Asian Pacific Journal of Reproduction
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    2021(2):49-55, DOI: 10.4103/2305-0500.311608
    Abstract:
    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.
    2021(2):56-67, DOI: 10.4103/2305-0500.311609
    Abstract:
    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) was 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.
    2021(2):68-74, DOI: 10.4103/2305-0500.311610
    Abstract:
    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.
    2021(2):75-81, DOI: 10.4103/2305-0500.311615
    Abstract:
    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.
    2021(2):82-89, DOI: 10.4103/2305-0500.311616
    Abstract:
    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.
    2021(2):90-96, DOI: 10.4103/2305-0500.311618
    Abstract:
    Objective: To evaluate the effect of human chorionic gonadotropins (hCG) and equine chorionic gonadotropins (eCG) onin 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 ℃ for 44 h. Maturation of the oocytes to metaphaseⅡ 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 metaphaseⅡ 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.
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    2019(2):63-69, DOI: 10.4103/2305-0500.254647
    Abstract:
    Objective: To investigate ameliorative effects of Vitex agnus-castus (VAC) and VAC containing pharmaceutical supplement (VPS) against polycystic ovary syndrome (PCOS). Methods: PCOS in the rats was induced by daily administration of letrozole at 1 mg/kg body weight concentration for 21 d. PCOS rats were then treated daily either with metformin, VAC plant extract or VPS at 70, 8 or 8 mg/kg body weight concentration for 15 d. Rats that received none of these treatments were considered as control. Blood and ovaries were collected from all the rats. Serum glucose, cholesterol, triglyceride and high density lipoprotein-cholesterol were measured spectrophotometrically. Serum insulin, estrogen, progesterone, testosterone, luteinizing hormone, follicle-stimulation hormone, catalase, superoxide dismutase, malondialdehide and reduced glutathione were measured using enzyme-linked immunosorbent assay. Results: Rats treated with letrozole demonstrated a significant increase in serum testosterone, estrogen, cholesterol, luteinizing hormone, triglycerides, glucose, insulin, and malondialdehide levels, and a significant decline in progesterone, follicle-stimulating hormone, high density lipoprotein-cholesterol, catalase and reduced glutathione levels compared to control. Contrarily, no significant change in superoxide dismutase was noted in response to letrozole treatment. Rats treated with metformin, VAC or VPS showed a remarkable reversal in the levels of parameters affected by letrozole treatment. Conclusions: Data indicate that VAC and VPS exert potential ameliorative effects against PCOS through the modulation of hormonal and lipid profile as well as oxidative stress. Moreover, the favorable effects of these compounds are comparable to that of metformin.
    2019(2):70-74, DOI: 10.4103/2305-0500.254648
    Abstract:
    Objective: To evaluate effect of Phaseolus vulgaris (P. vulgaris) on the breastfeeding in female rats. Methods: This experimental study was done from May 2018 to December 2018 in the Anatomical Department of Medical School in Kermanshah University of Medical Sciences in Iran. In this study, after one-week adaptation and fertilization by male, 40 female rats within 20 days of pregnancy (on average, every mother had 10 newborns) were equally separated into four groups (animals were administrated after delivery of offspring). Group 1 was control group receiving normal saline interaperitoneally, and groups 2, 3, 4 were treatment groups receiving the dose of 20, 50, 100 mg/kg of P. vulgaris interaperitoneally respectively once a day for 60 days. The prolactin hormone was measured by radio immune assay, number and diameter of alveoli via histological and morphometrical examinations, and receptor prolactin gene expression by real-time polymerase chain reaction method. Results: P. vulgaris significantly improved alveoli’s number and diameter, prolactin hormone and receptor prolactin expression when compared to the control group (P<0.001). Conclusions: P. vulgaris is helpful to improve the breastfeeding parameters of rats’ mammary glands.
