M.O. Marques

Equine chorionic gonadotropin and gonadotropin-releasing hormone enhance fertility in a norgestomet-based, timed artificial insemination protocol in suckled Nelore (Bos indicus) cows

Two experiments were conducted to investigate the effects of equine chorionic gonadotropin (eCG) at progestin removal and gonadotropin-releasing hormone (GnRH) at timed artificial insemination (TAI) on ovarian follicular dynamics (Experiment 1) and pregnancy rates (Experiment 2) in suckled Nelore (Bos indicus) cows. Both experiments were 2x2 factorials (eCG or No eCG, and GnRH or No GnRH), with identical treatments. In Experiment 1, 50 anestrous cows, 134.5+/-2.3 d postpartum, received a 3mg norgestomet ear implant sc, plus 3mg norgestomet and 5mg estradiol valerate im on Day 0. The implant was removed on Day 9, with TAI 54 h later. Cows received 400 IU eCG or no further treatment on Day 9 and GnRH (100 microg gonadorelin) or no further treatment at TAI. Treatment with eCG increased the growth rate of the largest follicle from Days 9 to 11 (means+/-SEM, 1.53+/-0.1 vs. 0.48+/-0.1mm/d; P<0.0001), its diameter on Day 11 (11.4+/-0.6 vs. 9.3+/-0.7 mm; P=0.03), as well as ovulation rate (80.8% vs. 50.0%, P=0.02), whereas GnRH improved the synchrony of ovulation (72.0+/-1.1 vs. 71.1+/-2.0 h). In Experiment 2 (n=599 cows, 40 to 120 d postpartum), pregnancy rates differed (P=0.004) among groups (27.6%, 40.1%, 47.7%, and 55.7% for Control, GnRH, eCG, and eCG+GnRH groups). Both eCG and GnRH improved pregnancy rates (51.7% vs. 33.8%, P=0.002; and 48.0% vs 37.6%, P=0.02, respectively), although their effects were not additive (no significant interaction). In conclusion, eCG at norgestomet implant removal increased the growth rate of the largest follicle (LF) from implant removal to TAI, the diameter of the LF at TAI, and rates of ovulation and pregnancy rates. Furthermore, GnRH at TAI improved the synchrony of ovulations and pregnancy rates in postpartum Nelore cows treated with a norgestomet-based TAI protocol.

Lecirelin and Buserelin (Gonadotrophin releasing hormone agonists) are equally effective for fixed time insemination in buffalo

Buffalo has peculiar reproductive patterns, which make artificial insemination programs a hard and expensive task. Artificial insemination in fixed time is advantaged because females show low incidence of homosexual behaviour and strong dominance relationships, which leads to a poor accuracy in estrus detection. The aim of this experiment was to compare the efficiency of two different GnRH agonists in the GnRH/PGF2a/GnRH protocol (Buserelin vs Lecirelin). Two hundred and seventy buffaloes with 45 to 60 days postpartum were synchronized and fixed-time inseminated. The animals were kept on pasture in two farms at Sao Paulo and Mato Grosso do Sul (Brazil). Cows in Group 1 (n = 132) received, intramuscularly, 20 µg of Buserelin at a random day of the estrous cycle and, seven days later, 15 mg of prostaglandin F2a. Two days after prostaglandin administration, 10 µg Buserelin were intramuscularly injected. Cows in Group 2 (n = 138) were treated with the same protocol, but with intra-muscular administrations of Lecirelin (50 µg in the first administration and 25 µg in the second). Artificial insemination was performed 16 hours after the last injection in both groups. Pregnancies were diagnosed by ultrasonography (Pie Medical 480, 5.0 and 7.5 MHz linear probe), 30 days after artificial insemination. Conception rates were not influenced by farm (P > 0.05) and were similar in both groups [Group 1: 47.0% (62/132); Group 2: 50.0% (69/138); P > 0.05]. Results show that Lecirelin is as efficient as Buserelin to synchronize ovulation for fixed-time artificial insemination in buffaloes.

