Sarvajeet Yadav

Effect of zinc and selenium supplementation on antioxidative status of seminal plasma and testosterone, T4 and T3 level in goat blood serum

The present study was conducted to investigate the effect of dietary supplementation of zinc (Zn) and selenium (Se) on antioxidative status and testosterone, T4 and T3 levels in 12 bucks aged between 2 and 4 years, weighing 25–35 kg. The experimental animals were divided in control and test groups containing six animals each. The animals of the control group were not given any supplementation, whereas test group were given supplementation of Zn and Se in the form of 150-ppm zinc sulphate and 0.50-ppm sodium selenite, respectively, for 105 days. Zn and Se levels in serum and seminal plasma, testosterone, T4 and T3 levels in serum and antioxidative status of seminal plasma were estimated at 0, 60, 75, 90 and 105 days of the experiment. After 60 days of supplementation, concentration of Zn and Se in seminal plasma and serum increased significantly (P<0.05) as compared to control. After 60 days, superoxide dismutase (SOD), glutathione peroxidase (GSHPx), glutathione reductase (GSHR) and catalase (CAT) activity in seminal plasma of test group increased significantly (P<0.05), whereas lipid peroxidation (LPO) decreased significantly (P<0.05) as compared to control. After 60 days, serum testosterone, T4 and T3 concentration in test group increased significantly (P<0.05) as compared to control. It may be concluded that Zn and Se supplementation can improve the antioxidative status and hormone levels by increasing the Zn and Se level in seminal plasma and serum, respectively.

Effect of misting and wallowing cooling systems on milk yield, blood and physiological variables during heat stress in lactating Murrah buffalo

BackgroundHeat stress adversely affects the physiological and metabolic status, and the productive performance of buffalo.MethodsThe present study was conducted to explicate the effect of misting and wallowing cooling strategies during heat stress in lactating Murrah buffalo. The study was conducted for three months (May–July) of which first two months were hot dry and last month was hot humid. Eighteen lactating buffaloes, offered the same basal diet, were blocked by days in milk, milk yield and parity, and then randomly allocated to three treatments: negative control (no cooling), cooling by misting, and cooling by wallowing.ResultsThe results showed higher (P < 0.05) milk yield in buffaloes of misting and wallowing group compared to control during the experimental period however wallowing was found more (P < 0.05) effective during July (hot humid period). Both the treatments resulted into significant (P < 0.05) reduction in rectal temperature (RT) and respiratory rate (RR) compared to control animals during study period whereas wallowing was found to be effective on pulse rate (PR) only during July. Both treatments were resulted in mitigating the heat stress mediated decrease in packed cell volume (PCV), lymphocytopnoea and neutrophilia whereas decrease in total erythrocyte count (TEC) and monocytes was only mitigated by wallowing. Heat load induced alteration in serum creatinine and sodium concentration was significantly (P < 0.05) ameliorated by misting and wallowing whereas aspartate aminotransferase, alkaline phosphatase and superoxide dismutase activity, and reactive oxygen species concentration could be normalized neither by misting nor by wallowing. The significant (P < 0.05) increment in serum cortisol and prolactin levels observed in June and July period in control animals was significantly (P < 0.05) prevented by misting and wallowing.ConclusionsIt can be concluded that misting and wallowing were equally effective in May and June (hot dry period) whereas wallowing was more effective during hot humid period in preventing a decline in milk production and maintaining physiological, metabolic, endocrine and redox homeostasis.

Effect of reduced glutathione supplementation in semen extender on tyrosine phosphorylation and apoptosis like changes in frozen thawed Hariana bull spermatozoa

