Morphometric, subcellular, in vitro fertilisation and embryonic developmental assessment of mouse oocytes produced by anti-inhibin serum or pregnant mare serum gonadotrophin superovulation

This study compared the morphometric, subcellular characteristics, in vitro fertilisation (IVF) and embryonic developmental potential of metaphase II (MII) mouse oocytes obtained from females superovulated with either anti-inhibin serum–human chorionic gonadotrophin (AIS-hCG) or pregnant mare serum gonadotrophin (PMSG)-hCG. The oocyte’s quantity, quality, zona pellucida (ZP) thickness, perivitelline space (PVS), diameter, microtubules, F-actin, cortical granules (CGs) and mitochondrial distribution were determined. Superovulation using AIS-hCG resulted in a higher numbers of oocyte/donor compared with PMSG-hCG (P=0.002). There was no difference in morphologically normal and abnormal oocytes between AIS-hCG and PMSG-hCG (P=0.425 and P=0.194, respectively). The morphometric measurements showed no difference in oocyte diameter between AIS-hCG and PMSG-hCG (P=0.289). However, the thickness of the ZP of oocytes from AIS-hCG females was decreased compared with PMSG-hCG (P<0.001). The PVS of oocytes from the AIS-hCG was larger than with PMSG-hCG (P<0.001). The microtubules of oocytes from both AIS-hCG and PMSG-hCG were normal, although there was an increased fluorescence intensity in the AIS-hCG oocytes (P<0.001). The F-actin and CGs distribution in oocytes from both AIS-hCG and PMSG-hCG were similar (P=0.330 and P=0.13, respectively). Although the oocytes from PMSG-hCG females had homogenously distributed mitochondria, AIS-hCG oocytes showed more peripheral distribution with no differences in fluorescence intensity (P=0.137). The blastocyst development rates after IVF with fresh sperm showed no difference between AIS-hCG and PMSG-hCG (P=0.235). These data suggested that AIS-hCG superovulation produces high numbers of morphologically normal oocytes that also possess normal subcellular structures, good morphological characteristics and had high invitro embryonic developmental potential.

Improving the Quality of Oocytes of Old and Productive Ages White Rats (Rattus norvegicus) Using Pregnant Mare Serum Gonadotrophin

This study was conducted to improve the quality of oocytes in old-female rats by using pregnant mare serum gonadotrophin (PMSG). Female rats at productive age were used as a control. The experimental rats were injected with 4 doses of PMSG i.e., 0, 2.5, 5.0, and 10 IU PMSG. After 2 weeks of acclimation to the experimantal condition, the experimental rats were injected with PGF2α at a dose of 25µg/g BW two times with 2 days interval to synchronize estrous cycle. PMSG injections were conducted at the same time with the second PGF2α injection. After PMSG injection, the experimental rats were divided into two groups rats, i.e., rats without mating for masurement of oocyte qualities and rats mated for measurement of offspring qualities. Therefore, 16 experimental rats from each age group were sacrifized for maeasurement of oocyte qualities. The other 16 experimental rats for each age group were mated for measurement of offspring qualities. Parameters measured were hematological profile, uterine and ovarian weights, the qualities of oocytes, the qualities of the offsspring born by using swimming test and rat maze test. The collected data were analyzed by analysis of variance (ANOVA) and continued with Duncan test with a 95% confidence level. The data were analyzed using SSPS. The results showed that the improvement in the quality of oocytes in old female rats using the PMSG hormone in this study showed an increase in the quality of oocytes in old and productive age rats. The highest number of oocyte quality was found in rats of productive age (3.25) and significantly different (P< 0.05) from the other oocyte qualities. The quality of offspring born to old age and productive age rats injected with PMSG were improved. It was concluded that the improvement of oocyte quality by using PMSG also improves offspring qualities.

Comparative effect of the aqueous extracts of Aloe buettneri, Dicliptera verticillata, Hibiscus macranthus and Justicia insularis on the sexual maturation of pregnant mare serum gonadotrophin-primed immature female rats

Background Aloe buettneri, Dicliptera verticillata, Hibiscus macranthus and Justicia insularis are medicinal plants used in the western region of Cameroon to cure infertility among women. This experiment was designed to compare the effect of the aqueous extracts of A. buettneri (AAB), D. verticillata (ADV), H. macranthus (AHM), J. insularis (AJI), their mixture (AME) and the extract obtained from the mixture of their powders (AMP) on some parameters of reproduction. Materials and methods High-performance liquid chromatography coupled with the mass spectrometry (HPLC-MS) was used to detect the constituents of different extracts. The extracts (50 mg/kg) were administered to PMSG-primed immature rats (10 rats/group) for 5 days. At the end of the treatment, five animals of each group were sacrificed and various biological markers of reproduction were recorded. The remaining animals were injected with 10 IU of human chorionic gonadotropin (hCG) sacrificed 48 h later and the number of ovarian hemorrhagic points counted. Results Fifty-four compounds were found in the various extracts with 37 individually distributed among them and the remaining found in at least two extracts. As compared with the control, the AMP and AJI increased serum estradiol level by 2.36 and 2.23 times, respectively. The AMP was the only extract whose administration resulted in a significant increase (p<0.001) in the serum levels of follicle stimulating hormone (FSH) and luteinizing hormone (LH). Compared with the other extract-treated rats, the AMP-treated rats showed the highest number of hemorrhagic points. Conclusions These findings provided evidence on the synergistic effect of compounds present in these plant extracts and supported the usage of the mixture in traditional medicine.

