Asian Pacific Journal of Reproduction
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    2020(1):1-8, DOI: 10.4103/2305-0500.275522
    Abstract:
    Objective: To find the association between testosterone serum levels and severity of late onset pre-eclampsia. Methods: This case-control study involved 34 patients with pre-eclampsia and 24 healthy control pregnant women matched for gestational and maternal age. The recruited pregnant women were divided into two groups: the pre-eclampsia group (34 pregnant women with pre-eclampsia) and the control group (24 healthy pregnant women). Lipid profile, blood urea, serum creatinine, proteinuria, total serum testosterone, sex hormone binding globulin and free androgen index were evaluated. Moreover, body mass index and blood pressure profile were measured. Results: There were high systolic and diastolic blood pressures in the pregnant women with late-onset pre-eclampsia when compared with healthy pregnant women (P<0.01). Total serum testosterone was higher in women with pre-eclampsia compared with healthy pregnant women (P=0.001). The free androgen index was higher in women with pre-eclampsia compared with healthy pregnant women (P=0.001). Sex hormone binding globulin level was low in women with pre-eclampsia compared with healthy pregnant women (P=0.001). High total serum testosterone was significantly correlated with all measured variables (P=0.001), except for body mass index and pulse pressure (both P=0.070). Smoking habit was low in those patients compared with healthy pregnant women. Meanwhile, total serum testosterone serum level was significantly correlated with number of cesarean sections (r=0.86, P<0.01) and nulliparty (r=0.56, P<0.01). Conclusions: Late onset pre-eclampsia in pregnant women is associated with high serum levels of total serum testosterone, free androgen index, low sex hormone binding globulin, low smoking habit with positive history for nulliparity and caesarean sections that are correlated with high blood pressure profiles. Therefore, high total serum testosterone which is correlated with most of risk factors of late-onset pre-eclampsia is regarded as a proxy biomarker reflecting the severity of late onset pre-eclampsia.
    2020(1):9-15, DOI: 10.4103/2305-0500.275523
    Abstract:
    Objective: To determine the antioxidant status of females with polycystic ovarian syndrome. Methods: Blood samples of 85 females (45 newly diagnosed polycystic ovarian syndrome patients and 40 apparently healthy subjects) between 25 and 45 years of age were obtained from Ekiti State University Teaching Hospital, Ado Ekiti, Ekiti State, Nigeria. Spectrophotometry was used to evaluate the plasma concentrations of malondialdehyde, vitamin A, C and E, reduced glutathione and activities of glutathione peroxidase, catalase and superoxide dismutase. Results: The concentrations of malondialdehyde and glutathione peroxidase activity significantly increased (P<0.05) in patients with polycystic ovarian syndrome compared with the healthy controls, while concentrations of reduced glutathione, vitamins A, C, E and activities of catalase and superoxide dismutase of patients with polycystic ovarian syndrome showed significant decrease (P<0.05) compared to the healthy controls. This study showed that oxidative stress may assume a role in the pathogenesis of polycystic ovarian syndrome. There were significant negative correlations between malondialdehyde and superoxide dismutase, catalase (P<0.01) and vitamin A (P<0.05), while there was a significant positive correlation (P<0.01) between malondialdehyde and glutathione peroxidase. In addition, vitamins A (P<0.05), C (P<0.01) and E (P<0.01) showed significant positive correlations with catalase antioxidant enzyme. However, vitamins C and E showed significant positive correlation (P<0.05) among each other. Conclusions: The involvement of antioxidants in the management of polycystic ovarian syndrome may be helpful as secondary therapy to prevent oxidative damage and may be used as a potential approach to overcome metabolic as well as reproductive disorders associated with infertility in polycystic ovarian syndrome.
    2020(1):16-21, DOI: 10.4103/2305-0500.275524
    Abstract:
    Objective: To assess the association between genetic variants of transferrin receptor 2 (TFR2) exon 4 and anemia status and to describe the expression levels of several cytokines, hepcidin, soluble transferrin receptor and erythropoietin. Methods: Institutional based comparative study was done randomly to recruit 106 pregnant women who attended antenatal care in three different health centers in Boyolali Regency, Central Java from May 2015 to September 2015. DNA was extracted from peripheral blood samples of selected pregnant women and sequencing was done for TFR2 exon 4. Furthermore, enzyme-linked immunosorbent assay was conducted to measure the expression levels of interleukin 6, interleukin 4, transforming growth factor β and iron-metabolism related proteins such as hepcidin, soluble transferrin receptor, and erythropoietin. Gene alignment was performed by using a CLUSTAL W program. Collected data were analyzed statistically by using parametric and nonparametric tests with Statistical Product and Service Solutions (SPSS) 20.0 for Windows. Results: Three novel genetic variants from TFR2 exon 4 (position 603, 605 and 606) were associated with anemia status. Moreover, the expression levels of interleukin 6, interleukin 4, transforming growth factor β and erythropoietin were higher in anemic pregnant women than those of nonanemic pregnant women but only erythropoietin level reached statistical significance. These results were followed by decreases of hepcidin and soluble transferrin receptor levels. Conclusions: Various factors contribute to anemia prevalence among pregnant women in Boyoali Regency, Central Java, Indonesia. Our novel findings showed that TFR2 exon 4 has 3 mutational sites in position 603, 605 and 606. These novel genetic variants may provide a new insight into the role of TFR2 in anemia.
