Asian Pacific Journal of Reproduction

241-247

Authors: GN Purohit, Atul Shanker Arora, Tilok Gocher, Mitesh Gaur, Chandra Shekher Saraswat, Pankaj Mishra

Uterine prolapse is an emergency postpartum problem occurring within 24 h of parturition and resulting in death or serious complications in unattended cases. Poor myometrial contractions during the post-partum period and traction during difficult births are two postulated etiologies and low serum calcium appear to be a significant risk factor for uterine prolapse in buffaloes. Shortly after eversion the uterus gets inflamed and edematous and shock may ensue in cases with excessive bleeding. Prompt replacement of prolapsed uterus with sufficient care assures good prognosis. The etiology, risk factors, clinical findings and approaches for therapy of uterine prolapse in buffaloes are mentioned in this review.

254-260

Authors: Asmaa A. Mostafa, Mohamed. S. El-Belely, Sayed. T. Ismail , Reda. I. El-Sheshtawy, Mohamed I. Shahba

Objective: To estimate the result of egg yolk replacement with alternative cryopreservatives such as plant-derived lecithin from soybean on sperm quality parameters pre and post freezing in buffalo bulls. Methods: The control cryopreservation extender was tris-citric acid-fructose-egg yolk-glycerol (TCFYG) diluent. Semen samples were extended gradually 1:10 with TCFYG control extender and tris-citric acid-fructose-glycerol (TCFG) extender plus variable concentrations of soybean lecithin (0.5%, 1.0%, 1.5%, 2.0%, 2.5% and 3.0%) to ensure 60 million active spermatozoa/mL of the extended semen. The diluted semen samples were refrigerated slowly (roughly for 2 h) up to 5 ℃ and equilibrated for 2 h. Semen was filled into 0.25 mL polyvinyl French straws (IMV, France). After equilibration period, the straws were placed horizontally on a rack and frozen in a vapor 4 cm above liquid nitrogen for 10 min and were then dipped stored in liquid nitrogen at -196 ℃ . Results: The respective overall percentages of forward motile spermatozoa, live spermatozoa, morphologically normal spermatozoa, acrosome integrity and hypo-osmotic swelling reactivity observed primarily in fresh semen, after equilibration (pre-freeze stage) and post freezing (post-thaw stage) in TCFYG (control) extended semen declined progressively and statically (P<0.01) during these periods of study. Pre-freezing stage: replacement of egg yolk into TCFG with soybean lecithin at concentrations of 1.0% and 1.5% significantly (P<0.01) ameliorated the maintenance of (motility, viability, acrosome and membrane integrity %), meanwhile it had significantly (P<0.01) reduced the abnormality % of spermatozoa to the lowest value compared to control TCFYG and to some other concentrations in use. Post-thaw stage: the replacement of egg yolk with 1.0% soybean lecithin (SL) showed significantly (P<0.01) higher percentage of sperm progressive motility compared to 1.5% SL and TCFYG control. These values were significantly (P<0.01) higher than 0.5%, 2.0%, 2.5% and 3.0% SL. The post thawing live sperm percentage mean values were significantly (P<0.01) higher in 1.0% SL and 1.5% SL compared to control. These values were significantly (P<0.01) higher than in 0.5%, 2.0%, 2.5% and 3.0% SL. The mean values of post-thaw morphological normal sperm percentage did not differ between 1.0% SL and control groups but significantly (P<0.01) higher than 0.5%, 1.5%, 2.0%, 2.5% and 3.0% SL. The respective percentage mean values of post-thaw sperm with head, mid-piece and tail abnormalities were significantly (P<0.01) lower in 1.0% SL than all other SL concentrations. Concerning the post-thaw percentages of acrosome and sperm membrane integrity, the respective mean values were significantly (P<0.01) higher in 1.0% SL and 1.5% SL as compared to control. Mean values of both parameters in the 0.5% SL were intermediate between 1.0% and 1.5% SL versus control groups. The previously mentioned mean values in acrosome/membrane integrity were significantly (P<0.01) higher than 2.0% SL, 2.5% SL and 3.0% SL. Conclusions: Lecithin-based diluent can be a potent proper alternative extender for preservation of spermatozoa during pre- and post-freezing process. SL 1.5% extenders have supplied an optimal environment and condition for ameliorating the quality of pre-freezing and post- thaw buffalo spermatozoa by means of improved motility, viability,