Resynchronization with unknown pregnancy status using progestin- based timed artificial insemination protocol in beef cattle

Two experiments were designed to evaluate the use of resynchronization (RESYNCH) protocols using a progestin-based timed artificial insemination (TAI) protocol in beef cattle. In experiment 1, 475 cyclic Nelore heifers were resynchronized 22 days after the first TAI using two different inducers of new follicular wave emergence (estradiol benzoate [EB; n = 241] or GnRH [n = 234]) with the insertion of a norgestomet ear implant. At ear implant removal (7 days later), a pregnancy test was performed, and nonpregnant heifers received a dose of prostaglandin plus 0.5 mg of estradiol cypionate, with a timed insemination 48 hours later. The pregnancy rate after the first TAI was similar (P = 0.97) between treatments (EB [41.9%] vs. GnRH [41.5%]). However, EB-treated heifers (49.3%) had a greater (P = 0.04) pregnancy per AI (P/AI) after the resynchronization than the GnRH-treated heifers (37.2%). In experiment 2, the pregnancy loss in 664 zebu females (344 nonlactating cows and 320 cyclic heifers) between 30 and 60 days after resynchronization was evaluated. Females were randomly assigned to one of two groups (RESYNCH 22 days after the first TAI [n = 317] or submitted only to natural mating [NM; n = 347]). Females from the NM group were maintained with bulls from 15 to 30 days after the first TAI. The RESYNC-treated females were resynchronized 22 days after the first TAI using 1 mg of EB on the first day of the resynchronization, similar to experiment 1. No difference was found in P/AI (NM [57.1%] vs. RESYNC [61.5%]; P = 0.32) or pregnancy loss (NM [2.0%] vs. RESYNC [4.1%]; P = 0.21) after the first TAI. Moreover, the overall P/AI after the RESYNCH protocol was 47.5%. Thus, the administration of 1 mg of EB on day 22 after the first TAI, when the pregnancy status was undetermined, promotes a higher P/AI in the resynchronized TAI than the use of GnRH. Also, the administration of 1 mg of EB 22 days after the TAI did not affect the preestablished pregnancy.

Influence of category-heifers, primiparous and multiparous lactating cows-in a large-scale resynchronization fixed-time artificial insemination program

This study was conducted to evaluate the influence of category (heifers, primiparous or multiparous cows) on pregnancy rates in a large scale resynchronization ovulation program. Nelore heifers (n = 903), primiparous lactating cows (n = 338) and multiparous lactating cows (n = 1,223) were synchronized using a conventional protocol of estradiol/P4-based fixed-time artificial insemination (FTAI). Thirty days after ultrasonography, females who failed the first FTAI were resynchronized with the same hormonal protocol prior to a second FTAI. The pregnancy status of each cohort was evaluated by ultrasonography 30 days after each FTAI. The average conception rate after the first FTAI and resynchronization was 80.5%. Heifers had a higher conception rate (85%) than primiparous (76%) or multiparous cows (78%; p = 0.0001). The conception rate after the first FTAI was similar among heifers (57%), primiparous cows (51%) and multiparous cows (56%; p = 0.193). After the second FTAI, heifers exhibited a higher conception rate (66%) than primiparous or multiparous cows (51%; p = 0.0001). These results demonstrate the feasibility of resynchronization in large beef herds for providing consistent pregnancy rates in a short period of time. We also demonstrated that ovulation resynchronization 30 days after FTAI is particularly effective for heifers, providing a conception rate of up to 66%.

Impact of progesterone and estradiol treatment before the onset of the breeding period on reproductive performance of Bos indicus beef heifers.