    2020(1):22-30, DOI: 10.4103/2305-0500.275525
    Abstract:
    Objective: To evaluate changes of feline (Felis catus) oocytes proteins during in vitro maturation by using the proteomic approach. Methods: Immature oocytes (germinal vesicle) isolated from female cats were cultured and collected at 0 h and 24 h. After collection, oocytes were investigated into immature (germinal vesicle) and mature (metaphase Ⅱ) stages. The qualitative profiles of the proteins at the immature and mature stages were determined by one-dimensional electrophoresis and liquid chromatography-mass spectrometry. Results: Our data revealed that following 24 h in vitro maturation the maturation rate (metaphase Ⅱ stage) was 58.7%. Eighty-one of the 260 proteins analyzed were differentially expressed between the germinal vesicle stage and the metaphase Ⅱ-arrest stage. Proteomic analysis of germinal vesicle and metaphase Ⅱ oocytes showed abundant expression of proteins involved in transportation (10%), indicating that this was a major characteristic of germinal vesicle oocytes. Similarly, analysis of the proteome of metaphase Ⅱ oocytes indicated that cell cycle proteins were overexpressed. Interestingly, proteins involved in DNA repair and apoptosis were only expressed in germinal vesicle oocytes and proteins involved in fertilization were only expressed in metaphase Ⅱ oocytes. Conclusions: The overexpression of certain proteins in germinal vesicle and metaphase Ⅱ is necessary for oocyte development and maturation. Our findings provide a valuable resource for further investigations into protein expression in oocytes at different developmental stages.
    2020(1):31-36, DOI: 10.4103/2305-0500.275526
    Abstract:
    Objective: To assess the estrogenic activity of hydroalcoholic extract of Clitoria (C.) ternatea leaves in female Wistar rats. Methods: Hydroalcoholic extract of C. ternatea leaves prepared by using cold maceration method was tested for estrogenic activity. An acute toxicity study was conducted to estimate the safe dose using OECD 423 guidelines. For estrogenic activity, ovariectomized female rats were divided into four groups, with 6 rats in each group. The control and standard groups were administered with 1% carboxymethyl cellulose orally and estradiol valerate at 1 μg/rat/day subcutaneously, respectively. The third group was administered with hydroalcoholic extract of C. ternatea at the dose 500 mg/kg body weight orally and the fourth group was administered with hydroalcoholic extract of Clitoria (C.) ternatea at the dose 500 mg/kg body weight orally along with estradiol valerate at dose 1 μg/rat/day subcutaneously. All treatments lasted for 7 consecutive days and estrogenic activity was assessed by observing vaginal cornfication. On day 8, all animals were sacrificed and uterine horns were dissected out. Utrine weight was measured and blood serum was further processed for the estimation of biochemical parameters like cholesterol, total proteins, alkaline phosphatase and estrogen by autoanylser. Histological studies of uterus were also carried out. Results: Acute toxicity studies indicated the hydroalcoholic extract of C. ternatea leave was found to be safe up to the dose level of 2 000 mg/kg. Oral administration of C. ternatea extract at the dose 500 mg/kg body weight and and estradiol valerate (1 μg/rat/day) caused morphological changes i.e. increase in uterine weight, vaginal opening and cornification of cells; biochemical changes i.e. increase in cholesterol, total protein, alkaline phosphatase and estrogen contents; histological changes i.e. increase in uterine diameter, thickness and height of endometrium. Simultaneous administration of C. ternatea extract with estradiol valerate showed a synergistic effect. Histological investigations further confirmed the strong estrogenic nature of C. ternatea extract. Conclusions: C. ternatea extract (500 mg/kg) showed a significant estrogenic activity which is also supported by biochemical and histological studies. So, on the basis of these findings, it can be concluded that C. ternatea can be used as an alternative to synthetic oral contraceptives.