266-269

Authors: Caner Öztürk, Mehmet Bozkurt Ataman

Objective: To examine the effects of paclitaxel and resveratrol on rabbit semen. Methods: This study consisted of four groups: control group (40 mL saline), paclitaxel group (5 mg/kg paclitaxel), resveratrol group (4 mg/kg resveratrol) and paclitaxel+resveratrol group (5 mg/kg paclitaxel+4 mg/kg resveratrol). Administrations were i.v. (in 40 mL saline) and continued 8 weeks. Sperm motility was evaluated using phase-contrast microscopy. Mitochondrial activity, membrane and acrosome integrity were performed by fluorescence staining. Lipid peroxidation, total glutathione and antioxidant potential levels were determined by spectrophotometry. Results: Paclitaxel decreased the sperm motility and fluorescence staining results compared to the control (P<0.05). The paclitaxel and resveratrol group showed better results of the same parameters compared to the paclitaxel group (P<0.05). No significant difference was observed in lipid peroxidation, total glutathione, antioxidant potential and fertility results (P>0.05). Results of this study showed that paclitaxel decreased semen parameters and resveratrol had a protective effect on these parameters. Conclusions: Paclitaxel has negative effects on spermatological indicators and biochemical assays, while resveratrol prevents these negative effects of paclitaxel.

274-279

Authors: Grace A. Essiet, Godwin C. Akuodor, Daniel OJ Aja, Mathew O. Nwokike, Desmond O. Eke, Anuli N. Chukwumobi

Objective: To evaluate the effect of Salacia lehmbachii (S. lehmbachii) ethanol root bark extract on estrous cycle and sex hormones in female rats. Methods: Forty-eight virgin rats with regular 4-day cycle were grouped into four and each group was further subdivided into ‘a’ and ‘b’ (n=6). Each group was orally treated for 28 days with 2 mL of distilled water (control), ethanol root bark extract of S. lehmbachii in doses of 250, 500 and 750 mg/kg/body weight (groups 2, 3, 4, respectively). Estrous cycle was determined daily using the vaginal smear method. Rats in ‘a’ subgroups were weighed and sacrificed on the 29th day, and blood was collected for serum generation which was used for hormonal assay and sex organs were removed and weighed. Rats in ‘b’ subgroups were discontinued from treatment for 2 weeks and the parameters above were reassessed. Results: The mean length of estrous cycle and duration of diestrous of treated rats were prolonged dose dependently compared to control. The increase was significant (P<0.05) at 500 and 750 mg/kg. The other estrous phases were shortened in the same pattern. Relative weights of sex organs were reduced significantly (P<0.05) at the highest dose. Sex hormones levels were significantly (P<0.05) reduced compared to control. The above changes reverted towards the control values two weeks post treatments. Conclusions: Ethanol root bark extract of S. lehmbachii (high doses) has antifertility effect in female rats as it prolongs the estrous and diestrous cycle, and reduces serum sex hormones levels. The observed alterations were reversible.

193-205

Authors: Kamel M.E. Mohammed

Many countries in the world consider the buffalo as a priority animal for the future, since it plays a pivotal role in human food sustainability. Even though Food and Agriculture Organization has termed the buffalo as an important undervalued asset, this species has yet to drive the same attention as cattle. Egypt has a wealth of buffaloes dispersed in small herds all over the country, so the efforts that have been made to improve their genetic background show little return. Contrarily, other countries concerned with buffalo improvement have already used a data recording system in buffalo herds, allowing to achieve a much faster improvement progress. This review intends to survey the existing information on the application of assisted reproduction techniques to improve buffalo productivity. The strength points that may help to improve buffalo production are identified, and the obstacles hindering the genetic improvement of Egyptian buffalo are characterized. Therefore, this work will gather information related to buffalo and compile it for an audience of researchers and specialists to enforce international collaboration for the development of buffalo production. Also, it will open the way for people interested in developing a future vision for buffalo potential, which will be helpful to close or minimize the biological gaps of buffaloes’ researches.

