Asian Pacific Journal of Reproduction

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Objective: To investigate the impact of ethanol extract of Spondias (S.) mombin leaves on the biochemical parameters of testicular function, hormonal profile, sperm profile and reproductive outcomes of guinea pigs. Methods: Sexually matured male [(1000.40±8.12) g] and female [(810.00±7.09) g] guinea pigs were used. In testicular function study, a total of 24 male guinea pigs were randomised into 4 groups of 6 guinea pigs each. Group A (control) were orally administered 1 mL of physiological saline, once daily for 60 days while groups B, C and D were treated like the control group except that they were orally administered 100, 250 and 500 mg/kg body weight of ethanol extract of S. mombin leaves. For the fertility study, the same animal groupings and treatments in the testicular function study were adopted. The male guinea pigs were paired with the females (1:1) and afterwards examined for pregnancy outcomes. Results: The ethanol extract of S. mombin leaves contained saponins, alkaloids, flavonoids, tannins, steroids, phenolics, phlobatannins, cardiac glycosides, cardenolides and dienolides with saponins (4.80 mg/mL) occurring the most whilst cardenolides and dienolides (0.08 mg/mL) were the least. The ethanol extract of S. mombin leaves significantly and dose dependently reduced the activities of alkaline phosphatase, glutamate dehydrogenase, 3-hydroxy-3-methylglutaryl coenzyme A reductase, malic enzyme, 17-β-hydroxysteroid dehydrogenase, lactate dehydrogenase, catalase and superoxide dismutase as well as levels of testosterone, glycogen, total protein and ascorbic acid in the testes when compared with the control group (P<0.05). All the doses of ethanol extract of S. mombin leaves also reduced the levels of sorbitol dehydrogenase, 3-β-hydroxysteroid dehydrogenase, glucose-6-phosphate dehydrogenase and sialic acid whereas the levels of testicular acid phosphatase, gamma glutamyl transferase and cholesterol increased dose dependently (P<0.05). The serum luteinising hormone, testosterone and estradiol were reduced after the administration of ethanol extract of S. mombin leaves whereas levels of serum follicle stimulating hormone increased significantly. The 100 and 200 mg/kg body weight of ethanol extract of S. mombin leaves increased the testosterone/estradiol ratios whilst the 500 mg/kg body weight of ethanol extract of S. mombin leaves decreased it. The sperm motility, sperm count, normal sperm morphology, sperm density, sperm viability and semen viscosity were significantly reduced in the ethanol extract of S. mombin leaves-treated guinea pigs (P<0.05) whereas the head-, tail- and neck-defects increased significantly when compared with the control group (P<0.05). In contrast, the semen volume and pH were not significantly altered by the ethanol extract of S. mombin leaves (P>0.05). The ethanol extract of S. mombin leaves at both 100 and 250 mg/kg body weight significantly reduced the total number, circumference, weight and length of the pups whereas the 500 mg/kg body weight of ethanol extract of S. mombin leaves-treated rats did not produce any pup. The 250 and 500 mg/kg body weight of ethanol extract of S. mombin leaves induced degenerative and necrotic changes in the seminiferous tubules with vacuoles in the germinal epithelium and a few to complete absence of spermatozoa. In all of these, the 500 mg/kg body weight of ethanol extract of S. mombin leaves produced the most pronounced alteration on the parameters. Conclusions: S. mombin leaves have induced infertility in the male guinea pigs via endocrine dysregulation, anti-spermatogenic activity, testicular dysfunction and oxidative stress and made possible by the presence of saponins, alkaloids, flavonoids, tannins, steroids, phenolics, and cardiac glycosides.

