Search

Ruminant placental lactogens (PL) are members of the growth factor/prolactin (GH/PRL) family. They are synthesized by trophectodermal binucleate cells. There is evidence to suggest that PL is involved in control of fetal growth, through actions in both the maternal and fetal compartments, as well as in influencing mammary growth during pregnancy (Byatt JC et al.1992 J. Anim. Sci. 70, 2911–2923). The structure and biology of PL have been studied in the cow, sheep, goat, human, and mice. The maternal concentration of PL is 100- to 1 000-fold greater in pregnant sheep and goats than in cows but no information exists about PL concentration in buffalo. The aim of the present study was to evaluate the ability to detect PL in buffalo fluids by using bovine PL antibody. Samples were collected in the slaughterhouse immediately after animal slaughter. The fetuses were measured after heart blood collection. A bPL RIA system was used to determine the bPL concentrations in the buffalo samples (Alvarez-Oxiley AV et al.2007 Reprod. Fertil. Dev. 19, 877–885). The rbPL molecules were radio-iodinated with [125]I-Na by using the lactoperoxidase method (Thorell JI and Johansson BG 1971 Biochim. Biophys. Acta 251, 363–369). Concentrations of buffalo PL are presented in Table 1. In this RIA system, the minimum detected value was 0.068 ng mL-1, and the binding competition curves of bovine PL standard and buffalo fluids dilution using bovine PL antibody were paralleled in all kinds of samples. The lowest concentration was detected in allantoid fluid and the greatest concentration in fetal plasma (P< 0.05). Study of the biology of PL in buffalo has proved difficult because the concentration of PL in all buffalo fluids is very low. Furthermore, the research concerning buffalo PL function required in vivoexperiments. Existing data suggest that at least the concentration of buffalo PL is different from cattle and other smaller domestic ruminants. In conclusion, our results provide preliminary information about concentrations of PL in buffalo fluids.Table 1.Concentration of placental lactogen in buffalo fluidsThis work was supported by a grant from the Belgian Technical Cooperation. [ABSTRACT FROM AUTHOR]

Reproductive activity in swamp buffalo is characterized by a clearly demonstrated anestrus season. The aim of the present study was to evaluate season effect on the oocyte collection, in vitro maturation, and somatic cell nuclear transfer. The ovaries collected from a slaughterhouse were divided into 3 groups according to the collection period: (1) G1: from January to April; G2: from May to August, which is characterized by higher climate temperature and low reproductive activity; and G3: from September to December. Cumulus–oocyte complexes (COCs) were aspirated from follicles 2-6 mm in diameter using an 18-gauge needle, washed in HEPES-buffered TCM-199 (Sigma-Aldrich, St Louis, MO, USA), and classified following 3 different quality levels: A (with 4–6 layers of cumulus cells), B (with 2–3 layers of cumulus cells), and C (few or without cumulus cells). The oocytes of A and B categories were used for IVM in maturation media currently used in cattle (TCM-199 medium + 10% fetal bovine serum) with an increase of FSH concentration (30 g mL-1) and estradiol-17? (3 g mL-1). Maturation was carried out at 39C in a water-saturated incubator, under 5% CO2 for 22 h. The oocytes were observed for the cumulus expanding and the presence of polar body (PB). The oocytes with PB were used for further enucleation and cell nuclear transfer using buffalo quiescent fibroblast cells and the technique described previously (Nguyen et al. 2000 Theriogenology 53, 235). The percentages of intact and fused oocytes as well as reconstructed embryos developed to blastocyst stage were compared for the oocytes from G1 and G2. The results indicated that the average number of good quality COCs collected per ovary for the G1, G2, and G3 period were 6.00 4.08 (n = 426), 2.93 2.55 (n = 346), and 4.78 1.05 (n = 445), respectively. The percentages of A and B oocytes were 62.4% (1.58 0.51 vs. 2.17 1.54), 63.2% (0.90 0.32 vs. 0.95 0.50), and 54.7% (1.12 0.25 vs. 1.49 0.53), respectively; the maturation rate was 55.08%, 56.28%, and 52.16%, respectively. There were no significant differences between G1 and G2 in the percentage of intact and fused oocytes (93.7% and 59% for G1; 100% and 60% for G2, respectively), but the rate of embryos developed to blastocyst stage was higher for oocytes from G1 (18.5% vs. 10.2%). In conclusion, in swamp buffalo, the hot season affected significantly the number of oocytes collected per animal and the subsequent results of somatic cell nuclear transfer. The optimal period for working with buffalo oocyte is from January to April.This work was aupported by a grant from the Vietnam-Italy 3AB3 Project. [ABSTRACT FROM AUTHOR]