The present aimed to develop a hormonal therapy based on the insertion of a progesterone (P4) insert (PI) during 10 d plus an estradiol injection (E2) at PI removal before the onset of breeding of beef heifers. In Exp. 1, the plasma P4 profile of prepubertal heifers showed that the insertion of a PI or 24 d previously used P4 insert (UPI) sustained plasma P4 above 1 ng/mL for at least the first 7 d of the treatment. In Exp. 2 there was no positive effect of additional estradiol benzoate (EB) administered at the insertion of UPI on the proportion of heifers with a corpus luteum (CL/Treated) 30 d after UPI removal [UPI + EB = 85.3%(a) (n = 134); EB + UPI + EB = 80.8%(a) (n = 125)]; however, both were greater (P < 0.0001) than the Control group [60.3%(b) (n = 129)]. In Exp. 3, a positive effect (P=0.01) of UPI treatment and both E2 supplementations [EB and estradiol cypionate (EC); P = 0.10] at the UPI removal was observed on CL/Treated [Control = 42.5%(b) (n = 94); UPI = 58.5%(a) (n= 130); UPI + EB = 64.0%(a) (n = 128); UPI + EC = 67.2%(a) (n = 128)]. However, greater pregnancy per treated heifer (P/Treated) following artificial insemination (AI) upon estrus detection was achieved when EC was applied [Control = 20.2%(b); UPI = 29.2%(ab); UPI + EB = 26.6%(b); UPI + EC = 36.7%(a)]. In Exp. 4, the treatment prior to the timed AI (TAI) tended to improve pregnancy per TAI [P/AI; Control 43.6% (n = 298) vs. UPI+EC 51.9% (n = 342); P = 0.08], but increased P/Treated [26.5% vs. 43.3%; P < 0.001]. Thus, UPI + EC treatment were efficient in increasing the CL/Treated, tended to improve the P/AI and consequently enhanced P/Treated of zebu beef heifers.

Prostaglandin treatment at the onset of norgestomet and estradiol-based synchronization protocols did not alter the ovarian follicular dynamics or pregnancy per timed artificial insemination in cyclic Bos indicus heifers

The aim of the present study was to evaluate the effects of the PGF2α treatment given at the onset of a synchronization of ovulation protocol using a norgestomet (NORG) ear implant on ovarian follicular dynamics (Experiment 1) and pregnancy per AI (P/AI; Experiment 2) in cyclic (CL present) Bos indicus heifers. In Experiment 1, a total of 46 heifers were presynchronized using two consecutive doses of PGF2α 12 days apart. At first day of the synchronization protocol the heifers received implants containing 3mg of NORG and 2 mg of estradiol benzoate (EB). At the same time, heifers were randomly assigned to receive 150 mg of D-cloprostenol (n=23; PGF2α) or no additional treatment (n=23; Control). When the ear implants were removed 8 days later, all heifers received a PGF2α treatment and 1 mg of EB was given 24 h later. The follicular diameter and interval to ovulation were determined by transrectal ultrasonography. No effects of PGF2α treatment on the diameter of the largest follicle present were observed at implant removal (PGF2α=9.8±0.4 vs. Control=10.0±0.3 mm; P=0.73) or after 24 h (PGF2α=11.1±0.4 vs. Control=11.0±0.4 mm; P=0.83). No differences in the time of ovulation after ear implant removal (PGF2α=70.8±1.2 vs. Control=73.3±0.9 h; P=0.10) or in the ovulation rate (PGF2α=87.0 vs. Control=82.6%; P=0.64) between treatments were observed. In Experiment 2, 280 cyclic heifers were synchronized using the same experimental design described above (PGF2α; n=143 and Control; n=137), at random day of the estrous cycle. All heifers received 300 IU of equine chorionic gonadotropin (eCG) and 0.5 mg of estradiol cypionate (as ovulatory stimulus) when the NORG ear implants were removed. Timed artificial insemination (TAI) was performed 48 h after implant removal and the pregnancy diagnosis was conducted 30 days later. No effects on the P/AI due to PGF2α treatment were observed (PGF2α=51.7 vs. Control=57.7%; P=0.29). In conclusion, PGF2α treatment at the onset of NORG-based protocols for the synchronization of ovulation did not alter the ovarian follicular responses or the P/AI in cyclic Bos indicus beef heifers synchronized for TAI.