Optimizing ReBreed21 II: Fertility and reproductive efficiency in different parities during a shortened breeding season in beef cattle.

This study evaluated the efficiency of a rapid reinsemination program allowing timed AI (TAI) every 21d (ReBreed21) in a commercial beef cow-calf operation. Nelore females from different parities (n = 2085) were synchronized for first TAI (D0 = TAI) using an estradiol/progesterone (E2/P4) protocol and assigned to one of three reinsemination programs: Resynch33 (n = 753), traditional resynch program with second TAI at D42 after first TAI; ReBreed21 (n = 687); or ReBreed21+EC (n = 670). The ReBreed females (n = 1357) received intravaginal P4 insert on D12, on D19 P4 was removed, and a dose of equine chorionic gonadotropin (eCG) was administered, then, ReBreed21 females received 0.6 mg of EC (ReBreed21+EC) or nothing (ReBreed21) and on D21, nonpregnancy (NP) was determined using Doppler ultrasound to detect corpus luteum (CL) blood flow (BF) (NP: <25 % BF pixels of total CL area) and NP cows received immediate TAI and GnRH to induce ovulation. Pregnancy diagnosis was performed at D33 after TAI following all TAIs. Cows considered pregnant at D21, based on CL BF, but NP on D33 were designated False-Positives (FP) and false negatives (FN) were number of nonpregnant cows/heifers on d21 based on the CL BF found to subsequently be pregnant on D33 divided by the total number pregnant. Pregnancy/AI (P/AI) did not differ for the first TAI (55.1 %) among the treatments. Heifers had similar P/AI at the second AI in all groups and similar to the first AI. Primiparous had greater P/AI in ReBreed21+EC and Resynch33 at s TAI compared to ReBreed21, 51.7 %, 55.8 %, 34.2 %, respectively. Multiparous had greater P/AI at second TAI in Resynch33 (60.9 %) than ReBreed21 programs (34.7 %). The percentage FP and FN among ReBreed21 programs did not differ, 13.8 and 0.2 %, respectively. Overall accumulative pregnancies on D21 of the breeding season were greater for ReBreed21 and ReBreed21+EC than Resynch33 (69.7 %, 71.6 %, and 55.5 %, respectively). However, on D42 of the breeding season, only heifers had greater pregnancies in ReBreed21 programs than Resynch33 (73.3 %, 74.3.6 %, and 63.2 %, respectively). Average days to pregnancy were less (P = 0.01) for ReBreed21 and ReBreed21+EC than Resynch33. Thus, the ReBreed21 strategy can improve the efficiency of TAI programs in beef cattle. Of interest, ReBreed21 was particularly effective in nulliparous, somewhat effective in primiparous when EC was added to the program, but relatively ineffective in multiparous beef cattle.

Optimizing ReBreed21 I: Evaluation of endocrine and ovarian dynamics in non-bred Bos indicus heifers.

The present study evaluated follicular and endocrine dynamics during ReBreed21, a reproductive strategy that allows resynchronization of ovulation every 21 days in Bos indicus (Nelore) heifers. A synchronized estrous cycle was induced using a standard timed ovulation protocol (d -10: P4 implant inserted + 2 mg estradiol benzoate; d -2: P4 removed+ 0.5 mg cloprostenol + 0.6 mg estradiol cypionate + 200 IU equine chorionic gonadotropin (eCG); d0: 8.4 µg buserelin) without AI to ensure nonpregnancy in heifers. Day of GnRH was designated d0 of estrous cycle. On d12, heifers (n = 80) were randomized into three experimental groups: (1) ReBreed21 (n = 28) d12 P4 device inserted, d19 P4 device withdrawal plus 200 IU eCG, and d21 8.4 µg buserelin (GnRH); (2) ReBreed21+G (n = 26) same as ReBreed21 plus GnRH (16.8 µg) treatment on d12; and (3) Control (n = 26) no treatment. ReBreed21+G increased two-fold (62.9%; 18/26) percentage of heifers with synchronized follicular wave emergence compared to Control (34.6%; 9/26) whereas ReBreed21 (53.6%; 15/28) was intermediate. The ReBreeed21 groups (eCG on d19) increased (P < 0.01) follicular growth between d19 and d21 in ReBreed21 (2.3 ± 0.2 mm) and ReBreed21+G (3.4 ± 0.2 mm) compared with Control (1.2 ± 0.3 mm), resulting in greater (P < 0.01) follicle diameter on d21 for ReBreed21 (10.7 ± 0.4 mm) and ReBreed21+G (10.8 ± 0.4 mm) compared with Control (9.1 ± 0.5 mm). Structural luteolysis was similar among groups (P = 0.51), although the average day when P4 was <1 ng/mL was later (P < 0.01) for ReBreed21 (20.5 ± 0.2) and ReBreed21+G (20.7 ± 0.2) compared to Control (19.2 ± 0.4). Overall ovulation at the end of the estrous cycle was increased (P = 0.03) for ReBreed21 groups (83.3%; 45/54) compared with Control (57.7%; 15/26). Synchronized ovulation on day 22-23 was greater (P < 0.01) for ReBreed21 (78.6%; 22/28) and ReBreed21+G (76.9%; 20/26) compared with Control (30.8%; 8/26). Thus, the ReBreed21 resynchronization program produced acceptable endocrine and follicular dynamics, including synchronized ovulation at the end of the protocol in nonpregnant heifers providing good rationale for testing the fertility and practical implementation of this protocol under field conditions.