214-219

Authors: Farzaneh Mahmoudi-Lafout, Fahimeh Mohammadghasemi

Objective: To evaluate the effect of graded doses of acetylsalicylic acid (ASA) on sperm chromatin integrity and sex hormones in the adult male mouse. Methods: Forty-nine male adult mice were divided into seven groups. Control group received no drug, sham group received ASA solvent (dimethylsulfoxide 0.1%), groups 3 to 7 received different doses of 0.05, 0.10, 0.50, 1.00 and 5.00 mg ASA, daily for 14 d, respectively. On day 15, evaluations were made by sperm chromatin dispersion test for the study of DNA sperm integrity, radioimmunoassay for the study of testosterone and luteinizing hormone (LH) level and histopathology of testis for Jhonson’s scoring. Results: ASA in groups 0.50, 1.00 and 5.00 mg reduced big halo sperms. DNA fragmentation significantly increased just in 5 mg group. Serum level of testosterone in doses of 0.50, 1.00 and 5.00 mg groups reduced significantly (P<0.01) while LH level was not affected. Johnson’s score reduced in all ASA treated groups. Conclusions: Administration of ASA over the 14 days in dose of 5.00 mg increases sperm DNA fragmentation index and therefore reduces DNA integrity and in doses of 0.50, 1.00, 5.00 mg reduces serum testosterone level with no effect on LH. Generally, ASA has deleterious effects on the male reproductive indices even in low doses.

225-228

Authors: Seyed Mansour Moosavi, Hossein Barzegar Ganji, Hajar Ramezanikhah, Mitra Arianmanesh

Objective: To investigate the effect of human chorionic gonadotropin (hCG) on the motility of rat sperm and the fertilization rate following in vitro fertilization. Methods: hCG concentrations of 25, 50, 75 and 100 ng/mL with incubation time of 1 h and 2 h were used in medium to study sperm motility. Then, 25 and 100 ng/mL of hCG with incubation time of 2 h were applied for in vitro fertilization. After 6 h, the number of two pronuclei was counted. Obtained data was subjected to one way ANOVA and Bonferroni-Dun hoc-post test. Results: Total motility and progressive motility of sperms were increased significantly (P<0.05) in hCG experimental group with concentration of 25 ng/L and incubation time of 2 h compared to control group, however, total motility and progressive motility of sperms were decreased significantly (P<0.05) with increasing dose of hCG. Fertilization rate was decreased significantly (P<0.05) in hCG experimental group with dose of 100 ng/mL compared to 25 ng/mL and control group. Conclusions: The effect of hCG on the rat sperm motility and the rate of their fertility is dose-dependent so that hCG with dose of 25 ng/mL leads to an increase in rat sperm motility and to some extent fertilization rate.

236-238

Authors: Chike F. Oguejiofor

Cases of acquired infertility in dogs often have poor prognosis and can be frustrating to clinicians in canine practice. In this case, a 3.5-year-old male Caucasian dog was presented for infertility evaluation. He had no record of successful breeding despite being mated with multiple bitches over a two-year period. Breeding soundness examination was performed including history evaluation, general physical and reproductive system examination, hematology, libido determination, and semen collection, evaluation and screening for infectious organisms. The presence of copious growth of bacteria (Escherichia coli) in conjunction with significant sperm abnormalities and leucospermia was applied to diagnose infertility due to a sub-chronic or chronic bacterial infection of the reproductive tract. Bacterial infection caused significant sperm abnormalities including low sperm motility, increased number of dead sperm and sperm with structural abnormalities such as detached heads, fractured necks, looped tails and coiled tails. Treatment was effected by administering ciprofloxacin orally for 4 weeks.