    2019(2):75-82, DOI: 10.4103/2305-0500.254649
    Abstract:
    Objective: To investigate the effect of different fractions from the aqueous methanolic root bark of Adansonia digitata (Linn) on the estrous cycle of female Wistar rats and to elucidate their phytochemical content. Methods: Crude aqueous methanol extract of Adansonia digitata (Linn) root bark was fractionated by column chromatography using various ratios of hexane, acetyl acetate, methanol and distilled water. Fractions with similar trails on a thin layer chromatography were pooled into four fractions. Each fraction was then screened for its phytochemical content. Forty mature Wistar female rats weighing (135±28) g with regular estrous cycles were given each fraction at two dose levels while a control group received distilled water of equal volume. Treatments were given per os for 15 days starting from the day of estrus. Vaginal smears were observed daily for the duration of each cycle, frequency of occurrence of each stage of the estrous cycle and number of cycles for 15 days before and during treatment. At the end of treatment, rats were humanely sacrificed. Ovaries and uteri were weighed and processed for histopathology. Results: Hexane acetyl acetate and acetyl acetate methanol fractions at 150 mg/kg body weight and 300 mg/kg body weight prolonged the occurrence of proestrus and diestrus and produced atretic cyst-like ovaries at 300 mg/kg body weight. Acetyl acetate methanol treated rats also had low relative ovarian weights when compared to the control. Conclusions: Sterols and triterpenes in hexane acetyl acetate and acetyl acetate methanol fractions of the root bark may be responsible for the disruption of estrous cycle in female Wistar rats and may have a contraceptive effect.
    2019(2):45-55, DOI: 10.4103/2305-0500.254645
    Abstract:
    Pyometra is one of the most important and common disorders in canine. The disease results in life threatening condition associated with septicemia and toxemia. The condition commonly occurs during luteal phase of estrous cycle, generally 4 weeks to 4 months of estrous cycle. The age, parity, breeds, administration of hormones are some predisposing factors of the condition. However, interaction between potentially pathogenic bacteria and hormonally primed uterus is believed to result in pyometra. The disease is initiated in the form of cystic endometrial hyperplasia, which later progresses into purulent uterine content fo llowing bacterial infection resulting in pyometra. The disturbance in antioxidant and prooxidant balance succeeding bacterial infection results in oxidative stress. The resultant stress further induces endometrial degeneration, immunosuppression and additionally aggravates the condition. An important clinical sign is malodorous, sanguineous or mucopurulent vaginal discharge with general symptoms such as vomiting, polyuria, polydipsia, depression, anorexia and occasionally fever. Till date, ultrasonography is the best diagnostic method while radiology and serological test are also helpful. Recently, the estimation of inflammatory mediators (acute phase proteins and prostaglandin metabolites) has appeared as marker for diagnosis of the pyometra. Determination of levels of inflammatory mediators has emerged as an important diagnostic method because it can be helpful in prognosis of the condition. The pyometra can be best treated by surgical manipulation (ovariohysterectomy);however, hormonal treatment can be given in less severe cases which aimed at reducing the effect of progesterone on the reproductive tract. Together, provision of antioxidants could be helpful in disease recovery along with the determined treatment. In this review, incidence,predisposing factors, etio-pathogenesis, oxidative stress associated with condition, diagnostic methods and treatment are discussed.
    2019(2):56-62, DOI: 10.4103/2305-0500.254646
    Abstract:
    Objective: To study the effects of co-administration of Gynura procumbens (GP) and kelulut honey (KH) on male fertility and libido in diabetes-induced rats. Methods: A total 42 males Sprague Dawley rats aged 8 weeks were randomly and equally divided into six different groups. All groups except a normal control group were induced with 50 mg/kg of streptozotocin (STZ) intravenously to induce diabetes. A positive control group was treated with an antidiabetic drug, metformin (500 mg/kg) whereas a negative control group remained untreated throughout the experiment. Meanwhile, another three treatments on diabetic rat groups were performed and categorised as Group 1 (450 mg/kg GP + 300 mg/kg KH), Group 2 (450 mg/kg GP + 600 mg/kg KH) and Group 3 (450 mg/kg GP + 1 200 mg/kg KH). Treatments were given for seven consecutive days through oral gavage and all rats were euthanized on day 8th for fasting blood glucose analysis, sperm quality, spermatogenesis, sexual behaviour and implantation sites analysis. Results: Fasting blood glucose levels were significantly reduced after treatment of GP and KH, compared to negative and positive controls. The treated groups showed significant increment in sperm quality compared to all control groups. Testes histology illustrated significant damages on leydig and sertoli cells for both negative and positive controls. On the contrary, co-administration of GP and KH displayed regeneration of leydig and sertoli cells in the testes. Additionally, the number of implantation sites significantly increased in females copulated with treated groups, compared to controls. Besides, the libido analysis displayed improvement of libido in treated groups, compared to all controls. Throughout the study, insignificant variances were recorded between the treated groups, indicating that treatment in Group 1 was sufficient to trigger significant improvement on fasting blood glucose level, fertility, and libido in diabetic male rats. Conclusions: Co-administration of GP and KH has great potential to serve as a pro-fertility agent amongst diabetic patients.