To provide new insights into the mechanisms through which reduced glutathione (GSH) is able to protect spermatozoa, we tested the hypothesis that cryocapacitation and apoptosis like changes can contribute to the negative effect of freezing and thawing on bull spermatozoa, and that GSH prevent this damage. Having known protective effects of GSH in terms of a potent antioxidant, we evaluated capacitation, tyrosine phosphorylation and apoptosis like changes in bull spermatozoa after freezing and thawing in egg yolk tris glycerol extender containing (0.5m M-GSH-T1 & 1mM GSH-T2) and without GSH serving as the control (C). Forty ejaculates were collected from four Hariana bulls and were pooled due to non significant variations among the bull ejaculates for the evaluation of sperm attributes. Capacitation like changes, tyrosine phosphorylation, localization of tyrosine phosphorylated proteins, apoptosis like changes in terms of mitochondrial transmembrane potential and DNA fragmentation after final dilution, 4h of equilibration at 4°C and 24h after freezing and thawing were evaluated. GSH supplementation at 0.5mM showed significant reduction in B- and AR- pattern spermatozoa during all stages of semen freezing and thawing. Immunoblot revealed six proteins which were tyrosine phosphorylated and protein of 30 and 75kDa (p30, p75) were the major tyrosine phosphorylted proteins. On further analysis, the p30 showed differential variation in intensity in all the three groups after freezing and thawing. Positive immune reactivity for tyrosine phosphorylated proteins was found in neck, middle piece and post-acrosomal regions of spermatozoa. Addition of 0.5mM GSH decreased percentage of spermatozoa showing fragmented DNA and increased the percentage of spermatozoa having high transmembrane mitochondrial potential (P<0.05). This study demonstrates that GSH favours survival of bull spermatozoa by interfering with apoptotic and cryocapacitation pathways, and thereby protects the spermatozoa from deleterious effects of cryopreservation. The findings of the study indicated that GSH at 0.5mM can be effectively used as an additive in bull semen extender for freezing and thawing.

Effect of zinc and selenium supplementation on semen quality of Barbari bucks.

The present study was conducted to investigate the effect of supplementation of zinc and selenium on semen quality in twelve Barbari bucks of proven fertility. The experimental animals were randomly divided into two groups of six and fed without any supplementation (control) or with 150-ppm zinc sulfate and 0.50-ppm sodium selenate (test group). Semen was collected on days 0, 60, 75, 90 and 105 to study its characteristics. In control group, no significant change was observed in semen quality during the study period. In test group, semen quality was improved in terms of a significant (P< 0.05) increase in semen volume, progressive motility, sperm count, percent live spermatozoa, acrosomal integrity and hypo osomotic swelling (HOS) responding spermatozoa and decrease in abnormal spermatozoa after 60 days of supplementation. Mass motility increased significantly (P< 0.05) only at 105 days as compared to 0 day. It is concluded that the supplementation of Zn and Se can improve semen quality in goats.

The functional dynamics of neutrophils during different seasons in zebu cattle

Abstract The present study was conducted in Hariana and Sahiwal cows to study neutrophil functional dynamics during different seasons (summer, rainy and winter) in terms of expression of toll like receptor 2 & 4, neutrophils’ apoptosis in terms of caspase 3, 7 activities, and transmembrane mitochondrial potential. Blood samples were collected from Hariana (n = 10) and Sahiwal cows (n = 10) throughout the year to evaluate the effects of season on neutrophil dynamics. In each season, eight blood samples were collected from each animal (twice in each month). Toll-like receptor 2 and 4 expressions were significantly (p < 0.05) decreased in neutrophils during summer season as compared to rainy and winter seasons. Significantly, (p < 0.05) higher caspase 3 & 7 activities and significantly higher per cent of neutrophils with low transmembrane mitochondrial potential indicating its enhanced apoptosis were found during extreme summer and lowest during winter. Both Sahiwal and Hariana breeds exhibited similar trends in neutrophil apoptosis. The results of the study evidently indicate that neutrophil competence varies from season to season in zebu cattle and could determine the possible incidences of infections in these cows. Hot-humid season could be conducive for microbial infections owing to accelerated apoptosis of neutrophils and down regulation of TLR2 and TLR4 of immune cells of zebu cattle.

Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis.

Aim: The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. Materials and Methods: A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck). Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Results: Significantly (p<0.01) higher value of live percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (p<0.01) higher followed by slow and rapid progression. Conclusion: This study showed that the best results for motion, vitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats.

