Effects of sodium selenite, cysteamine, bacterially synthesized Se-NPs, and cysteamine loaded on Se-NPs on ram sperm cryopreservation.

During the cryopreservation of sperm, the production of highly reactive oxygen species (ROS) can reduce their viability and fertility. However, the addition of antioxidants can help reduce the harmful effects of ROS. One such antioxidant is selenium, which is a co-factor of the glutathione peroxidase enzyme that is effective in scavenging ROS. Cysteamine can also take part in the structure of this enzyme. The use of nanoparticles can be less toxic to cells than their salt form. To this end, researchers synthesized Se-NPs using the streptococcus bacteria and loaded cysteamine onto the synthesized Se-NPs. The biosynthesis of Se-NPs and cysteamine loaded on Se-NPs was confirmed by UV-visible spectroscopy, X-ray diffraction (EDX), Fourier transforms infrared (FTIR) spectroscopy, and Field Emission Scanning Electron Microscope (FE-SEM). For cryopreservation, ram semen samples were diluted, and different concentrations (0, 1, 5, 25, and 125 µg/mL) of cysteamine, Se-NPs, cysteamine loaded on Se-NPs, and sodium selenite were added. An extender containing no supplement was considered as control group. After cooling the semen samples, they were frozen and stored in liquid nitrogen for evaluation. The samples were thawed and analyzed for mobility, viability, membrane and DNA integrity, and sperm abnormalities, as well as malondialdehyde level (MDA) and superoxide dismutase (SOD). The data was processed using SPSS, and a significance level of p < 0.05 was considered. The results of this experiment showed that adding 1 µg/mL of cysteamine loaded on Se-NPs to the diluent significantly increased the motility, viability, and membrane integrity and SOD of spermatozoa compared to the other treatment groups and control group, and reduced the abnormality, apoptosis, and MDA level of spermatozoa in comparison with the other treatment groups and control group (p < 0.05). In conclusion, the addition of cysteamine loaded on Se-NPs was found to improve the quality of ram sperm after cryopreservation.

Pre-treatment of ram semen extender with magnetic nanoparticles on freeze-thawed spermatozoa.

BACKGROUND: Extensive use of different nanoparticles caused significant concerns about their biological safety. OBJECTIVE: This study aimed to evaluate the effects of cryopreservation on ram semen after adding magnetic nanoparticles (MNPs) to separate X and Y chromosome-bearing spermatozoa. METHODS: The experimental ram sperms in this research included treated spermatozoa (50 µg/ml MNPs) and non-treated spermatozoa. DNA damage of spermatozoa was examined using an acridine orange (AO) assay. Sperm viability, membrane functionality, abnormality and malondialdehyde (MDA) level were also measured. RESULTS: Results indicated that the pre-treatment of ram semen extender with MNPs did not significantly affect the semen parameters such as viability, membrane functionality, abnormality, as well as lipid peroxidation (LPO) levels and DNA integrity in comparison with the control group (p < 0.05). CONCLUSIONS: These observations suggest that pre-treatment of ram semen extender with MNPs after semen sexing did not have adverse effects on different semen parameters after cryopreservation.

Effects of different concentrations of Chir98014 as an activator of Wnt/beta-catenin signaling pathway on oocyte in-vitro maturation and subsequent embryonic development in Sanjabi ewes.

The present study was conducted to investigate the effects of the activator factor of the WNT pathway, chir98014, leading to the in vitro sheep oocyte maturation medium, on the cumulus cell development, different nuclear maturation stages and the following process of embryonic development. Experiments included (a) addition of different concentrations (0, 0.1, 0.5, 1 µm) of chir98014 to the maturation medium and evaluation of the cumulus cell expansion, (b) addition of different concentrations of chir98014 to the maturation medium and investigation of different nuclear maturation stages, (c) addition of different concentrations of chir98014 to the maturation medium and examination of the subsequent embryonic maturation process and (d) addition of different concentrations of chir98014 to the embryonic development culture medium (the first 48 hr) and investigation of the subsequent embryonic development process. The extracted data were analysed using the SPSS software, considering the significance level of p < .05 and making the mean comparisons. The results showed that the addition of the 0.1 µM concentration of chir98014 to the maturation medium had no significant effects on the oocyte maturation and embryo development post-fertilization but it enhanced the Cumulus-oocyte complexes (COCs) expansion. In the fourth experiment, the low concentration of chir98014 in the embryo culture media improved the embryo development process, whereas the high one had a detrimental effect on it, as compared to the control group. Thus, the presence of the lower concentrations of this compound in the embryonic culture medium had favourable effects on the development of embryos.

