No abstract

The developmental competence of male and female gametes is frequently reduced under in vitro conditions, mainly due to oxidative stress during handling. The amino-acid derived hormone melatonin has emerged as a potent non-enzymatic antioxidant in many biological systems. The goal of the present study was to evaluate the effects of melatonin on post-thaw sperm quality, fertilizing ability, and embryo development and competence in vitro after in vitro fertilization. Frozen-thawed bovine spermatozoa were incubated either in the presence of 10−11 M melatonin (MT), or its solvent (ethanol; Sham-Control), or plain Tyrode’s Albumin Lactate Pyruvate medium (TALP, Control). Computer-Assisted Sperm Analysis (CASA) and flow cytometry data after 30 min, 120 min, and 180 min incubation did not reveal any significant effects of melatonin on average motility parameters, sperm subpopulation structure as determined by hierarchical cluster, or on the percentage of viable, acrosome intact sperm, or viable sperm with active mitochondria. Nevertheless, in vitro matured cumulus-oocyte-complexes fertilized with spermatozoa which had been preincubated with 10−11 M melatonin (MT-Sperm) showed higher (P < 0.01) rates of monospermic fertilization, reduced (P < 0.05) polyspermy and enhanced (P < 0.05) embryo development compared to the Control group. Moreover, the relative abundance of MAPK13 in the in vitro-derived blastocysts was greater (P < 0.05) than observed in the Control group. In conclusion, adding melatonin to the sperm-preparation protocol for bovine IVF improved proper fertilization and enhanced embryonic development and competence in vitro.

This study evaluated the pregnancy rate (PR) after timed artificial insemination (TAI) in water buffalo (Bubalus bubalis) during both non-breeding and breeding season, using either a new or reused intravaginal device (IVD) with two different progesterone concentrations. A total of 247 dairy buffalo cows were randomly assigned using a two-by-three factorial design and four replicates to the following groups: (1) new intravaginal device (IVD-New: DIB®, 1.0 g of P, n = 51 or CIDR®, 1.38 g of P, n = 55); (2) intravaginal device previously used once (9 days) (IVD-Used1x: DIB, n = 40 or CIDR, n = 51); or (3) intravaginal device previously used twice (18 days) (IVD-Used2x: DIB, n = 27 or CIDR, n = 23). On day 0, animals received the IVD plus 10.5 μg of buserelin acetate (GnRH) intramuscularly. On day 9, the devices were removed and 25 mg of PGFα plus 500 IU of eCG was given intramuscularly. On day 11 (48 h after IVD withdrawal), animals received 10.5 μg of GnRH and were artificially inseminated 8-12 h later. Data were analyzed using Proc Logistic of SAS®. Animals that received IVD-New-DIB, had a significantly higher PR (62.7%; P = 0.0193) compared to animals that received IVD-New-CIDR (40%). Pregnancy rate was not negatively affected by reusing both types of IVD. Overall PR (new and reused devices) was higher (P = 0.0055) in the DIB group (62.7%) compared to the CIDR group (45%). In conclusion, PR was higher in buffaloes treated with devices containing 1.0 g of P (DIB®) compared to those receiving 1.38 g of P (CIDR®). Reusing the intravaginal devices did not affect negatively PR/TAI, suggesting that P concentrations within the TAI protocols in water buffaloes could be reduced, without impairing their fertility.

To study the effectiveness of a treatment with an intravaginal progestagen in combination with estradiol-17beta (E-17beta) on estrus induction, prevention of prolonged anestrus and fertility in noncycling crossbred zebu cows, a trial was conducted in a farm in Zulia state, Venezuela. Fifty noncycling suckled crossbred cows (B. taurus x B. indicus) with 60 dpp were randomly allotted to one of the following treatments: MAP+E-17beta (n=22), intravaginal device (250 mg of MAP) during 7 days + 50 mg of MAP and 5 mg of E-17beta im (day 0) and 1.5 mg of E-17beta im (day 8) (Pregnaheat-E; VIATECA-Venezuela); CG (n=28), control group without hormonal treatment. The MAP+E-17beta group showed higher (P<0.05) estrus and synchronization rates (59.1 and 36.4 %) than CG (25.0 and 3.5 %). Cows receiving MAP had higher (P<0.05) overall pregnancy rate than CG (45.5 and 17.8 %, respectively). A decreased (P<0.05) anestrus rate was observed in MAP+E-17beta (31.8%) compared to CG (57.1%). Calving-conception interval was shorter (P<0.05) in MAP+E-17beta (132.5+/-16.8 days) than in CG (178.9+/-14.7 days). In conclusion, treatment with MAP+E-17beta at 60 days postpartum was effective to induce estrus, prevent prolonged anestrus, and reduce the calving-conception interval, improving reproductive performance in dual purpose crossbred Zebu cows.