Asian Pacific Journal of Reproduction
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    2020(6):255-260, DOI: 10.4103/2305-0500.298772
    Abstract:
    Early diagnosis of pregnancy plays an important role to minimize reproductive losses in farm animals. There are several methods for pregnancy diagnosis like profiling of reproductive hormones (such as progesterone and estrone sulfate), but sometimes they provide false-positive results. Embryo specific pregnancy markers, which delineate the presence and viability of the embryo, are considered as perfect for pregnancy determination. Pregnancy-associated glycoproteins are distinguished as the best indicator for the determination of early pregnancy, fetal number, and birth weight of kids. Pregnancy-associated glycoproteins are structurally correlated to aspartic proteinase and are communicated in the external epithelial cell layer of the placenta. They have been found to share about half amino acid sequence identity with pepsinogen, pepsin, cathepsin D and E. Dislike different individuals from aspartic proteinase family, numerous pregnancy-associated glycoproteins appear to be latent compound as a result of amino acid substitutions in and around the catalytic site. This review is to discuss the scope and prospects of pregnancy-associated glycoproteins as a pregnancy marker in farm animals, more specifically in goats.
    2020(6):261-266, DOI: 10.4103/2305-0500.298773
    Abstract:
    Objective: To investigate morphokinetic characteristics of embryos displaying either reverse cleavage or direct cleavage during the first, second or third cleavage cycle. Methods: A total of 167 in vitro fertilization and/or intracytoplasmic sperm injection treatment cycles undertaken by 167 women [aged (35.0±4.6) years] were included for reverse cleavage analysis, and a total of 167 in vitro fertilization and/or intracytoplasmic sperm injection treatment cycles undertaken by 167 women [aged (33.8 ±4.3) years] were included for direct cleavage analysis in this study. Using a sibling-embryo design, morphokinetic profiles (both before and after the onset of abnormal event) of embryos displaying reverse cleavage (n=241) or direct cleavage (n=244) were compared with their unaffected siblings (the controls) in the first, second and third cell cycles (n=32, n=142, n=562; n=195, n=412,n=205, respectively), at different developmental stages up to day 3. Results: Direct cleavage embryos demonstrated significantly delayed cleavage rates prior to the event regardless of developmental stage of the occurrence, while reverse cleavage embryos showed similar cleavage rates to their unaffected siblings. Post event, direct cleavage embryos sped up cleavage rates while reverse cleavage embryos slowed down. Conclusions: Altered morphokinetic profiles are displayed by direct cleavage embryos both before and after their occurrence and reverse cleavage embryos after the occurrence, which could potentially confound morphokinetic comparisons if not separated from their unaffected sibling embryos. Further study is warranted in order to fully understand the biological mechanisms of such events.
    2020(6):267-274, DOI: 10.4103/2305-0500.298774
    Abstract:
    Objective: To investigate the effect of Camellia (C.) sinensis in mitigating oxidative damage and reproductive toxicity in testis induced by sodium fluoride in a rat model. Methods: Twenty-four adult male Wister rats were divided into 4 groups, with 6 rats in each group. Group 1 orally received distilled water (1 mL/100 g body weight) daily and served as the control group, while group 2 received drinking water with 100 ppm sodium fluoride per day for 21 consecutive days, group 3 was administered with only C. sinensis extract by gavage at a dose of 100 mg/kg body weight and group 4 received drinking water with 100 ppm sodium fluoride and 100 mg/kg body weight C. sinensis leaf extract per day for 21 consecutive days. At the end of the treatment, the rats were sacrificed under light ether anesthesia. The gonado-somatic index, sperm count and motility, serum level of luteinizing hormone and testosterone were assayed. Lipid peroxidation [malondialdehyde (MDA) level], nitric oxide (NO) production, and activities of antioxidant enzymes - superoxide dismutase (SOD), catalase, and reduced glutathione level (GSH) were also analysed. Results: Sodium fluoride treatment significantly decreased gonadosomatic index, sperm count and motility as well as the serum level of luteinizing hormone and testosterone (P<0.05). The histological examination of testes revealed atrophy and degenerative changes in several seminiferous tubules, along with enhanced interstitial space and a reduced number of Leydig cells. There was a highly significant increase in NO and MDA production (P<0.05), while SOD, catalase activities and GSH level decreased significantly (P<0.05). However, C. sinensis significantly restored testicular weight, sperm parameter, hormonal level (P<0.05), and also reversed MDA and NO generation and antioxidant enzymes activities in the testicular tissue (P<0.05). Conclusions: C. sinensis may have an ameliorative role against sodium fluoride-induced oxidative damage in the testis probably because of its antioxidant property.
    2020(6):275-282, DOI: 10.4103/2305-0500.298775
    Abstract:
    Objective: To identify the alteration of tyrosine phosphorylated protein expression in rats with polycystic ovary syndrome (PCOS). Methods: Sixteen female Sprague-Dawley rats were divided into the control and letrozole-induced PCOS groups. The oestrus cycle of rats was performed by vaginal smear. Sex hormones and morphology of the ovary, oviduct, and uterus were observed. Expressions and intensity of androgen receptor and tyrosine phosphorylated proteins of reproductive organs were investigated by Western blot. Results: Various polycysts and increased androgen receptor expression were present in the ovary of the PCOS group. The levels of follicle-stimulating hormone and testosteone were significantly higher in the PCOS group while progesterone and estradiol levels were significantly decreased as compared with the control group (P<0.05). Only the size of uterus in the PCOS group was significantly smaller than the control group. However, the density of collagen fibers observed in PCOS uterus was greater than the control group. Moreover, tyrosine phosphorylated proteins were significantly overexpressed in ovary (52, 42, and 28 kDa), oviduct (72, 56, 42, and 28 kDa), and uterus (53 and 42 kDa) of the PCOS group compared to the control group. Conclusions: Presence of tyrosine phosphorylated proteins in the ovary, oviduct and uterus suggests that overexpression of tyrosine phosphorylated proteins may be involved in potential mechanism of female infertility especially in PCOS.