Objective: To determine the correlation of different serum estradiol levels on the trigger day with the clinical and laboratory outcomes of in-vitro fertilization (IVF) cycles comprising a single fresh top-quality blastocyst transfer. Methods: This was a retrospective observational study performed in Morula IVF Clinic Jakarta. Five hundred forty-two women were recruited and grouped according to their serum estradiol levels on the trigger day of follicular maturation as follows: <2 000 pg/mL, 2 000-2 999 pg/mL, 3 000-3 999 pg/mL, and ≥ 4 000 pg/mL. Clinical pregnancy and miscarriage rates were evaluated as the primary outcomes and embryology laboratory results as the secondary outcomes which consisted of the number of retrieved, mature, and fertilized oocytes, the total sum of derived embryos, and top-quality embryos at cleavage and blastocyst stage. Results: Clinical pregnancy and miscarriage rates did not differ among the groups (P>0.05). Nonetheless, the study demonstrated a positive correlation of the serum estradiol levels with the overall laboratory outcomes including the number of retrieved, mature, and fertilized oocytes, the total sum of derived embryos, and top-quality embryos at cleavage and blastocyst stage (P<0.001). The subject group with estradiol level of ≥4 000 pg/mL was superior to the other groups in its respective median number of retrieved, mature, fertilized oocytes, total derived embryos, and top-quality cleavage- and blastocyst-stage embryos. Conclusions: Although an apparent positive correlation is observed between estradiol levels and laboratory outcomes, serum estradiol level on hCG trigger day is not associated with the clinical outcomes of IVF.

Objective: To investigate the combined therapeutic potential of Gymnema (G.) sylvestre and Pergularia (P.) daemia on letrozole-induced polycystic ovarian syndrome (PCOS) in rats. Methods: Thirty six healthy female Wistar rats with regular estrus cycles were randomly divided into six groups each of 6. Group I received 1 mL of 0.5% carboxyl methyl cellulose orally and served as the vehicle control group, while groups II to VI were treated with letrozole (1 mg/kg body weight p. o.) for 21 days to induce PCOS. After induction of PCOS, group II served as the PCOS control group, without treatment; group III received metformin (20 mg/kg body weight p. o.) as the standard group, and groups IV to VI received G. sylvestre (100 mg/kg body weight p. o.), P. daemia (300 mg/kg body weight p. o.), and the combination of G. sylvestre and P. daemia, respectively, for 28 days. Vaginal smears were collected from all rats daily throughout the study to determine the phases of the estrus cycle. After completing the treatment schedule, oral glucose tolerance test, serum lipid profile and reproductive hormonal analysis were carried out. Subsequently, the rats were sacrificed to collect ovary and uterus for histopathological examination. Results: The PCOS control rats showed a significant irregularity in the estrus cycle, hyperglycemia, and the altered serum lipid profile such as the increased low and very low density lipoprotein, triglyceride, and decreased high density lipoproteins. In addition, the PCOS control rats showed a significant increase in serum luteinizing hormone, testosterone, and estrogen, and decrease in follicle stimulating hormone and progesterone. These changes were significantly revoked in all the treatment groups. The test drugs also significantly reduced the gained ovary weight (P<0.001), and histopathology of the ovary showed almost normal ovary. Among the treatment groups, the group of combination treatment of G. sylvestre and P. daemia showed superior ameliorative results in PCOS parameters. Conclusions: Combination of G. sylvestre and P. daemia presents potent synergistic activity against hyperandrogenism, hyperinsulinemia, anovulation and follicular cysts in letrozole-induced PCOS rats.

Objective: To study the aphrodisiac potential of Polyalthia (P.) bullata in fowl. Methods: In this study, testosterone, as an indicator of the aphrodisiac potential of P. bullata, was investigated for its release from TM3 Leydig cells grown in vitro and in 4 fowls given capsules containing P. bullata at a dose of 10 mg in each capsule twice a day, for 50 days. In the latter in vivo evaluation, mating behaviours were additionally determined after the treated fowls were released to the individual hens, and their testes and liver were dissected for histological examinations. Blood drawn from the fowls was assessed for any changes in diagnostic parameters. Results: In the in vitro test (TM3 Leydig cells), P. bullata was able to increase testosterone to 0.48 nmol/L within 72 h of incubation, compared to the untreated control with only 0.18 nmol/L, i.e., an increase of 170%. In the in vivo test, outcomes in the fowls dosed with P. bullata showed similar positive elevations of testosterone to (9.72±1.10) nmol/L in comparison to the controls that showed a level of only (4.05±0.84) nmol/L. Total frequencies of mating behaviours were observed (wing flapping, body shakes, crowing and beak pecking) to be 23 counts for the test compared to only 15 for the control fowls. Histological examination of the male reproductive organs provided evidence of testosterone boosting based on an observable increase in the activity at the seminiferous tubules of testis tissues without any damaging effects, compared to the controls. In the nine diagnostic blood parameters assessed, including alanine aminotransferase, aspartate aminotransferase, and gamma glutamyltransferase, none was remarkably elevated compared to the controls. The histological changes in the liver were not severe and mainly consisted of only localized moderate but recoverable obstructions and swellings of the vessels and tubules. Conclusions: P. bullata is able to boost testosterone both in vitro and in vivo, with no acute toxicities.

