Background: Sperm selection method is usually used to collect these cells for in vitroassisted reproduction. Few studies reported the relationship of in vivo fertility and semen parameters after sperm selection; hence, the present study attempted to assess different semen parameters after post-thaw or sperm selection, using density gradient separation BoviPure®, to predict in vivo fertility.Materials and Methods: In this experimental study, frozen semen quality of four Montbeliarde bulls were assessed after post-thaw (PT) or after sperm selection (SSp), using density gradient separation BoviPure®, to predict the fertility rate in vivo. In addition to PT or SSp, semen was examined for concentration, motility, morphology abnormalities, viability, acrosome and plasma membrane integrities. Fertility was measured as non-return rates within 56 days after the first insemination (NRR) or as corrected NRR, expressed as CNRR, to the factors influencing fertility using linear mixed model. Non-parametric Kruskal-Wallis test was performed to compare semen parameter variables. Fertility rates were compared using Chi-square test. Pearson correlation analysis was used to test the relationship between CNRR and semen parameters. Data was analysed using SPSS package program, version 21.0.Results: Most of the examined bulls exhibited a high fertility rate (3/4 bulls, 62.1-81.8% for NRR or 67.2-98.5% for CNRR). Fertility rate, expressed as CNRR, was significantly related to semen parameters after SSp, but not after PT. Thus, CNRR was increased with decrease of total motility, progressive spermatozoa and abaxial implantation frequencies after SSp (r=-0.999, P=0.001; r=-0.990, P=0.010; r=-0.988, P= 0.012, respectively); while, CNRR was decreased with decrease of SSp immotile spermatozoa (r=+0.995, P=0.005), underlying that maximal limit of determined immotile spermatozoa is 47%.Conclusion: High frequencies of total and progressive motility spermatozoa, and abaxial implantation in gradient selected sperm appear to be not favorable for fertility in vivo.

Oxygen metabolites as reactive oxygen species (ROS) are considered as major factors in male infertility. In the present investigation, we aimed to study the effect of different concentrations of ascorbate/vitamin C (300-2000 micromolar), on membrane integrity, acrosome reaction, and viability of bull (Holstein) spermatozoa in presence of iron (ferrous) ions as lipid peroxidation (LPO) promoter. LPO is one of the manifestations of ROS attack in biological membranes. LPO impinges on membrane integrity, triggers inactivation of membrane bound enzymes which involve in sperm motility. Ascorbate in concentrations below 1000 micromolar protects spermatozoa from free radical damages as evidenced from improvement in their membrane integrity (data of LPO test), elevated acrosome reactions, and increased viability. Concomitantly, there is also witnessed depletion of malondialdehyde (MDA) (an end product of LPO) generation following ascorbate supplementation. Ascorbate at 1000 micromolar concentration and above, however, is not protective, as evidenced by abrupt fall in sperm membrane integrity and rate of acrosome reactions and lowered % live sperm cells. Collectively, ascorbate acts as a double-edged sward in different concentrations and researchers must keep in mind this negative side of ascorbate application in their research fields like in vitro fertilization, cell culture, and preservation of gametes to minimize its deleterious impacts on biological systems.

Many in vitro methods of semen quality evaluation have been developed for predicting fertility of bull semen in routine AI practice. Conventional semen evaluation has some limitations due to the difficulty to detect some functional sperm cell impairments, which are responsible for decreased fertility (Aitken, 2006). Bavister (1990) and Rowe Semen quality and its relationship to fertility are major concern in animal production. The aim of this study was to assess the relationship between frozen thawed sperm characteristics and fertility following in vivo fertility through Artificial Insemination (AI). Semen samples were collected from four buffalo bulls. Semen volume and sperm concentration appeared to be significantly different (P<0.05) among bulls, while sperm motility and live sperm percentage did not vary in fresh semen. Frozen thawed semen was evaluated for motility, viability, sperm abnormalities, membrane integrity and in vivo fertility. A significant variation was found in semen parameters among bulls after thawing. Highly significant (P<0.001) differences in membrane integrity between fresh vs. frozen semen samples were noticed. Pregnancy rate was significantly (P<0.05) different among bulls. Significant correlations were found between motility and sperm abnormalities (r=-0.64; P<0.05) and membrane integrity (r=0.64; P<0.05). A significant negative correlation (r=-0.73; P<0.01) has been reported between sperm abnormalities and membrane integrity. In addition, a correlation between pregnancy rate and live sperm percent-ages (r=0.65; P<0.05), has also been reported. In conclusion, in this study motility was correlated with sperm abnormalities and membrane integrity. Live sperm percentage was the only parameter correlated with fertility. Motility and live sperm percentage can be used as a predicative measure in semen evaluation.