    2020(1):37-43, DOI: 10.4103/2305-0500.275527
    Abstract:
    Objective: To examine sex reversal both by oral and by immersion using 17α-methyltestosterone on the methyltestosterone residual concentration and the organ histopathology of tilapia fish. Methods: This study used oral and immersion treatment methods for sex reversal of tilapia fish and used normal fish as the control and each treatment was repeated 4 times. 17α-methyltestosterone at dosages of 60 mg/kg feed and 0.5 mg/L were used for oral and immersion methods, respectively. In the first step, tilapia fry were reared at 100 L aquaria, with a density of 1 fish/L for 2 months. In the next step, male tilapias were reared at happa (net cage) of (2×1×1) m3 size in the controlled pond, with a density of 30 fish/happa for 3 months. The methyltestosterone residual concentrations were analyzed by one-way analysis of variance and Duncan’s multiple range tests, while organ histopathology was analyzed by descriptive method. Results: Residual concentrations in the serum of methyltestosterone-treated fish were significantly lower than that in normal fish, especially in 4- and 5-month-old tilapias with averages of less than 5 µg/L, while in normal fish was more than 5 µg/L. In the flesh, methyltestosterone residual concentrations showed relatively no significant differences between the oral and immersion treatment groups and methyltestosterone-treated fish remained lower compared to normal fish, except in 5-month-old tilapia. Methyltestosterone-treated tilapia exhibited histopathological changes on gill, liver, kidneys, and intestine organs. Conclusions: Sex reversal either by oral or by immersion has methyltestosterone residual concentration, but does not exceed the limits (5 µg/L or 5 µg/kg) of synthetic steroid on the fish body, although methyltestosterone causes histopathological changes on gill, liver, kidneys, and intestine.
    2020(1):44-48, DOI: 10.4103/2305-0500.275528
    Abstract:
    Objective: To investigate the bacterial infections in repeat breeder dairy cows and to evaluate the treatment effects of hyperimmune serum against two main endometritis bacteria, Escherichia (E.) coli and Trueperella pyogenes. Methods: A total of 29 Holstein multiparous cows with three or more unsuccessful artificial inseminations were carried out and examined to confirm the absence of reproductive tract abnormality and vaginal discharges. Uterine lavage was performed to collect uterine samples for bacterial and fungal cultures. In addition, polymerase chain reaction (PCR) was done to detect Trueperella pyogenes, E. coli , Fusobacterium necrophorum, and Prevotella melaninogenicus. The cytological study was done on cervical mucus. The hyperimmune serums produced against Trueperella pyogenes and E. coli were infused into the uterus of repeat breeder cows and two doses of prostaglandin F2α were administrated intramuscularly within 14 days' interval. Results: There were 10 positive samples in the bacterial culture with 19 isolations and no growth of the main causative bacteria of endometritis. In the PCR method, no Trueperella pyogenes, Fusobacterium necrophorum, and Prevotella melaninogenicus were found. However, 11 positive samples of E. coli were identified by PCR. Cladosporium was detected in one case of repeat breeder cows. The median (interquartile range) of neutrophils in vaginal discharge was 12 (22.5). Eighteen from twenty-nine (62.1%) repeat breeder and eight from eleven (72%) cows with E. coli infections in PCR were made pregnant in the first artificial insemination after treatment by intrauterine infusion of hyperimmune serum and prostaglandin F2α . Conclusions: Intrauterine administration of hyperimmune serum could be an alternative to antibiotics for the treatment of repeat breeder cows.
    2020(1):49-54, DOI: 10.4103/2305-0500.275635
    Abstract:
    Objective: To establish reference values for pixel intensity of testicular ultrasonogram of rams in breeding and non-breeding seasons, and to investigate if the seasonal changes in testicular echogenicity and heterogeneity are associated with semen characteristics. Methods: Five Awassi rams aged 3-5 years old and weighing 45-60 kg were subjected to ultrasonographic scanning of the testes twice monthly for one year (from January 2018 to December 2018), together with semen collection and evaluation of ejaculate volume, sperm motility, sperm concentration, sperm morphology and viability. The ejaculate volume was determined in a graduated collection tube (scale of 0.1 mL). Mass and individual sperm motility was expressed in percentage of motile spermatozoa under optical microscope equipped with a warm stage. Sperm concentration was determined by using a Neubauer chamber. Semen smears stained with eosin-nigrosin were used to determine the percentage of live spermatozoa and sperm cell morphology by using a light microscope. Results: The mean testicular pixel intensity was the lowest in winter and increased gradually from breeding season to non-breeding season, reaching its maximum value in summer (P<0.05). Pixel intensity was found to have a significant negative correlation with progressive motility (r=-0.605, P<0.05), and sperm concentration (r=-0.619, P<0.05). It is also positively correlated with the percentage of sperm morphological abnormalities (r=0.666, P<0.05). Conclusions: Pixel intensity values of testicular ultrasonogram in rams undergo marked seasonal changes that are associated with fluctuations in photoperiod and ambient temperature. The resulting values of testicular echogenicity (pixel intensity) throughout the year provide useful reference values for predicting the testicular function in Awassi rams.
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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):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):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):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(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):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.
    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.
    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):54-57, DOI: 10.12980/apjr.6.20170202
    Abstract:
    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.
    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.
    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.
    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.
    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 ) 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 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.
    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.
    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.
    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(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.

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

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