49-55

Authors: Hossein Yazdekhasti, Jalil Hosseini, Zahra Rajabi, Maryam Hosseinzadeh Shirzeyli, Fereshte Aliakbari

Previous attempts have indicated that mesenchymal stem cells (MSCs) are a valuable source and candidate and new approach for tissue engineering and reproductive medicine. MSCs have this potential to be induced and differentiated in an appropriate in vivo and in vitro condition toward various cell lineages and then they can be applied in cell therapies and clinical applications. During recent two decades, various sources have demonstrated they are a great source for MSCs, including bone marrow, the human umbilical cord as well as Wharton’s jelly. Due to discarding after birth, easily accessible cells and less ethical concerns, these cells have attracted more and more scientists’ attention. Infertility and reproduction diseases have provided special opportunity to examine the efficiency of MSCs in this kind of application. Based on recent investigations, MSCs embedded in Wharton’s jelly tissue are more appealing for cell therapies, especially in infertility treatment purposes. So, differentiation of MSCs embedded in Wharton’s jelly tissue into germ layer cells for cell-based therapy purposes is now under intensive study.

62-71

Authors: Hammad Ahmad Gakhar, Ishrat Waheed, Taseer Ahmad, Naeem-ur-rahman

Objective: To test the effect of voltage-gated sodium channels (VGSCs) blockers on the motility and viability of human sperm in-vitro and to evaluate the tested compounds as potential contact spermicidal. Methods: Sperm samples were obtained from healthy nonsmoking volunteers of age 25-30 years who had not taken any drug 3 months before and during the course of the study. The effect of VGSCs blockers evaluated from two pharmacological classes including antiarrhythmic (amiodarone, procainamide and disopyramide) and antiepileptic (carbamazepine, oxcarbazepine, phenytoin, and lamotrigine) drugs. They were tested on the in-vitro motility and viability of human sperm using Computer Assisted Semen Analyzer. Results: All tested drugs except oxcarbazepine showed dose dependent inhibition of total motility with significant reduction (P<0.05) at the maximum concentration of 200 μM when compared with the control. The concentrations of drugs that reduced total sperm motility to 50% of control (half maximal inhibitory concentration) were 2.76, 14.16 and 20.29 μM for phenytoin, lamotrigine and carbamazepine, respectively; and 2.53, 5.32 and 0.37 μM for amiodarone, procainamide and disopyramide, respectively. The anti-motility effects were reversible to various degrees. There was statistically insignificant difference in the inhibition of sperm viability among amiodarone, procainamide and disopyramide. Phenytoin demonstrated the most potent spermicidal action. Conclusions: VGSCs blockers have significant adverse effects on in-vitro motility of human spermatozoa. So in-vivo studies are required to determine their potential toxicological effects on human semen quality, which is an important factor regarding fertility. Moreover, these drugs have the potential to be developed into contact spermicidal.

79-86

Authors: Madhusmita Panda, Asmita, Sandeep Kumar, Purusottam Mishra, Mahesh Chandra Sahu, Sachinandan De, Tirtha Kumar Datta, Rakesh Kumar