Accessory corpus luteum regression during pregnancy II: reproductive outcomes.

The objective of this study was to evaluate the effect of accessory corpus luteum (CL) induction on fertility in dairy cows. On day 5 after artificial insemination (AI), lactating Holstein cows were assigned unequally to receive gonadotrophin-releasing hormone treatment (GnRH) (n = 641) or no treatment (control; n = 289). Cows had their blood sampled for progesterone (P4), and ovaries were scanned by ultrasound on days 5, 12, 19, 26, 33, 47, and 61 after AI. Pregnancy diagnosis was performed on days 26, 33, 47, and 61. On day 12, cows treated with GnRH were allocated to ipsilateral (n = 239) or contralateral (n = 241) groups based on the side of accessory CL formation relative to previous ovulation. Accessory CL cows had greater P4 than controls. In total, 52.7% (78/148) of pregnant cows in contralateral group had accessory CL regression earlier (

Effects of feeding rumen-protected methionine pre- and postpartum on reproductive outcomes of multiparous Holstein cows.

Our primary objective was to evaluate the effect of feeding rumen-protected Met (RPM) in the pre- and postpartum total mixed ration (TMR) on pregnancy per artificial insemination (AI) and pregnancy loss in multiparous Holstein cows. We also evaluated multiple secondary reproductive physiological outcomes before and after AI, including uterine health, ovarian cyclicity, response to synchronization of ovulation, and markers of embryo development and size. A total of 470 multiparous Holstein cows [235 at the University of Wisconsin (UW) and 235 at Cornell University (CU)] were used for this experiment. Experimental treatment diets were applied at the pen level (2 and 4 close-up pens at CU and UW, respectively, and 12 and 6 postfresh pens at CU and UW, respectively); thus, pen was the experimental unit, and cow was the observational unit. Cows were enrolled and randomly assigned to be fed the experimental treatment diets at approximately 4 wk before parturition until 67 d of gestation [147 d in milk (DIM)] after their first service. Close-up dry cow and replicated lactation pens were randomly assigned to treatment diets: RPM, prepartum = 2.83% (UW) and 2.85% (CU), postpartum = 2.58% (UW) and 2.65% (CU); and control (CON), prepartum = 2.30% (UW) and 2.22% (CU), postpartum = 2.09% (UW) and 2.19% (CU; Met as percentage of metabolizable protein). Vaginal discharge and uterine cytology (percentage of polymorphonuclear leucocytes) were evaluated at 35 ± 3 DIM. Cows received timed AI (TAI) at 80 ± 3 DIM after synchronization of ovulation with the Double-Ovsynch protocol. Ovarian cyclicity status, response to synchronization of ovulation, and luteal function were determined by measuring circulating concentrations of progesterone at 35 and 49 ± 3 DIM, 48 and 24 h before TAI, and 8, 18, 22, 25, and 29 d after TAI. Interferon-stimulated gene expression in white blood cells were compared on 18 d after TAI (CU only) and pregnancy-specific protein B concentrations at 22, 25, 29, 32, and 67 d after TAI. Pregnancy status was determined using pregnancy-specific protein B at 25 and 29 d after TAI, and by transrectal ultrasonography at 32, 39, and 67 d after TAI. Embryo and amniotic vesicle size were determined at 32 and 39 d after TAI. Pregnancy per AI (25 d: 64.7 vs. 64.0%, 32 d: 54.3 vs. 55.1% for CON and RPM, respectively) and pregnancy loss (25 to 67 d: 22.6 vs. 19.2% for CON and RPM, respectively) for synchronized cows did not differ. The proportion of cows with purulent vaginal discharge (CON = 7.7 vs. RPM = 4.6%) and cytological endometritis (CON = 20.8 vs. RPM = 23.6%) did not differ. Cyclicity status, ovarian responses to the synchronization protocol, and synchronization rate also did not differ. In addition, fold change for interferon-stimulated genes, concentrations of pregnancy-specific protein B, and embryo size were not affected by treatments. In conclusion, feeding RPM in the pre- and postpartum TMR at the amounts used in this experiment did not affect uterine health, cyclicity, embryo development, or reproductive efficiency in dairy cows.