Evaluation of adaptability to different seasons in goat breeds of semi-arid region in India through differential expression pattern of heat shock protein genes

Abstract The present study was conducted to examine differential expression pattern of HSP genes and adaptability in Indian goat breeds of semi-arid region. The study was conducted in five animals from each breed viz. Barbari, Sirohi, and Jhakrana during winter, thermo-neutral and summer seasons. The respiratory rate (RR) and rectal temperature (RT) of the goats were recorded at 09:00 h during the study period. The blood samples were collected for RNA isolation, cDNA synthesis, and quantitative analysis of HSP genes expression by quantitative RT-PCR. The RR increased significantly (p < 0.01) during summer as compared to winter and thermo-neutral season however, RT did not change (p > 0.05) during different seasons. The expression of HSP genes was significantly (p < 0.01) increased during summer (high THI) as compared to thermo-neutral season in all the goat breeds. Among HSPs, only HSP90 was upregulated (p < 0.01) in Jhakrana goats during winter as compared to thermo-neutral season. The deviation in expression of HSP genes during summer and winter with respect to thermo-neutral season was minimum in Barbari goats. Therefore, it can be concluded that Barbari goats possessed better adaptability during summer and winter as compared to Sirohi and Jhakrana goats in semi-arid climatic conditions of India.

Effect of egg yolk concentration and washing on sperm quality following cryopreservation in Barbari buck semen

ABSTRACT The present study was conducted to evaluate the effect of egg yolk concentration and washing on sperm quality in cryopreserved Barbari buck semen at higher dilution (100 × 106 cells/ml). Five healthy Barbari bucks of similar age and weight were selected as semen donor. Six ejaculates were collected from each buck during the experiment. Collected semen samples were pooled and diluted with Tris-based semen extender containing 6% glycerol and egg yolk according to the treatments – T-1, that is, 3% egg yolk, T-2, that is, 20% egg yolk and T-3, that is, 20% egg yolk after washing and cryopreserved using the slow freezing technique. Semen was evaluated at three different steps, namely, after dilution, after equilibration and after freeze–thawing. Significantly (p ≤ .01) higher values for motility and viability were observed in T-3 just after dilution while significantly (p ≤ .01) higher values for acrosomal integrity and pattern F (uncapacitated sperm) was observed in T-2. Significantly (p ≤ .01) higher values were observed in T-2 after equilibration and thawing. It may be concluded that incorporation of 20% egg yolk compared to 3% egg yolk or 20% egg yolk after washing in extender confer better cryoprotection to Barbari buck spermatozoa as reflected in semen quality parameters studied.

Effect of glutamine supplementation and replacement of tris-egg yolk based extender with defatted cow milk on spermatozoa quality after equilibration and thawing

Aim: The present study was designed to evaluate the effect of supplementation of glutamine and replacement of Tris-egg yolk (TE) based buffer with defatted cow milk on the spermatozoa quality after equilibration and thawing. Materials and Methods: Semen was collected from five Bhadawari bulls biweekly, and a total of 30 ejaculates were taken. The semen ejaculates were pooled and divided into three equal parts. The pooled semen was diluted by TE based extender (control), TE + glutamine (8 mM) (T1) and 50% TE + 50% deffated cow milk + glutamine (8 mM) (T2). At two stages viz. after equilibration and after 12 h of cryopreservation (thawed samples), progressive motility, percent live, and percent acrosomal damage of the spermatozoa was assessed. Results: Supplementation of glutamine improved (p<0.05) the spermatozoa quality with respect to the progressive motility, percent live and acrosomal damage both post-equilibration and post-thaw. T2 improved (p<0.05) the spermatozoa quality as compared to control, however; it was less (p<0.05) effective as compared T1 both post-equilibration and post-thaw. Conclusion: From the results of present study it can be concluded that glutamine supplementation was effective in maintaining post-equilibration and post-thaw spermatozoa quality whereas defatted cow milk was not as effective as TE based buffer in the extender in improving the spermatozoa quality.

Insights into pH regulatory mechanisms in mediating spermatozoa functions

Regulation of pH in spermatozoa is a complex and dynamic process as sperm cells encounter different pH gradients during their journey from testes to the site of fertilization in female genital tract. The precise regulations of pH in sperm cells regulate the sperm functions such as motility, hyperactivity, capacitation, and acrosome reaction. Electrophysiological, pharmacological, and molecular studies have revealed the presence of different ion channels and exchanger systems which regulate intracellular pH in sperm cells as well as regulate sperm functions. Recent studies also have shown the potential involvement of pH in the regulation of fertility competence of sperm cells, and alterations in pH have shown to impede sperm functions. This mini-review discusses the probable mechanisms involved in pH regulation in sperm cells and how pH is involved in regulation of various sperm functions.