    2019(2):83-87, DOI: 10.4103/2305-0500.254650
    Abstract:
    Objective: To compare the levels of p38 mitogen-activated protein kinase (MAPK), soluble endoglin and endothelin-1 in the serum of patients with severe preeclampsia, hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome and normal pregnancies. Methods: This study was an observational analytic cross-sectional study performed at Wahidin Sudirohusodo Hospital, Makassar, Indonesia, in the period of 5th February 2016 to 20th January 2017. P38 MAPK, soluble endoglin and endothelin-1 levels of patients with normal pregnancies, severe preeclampsia and HELLP syndrome were measured by enzymelinked immunoabsorbentassay technique, using kits of human soluble endoglin, endothelin-1 and p38 MAPK, Quantikine immunoassay: R&D System Inc. Results: Level of serum p38 MAPK in HELLP syndrome group was higher than in severe preeclampsia and normal pregnancy groups. Soluble endoglin and endothelin-1 levels in pregnancies with severe preeclampsia and HELLP syndrome were higher than normal pregnancy but there was no significant difference between these two groups (P>0.05). Levels of p38 MAPK, soluble endoglin and endothelin-1 also had a positive linear correlation with systolic and diastolic blood pressures (P<0.05). Conclusions: P38 MAPK in serum may be a marker for evidence of the severe hypoxia and its application may be considered for the diagnosis of HELLP syndrome.
    2019(2):88-92, DOI: 10.4103/2305-0500.254651
    Abstract:
    Objective: To assess the secondary sex ratio (SSR) of assisted reproduction technology babies within a fertility clinic in Melbourne, Australia. Methods: A retrospective cohort study was conducted on 3 369 babies who were born following single embryo transfer between 2011 and 2016. Variables examined included embryo creation date, maternal and paternal ages, maternal body mass index (BMI), type of infertility, sperm parameters, fertilization method, type of embryo and stage of transfer. Multivariate regression analysis was performed on the data set using STRATA V9.2. Results: More males were found to be born to embryos created in summer (adjusted odds ratio=1.46, 95% confidence interval: 1.08-1.95; P=0.01). Lower BMI of mother (30 kg/m2) and younger age of parents (35 years) were associated with an increase in SSR (50.1% vs. 47.1%; 50.6% vs. 48.8%; 50.0% vs. 48.9%) respectively. Decreased SSR value was associated with primary subfertility, using in-vitro fertilization procedure and fresh embryo compared to those with secondary subfertility, using intracytoplasmic sperm injection procedure and thawed embryo (49.3% vs. 50.6%; 47.1% vs. 50.8%; 49.4% vs. 50.1%) respectively. Conclusions: The fertilization methods, type of embryo, stage of embryo transfer, parameters of the sperm and status of subfertility do not significantly affect the SSR. SSR is affected by the seasonality, maternal age, and BMI.
    2019(3):93-101, DOI: 10.4103/2305-0500.259166
    Abstract:
    Among resources of mesenchymal stem cells, human umbilical cord appears to be a rising source capable of differentiating into all germ layers, reaching and repairing lesion areas, and promoting wound repair, and it has also the capacity to influence the immune response. Human umbilical cord-derived mesenchymal stem cells are considered to be an optimal resource compared with other mesenchymal stem cells sources because they require a noninvasive recovery. All these characteristics allow their use in heterogeneous applications. Human umbilical cord-derived mesenchymal stem cells can regenerate tissues, stimulate angiogenesis, modulate inflammatory pathway signals and recruit endogenous stem cell. Human umbilical cord-derived mesenchymal stem cells suppress mitogen-induced signals and modulate the activation and proliferation of several immune cells, modifying lymphocyte phenotypes activity. In culture, human umbilical cord-derived mesenchymal stem cellss show the capacity to create several tissues such as bone, cartilage, and fat. Human umbilical cord-derived mesenchymal stem cells can be isolated from the different compartments of umbilical cord and processed by using different techniques. Clinical applications of human umbilical cord-derived mesenchymal stem cells include graft-versus-host disease, autoimmune diseases such as Sj.gren's syndrome and diabetes mellitus types 1 and 2, gynecological disorders like endometriosis. Recent studies have shown possible application on rheumatoid arthritis, osteoarthritis, and neuronal degenerative diseases. This review is focused on the resources, molecular profiles, propriety, in vitro characterizations, clinical applications and possible future usage of human umbilical cord-derived mesenchymal stem cells.
