Estrus synchronization, artificial insemination, and pregnancy diagnosis in water buffaloes (Bubalus bubalis)

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 PGF2α (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 Blood samples were withdrawn by jugular venipuncture (heparinized Vacutainer tubes) and kept in a cool box until centrifugation (500g, 10 min).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 -20°C until assayed for PAG. Pregnancy diagnosis was considered negative, doubtful, or positive according to the PAG concentrations (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