Kondisi Neonatus Anak Babi yang Dilahirkan oleh Induk yang Disuntik Gonadotropin Sebelum Pengawinan (THE NEONATE CONDITIONS OF PIGLETS BORN FROM SOW INJECTED WITH GONADOTROPIN PRIOR TO MATING)

Injections of sows with pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG) prior to mating have been shown to improve endogenous secretions of pregnant hormones that improve and optimize the uterine environment and condition during pregnancy that eventually lead to give birth to superior piglets. This experiment was designed to study the effects of PMSG and hCG injections of sows prior to mating on prenatal growth as indicated by the conditions of the the neonate piglets. Thirty mature female Landrace pigs were divided into 2 groups i.e., 15 pigs were injected with PMSG and hCG prior to mating, and the other 15 pigs were injected with physiological NaCl solution as a control. Before mating, the estrus cycles of the experimental sows were synchronized by injecting prostaglandin two times with 14 days interval. The experimental pigs were injected with PMSG and hCG or 0.90% NaCl solution at the same time with the second prostaglandin injection and were further mated naturally. During pregnancy, the experimental sows were maintained with the normal management condition. The results showed that piglest born to sows injected with PMSG and hCG prior to mating had shorter and faster birth process with higher activities and survivals as compared to those born to control sows. Piglets born by sow that were injected with PMSG and hCG before mating had a more optimum birth weight accompanied by greater body length, limb height, and higher rear limb height. Piglest born to by sows that were injected with PMSG and hCG prior to mating showed superior conditions that would contribute to optimum and superior preweaning and postweaning growths until maturity.

Different estrous induction methods during the non-breeding season in Kivircik ewes

The efficiency of medroxyprogesterone acetate (MAP) sponges in combination with either pregnant mare serum gonadotrophin (PMSG) or cloprostenol (PGF_2&alpha;) for inducing and synchronizing the estrous cycle in non-lactating Kivircik ewes was investigated during the natural non-breeding season. All ewes (n = 69) were treated with 60 mg MAP sponges for 12 days. In addition, each ewe received an intramuscular injection of either 1.5 ml sterile saline solution (n = 18); 125 &mu;g PGF_2&alpha; (n = 14); 500 IU PMSG (n = 18) or 500 IU PMSG and 125 &mu;g PGF_2&alpha; (n = 19), 48 h before the sponge removal. Cervical artificial insemination (AI) with diluted fresh semen was performed at a fixed time (48 and 60 h) following progestagen withdrawal. The different groups estrous response for the first 24 &plusmn; 6 h and within 120 h, time to onset and duration of the induced estrous, and pregnancy rate was found to be 36.2%, 81.6%, 41.7 &plusmn; 2.3 h, 29.6 &plusmn; 1.5 h, and 54.5%, respectively. There were significant differences between groups MAP and MAP/PGF_2&alpha; and their with the two latter groups (MAP/PMSG, MAP/PMSG/PGF_2&alpha;) in terms of the onset of induced estrous (P &lt; 0.05) and between groups MAP and MAP/PGF_2&alpha; in terms of the duration of induced estrous (P &lt; 0.05) and between the first two groups (MAP, MAP/PGF_2&alpha;) and the latter two groups (MAP/PMSG, MAP/PMSG/PGF_2&alpha;) in terms of estrous response at the first 24 &plusmn; 6 h (P &lt; 0.05). These results indicate that, the use of MAP/PMSG, rather than MAP or MAP/PGF_2&alpha;, was effective in the attainment of early and compact induction and synchronization of estrous in non-lactating Kivircik ewes during the natural non-breeding season.

Uterine and vaginal insemination optimised in brushtail possums (Trichosurus vulpecula) superovulated with pregnant mare serum gonadotrophin and porcine luteinising hormone

Artificial insemination of brushtail possums (Trichosurus vulpecula) is being developed as an assisted breeding model for endangered marsupials, as well as a bioassay for testing fertility control vaccines to manage overabundant populations. Procedures were optimised in animals superovulated with pregnant mare serum gonadotrophin (PMSG) and porcine luteinising hormone (pLH). Of three intervals examined, yields were maximal following uterine insemination at 27–29.5 h after pLH treatment (four eggs, two to three embryos per female). Compared with no insemination, uterine-inseminated animals ovulated 30–36 h rather than 28–34 h after pLH treatment. For the vaginal route, yields were maximal following insemination at 10–13 h after pLH treatment (six to seven eggs, four embryos per female) than at five other intervals, and when using acclimatised females during the autumn breeding season. This protocol was suitable for testing fertility control vaccines in April–June and was influenced by the housing location of animals, the presence of an active corpus luteum and PMSG batch, but not other factors (year of trial, Freund’s adjuvant treatment, changes in bodyweight, dose of PMSG kg–1). Embryos developed to the eight- to 16-cell or unilaminar blastocyst stage after uterine or vaginal insemination, respectively. With the timing of artificial insemination optimised, new methods to synchronise or induce oestrus and ovulation are required to achieve year-round testing of fertility control vaccines or birth of offspring.

Porcine oocyte maturation in vitro under different conditions

The effects of hormone additions at various stages and different basic media, with or without serum, on porcine oocyte maturation in vitro were studied. The results showed that the rate of maturation was not significantly different with three different stages of hormone supplement; the rate of maturation on modified TCM199 medium (54.01%) was higher than that on TCM199 (46.16%) and (47.14%), but these differences were not significant; and the rate of maturation on serum-free medium (67.10%) was significantly higher than that on medium plus serum (52.22%). Therefore, modifed tissue culture medium 199 (mTCM199)+10I U/ml pregnant mare serum gonadotrophin (PMSG)+10I U/ml human chorionic gonadotrophin (hCG)+2.5 IU/ml follicle stimulating hormone (FSH) was a suitable medium for culture of porcine oocytes in vitro, and the rate of maturation was 67.10%.