    2016(3):240-246, DOI: 10.1016/j.apjr.2016.03.002
    Abstract:
    ObjectiveTo determine the effects of the water quality of Manila Bay on allometric parameters and histological biomarkers of selected organs of P. viridis. Methods Green mussels were collected from two coastal sites of Manila Bay, Las Piñas – Parañaque (LPP) and Bacoor, Cavite (BC). Twenty-four green mussels from each site were used for the assessment of allometric parameters, and six green mussels from LPP and eight from BC were used for the assessment of histological structures of gonads, gut, and digestive glands. Gonad development was categorized into five stages, whereas gut and digestive glands were scored into four categories. Results Allometric parameters that include shell height, weight, and total wet and dry soft tissue weight were significantly different between LPP and BC. It was also observed that exposure to the pollutants in Manila Bay resulted to delays in gonadal development, and detrimental changes and lesions in the histostructure of digestive gland and gut. Conclusions Pollutants in Manila Bay have detrimental effects to the growth, reproductive development, and histological structure of digestive organs of P. viridis.
    2019(2):88-92, DOI: 10.4103/2305-0500.254651
    Abstract:
    Objective: To assess the secondary sex ratio (SSR) of assisted reproduction technology babieswithin a fertility clinic in Melbourne, Australia.Methods: A retrospective cohort study was conducted on 3 369 babies who were born following single embryo transfer between 2011 and 2016. Variables examined included embryo creation date, maternal and paternal ages, maternal body mass index (BMI), type of infertility,sperm parameters, fertilization method, type of embryo and stage of transfer. Multivariate regression analysis was performed on the data set using STRATA V9.2. Results: More males were found to be born to embryos created in summer (adjusted odds ratio=1.46, 95% confidence interval: 1.08-1.95; P=0.01). Lower BMI of mother (30 kg/m2)and younger age of parents (35 years) were associated with an increase in SSR (50.1% vs.47.1%; 50.6% vs. 48.8%; 50.0% vs. 48.9%) respectively. Decreased SSR value was associated with primary subfertility, using in-vitro fertilization procedure and fresh embryo compared to those with secondary subfertility, using intracytoplasmic sperm injection procedure and thawed embryo (49.3% vs. 50.6%; 47.1% vs. 50.8%; 49.4% vs. 50.1%) respectively.Conclusions: The fertilization methods, type of embryo, stage of embryo transfer, parameters of the sperm and status of subfertility do not significantly affect the SSR. SSR is affected by the seasonality, maternal age, and BMI.
    2018(4):167-171, DOI: 10.4103/2305-0500.237054
    Abstract:
    Objective: To study the methods of increasing the reproductive qualities and indices of viability in the offspring of Holstein cows under conditions of intensive milk production. Methods: Studies were conducted on 3 groups of Holstein cows with 30 heads in each group. Animals of the experimental groups had a different length physiological period. The study focused on experimental groups of animals: the reproductive indicators, the viability of newborn calves, and their growth rate. All the digital materials of the experimental data were processed by the method of variation statistics for the significance of the difference of the compared parameters using the Student’s criterion, adopted in biology and zootechnics. Results: Studies found that reduction of the duration of the service period, along with a simultaneous increase in the inter-lactation period to 80–90 days, increased the reproductive capacity of the cows, and enhances the viability of the offspring compared with their peers. Conclusions: The study revealed the optimal duration of the physiological periods in highproducing cows under conditions of intensive milk production technology.