Objective: To understand the level of progesterone (P4) in different quality of buffalo cumulus oocyte complexes (COCs) and further to evaluate the effect of exogenous P4 supplementation on maturation and subsequent developmental ability of poor quality brilliant cresyl blue (BCB- ) COCs. Methods: Progesterone secreted by different quality of buffalo oocytes was estimated by enzyme linked immunosorbent assay and the concentration differences were translated into P4 doses to be incorporated in the maturation medium of BCB-ve COCs followed by expression analysis of genes involved in the cumulus expansion, extracellular matrix disintegration and progesterone receptor signalling. In addition, the study also evaluated the effect of exogenous P4 on sperm-cumulus interaction. Results: More than 10-fold upregulated expression of progesterone receptor in P4 supplemented oocytes signified that P4 might be acting predominantly through this receptor. Also, exogenous P4 supplementation had significant effect on transcatheter arterial chemoembolization protease regulated by P4- progesterone receptor pathway which in turn had an important role in extracellular matrix disintegration. On the contrary, cumulus expansion genes HAS2, TNFAIP6, AREG were not altered upon P4 supplementation. Also, it was observed that P4 addition did facilitate passage of significantly more number of spermatozoa through P4 treated cumulus cells. Further, incorporation of different doses of P4 did not improve significantly the cleavage and blastocyst rates of BCB-ve COCs. Conclusions: Different qualities of buffalo COCs secrete substantially diverse levels of P4, and its supplementation has a role in oocyte maturation via modulation of cumulus characteristics but perhaps not fertilization.

93-96

Authors: El-Sheshtawy RI, El-Nattat WS

Objective: To evaluate effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability. Methods: Pooled bull semen were extended with tris-citrate-fructose egg yolk diluent (control, 0% strawberry) and various concentrations of tris strawberry (TSB) (1%-6%) to achieve 60 million motile spermatozoa per milliliter. Extended semen were subjected to semen freezing protocol. Semen assessment including motility, alive%, abnormality%, intact sperm membrane (hypo-osmotic swelling test) and conception rate were carried out for both chilled and frozen semen. Results: Results showed that sperm motility after chilling was enhanced in groups treated with various concentrations of TSB from 1% to 5% and exhibited higher significance (P<0.000 1) at 6-day post-chilling. In frozen semen, 3%, 4%, 5% and 6% concentrations gave the best significance (P<0.000 1) on sperm motility in comparison with the control. Concentration 1% revealed the highest significance (P<0.000 1) on alive% as compared to the control. Hypoosmotic swelling test was maintained as the control. Concentration 3% gave the lowest significance (P<0.000 1) considering abnormality%. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 5% and 6% in cattle. Conclusions: It is concluded that 1%-5% concentrations of TSB ameliorate bull semen characteristics after chilling, and 3%-6% concentrations of TSB improve bull semen characteristics after freezing. Higher conception rate exists in 5% and 6% concentration of TSB.

145-150

Authors: S. Bhargavi, Subhalaxmi Swain, Abhisek Mishra, Arun Kumar Pradhan

In vitro fertilization (IVF) is one of the assisted reproductive technologies in the field of medical sciences. Fusion of collected egg from female and sperm from male done in a culture media under aseptic condition in laboratory is called as IVF. This technique is one of the gifts of science towards the human society and mainly those who are facing problem in reproduction due to having either male or female defective reproductive systems. In this present study, factors to consider during IVF, steps followed in this technology, need of IVF are discussed. Risk of multiple birth and phenotypic changes to the newborn due to IVF are also well reviewed. And different types of instruments used during this process are focused. Few ethical and legal issues arising during this IVF process are shown as well. We also emphasize that part of IVF.

155-160

Authors: Oluwatosin V. Imade, Wuraola A. Erinfolami, Rasheed A. Ajadi, Monsuru O. Abioja, Samson A. Rahman, Olusiji F. Smith, Oladele S. Gazal