    2020(6):283-290, DOI: 10.4103/2305-0500.298776
    Abstract:
    Objective: To investigate the hepatic and reproductive toxicity of dichlorvos and lead acetate on male Wistar rats. Methods: Fifteen adult male Wistar rats (170-190 g) were randomly divided into three groups, with 5 rats in each group. Group 1 received 0.5 mL distilled water orally and served as the control group, while groups 2 and 3 were orally treated with 2 mg/kg body weight (b.w.) dichlorvos and 10 mg/kg b.w. lead acetate, respectively, for 55 days. Epididymal sperm, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone concentrations, testicular 17β-hydroxysteroid dehydrogenase activity (17β-HSD), androgen receptor expression, aspartate aminotransferase (AST), alanine aminotransferase (ALT), testicular oxidant and antioxidant enzymes were evaluated with standard methods. Results: Sperm count, motility, morphology, FSH, LH, testosterone levels, 17β-HSD, androgen receptor expression, and catalase activity were significantly reduced in the dichlorvos and lead acetate treated groups as compared with the control group (P<0.05). The liver AST, ALT activities and malondialdehyde concentration were significantly increased in the dichlorvos and lead acetate treated groups as compared with the control group (P<0.05). Conclusions: The reproductive and hepatic toxicity activities of dichlorvos and lead acetate in male Wistar rats are similar.
    2020(6):291-297, DOI: 10.4103/2305-0500.298777
    Abstract:
    Objective: To evaluate the development of ovarian follicles in female albino rats following vitamin D3 supplementation. Methods: Eighteen prepubertal female albino rats, aged 3-4 weeks, weighing (70.25±9.16) g, were assigned to three groups (n=6 in each group). Group A was treated with 5.00 mL/kg of distilled water and served as the control group, group B was treated with 0.025 mg/kg of vitamin D3 dissolved in distilled water, and group C was treated with 0.125 mg/kg of vitamin D3 dissolved in distilled water. All treatments were administered orally, twice weekly for 28 days. Blood and ovaries were harvested under anaesthesia. Serum vitamin D3 levels were determined by using spectrophotometric method. Ovaries were processed for histology and every10th hematoxylin and eosin stained-section was selected for histomorphometry. The number of follicles at each developmental stage was estimated. Results: Both 0.025 mg/kg and 0.125 mg/kg of vitamin D3 significantly increased serum concentrations of vitamin D3 and calcium (P<0.05), but did not alter inorganic phosphorus concentration (P>0.05). The control group had fewer growing follicles (primary, secondary and antral follicles) and more nongrowing follicles (primordial and atretic follicles) when compared with the vitamin D3-supplemented groups (P<0.05). Vitamin D3 at 0.025 mg/kg significantly increased antral follicles and corpora lutea counts (P<0.05). Vitamin D3 at 0.125 mg/kg significantly increased total, primordial and atretic follicles counts (P<0.05), but significantly decreased primary, secondary, antral follicles count, ovarian weight, relative ovarian weight, and ovarian surface area when compared with the control group and rats treated with 0.025 mg/kg of vitamin D3 (P<0.05). Conclusions: Vitamin D3 supplementation at 0.025 mg/kg can enhance optimal ovarian follicle recruitment and development in female rats.
    2020(6):298-304, DOI: 10.4103/2305-0500.298778
    Abstract:
    Objective: To investigate the effects of soft agar on in-vitro proliferation of neonate mouse spermatogonial stem cells cocultured with Sertoli cells. Methods: Tissues of neonate NMRI male mice testes were used for harvesting spermatogonial stem cells and Sertoli cells. After cell harvest, flow cytometry using promyelocytic leukemia zinc-finger (PLZF) protein antibody was used to assess the purity of the cells. The isolated testicular cells were cultured in the absence (the control group) or presence of soft agar-coated dishes (the experimental group) supplemented with leukemia inhibitory factor and glia cell line–derived neurotrophic factor for two weeks. Alkaline phosphatase activity was assessed in the colonies formed after two weeks of culture by alkaline phosphatase staining. On day 14 of culture, the expression levels of DNA-binding protein inhibitor (ID-4) and PLZF genes in the undifferentiated cells were evaluated by the detection of PLZF protein antibody using real-time PCR and immunocytochemistry techniques. The number and diameter of the colonies of spermatogonial stem cells were assessed by ImageJ software. Results: In the experimental group, the number and the diameter of colonies significantly increased as compared with those in the control group (P<0.05, P<0.01, respectively). In addition, the level of expression of ID-4 and PLZF genes in the undifferentiated cells significantly increased in the experimental group as compared with the control group (P<0.01). However, the expression of tyrosineprotein kinase kit (c-kit) gene in differentiated cells decreased in the experimental group as compared with the control group, but there was no significant difference between the two groups. Conclusions: Spermatogonial stem cells can be efficiently proliferated by culturing the stem cells in soft agar-coated dishes. The new protocol used in this study can be a valuable method for future studies.
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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):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):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 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.
    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(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.
    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(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(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(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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