    2017(2):77-80, DOI: 10.12980/apjr.6.20170205
    Abstract:
    Objective: To explore the effect of BHT on cattle spermatozoa during cooling and cryopreservation. Methods: Pooled bull semen were diluted by Tris-Citrate-Fructose egg yolk (TCFY) diluent considered as control (0 BHT) and different concentrations of BHT (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mM were prepared in ethanol in prewarmed (37°C) test tubes. The ethanol was allowed to evaporate so that, a thin crystallized layer of BHT was deposited on the inner surface of the tubes. Then extended semen was added into the tubes and incubated at 37°C for 5 min to allow uptake of BHT by spermatozoa. The tubes were cooled slowly (approximately for 2 h) up to 5 and equilibrated for 4 h. After equilibration, semen freezing process was carried out. Extended semen was subjected to evaluation (motility, alive sperm, intact sperm membrane (HOST) % and acrosome integrity) in both cooled and cryopreserved semen. Results: The result revealed that sperm motility of post-cooled spermatozoa improved (P<0.05) by the use of BHT concentrations (1, 2 and 3 mM) in Tris semen extender if compared to the control (85.00±1.09), (83.33± 0.63), (81.67± 0.63) and (78.33± 0.63), respectively. Alive sperm percent was significantly higher in all concentrations of BHT. Sperm abnormalities percent were significantly lower in concentrations of BHT 1 and 2 (11.2±0.2), (11.8±0.2)and (13.4±0.4), respectively. Sperm membrane integrity were significantly higher in BHT concentrations (1, 2, 3, 4 and 5 mM). It is exhibited that improved sperm motility in post-thawed frozen semen in the concentrations of BHT (1, 2, 3 and 4 mM) if compared to the control. The sperm membrane integrity were significantly improved at all concentrations of BHT. Acrosome integrity was significantly higher at BHT concentration 1 mM (81.80±0.57) and (76.00±2.05), respectively. Conclusions: It could be concluded that some concentrations of BHT improved bull semen quality post-cooling and post-freezing.
    2019(2):63-69, DOI: 10.4103/2305-0500.254647
    Abstract:
    Objective: To investigate ameliorative effects of Vitex agnus-castus (VAC) and VAC containing pharmaceutical supplement (VPS) against polycystic ovary syndrome (PCOS). Methods: PCOS in the rats was induced by daily administration of letrozole at 1 mg/kg body weight concentration for 21 d. PCOS rats were then treated daily either with metformin, VAC plant extract or VPS at 70, 8 or 8 mg/kg body weight concentration for 15 d. Rats that received none of these treatments were considered as control. Blood and ovaries were collected from all the rats. Serum glucose, cholesterol, triglyceride and high density lipoprotein-cholesterol were measured spectrophotometrically. Serum insulin, estrogen, progesterone, testosterone, luteinizing hormone, follicle-stimulation hormone, catalase, superoxide dismutase, malondialdehide and reduced glutathione were measured using enzyme-linked immunosorbent assay. Results: Rats treated with letrozole demonstrated a significant increase in serum testosterone, estrogen, cholesterol, luteinizing hormone, triglycerides, glucose, insulin, and malondialdehide levels, and a significant decline in progesterone, follicle-stimulating hormone, high density lipoprotein-cholesterol, catalase and reduced glutathione levels compared to control. Contrarily, no significant change in superoxide dismutase was noted in response to letrozole treatment. Rats treated with metformin, VAC or VPS showed a remarkable reversal in the levels of parameters affected by letrozole treatment. Conclusions: Data indicate that VAC and VPS exert potential ameliorative effects against PCOS through the modulation of hormonal and lipid profile as well as oxidative stress. Moreover, the favorable effects of these compounds are comparable to that of metformin.