Objective: To characterize the semen of three genetic types of boars (local, improved and Large White) reared in Benin. Methods: Semen of local, improved and Large White boars reared in Benin were collected using the gloved hand method and analyzed to determine volume, pH, concentration, mobility, motility, and morphology. The effect of the genetic type of boar on semen characteristics was aslo studied. Results: Duration of ejaculation and semen volume of Large White boar were significantly higher than those of local and improved boars (P<0.05). The semen of improved boars had a higher motility score than that of Large White and local boars (P<0.001). The semen of local boars was more concentrated in the spermatozoa than that of improved and Large White boars (P<0.05). The proportion of spermatozoa of improved boars with normal morphology (93.6%) was significantly higher than that of local (82.2%) and Large White boars (81.6%) (P<0.001). The proportion of spermatozoa with folded tails in the semen of Large White boars (9.2%) was significantly higher than that observed in improved (1.8%) and local (5.0%) boars (P<0.001). The proportion of spermatozoa with proximal cytoplasmic droplets in semen of improved boars (2.7%) was significantly lower than that in Large White (6.8%) and local (9.7%) boars (P<0.001). The local (1.5%) and Large White boars (1.1%) showed more spermatozoa with distal cytoplasmic droplets in their semen compared to the improved boars (0.4%). Conclusions: The semen characteristics of pigs reared in Benin vary from one genetic type to another. Each genetic type has a strong point. The Large White boar produces larger semen, the local boar produces more concentrated semen and the improved boar produces spermatozoa that are morphologically better. The semen of these three genetic types can be used in artificial insemination but the improved boar’s semen is more recommended.

Objective: To evaluate the effect of human chorionic gonadotropins (hCG) and equine chorionic gonadotropins (eCG) on in vitro gilt oocyte maturation and embryonic development, using frozen semen for fertilization. Methods: Two independent experiments (6 replicates each) were carried out to evaluate gilt oocyte maturation, and fertilization and embryonic development by using ovaries from a local abattoir. Totally, 712 oocytes were randomly distributed in four-well dishes to receive Novormon (eCG 5.0 IU), PG600 (eCG 5.0 IU and hCG 2.5 IU), Chorulon (hCG 5.0 IU), or no hormones. Oocytes were incubated with 5% CO₂, 95% air and saturation humidity at 39 °C for 44 h. Maturation of the oocytes to metaphase II was assessed by using the aceto-orcein technique. In addition, 741 oocytes were used and randomly distributed in four-well dishes, and then oocyte maturation was carried out as mentioned, but matured oocytes were washed and placed in fertilization medium with frozen-thawed sperm. Gametes were co-incubated for 7 h, and then washed and placed in development medium, and incubated for further 7 days, at which time embryonic development was evaluated. Fertilization and embryo development media were not supplemented with the studied hormones. Results: Novormon (eCG) and PG600 (eCG+hCG) treatments significantly improved the percentages of metaphase II oocytes compared to the control group (P<0.05). Furthermore, a significant increase was also observed in the young blastocyst stage between the control group and the PG600 treatment group (P<0.05). Conclusions: Hormonal products Novormon (eCG) and PG600 (eCG+hCG) can obtain the highest percentages of in vitro maturation in gilt oocytes; however, this effect is not transferred to fertilization rates.

Online since 14^th Aug 2017.