The Ban minipig is a local breed characterized by small ovaries with a scant number of follicles available for in vitro maturation (IVM). The combination of eCG and hCG has been used successfully to control estrus in pig breeding programs. In this paper we present the first results of IVF in this breed in comparison with 2 types of oocyte preparation: (1) from animals not receiving gonatotropin treatment (group 1, n = 9); and (2) from animals receiving an injection of mixed pregnant mare serum gonadotropin and hCG, 300 IU/animal, for 3 days before oocyte collection (group 2, n = 4). All animals were 1 to 3 years old and with body weights that varied from 8 to 12 kg. At the time of collection, the ovaries were observed for follicle development; the cumulus–oocyte complexes (COCs) were aspirated using a 18-gauge needle. COCs of categories A (with more than 4 layers of cumulus cells) and B (with 2 to 4 layers of cumulus cells) were collected and matured in vitro as described previously (Kikuchi et al. 2002 Biol. Reprod. 66, 1033–1041) at 39C under 5% CO2 in air. Matured oocytes with expanding cumulus cells were inseminated using male Ban minipig epididymal semen frozen by the methods reported by Kikuchi et al. (1998 Theriogenology 50, 615–623). The frozen–thawed spermatozoa were pre-incubated for 1 h in modified medium-199 adjusted to pH 7.8 in the incubator at 37C. The capacitated spermatozoa were diluted and added to drops of fertilization medium (Fig-FM; Suzuki et al. 2002 Int. J. Androl. 123, 135–142) containing oocytes; the final concentration of sperm was 106/mL. After 3 h of co-incubation, attached spermatozoa and cumulus cells were removed from oocytes and the oocytes were the cultured in vitro as described previously (Kikuchi et al. 2002). The results obtained from 4 replicates showed that the number of follicles with a diameter larger than 2 mm and the rates of oocytes categorized as A and B were significantly lower (P < 0.05; ANOVA test) in the nontreated animals (0.0 and 67.5%, respectively) than in the treated group (25.5 and 87.1%, respectively). The rates of oocytes with a clearly expanding cumulus obtained after IVM were 78.6 (n = 136) and 88.1% (n = 101) for groups 1 and 2, respectively. The rates of cleaved embryos and embryos developed to the compact morula stage were 47.2 and 9.1% (n = 39), respectively, for group 1; and 89.1 and 18.8% (n = 101), respectively, for group 2. In conclusion, gonadotropin treatment before the collection of oocytes is recommended for application of IVM–IVF to local Ban minipigs.This work was supported by a grant from the VAST-Japan Society for the Promotion of Science Project. [ABSTRACT FROM AUTHOR]

One of the big obstacles in management of reproductive activity in water buffaloes is their silent estrus. This characteristic prevents the determination of time for artificial insemination (AI), resulting in lengthening of the calving interval (Nguyen et al. 1990 Proc. FAO Workshop, Bulgaria, 210?216). Pregnancy-associated glycoprotein (PAG) measurement in maternal blood is an effective method of early pregnancy diagnosis in ruminant species (Zoli et al. 1992 Biol. Reprod. 46, 83?92). The present study describes a preliminary attempt to improve buffalo reproductive performance by association of three different approaches: synchronization of estrus by hormonal treatment, followed by two AI, and finally pregnancy diagnosis based on PAG measurements. A total of 33 buffalo females (4 to 15 years old), with no apparent estrus during a period of 9 to 36 months after last calving, were used. Each female received 2 injections of 5 mL of PGF2? (Intervet, France) 10 days apart (Day -12 and Day -2), one injection of 500 IU of PMSG (SABC, Vietnam) at Day -5, and 1000 IU of hCG (Intervet, France) on the day of AI (Day 0). A second AI was performed 8 h after hCG injection. Blood samples (9 mL) were collected from each female at Days 20, 25, 40, and 45 after AI. Blood samples were withdrawn by jugular venipuncture (heparinized Vacutainer tubes) and kept in a cool box until centrifugation (500g, 10 min). Plasma samples were stored at -20C until assayed for PAG. Pregnancy diagnosis was considered negative, doubtful, or positive according to the PAG concentrations (<0.6 ng mL-1, 0.6 to 0.8 ng mL-1, and >0.8 ng mL-1, respectively). Clinical observation of females after insemination showed that the total rate of animals that exhibited signs of estrus until Day 45 was 9.1? (3/33). These 3 females were not considered for pregnancy diagnosis. Table 1 shows the results of pregnancy diagnosis in 30 water buffalo females. These results show that fertilization is easily obtained after estrus synchronization in buffaloes and that measurement of PAG concentrations in maternal plasma can be used for pregnancy diagnosis from Day 40 after AI. [ABSTRACT FROM AUTHOR]