Objective: To test the effects of dietary supplementation of Lepidium sativum (LS) seed powder on growth performance and gonadotropins secretion in ovariectomized, estradiol- implanted rabbits. Methods: Ovariectomized, estradiol-implanted Chinchilla rabbits were assigned into four experimental groups: LS seed powder was included into normal rabbit chow at 0% (control), 5% (low), 7% (mid) and 10% (high) w/w. Experimental feed and water were given ad-libitum for 3 weeks. Weekly body weights and daily feed intake of rabbits were recorded. Twenty-one days post-feeding, blood samples were collected at 15-minute interval for 3 h (Period Ⅰ) after which 2.5 μg gonadotropin-releasing hormone (GnRH) was injected intravenously and the sampling continued for another hour (Period Ⅱ). Plasma was harvested and analyzed for luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by radioimmunoassay. Results: Feed intake was significantly (P<0.05) increased in LS- supplemented rabbits. However, the increase in feed intake did not result in significant body weight gain. LS seed supplementation significantly (P<0.001) increased mean plasma LH dose-dependently from the low- to the mid-LS level and then decreased LH at the high-LS level. LS supplementation increased (P<0.001) plasma FSH secretion. Injection of GnRH had no effect on plasma LH, however significantly (P<0.05) decreased overall plasma FSH secretion. Conclusions: LS seed supplementation stimulates feed intake and gonadotropins secretion in rabbits. Gonadotropins effect may be mediated through LS seeds phytosterols through the activation of estrogen receptors thereby producing agonistic effects resulting in LH and FSH secretion. The differential responses of gonadotropins to GnRH in LS- supplemented rabbits suggest differential regulation of the synthesis and secretion of these gonadotropins.

167-171

Authors: Khamidulla B. Baimishev, Murat H. Baimishev, Vasily S. Grigoryev, Alexander P. Kokhanov, Inna V. Uskova, Ismagil N. Khakimov

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 high- producing cows under conditions of intensive milk production technology.

178-184

Authors: P. Perumal

Objective: To assess the deleterious effects of foot and mouth disease vaccination on antioxidant profiles as well as oxidative stress in the semen of breeding mithun bulls. Methods: A total of 160 semen samples were collected from 8 adult healthy mithun bulls with good body condition score (5-6), which were maintained at semen collection centre, ICAR-National Research Centre on Mithun, Medziphema, Nagaland, India, twice a week, 4 weeks before vaccination (pre-vaccination stage) and 12 weeks after vaccination (post-vaccination stage) to understand and know the influence of vaccine stress on seminal antioxidant as well as oxidative stress profiles in mithun. Vaccine was injected at the end of week 4 and semen samples were collected & analysed up to week 16 of the investigation period. Results: Data analysis showed that foot and mouth disease vaccination has significantly (P<0.05) altered the seminal antioxidant, biochemical and oxidative stress profiles up to week 10th of vaccine administration. However, the immunised bulls were recovered in physical health status, semen production and its antioxidant profiles gradually. Conclusions: The results of the present study suggest that the semen sample collection as well as preservation should to be suspended up to week 10th of post immunization to get antioxidant and biochemical profiles in normal level in the semen to preserve normal conception rate in artificial breeding programme by using such semen in mithun species.

191-192

Authors: Sulagna Dutta, Pallav Sengupta

No

97-102

Authors: Adil Rasool Paray, Mukesh Bhakat, Shabir Ahmad Lone, Tushar Kumar Mohanty, Ranjana Sinha, Javid Ur Rahman, Zahid Bashir Khanday, Ziaullah Danish

Quality semen production remains the main focus and objective of semen processing laboratories throughout the world. Bacterial and other microbial contaminants affect the semen quality and hence the fertility, and also lead to reproductive disorders as well as lower conception rates and increased embryonic mortality, abortion and other complications in females. Microbial contamination affects the semen adversely, by exerting direct spermicidal effect, formation of reactive oxygen species, toxin production, adherence with spermatozoa, deriving nutrients and oxygen from the medium and thus competing with spermatozoa for the factors of growth and normal functioning. Despite hygienic measures, several ubiquitous and opportunistic microbes find their ways into semen during collection, processing, and storage of semen, and survive even during freezing. Stringent sanitary precautions are therefore required at every step of collecting semen and its processing. Preputial cavity is considered as main source of semen contaminating microorganisms. Flushing the preputial cavity with normal saline or any suitable liquid combination with antimicrobial activity, prior to semen collection reduces the microbial load and thereby improves the semen quality.