Recovery of sperms bearing X chromosomes with different concentrations of magnetic nanoparticles in ram.

Pre-conceptual sex selection is still a highly debatable process whereby X and Y chromosome bearing spermatozoa are isolated before oocyte fertilization. Recently, magnetic nanoparticles (MNP) have been used to determine X and Y chromosomes bearing spermatozoa as a result of searching for a cheap, highly efficient method using non-toxic materials. This study aimed to recover the sperm bearing X chromosomes in ram with different concentrations of MNP and then evaluate the success of this method using polymerase chain reaction (PCR). Ram sperms were divided into four groups, treated with 0 (control), 50, 100 and 200 µg/ml MNP, respectively. MNP was used to restore sperm cells bearing X chromosomes. Upon recovery, the PCR was performed to identify the X and Y sperms, Methyl ThiazoleTetrazolium (MTT), to assess MNP toxicity and sperm viability and acridine orange (AO) to evaluate sperm DNA integrity. The results of PCR revealed that the treatment of spermatozoa- bearing X chromosomes with 50 µg/ml MNP had the highest effects on the recovery of X sperm rather than the other concentrations of MNP. However, the concentrations of MNP did not have any toxic effects on spermatozoa, sperm viability and, DNA integrity, but the high concentration of MNP (200 µg/ml) significantly reduced DNA integrity. According to MTT and AO results, the concentrations of MNP used in this study had no toxic effects on spermatozoa and did not reduce the sperm viability and DNA integrity, except that 200 µg/ml MNP significantly reduced DNA integrity.

In vitro culture medium (IVC) supplementation with sericin improves developmental competence of ovine zygotes.

This study was carried out to investigate the effects of supplementation of potassium simplex optimized medium (KSOM-aa) with various sericin concentrations (0, 0.1, 0.5, 1 and 2.5%) on ovine zygotes. The results indicate that the supplementation of oocyte in vitro culture medium with optimal concentration of sericin (0.1 and 0.5%) may have beneficial effects on developmental competence of in vitro-derived ovine embryos.

In vivo and in vitro study of the function of the left and right bovine ovaries.

Inequality in function of the left and right bovine ovaries and uterine horns was evaluated in two separate experiments. In the first experiment (in vivo), the relationship between the left and right ovarian activities and reproductive indices was evaluated. Therefore, the total number of 1284 randomly chosen lactating dairy cows were examined from Day 50 to 60 postpartum, and according to the presence of an active CL on the ovaries, they were divided into 502 LCL3-cows and 782 RCL3-cows (cows with an active CL on the left [L] or right [R] ovary, respectively). To induce estrus synchronization and investigate the effects of PGF2α administration on the incidence of estrus in both LCL3-cows and RCL3-cows, the cows were treated with one luteolytic dose of PGF2α and were inseminated after observed estrus (via visual observation lasting at least 30 minutes three times a day). To investigate the effects of side of ovulation at the time of PGF2α administration on reproductive parameters, pregnancy diagnosis was performed 28 days after insemination (using ultrasound) and 42 days after insemination (using transrectal palpation). The results showed that the percentage of the RCL3-cows was greater than the LCL3-cows (60.9% vs. 39.1%, respectively). Furthermore, ovulations switching from the left to right ovary in two successive ovulations were greater than those that switched from the right to left ovary. On the other hand, the sex ratio (male percentage) in the right uterine horn was greater than that of the left one. In the second experiment (in vitro), the developmental potential of bovine oocytes derived from the left (L-oocytes) and right (R-oocytes) ovaries after in vitro embryo production and heterogeneity in the developmental competence of L-oocytes and R-oocytes using the brilliant cresyl blue staining test as a selection criterion were evaluated. Results of the in vitro experiment showed that the percentage of cleavage and blastocyst rate of R-oocytes were greater (P < 0.001) than those of L-oocytes. Moreover, it appears that the side of ovaries had greater effects on the developmental competence of oocytes than other factors associated with heterogeneity in the developmental competence of oocytes, which can be detected by the brilliant cresyl blue test. In conclusion, the results of the in vivo study confirmed the observations in previous studies in which the right ovarian response (distribution of ovulation) was superior to that of the left ones. Interestingly, the in vitro experiments for the first time clearly showed that more ovulation on the right side is not the only reason for this unequal activity. In fact, in cattle, the greater developmental potential of oocytes originating from right ovaries may cause superior activity of the right side, and the effect is even higher than the differences in ovulation response between the left and right ovaries.