The dead and damaged spermatozoa present in the fresh ejaculates are carried passively along with the live ones into the processed semen. Such dead and damaged spermatozoa are primarily responsible for the production of reactive oxygen species (ROS). The detrimental effect by way of oxidative stress on the contemporary live spermatozoa results in poor fertility (Roca, Martinez-Alborcia, Gil, Parrilla, & Martinez, 2013). Since the buffalo spermatozoa show high lipid peroxidation (LPO) rate (Nair, Brar, Ahuja, Sangha, & Chaudhary, 2006), comparatively reduced activity of existing antioxidant system (Nair et al., 2006), high content of membrane polyunsaturated fatty acids (Dawra, Sharma, & Makkar, 1983) and increased susceptibility to osmotic stress the damage owing to oxidative stress is exaggerated (Kadirvel, Kumar, & Kumaresan, 2009). The nuclear integrity as well as plasma membrane fluidity is also affected by LPO is another consequence of concern (Roca et al., 2013). In live spermatozoa, production of free radicals is of mitochondrial origin, mostly for the physiological processes. However, the imbalance of free radicals arise either due to excessive production intracellularly under physiological situations

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.

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Background: Oxidative stress is one of the leading factors responsible for poor postthaw 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