    2019(2):45-55, DOI: 10.4103/2305-0500.254645
    Abstract:
    Pyometra is one of the most important and common disorders in canine. The disease results in life threatening condition associated with septicemia and toxemia. The condition commonly occurs during luteal phase of estrous cycle, generally 4 weeks to 4 months of estrous cycle. The age, parity, breeds, administration of hormones are some predisposing factors of the condition. However, interaction between potentially pathogenic bacteria and hormonally primed uterus is believed to result in pyometra. The disease is initiated in the form of cystic endometrial hyperplasia, which later progresses into purulent uterine content following bacterial infection resulting in pyometra. The disturbance in antioxidant and pro-oxidant balance succeeding bacterial infection results in oxidative stress. The resultant stress further induces endometrial degeneration, immunosuppression and additionally aggravates the condition. An important clinical sign is malodorous, sanguineous or mucopurulent vaginal discharge with general symptoms such as vomiting, polyuria, polydipsia, depression, anorexia and occasionally fever. Till date, ultrasonography is the best diagnostic method while radiology and serological test are also helpful. Recently, the estimation of inflammatory mediators (acute phase proteins and prostaglandin metabolites) has appeared as marker for diagnosis of the pyometra. Determination of levels of inflammatory mediators has emerged as an important diagnostic method because it can be helpful in prognosis of the condition. The pyometra can be best treated by surgical manipulation (ovariohysterectomy); however, hormonal treatment can be given in less severe cases which aimed at reducing the effect of progesterone on the reproductive tract. Together, provision of antioxidants could be helpful in disease recovery along with the determined treatment. In this review, incidence, predisposing factors, etio-pathogenesis, oxidative stress associated with condition, diagnostic methods and treatment are discussed.
    2017(3):128-132, DOI: 10.12980/apjr.6.20170306
    Abstract:
    Objective: To explore the effect of Diospyros kaki on cattle spermatozoa during chilling and cryopreservation. Methods: Five milliliter of blended Persimmon (Diospyros kaki) flesh was added to 45 mL TCF to obtain 10% stock solution. Kaki enriched extender (KEE) was prepared by adding to TCF in concentrations 0.0/5.0 mL (control, 0%), 0.5/4.5 mL (1%), 1/4 mL (2%), 1.5/3.5 mL (3%), 2.0/3.0 mL (4%), 2.5/2.5 mL (5%), 3.0/2.0 mL (6%), 3.5/1.5 mL (7%), 4.0/1.0 mL (8%), 4.5/0.5 mL (9%) and 5.0/0.0 mL (10%) to obtain a final volume 5 mL in each tube. Whole egg yolk was added to each tube to obtain KEE with 20% egg yolk (KEEY), all tubes were centrifuged to get rid of debris. Semen was added to the supernatants in other tubes. Extended semen was subjected to evaluation [motility, alive sperm and intact sperm membrane (HOST) %] in both chilled and cryopreserved semen. Conception rate was carried out. Results: Sperm motility was significantly (P<0.000 1) kept high after 11 d of chilling with the concentration 1%, 2%, 3%, 4%, 5% as compared to the control (41.67±1.67, 41.67±1.67, 40.00±0.00, 41.67±1.67 and 41.67±1.67, respectively) and also non-significantly kept high at the other concentrations up to 9 d of chilling. Addition of KEE had significantly(P<0.003 3) improved post thawing sperm motility % with the concentrations 1, 2, 3, 4, 5 and 6% as compared to the control (51.67±5.27, 55.00±3.16, 48.33±1.05, 45.00±3.96, 57.00±2.50,55.00±5.00 and 43.33±5.11 respectively).While the other concentrations exhibit no effect.Addition of KEE maintained alive sperm%, abnormalities% and % of intact spermatozoa membranes (HOST%) as good as the control with all concentrations of kaki used in our study.The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 2%, 4% and 6 % KEE in cattle. Conclusion: It could be concluded that some concentrations of Diospyros kaki improved bull semen quality post-chilling and post-freezing.
    2017(3):97-103, DOI: 10.12980/apjr.6.20170301
    Abstract:
    This article presents a review pertains the laser irradiation effects and its possible mechanisms of action on spermatozoa functions in domestic animals. To improve artificial insemination, laser is sensitive and cost effective technique, when compared to other conventional methods. Laser may have both positive and negative effects on spermatozoa functions. Since the effects of light are mediated by reactive oxygen species, and the levels of these reactive oxygen species following irradiating spermatozoa with laser may be responsible for determining the effects of laser on sperm. Dose of laser may be regarded as of great significance and this dosage of laser may be responsible for determining its effects on spermatozoa. Optimum dosage of laser for improving seminal attributes may vary among various species and this need to be standardized in each of them. The beneficial effects include improving sperm livability, acrosomal integrity, hypo-osmotic swelling response, mitochondrial function and computer-aided sperm analysis parameters. The increase in cytochrome c oxidase activity, ATP levels and mitochondrial membrane potential, in laser irradiated cells may be responsible for enhanced sperm quality parameters. Improving fertility with laser irradiated spermatozoa has been reported in few species like boar and need to be elaborated in other species. In conclusion laser may be regarded as an easy, cheap and time saving technology for improving artificial insemination; in addition, laser may have various potential applications in the field of reproductive biotechnology as well as in livestock farms and veterinary polyclinics.
