Asian Pacific Journal of Reproduction

54-57

Authors: Dian Aby Restanty, Setyawati Soeharto, I Wayan Agung Indrawan

Objective: To prove the effect of administered orally lead acetate exposure on Bax expression and the apoptosis index of granulosa cells on antral follicle female albino rats Wistar strain (Rattus norvegicus). Methods: Post-test only control group, using female albino rats Wistar strain aged (10-12) wk (reproductive age) weighing (100-200) g. Twenty-four animal samples were classified into one control group and three groups exposed to lead acetate in doses of 30, 100, and 300 ppm, respectively. Lead acetate was administered orally through a feeding tube over 30 d. Bax expression of granulosa cells on antral follicle was checked using immunohistochemistry, and apoptosis index were examined using Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Results: Lead increases apoptosis index on doses of 300 ppm. The increase of the Bax expression granulosa cell on antral follicle was not statistically significant. Conclusion: The exposure of lead acetate administrated orally can increase apoptosis index of granulosa cell on antral follicle although it doesn't affect Bax expression.

68-76

Authors: Islam El-Sayed El-Seadawy, Walid Said El-Nattat, Magda Mohamed El-Tohamy, Samy Ali Hussein Aziza, Yakot Abdelfattah El-Senosy, Ahmed Said Hussein

Objective: To maintain semen quality of male rabbits during chilled storage by enrichment the tris based diluent with different concentrations of propolis ethanolic extracts. Methods: Total phenolic and total flavonoid contents, as well as antioxidant activity was determined in propolis ethanolic extract (PEE). The extract was analysed by HPLC for separation and identification of target metabolites. Semen was collected from 10 rabbit bucks, pooled, then divided into five aliquots (each of 500 μL) and diluted each in 5 mL Tris-citric acid-glucose-egg yolk extender (TCGY). The 1st aliquot served as control while PEE was added at concentration of 0.8, 1.2, 1.6 and 2.0 mg/5 mL tris extender in the aliquot 2, 3, 4 and 5 respectively. Diluted semen samples were subjected to cooling at 4 ℃ for 72 h. Sperm motility, sperm viability, sperm abnormality, sperm membrane integrity and acrosome integrity were evaluated in chilled semen allover the chilling period. Results: The resluts revealed presence of a considerable amount of total phenolic compounds (98.67 mg GAE/g extract) and total flavonoids (70.16 mg CE/g extract) which were parallel to an antioxidant activity assessed as ABTS, DPPH and FRAP (198.65, 180.18 and 306.17 mM TE/g extract respectively). The dominant phenolic acid was chlorogenic acids (3.959 mg/g extract). Other compounds were found in less amounts rosmarinic acid (3.959 mg/g extract), myrcetin (1.946 mg/g extract), kaempferol (1.089 mg/g extract) and apeginin-7-glucoside (1.113 mg/g extract). Obtained results clearly demonstrated that the addition of (1.2-1.6) mg PEE in the chilled extended rabbit semen proved to be beneficial for maintaining semen characteristics compared to control and the addition of 0.8 and 2 mg PEE. Conclusions: The enrichment of rabbit semen tris-basic extender with (1.2- 1.6) mg PEE/5 mL tris-extender (as the best and safe concentrations) maintain the sperm characteristics in good condition all over 72 h of chilling.

81-84

Authors: El-Sheshtawy RI, El-Nattat WS

Objective: To explore the effect of silymarin on bull spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk diluents, purified silymarin powder (obtained from the milk thistle silybum marianum), purchased from Unipharma, Al Obour city, Egypt, was soaked in Tris-citric acid-fructose diluent for 48 h at 10 ℃ making a stock solution (70 mg/ mL), from this stock solution we obtained concentrations of 0.18 mg/mL, 0.36 mg/mL, 0.54 mg/mL, 0.72 mg/mL, 0.90 mg/mL in addition to the control (0.0 mg/mL) reaching a final volume of 5 mL in each tube. Egg yolk was added to each tube to obtain silymarin enriched semen extender (SEE) with 20% egg yolk, cooled slowly up to 5 ℃ and equilibrated for 4 h. Semen was packed into 0.25 mL polyvinyl French straws (IMV, France). After equilibration periods, the straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen for 10 min and were then dipped in liquid nitrogen. Extended semen was subjected to evaluation (motility, alive %, abnormality %, intact sperm membrane (HOST)% and conception rate) in both chilled and frozen semen. Results: Table 1 revealed that Sperm motility of the concentrations 2, 3 and 4 after 8 d of chilling were significantly (P<0.02) higher than control. Sperm motility of the concentration 2 (45.00%±2.89%) after 9 d of chilling was higher than control and the other concentrations. Addition of SEE in concentration 1 and 2 gave post thawing sperm motility as high as the control (47.50±2.81 and 45.00±2.58, respectively) while other concentration have lower effects on motility as compared to the control. Addition of silymarin improved post thawing alive% and was significantly higher (P<0.0 001) than the control. SEE decreased significantly (P<0.0 001) the % of post thawing abnormal sperm in concentration 3 and 4 (11.83±0.65 and 16.00±0.58, respectively). SEE improved significantly (P<0.018) the % of post thawing intact spermatozoa membranes (HOST%) in concentrations 2, 4 and 5 (71.17±0.83, 71.83±0.91 and 75.00±3.42 respectively) (Table 2). Conclusion: It could be concluded that silymarin as a natural additive to semen extenders improved preservability in both chilled and frozen bull semen.

89-92

Authors: Samaher Alfaraj, Fatima Alzaher, Sahar Alshwaiaer, Anwar Ahmed

Objective: To compare the pregnancy rate between day 3 and day 5 transfer regardless grades of embryos and number of transferred embryo. Methods: Retrospective cohort, a total of seven hundred and four patients met our inclusion criteria, with 411 had day 3 embryo transfer and 293 had day 5 embryo transfer. The patients who were older than 40 years old were excluded. Embryo transfer was carried out in all patients in both transfer groups. Results: Both clinical pregnancy rate and implantation rate did not show any statistically significant difference between the day 3 and day five transfer groups. These were 44% vs. 45% with P=0.82 and 19% vs. 19% with P=0.99 respectively. An increase of miscarriage rate with day 5 embryo transferred compare with day 3 (12.0% vs. 4.4%, P=0.01), but no significant difference was found about biochemical pregnancy rate (P=0.52). Conclusions: Transferring embryo at day 5 may not provide any additional benefit over day 3 transfers to patients. In addition, it increases the risk of miscarriage. Further studies of this issue needed for confirming our findings.