Background: Embryo production in vitro requires three steps: in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC). The first step in the in vitro maturation of oocytes is germinal vesicle breakdown (GVBD), followed by completion of the 1st meiotic division and formation of the 1st polar body. These parameters are critical during ovary and oocyte selection. This study aimed to evaluate bovine ovary and oocyte collection for IVM. Methods: Ovaries collected from cows in a 0.9% NaCl saline solution at the central slaughterhouse were divided into two normal and abnormal ovaries according to their morphological appearance. The morphometric dimensions of bovine ovaries, such as weight and deamination (length, width and volume), were recorded. The oocyte was extruded and its deamination occurred before and after culture in maturation medium. Result: The mean ovary weight, volume, length and width were not significantly different between the two ovary types. Additionally, the dominant and subordinate follicle diameters in both ovary types showed no significant differences. Furthermore, the oocyte number per ovary of the normal and abnormal oocytes showed no significant differences. The mean cumulus oocyte complex before maturation showed no significant difference (52.73±7.23 mm for the normal ovary vs. 43.015±5.41 mm for the abnormal ovary). However, after maturation, a highly significant difference was found (P less than 0.001) between the normal ovaries before and after maturation (178.10±15.36 µm) and abnormal ovaries (10.45±7.99 µm). Additionally, data analysis of oocytes with or without 1st polar bodies revealed a highly significant difference (P less than 0.001) between oocytes of the normal ovary with (32.15±4.19) and without (10.95±1.59) 1st polar bodies and oocytes of the abnormal ovary with (7.0±078) and without 1st polar bodies (3.3±0.32). Thus, the critical point at which the normal ovary produces better in vitro matured follicles with good oocyte quality and produces the 1st polar body determines developmental competence. Therefore, the best selection of normal ovaries will enhance in vitro maturation for subsequent experiments, such as in vitro fertilization or cloning and in vitro embryo development.

Background: The utilization of in vitro fertilization (IVF) and in vitro embryo production (IVEP) has increased significantly as an effective tool of assisted reproductive technology. Sperm capacitation is a process of membrane maturation allowing the acrosome reaction to take place. Ionomycin is a calcium ionophore that induces the acrosome reaction by increasing calcium cycling across phospholipid membranes. The aim of this study was to enhance the cleavage rate and development of bovine embryos. Methods: Bovine ovaries were collected from the central slaughter house. Oocytes were obtained from follicles and were let to mature for 24 h. Frozen bull semen thawed in 37°C water and transferred to a discontinuous percoll gradient (Sigma GE17-0891-01). The spermatozoon pellet was separated into four groups after resuspension in bovine in vitro fertilization media (BO-IVF): control without additives; with DMSO and with ionomycin (25 and 50 nM) and kept in an incubator for three hours. The oocytes matured under commonly used media (TCM-199) for 24 h. The mature oocytes were then fertilized for 18-24h. The embryos were cultured in SOF medium until the seventh day to study cleavage rate and blastocyst rate. Result: The results indicated that 50 nM ionomycin supplementation considerably reduced cleavage while DMSO and 25 nM ionomycin supplementation had no effect on cleavage percentage. Moreover, when compared to the other groups, ionomycin at 25 nM significantly increased blastocyst development. In conclusion, the highest rate of blastocyst development was observed in the capacitation medium supplemented with 25 nM ionomycin, apparently due to the importance of calcium and metabolism in spermatozoa for fertilization and embryo development.

Background: Launaea angustifolia (LAN) is a wild plant grown in several Arabian countries in West Asia and North Africa. Launaea is a polymorphic inter- and intraspecific genus that includes about 54 species and 10 subspecies and is classified into eight sections. This study illustrates the effects of Launaea angustifo LAN (LAn) extract on male and female body weight and blood serum enzymes alanine aminotransferase (LAT) and Alkaline phosphatase LAN, with the histology of liver and kidneys used as an animal model for humans. Methods: A quantity of 100 mg of the extract powder was dissolved in 1 L of DMSO. Experimental rats were divided into five groups with 10 in each group (A, B, C, D, E: 100, 50 and 10 mg/kg and positive control and negative control, respectively). Result: No significant differences were observed between the treated and control-group females. However, the mean male body weight showed some variation within the 5th group (E) given the negative control (DMSO) and all treated groups presented lower body weights than the normal control (C). Analysis of the enzymatic activities of (ALT) alanine aminotransferase showed that the females in treatment group A (100 mg) and B (50 mg) had higher levels of ALT than those of group D females (10 mg) at P less than 0.05 and females in group E at P less than 0.001). The histology section of the liver and kidney and the histological structure of the rat kidney also showed no differences in all groups. Launaea angustifolia extract (LAn) was found to be safe, especially in the histology of the liver and kidney and for ALT and ALP blood parameters.

Background: Recent breakthroughs in vitro maturation, fertilization and culture technology have allowed for an increase in the number of offspring generated from genetically better females, but this progress is still hampered by critical factors affecting oocyte yield and quality. This study aims to evaluate the effect of the seasons on the rate of maturation and recovery of oocytes. Methods: The ovaries were brought from the slaughterhouse in Riyadh, Saudi Arabia, in a 0.9% NaCl saline solution and brought to the laboratory within 2-3 hours at a temperature of 25-35°C. Only the collected oocytes with two layers or more of cumulus cells (CCs) and uniform ooplasm were used. Five repetitions were made in each season of the year, followed by their ripening in the laboratory and a follow-up ripening after 24 hours. Observe the maturity rate after the stain and the expansion of cumulus cells and compare them with other seasons. Result: The mean maturation rates of MII and MI oocytes were not significantly different between all seasons. Additionally, there were no significant changes in the cumulus-oocyte complex before maturation in all seasons. However, after maturation, a highly significant difference (P less than 0.001) was found between the spring season before and after maturation (156.31±17.68 mm) and the rest of the seasons (summer; 169.89±19.96 mm; autumn; 176.66±20.14 mm; and winter; 188.84±24.50 mm). In order to improve in vitro maturation for following investigations, such as in vitro fertilization or cloning and in vitro embryo development, the ideal season should be chosen for collecting oocytes.