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The present experiment was conducted to study the impact of herbal treatment on post thaw semen quality and fertility in subfertile buffalo bulls. Three Murrah subfertile buffalo bulls maintained at bull station, Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, India (Latitude/Longitude, 30.55°N, 75.54° E) with the history of poor semen quality (pre-freeze individual motility < 60%, post-thaw individual motility < 40%, Viability < 70% and Abnormality > 20%) were orally supplemented with herbal mixture (Panax ginseng roots, Shilajit, Withania somnifera roots, Tribulus terrestris fruits, Turnera diffusa leaves, Ptychopetalum olacoides bark of each 400 mg/100 kg body weight and 300 mg/100 kg body weight of Pausinystalia yohimbe bark) daily for 60 days of treatment phase. Two semen ejaculates per week per bull were collected during pre-treatment, treatment and post-treatment phases of 60 days each and extended with Tris egg yolk extender. The freezing of extended semen was carried out in a biofreezer (4°C to -15°C @-30°C/min, -15°C to -140°C @-50°C/min) followed by plunging into liquid nitrogen for storage. Post thaw semen quality was assessed in terms of individual motility, viability, total sperm abnormalities. Present study revealed that the oral treatment of subfertile buffalo bulls with Panax ginseng roots, Shilajit, Withania somnifera roots, Tribulus terrestris fruits, Turnera diffusa leaves; Ptychopetalum olacoides bark each @ 400 mg/100 kg body weight and Pausinystalia yohimbe bark @ 300 mg/100 kg body weight for 60 days improved post thaw semen quality and fertility. [ABSTRACT FROM AUTHOR]

The quality of spermatozoa at post-thaw stage depends on many factors including the level of lipid per-oxidation (LPO) and the antioxidant profile of the ejaculates at fresh stage. Thus, the present investigation was done to evaluate discard rate of semen ejaculates at fresh and post-thaw stages in association with various semen quality parameters in Murrah buffalo. A total of 79 ejaculates from seven Murrah buffalo bulls formed part of the study. Following application of rigorous prescribed selection criteria it was observed that 17.72% (14/79) ejaculates were discarded at fresh stage due to poor quality. The initially selected ejaculates were processed for cryopreservation as per the minimum standard protocols. The study show satisfactory results for 31 (47.69%) samples for inclusion in the further AI programme (discard rate of 52.31% (34/65) at post thaw. The mean post-thaw motility (PTM; %) of the passed ejaculates was found to be 44.2±1.0. In conclusion, study results show efforts to minimize LPO and enhance total antioxidant capacity (TAC) at various processing stages shall play a vital role in reducing discard rate of the semen samples in buffalo bulls. [ABSTRACT FROM AUTHOR]

Background: Oxidative stress is one of the leading factors responsible for poor post‐thaw semen quality because of overproduction of reactive oxygen species (ROS) over neutralizing antioxidants present in semen. Mainly two ROS generation sites are present in spermatozoa, that is, mitochondria and plasma membrane. Therefore, the idea of targeting these specific sites for minimization of ROS production with the compounds having known mechanism of actions was built up as a core for this research. Objective: Present study was done to investigate the effects of Mito TEMPO and acetovanillone individually and in combination on freezability of buffalo spermatozoa. Materials and Methods: For the experiment, semen extender was supplemented with Mito TEMPO (50 μM), acetovanillone (50 μM), and a combination of Mito TEMPO + acetovanillone (50 μM+ 50 μM), designated as Group II, Group III, and Group IV, respectively. Control group without any supplementation was designated as Group I. A total of 24 ejaculates with individual progressive motility (IPM) of ≥70% were selected for the study. After final dilution, filling‐sealing of straws, equilibration, and freezing were done as per the standard procedure. Semen samples were evaluated for IPM, plasma membrane integrity, lipid peroxidation, total antioxidant capacity (TAC), and cholesterol to phospholipids (C/P) ratio at both fresh and post‐thaw stages. Evaluation of ROS, mitochondrial membrane potential (MMP), capacitation status (CTC assay), and in vitro fertility potential were conducted only on frozen‐thawed samples. Results: The addition of Mito TEMPO (50 μM) and acetovanillone (50 μM) individually and in combination significantly (p < 0.05) improved post‐thaw semen quality in terms of IPM, plasma membrane integrity, TAC, cholesterol content, C/P ratio, MMP, Chlortetracycline (CTC)‐Full (F) pattern, and zona binding ability of buffalo spermatozoa, while significantly (p < 0.05) reduced ROS production, lipid peroxidation, and capacitation like changes as compared to the control group. Discussion: As Mito TEMPO acts as an SOD mimetic and also detoxifies ferrous iron at the mitochondria level, it aids in neutralization of excessive ROS production and minimizes oxidative stress‐related damages that enhances the antioxidant potential of sperm mitochondria. Earlier studies also indicated improved post‐thaw semen quality in 50 μM supplemented group. The improvement observed in acetovanillone (50 μM) group might be because of inhibition of Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase as this enzyme activation by various physical/chemical inducers during cryopreservation process leads to activation of CatSper channel resulting in calcium influx, premature capacitation, and acrosomal reaction like changes through activation of adenylate cyclase and cAMP/PKA‐mediated tyrosine phosphorylation of sperm proteins. Acetovanillone also prevents NADPH oxidase–mediated inhibition of glutathione reductase activity, which has a vital role in protecting the structural and functional integrity of sperm plasma membrane. Conclusion: Results indicated beneficial effects of supplementation of Mito TEMPO and acetovanillone on sperm freezability and individual supplementation was as efficient as the combination group for sustaining post‐thaw semen quality. [ABSTRACT FROM AUTHOR]

The synthesis of iron oxide nanoparticles (IONPs)–antiubiquitin antibodies (Abs) complex for depletion of dead/damaged spermatozoa from buffalo semen was done. The IONPs synthesized were round in shape with size of 12.09 ± 0.91 nm. At the end of the two‐step functionalization, that is, silanization and pegylation of bare IONPs and bioconjugation of functionalized IOPNs, particles with the sizes of 19.15 ± 1.46, 20.72 ± 0.95, and 73.01 ± 7.56 nm, respectively, were obtained. Twenty‐four semen samples from four bulls with mean individual progressive motility (%) and sperm concentration (million/mL) of 77.1 ± 0.9 and 1,321.2 ± 84.7, respectively, were divided into Group I (control), and treatment groups viz. Groups II, III, and IV; with each group containing 150 ± 25 million dead/damaged spermatozoa. The IONPs–Abs complex was added at the ratio of 1:1 (0.5 μg/mL), 1:2 (1.0 μg/mL), and 1:4 (2.0 μg/mL), respectively, in the Groups II, III, and IV. The mean efficiency (%) of nanopurification was estimated to be greater in nanopurified semen with the increasing doses of the IONPs–Abs complex. A reduction of 29.3 ± 6.4%, 48.4 ± 5.3%, and 55.4 ± 4.4% in dead/damaged spermatozoa following nanopurification in Groups II, III, and IV, respectively, was observed. The study shows that in‐house synthesized IONPs–Abs complex can be successfully used to deplete dead/damaged spermatozoa from buffalo semen with improvement in quality. [ABSTRACT FROM AUTHOR]