117-122

Authors: Tomilola D. Olaolu, Damilare E. Rotimi, Ayotunde P. Olaolu

Objective: To examine the effect of Cissus populnea root on testicular function of male Wistar rats. Methods: Twenty albino male Wistar rats [(100.0 ±25.5) g] were selected into four groups randomly. The control group were administered 0.20 mL of distilled water while the other groups received 0.05 mL, 0.10 mL, and 0.20 mL/kg body weight of ethanol infusion. Oral administration was done using oropharyngeal cannula once daily for 14 days. Animals were thereafter sacrificed on day 15. Data obtained from this study were analysed using one- way analysis of variance (ANOVA) with multiple comparisons, post-hoc Tukey HSD was subsequently used and values at P<0.05 were considered statistically significant. Results: There were significant increases in serum testosterone, testes-body weight ratio, luteinising hormone and follicle stimulating hormone, respectively, compared with those in control group. Similarly, increase in acid phosphatase, alkaline phosphatase, protein, glycogen and cholesterol levels were more significant compared with those in control group. Conclusions: The findings of this study reveals enhanced secretions of the sex hormones and other indices at 0.05 mL/kg body weight administration of Cissus populnea root ethanol infusion. However, dosages exceeding 0.05 mL/kg body weight gradually depletes the level of reproductive indices assayed for in this current study. Therefore, caution must be taken when consuming ethanol infusion of Cissus populnea root above the dosages used in this current study.

129-135

Authors: Abdelgader Binyamen Abdelgader, Adil Salim Elsheikh

Objective: To determine the effects of Calotropis procera latex on pubertal traits of immature male Wistar rats. Methods: A total of 30 immature male Wistar rats aged 3 weeks old were grouped randomly into 5 groups: group A (control group) was offered distilled water as a placebo; group B was daily oral dosed with suspension of Calotropis procera latex at a dose rate of 5 mg/kg body weight (BW), group C 10 mg/kg BW; group D 15 mg/kg BW and group E 20 mg/kg BW. The rats were weighed daily to adjust the dose and record the BW changes, and the treatment continued for 4 weeks; thereafter, rats were sacrificed. Serum samples (n=30) were collected from all rats and kept frozen until assayed for reproductive hormones. Furthermore, the testes and epididymae were weighed; epididymal sperms were counted; sperm motility & abnormality were estimated; and histopathological sections of the testes were prepared. Results: The results of this study showed that oral dosing of immature male rats with Calotropis procera latex at doses rate 10 mg/kg BW significantly (P<0.05) reduced the growth rate, BW, testicular & epididymal weights, the level of most of reproductive hormones as well as the sperm traits examined; however, it significantly (P<0.05) augmented the abnormalities of spermatozoa and the seminiferous epithelium. Conclusions: Latex of Calotropis procera contains substances that have anti-androgenic activities and/or endocrine disrupting effects. If these substances are purified and identified, they can be used as male contraceptives.

139-142

Authors: Abdul Malik, Jaelani A, Neni Widaningsih, Gt Khairun Ni’mah, Raviani, Sakiman, Sasongko N

Objective: To explore the effect of fish oil at different concentrations on post-thawed semen of Kalang swamp buffalo. Methods: A total of 4 Kalang swamp buffalo bulls with 3-5 years of age and weighed about 340-360 kg were slected. Semen was regularly collected from these buffalo bulls once a week by an artificial vagina. Fish oil was supplementary at the dosages of 0 mg (control), 50 mg, 100 mg, 150 mg, and 200 mg to the extender (skim milk- egg yolk). Fresh, pre-freezing and frozen semen were thawed at 37 ℃ and evaluated for motility, viability, morphology, and plasma integrity of membrane. Results: The study results indicated that before freezing, supplementation of fish oil at the dose of 150 mg in the extender had significantly motility. And a significant (P<0.05) increase was observed in viability and motility of post-thawed semen at the dose of 150 mg fish oil, which was in difference with other treatment groups. Conclusions: Addition of 150 mg fish oil in the extender could be positive for the enhancement of the quality of post-thawed semen of Kalang swamp buffaloes.