Use of Purslane (Portulaca oleracea) Extracts and Cysteine in Diluents to Improve the Cryopreserved Quality of Goat Sperm.

Highlights Using cysteine and purslane extracts in extenders improved significantly the post-thaw sperm characteristics. Sperm viability, DNA integrity, and mitochondrial activity demonstrate an improvement in post-thaw sperm. Malondialdehyde production was decreased based on the positive effects of treated extenders. The obtained results demonstrate that supplementation of 50 µg/mL of purslane methanolic extract with cysteine to freezing extenders was significantly superior compared with other treatments.

How origin of ovaries influences the vitrification outcome of bovine ovarian tissue: effects of side of ovaries and corpus luteum.

Although cryopreservation of ovarian tissue has advanced greatly, it remains a challenge, and protocols should be optimized to handle the heterogeneous nature of ovarian samples. In an effort to address this factor, the present study evaluated the effects of corpus luteum (CL) and side of ovaries (right versus left) on cellular morphology and viability of vitrified bovine ovarian fragments in a closed system. The ovaries were categorized according to whether they had a CL and which side they were on, and then divided into six groups: 1) CL+ (with CL) group; 2) CL- (without CL) group; 3) right ovaries group; 4) left ovaries group; 5) fresh control group (ovaries without vitrification or culture that were not selected for CL or ovarian side) and 6) In vitro culture medium control group (non-vitrified ovaries that were not selected for the presence or absence of CL or side of the ovaries). The current study shows that the CL- and right groups had the greatest percentage of follicles with normal morphology compared to other vitrified-warmed groups. Furthermore, the levels of necrosis and tissue damage of the right cultured group were the lowest compared to other groups. It was shown that bovine ovarian tissues derived from right ovaries and ovaries without a corpus luteum can be functionally and morphologically preserved after vitrification. For the first time, the present study suggests that bovine ovarian tissue vitrification can be improved by considering the origin of the ovaries.

Evaluation of used Purslane extracts in Tris extenders on cryopreserved goat sperm.

This study aimed to evaluate the comparative effects of Purslane aqueous extract (PAE), Purslane methanolic extract (PME) and Purslane ethanolic extract (PEE on the quality of frozen-thawed goat spermatozoa. Collected semen with motility >75% and sperm concentration >1.0 × 109 sperm/ml was pooled and divided into 10 equal aliquots and supplemented by basic extender containing 25, 50 or 100 µg/ml of Purslane aqueous extract (PAE25µg/ml, PAE50µg/ml, PAE100µg/ml, respectively), basic extender containing 25, 50 or 100 µg/ml of Purslane methanolic extract (PME25µg/ml, PME50µg/ml, PME100µg/ml, respectively), basic extender containing 25, 50 or 100 µg/ml of Purslane ethanolic extract (PEE25µg/ml, PEE50µg/ml, PEE100µg/ml, respectively). Control diluent contained no additives. For the determination of sperm quality, frozen straws were thawed and then the sperm characteristics were assessed. Results indicated that higher (P < 0.05) percentages of total motility, viability, mitochondrial activity and lower percentages of malondialdehyde (MDA) for PAE50µg/ml, PME50µg/ml and PEE50µg/ml than those of the control. In addition, PME50µg/ml resulted in the highest) P < 0.05) total motility and the lowest (P < 0.05) MDA levels compared to other treatments. Compared to the control group, PME50µg/ml resulted in higher integrity (P < 0.05) of plasma membranes and in lower amounts of apoptotic and dead spermatozoa. PME50µg/ml and PAE50µg/ml showed higher (P < 0.05) percentages of progressive motility, DNA integrity and live post-thawed spermatozoa than those of the control. No significant differences in the motility, viability, mitochondrial activity and number of live sperms were observed between PME50µg/ml and PAE50µg/ml treatments. In conclusion, the results of this study indicated that 50 µg/ml purslane extracts could be used for the cryopreservation. However, the results of methanolic extract was more beneficial compared to other extracts.

Developmental competence of bovine oocytes selected based on follicle size and using the brilliant cresyl blue (BCB) test.

BACKGROUND: Many studies reported that follicle size has an essential role in developmental potential of oocytes. Also, the brilliant cresyl blue (BCB) test is one of the most important criteria in selection of more competent oocytes. OBJECTIVE: Selection of developmentally competent bovine oocytes. MATERIALS AND METHODS: A total of 1730 bovine cumulus oocyte complexes (COCs) were recovered from the ovaries by follicles isolation and classified into 3 categories according to the diameters of the follicles (small, <3 mm; medium 3-6 mm and large >6 mm). Oocytes were exposed to the BCB stain, diluted in Dulbecco's phosphate-buffered saline, modified with 0.4% bovine serum albumin (BSA) for 90 min. Oocytes with or without blue coloration of the cytoplasm were designated as BCB(+) and BCB(-), respectively. RESULTS: The BCB(+) and control oocytes originated from large and medium follicles exhibited a higher (p<0.0001) cleavage and blastocyst rate than BCB(-) oocytes. Furthermore, the BCB(+) oocytes from large and medium follicles had the highest (p<0.0001) proportion of blastocyst than other treatment groups. In contrast, the BCB(-) oocytes from small follicles had the lowest (p<0.0001) proportion of blastocyst than other treatment groups. Interestingly, the percentage of the BCB(+) oocytes from the large and medium ovarian follicles was significantly higher (p<0.0001), than the BCB(+) oocytes from the small follicles. CONCLUSION: Current results confirmed that each BCB(+) oocyte could not lead to perfect embryo development and the BCB test is not sufficient enough for the identification of oocytes that are competent for in vitro embryo development.