fat, and sweetener alleviated negative energy balance by stimulating feed intake and modulating hepatic lipid metabolism; and both of these additives improved the postpartum health (antioxidant status and immune function) of transition dairy cows.

Aim:This study was conducted with the objective to identify single nucleotide polymorphism (SNP) in protein phosphatase 1 regulatory subunit 11 (PPP1R11) gene in Murrah bulls.Materials and Methods:Genomic DNA was isolated by phenol–chloroform extraction method from the frozen semen samples of 65 Murrah bulls maintained at Artificial Breeding Research Centre, ICAR-National Dairy Research Institute, Karnal. The quality and concentration of DNA was checked by spectrophotometer reading and agarose gel electrophoresis. The target region of PPP1R11 gene was amplified using four sets of primer designed based on Bos taurus reference sequence. The amplified products were sequenced and aligned using Clustal Omega for identification of SNPs. Animals were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using EcoNI restriction enzyme.Results:The sequences in the NCBI accession number NW_005785016.1 for Bubalus bubalis were compared and aligned with the edited sequences of Murrah bulls with Clustal Omega software. A total of 10 SNPs were found, out of which 1 at 5’UTR, 3 at intron 1, and 6 at intron 2 region. PCR-RFLP using restriction enzyme EcoNI revealed only AA genotype indicating monomorphism in PPP1R11 gene of all Murrah animals included in the study.Conclusion:A total of 10 SNPs were found. PCR-RFLP revealed only AA genotype indicating monomorphism in PPP1R11 gene of all Murrah animals included in the study, due to which association analysis with conception rate was not feasible.

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