Aim and Background: Silver nano particles (NPs) or nano silver (NS) are clusters of silver atoms that range in diameter from 1 to 100nm and are attracting interest as antibacterial and antimicrobial agents of applications in medicine. Resistance to antimicrobial agents by pathogenic bacteria has emerged in recent years and is a major reason for replacing of antibiotics with nano silver, but yet nanoparticle hazardous and toxic effects have not been studied extensively.Materials and Methods: One of the agent of semen extenders is antibiotics. We replaced antibiotics with nano silver and investigated mobility, progressive of sperm. Also result of nano silver on the growth of semen’s bacteria was studied and them was compared with penicillin and streptomycin in similar conditions.Results: Semen was diluted with citrate sodium, Yolk extender and was divided into nine portions. The control (sham) semen extender solution contained extended semen and no antibiotic and nano silver, whereas streptomycin and penicillin solutions contained extended semen plus 1.0 mg/mL of streptomycin and penicillin. Also semen was diluted with Citrate sodium, Yolk extender supplemented with different concentrations of silver nano particle. Here, the effects of different concentration of silver NPs on bull sperm at frequent times after1, 3 hours was evaluated. Therefore, the Mean percentages of motile and progressively motile sperm did not differ significantly among control (sham) and antibiotic and nano silver -containing solutions after 1 hour. but after 3 hour, nano silver was cytotoxic on bull sperm cells in more than 0.1mL concentration .(p<0.001).Conclusion: Control extended semen samples from ejaculates of bulls were contaminated with gram pasitive bacteria. Samples of antibiotic and concentration of 0.005mL, 0.01mL nano silver had growth a few of bacteria. Nano silver has good antibacterial properties in semen extender in more than 0.05mL concentration. Thus, It can be suggested that antibiotics may be replaced with nano silver as antimicrobial agent in semen extenders in Some few concentration.

Aim: The purpose of this study was to evaluate the effects of sub-lethal osmotic stress (325, 350, 375 and 400 mOsm) during cryopreservation in commercially diluter medium (Bioexcell) on Holstein mail cow sperm qualitative characters after freezing-thawing.Material and Methods: Semen was collected from four Holstein mail cows using artificial vagina two times for a week. All of obtained semen mixed together and then were divided into five equal parts. Each part was freeze-melted according to the experimental treatments consisting of different osmotic stresses. 300 mOsm treatment medium was applied as control group. Sperms motility and progressive motility were assessed by computer assessment semen analysis. Also sperms viability, membrane integrity, mitochondria activity and membrane lipid peroxidation of frozen-thawed semen were assessed using Eosin-Nigrosin, hypo osmotic swelling test, Hankok, Rhodamin 123 and TBA procedures, respectively.Results: Results showed that 375 mOsm osmotic stress treatment had the most significant improvement of motility%, progressive motility and viability in comparison with other treatments. Also mitochondria activity in 350 and 375 mOsm osmotic pressures treatments was significantly higher than other experimental treatments (P<0.05). Different osmotic stresses treatments did not show significant effect on sperm morphology and membrane lipid proxidation.Conclusion: It is seemed that mild and sub-lethal osmotic stresses induction in cow sperm freezing diluters can significantly improved some of sperm qualitative characters such as their motility and viability.

This paper studied a potential hazard of titanium dioxide nanomaterials in the primary sreening methods on the sample of bull sperm. A method for stabilizing metal nanopowders and their derivatives for biomedical studies is proposed. It was found that the complex of titanium dioxide doped with silver (Ag-TiO2) and nanopowder of titanium dioxide (TiO2) at concentrations of 3 mg/ml initiate pathological changes in the bull sperm: the release of phospholipids as a result of the destruction of membranes and morphological abnormalities (abnormalities of the head, middle part, and tail, as well as the absence of acrosome, etc). The pathological effect of Ag-TiO2 nanocomposite was more pronounced. Doping with silver can increase the toxicity of nanotitanium dioxide, which requires further in-depth experimental studies using different concentrations by in vivo and in vitro methods.

The purpose of this study was to investigate the effect of different doses of trehalose and cysteine on bull frozen-thawed sperm quality. In this study, semen samples were collected from three healthy mature Holstein bulls by an artificial vagina. After primary evaluations, the samples were pooled to eliminate individual effects. Then, semen samples were divided into six equal parts and each part was extended and frozen with one of the following extenders. 1) without cysteine and trehalose (T0C0), 2) Five mM cysteine without trehalose (T0C5), 3) Ten mM cysteine without trehalose (T0C10), 4) A hundred mM trehalose without cysteine (T100C0), 5) Five mM cysteine with 100 mM trehalose (T100C5), 6) Ten mM cysteine with 100 mM trehalose (T100C10). Motility parameters, acrosome and membrane integrity and phosphatidylserine translocation assay were evaluated after thawing. All data were analyzed using a 2×3 factorial trail following data collection. The results of this study showed that cysteine had no effect on motion characteristics, apoptotic status, acrosome integrity and viability (P≥ 0. 05). Also motion parameters, viability, acrosome and membrane integrity, in group without trehalose was higher than group containing that (P≤ 0. 05). Regarding to interactive effects, the T0C10 group had higher amount of viability (P≤ 0. 05) and lower necrotic sperms compared to rest of the groups. Based on the results, it can be concluded that cysteine had no remarkable effect and addition of Trehalose could negatively affects post-thawed bull sperm quality.