Asian Pacific Journal of Reproduction

260-266

Authors: Minh Tam Le, Van Trung Nguyen, Thanh Tung Nguyen, M. Blake EVans, Thai Thanh Thi Nguyen, Tam An Thi Nguyen, Dinh Duong Le, Vu Quoc Huy Nguyen, Ngoc Thanh Cao, Micah J.Hill

Objective: To evaluate the impact of prolonged post-thaw embryos culture on pregnancy outcome during frozen embryo transfer cycles. Methods: This prospective cohort study evaluated 324 thaw transfer cycles with 819 embryos from 269 patients at the Center for Reproductive Endocrinology and Infertility of Hue University Hospital in Vietnam. These frozen embryo transfer cycles were divided into two groups at the time of thawing: the short culture group (2-hour post-thaw culture) and the overnight culture group (overnight culture for 18 h) before the embryo was transferred into the uterus. The rates of embryo intact, grade A embryo at frozen and transfer time and continuing cleavage were recorded. The clinical outcomes including serum beta-human chorionic gonadotropin, clinical pregnancy and implantation rate were evaluated after 14 days, 4 weeks, 6 weeks, respectively, after embryo transfer. Results: Human chorionic gonadotropin positive occurred in 39.5% of patients in the short culture group compared to 25.9% in the overnight culture group with risk difference (RD)=13.6%, relative risk (RR)=1.343, 95% confidence interval (CI) 1.085-1.663, P<0.01. Clinical pregnancy rate of the short culture group and overnight culture group was 33.3% and 24.1%, respectively (RD=9.2%, RR=1.242, 95% CI 0.996-1.549, P=0.06) and the implantation rate in the short culture group and overnight culture group was 16.5% and 11.0%, respectively (RD=5.5%, RR=1.244, 95% CI 1.046-1.479, P=0.01). In women of advanced age (≥35 years) and women who received 3 embryos, pregnancy outcomes were found to be significantly (P<0.05) higher in the short culture than in the overnight culture group. Conclusions: The prolonged post-thaw culture period does not increase pregnancy outcome in comparison with the short culture.

276-282

Authors: Ferdinand Ngoula, Margaret Mary Momo Chongsi, Omer Bébé Kenfack Ngouateu, Alexane Marquise Ndekeng Makona, Augustave Kenfack , Bertin Narcisse Vemo, Joseph Tchoumboue

Objective: To evaluate the effects of Crassocephalum (C.) bauchiense (Hutch) leaf aqueous extract on the biochemical markers of toxicity, oxidative stress indicators and reproductive parameters in rabbit does (Oryctolagus cuniculus) exposed to potassium dichromate. Methods: A total of 36 nulliparous and sexually mature female rabbit does, aged 8 months and weighing 2.8-3.0 kg, were divided into 6 groups of 6 animals. After mating, group T0 received distilled water (1 mL/kg body weight), while groups T0-, VC100, AE100, AE200 and AE400 were treated with 40 mg/mL/kg body weight of potassium dichromate. In addition, group VC100 received 100 mg/mL/kg body weight of vitamin C, while groups AE100, AE200 and AE400 received aqueous extract of C. bauchiense leaves at 100, 200 and 400 mg/mL/kg body weight, by oral gavage for 27 days, respectively. On the 28th day post-coï tum, animals were sacrificed. Biochemical indicators such as creatinine, urea, alanine amino tranferase, aspartate amino transferase, total protein and total cholesterol were measured by using a spectrophotometer. Follicle stimulating hormone, luteinizing hormone and progesterone were measured by enzyme- linked immuno sorbent assay. Oxidative stress markers like malondialdehyde, catalase, superoxide dismutase and total peroxidase in ovary homogenates were measured by spectrophotometer. Results: Serum concentrations of urea, creatinine, alanine aminotransferase, aspartate aminotransferase and total cholesterol were significantly higher (P<0.05), and total protein level was significantly lower in T0- group (receiving potassium dichromate only) compared to other groups. Administration of ethanolic extract of C. bauchiense significantly decreased serum concentrations of urea, creatinine, alanine aminotransferase, aspartate aminotransferase and total cholesterol, and significantly increased total protein level. However, ethanolic extract of C. bauchiense had no significant effect on the foetotoxic characteristics. The serum concentrations of follicle stimulating hormone and luteinizing hormone were significantly (P<0.05) lower in T0- group as compared to the control group while progesterone were comparable (P>0.05) among all groups; C. bauchiense leaf extracts significantly (P<0.05) increased the level of follicle stimulating hormone, but the increase in luteinizing hormone was not significant (P>0.05). Catalase and total peroxidase activities significantly decreased (P<0.05), and malondialdehyde significantly increased in T0- group compared to other groups. The administration of C. bauchiense leaf extracts significantly (P<0.05) increased the activities of catalase and peroxidase. Conclusions: Potassium dichromate-induced oxidative stress involves in hepatotoxicity, nephrotoxicity and reprotoxicity. The aqueous extract of C. bauchiense leaves could mitigate these adverse effects via antioxidant properties.

289-296

Authors: Diego F. Carrillo-González, Nélida Rodríguez-Osorio, Charles R. Long, Neil A. Vásquez-Araque, Juan G. Maldonado-Estrada

Objective: To assess the effect of L-carnitine supplementation during in vitro oocyte maturation and in vitro culture process of bovine oocytes. Methods: L-carnitine (3.8 mM) was added to maturation medium and the effect was assessed in the quality (Experiment 1) and in the cleavage and 4-cells stage (Experiment 2). Besides, the effect of L-carnitine addition on maturation medium (3.8 mM) and culture medium (1.5 mM) on embryo rate production was assessed. In Experiment 1, bovine oocytes from abattoir were randomly separated into two groups (the control group and L-carnitine group) for in vitro maturation. Matured oocytes were examined for cumulus cells expansion as an indicator of maturation, and the content of the mitochondrial activity, the presence of lipid droplets, the reduced glutathione, and the reactive oxygen species were measured by using specific fluorochromes. In Experiment 2, oocytes were matured as performed in Experiment 1, afterward fertilized and cultured until day 3, and cleavage rate and 4-cells stage rate were determinated. In Experiment 3, in vitro maturation and fertilization were done as performed in Experiment 2, but at day 3 of culture, each group of embryos was separated into two new groups, and L-carnitine (1.5 mM) was added in culture media until day 8. The cleavage and embryo development rate were determined on the basis with the oocytes put on maturation. Hatching rate was calculated from cleaved embryos. Results: The cumulus expansion rate at grade 栿 and mitochondrial activity were significantly higher in the L-carnitine group in comparison with the control group (P<0.05). However, the content from the other variables tested did not significantly differ from the control group. Furthermore, the cleavage rate and 4-cells stage rate were similar in both groups (P>0.05). In addition, cleavage and the proportion of embryo development and hatching rate were similar for all groups (P>0.05). Conclusions: L-carnitine as a supplement in culture media improves the cumulus expansion and increases the mitochondrial activity during in vitro maturation process but has no apparent effect on the cleavage and development of bovine embryos. Further investigations of L-carnitine addition on in vitro culture are needed to test their effect on embryo quality.