    2019(3):102-111, DOI: 10.4103/2305-0500.259167
    Abstract:
    Objective: To determine the effect of preparation to play a maternal role in mothers fertilized through in vitro fertilization on maternal anxiety. Methods: A total of 60 mothers undergoing in vitro fertilization were assigned into intervention group and control group. Pregnancy concerns and stress questionnaires were research tools. Questionnaires were completed in both groups before organizing preparation program. Then, the preparation program was held for four sessions for the mothers in the intervention group, and the subjects in both groups were immediately investigated again after completion of preparation program and one month later. Data analyses about pregnancy stress and concerns of mothers and its dimensions were performed by repeated measure (analysis of variance), Mann-Whitney, Friedman and Wilcoxon tests. Analysis of demographic variables was performed by using independent t-test and Chi-square test in SPSS 21. Results: The score of pregnancy stress and concerns before the intervention was not significantly different between the two groups. There was a significant difference in the intervention group in the score of pregnancy stress and concerns before and after the intervention (P<0.001), before and one month after the intervention (P<0.001), immediately after and one month after the intervention (P<0.001) which was not significant in the control group. Conclusions: Maternal preparation program can be effective in reducing maternal stress and concerns.
    2017(2):58-67, DOI: 10.12980/apjr.6.20170203
    Abstract:
    The present review article has described the prevalence of various pathological conditions of reproductive system of female buffaloes starting from ovary to vulva and vagina. Various pathological conditions were analyzed and tabulated as the total number of animal examined, number of the animals showed pathological lesions, percentage of animal showed various pathological lesions and percentage of individual pathological lesion in different parts of reproductive system of female buffaloes. The incidence of disorders of female genital organs of buffaloes has been reported by various authors at various percentages in different countries. The incidence of pathological conditions was recorded in clinically infertile cows after slaughtering or from apparently healthy buffaloes slaughtered for human consumption and/or based on postmortem examination. This review is comprehensively covering pathological conditions of female buffalo hitherto which was not previously described at one place. This review will provide a comprehensive knowledge about the prevalence of different pathological condition of different parts of reproductive tract of female buffaloes. The review has six numbers of tables described about the various pathological conditions from ovary to vulva and vagina in female buffaloes. The present review article will be very useful to the buffalo farmers, buffalo breeders and researchers are working in buffalo reproduction & breeding and pathology.
    2017(2):93-96, DOI: 10.12980/apjr.6.20170209
    Abstract:
    Objective: To determine the effect of different cervical dilators on cervical dilation and reproductive performance of fat-tailed ewes. Methods: In experiment 1 140 ewes were divided into seven groups with seven different treatments as following: 10 mL normal saline (control group), 100 IU oxytocin (OT group), 100 μg estradiol and 100 IU oxytocin (E2+OT group), 5 mL relaxin (R group), 2 mL sensiblex (SEN group), 200 μg misoprostol (MIS group) or 200 μg dinoprostone (DIN group). In experiment 2, artificial insemination was applied for evaluation of reproductive performance in experimental groups. Results: In experiment 1, the highest cervical dilation was observed in OT (90%) and E2+OT (100%) groups (P<0.05), while no significant differences was found among DIN, MIS, SEN and R groups (80%, 75%, 70% and 65%, respectively). In addition, the lowest cervical dilation was observed in control group. Experiment 2 found no significant differences among control, OT and E2+OT groups. The highest pregnancy rate, parturition rate and lambing rate were observed in OT groups (60%, 60% and 70%, respectively) and E2+OT groups (65%, 60% and 70%, respectively) compared to SEN, R, MIS and DIN groups (P<0.05). Conclusions: Oxytocin treatment alone or with estradiol could be used as a suitable dilator for improving reproductive efficiency during artificial insemination in fat-tailed ewes.

Asian Pacific Journal of Reproduction, a publication of Hainan Medical University, is a peer-reviewed print + online Bimonthly journal.

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