Hormonal combinations aiming to improve reproductive outcomes of Bos indicus cows submitted to estradiol/progesterone-based timed AI protocols.

The aim was to study reproductive outcomes of Nelore (Bos indicus) cows submitted to a 7-d estradiol (E2)/progesterone (P4)-based timed artificial insemination (TAI) protocol, receiving various combinations of doses and hormones. Primiparous (n = 962) and multiparous (n = 1935) cows were submitted to synchronization (n = 2012) and resynchronization (n = 885 non-pregnant cows at pregnancy diagnosis 30 d after TAI) protocols, following a 2 × 2 × 2 factorial arrangement of eight treatments. At the initiation of the TAI protocol (Day -9), all cows received a 1.0 g intravaginal P4 insert, 2.0 mg E2 benzoate and received (PGF1) or not (PGF0) 0.5 mg cloprostenol sodium (PGF). On Day -2, the P4 insert was removed, all cows received 0.5 mg PGF, 300 IU equine chorionic gonadotropin (eCG) and 0.5 (EC0.5) or 1.0 mg estradiol cypionate (EC1.0). On Day 0, cows were treated (G1) with 8.4 µg buserelin acetate (GnRH) or not (G0), concurrently with TAI. The eight treatments were generated: 1) PGF0-EC0.5-G0 (n = 364), 2) PGF0-EC0.5-G1 (n = 363), 3) PGF1-EC0.5-G0 (n = 363), 4) PGF1-EC0.5-G1 (n = 360), 5) PGF0-EC-1.0-G0 (n = 360), 6) PGF0-EC1.0-G1 (n = 363), 7) PGF1-EC1.0-G0 (n = 361), and 8) PGF1-EC1.0-G1 (n = 363). Pregnancy per AI (P/AI) was greater at first AI compared with resynchronization (58.9 [n = 2012] vs. 54.9% [n = 885]). Presence of CL on Day -9 resulted in more cows expressing estrus (81.3 [n = 680] vs. 67.1% [n = 2033]) and greater P/AI (66.0 [n = 692] vs. 54.9% [n = 2106]). There was no difference in P/AI between cows that received or not PGF on Day -9 (58.7 [n = 1447] vs. 56.6% [n = 1450]). In contrast, PGF tended to increase P/AI of cows with CL on Day -9 (with PGF = 69.1 [n = 375] vs. without PGF = 62.5% [n = 317]). Cows that received 1.0 mg EC expressed more estrus than those treated with 0.5 mg (73.8 [n = 1414] vs. 67.9% [n = 1398]) and had greater P/AI (60.2 [n = 1447] vs. 55.1% [n = 1450]). P/AI was greater in cows treated with GnRH at TAI (59.8 [n = 1449] vs. 55.5% [n = 1448]), particularly in cows that did not show estrus (52.7 [n = 393] vs. 38.1% [n = 420]). Moreover, GnRH on Day 0 increased P/AI in cows with BCS < 3.0 (57.1 [n = 723] vs. 48.6% [n = 698]), in primiparous (50.1 [n = 465] vs. 41.9% [n = 497]) and in cows that received 0.5 mg EC (58.9 [n = 723] vs. 51.3% [n = 727]). In conclusion, 1.0 mg of EC on Day -2 and GnRH at TAI improved P/AI, but the combination of a higher dose of EC and GnRH treatment at AI did not enhance this effect. Furthermore, GnRH improved P/AI especially in Bos indicus cows with lower expression of estrus, such as primiparous, thinner cows, and cows treated with 0.5 mg of EC.

Progesterone-based timed AI protocols for Bos indicus cattle III: Comparison of protocol lengths.

This study aimed to validate a 7 d progesterone (P4)-based fixed-time AI (FTAI) protocol for Bos indicus cattle by comparing to 8 and 9 d-type protocols. The first study compared 7 vs. 8 d protocols in Nelore heifers (Exp. 1.1; n = 742) and cows (Exp. 1.2; n = 2488), and the second study compared 7 vs. 9 d protocols in cows (Exp. 2; n = 1343). On experimental Day -10 and Day -11 the 8 and 9 d groups received an intravaginal P4 implant, 2.0 mg estradiol benzoate (EB) and 0.5 mg cloprostenol sodium (PGF). On Day -9 the 7 d group received the same treatments (P4, EB, and PGF). Then, on Day -2 all groups had the P4 implants removed, and PGF, 0.6 mg estradiol cypionate, and 300 IU equine chorionic gonadotropin (eCG) was administered. Fixed-time AI was performed 48 h later (Day 0) and 8.4 mg buserelin acetate (GnRH) was administered to 7d-G, 8d-G and 9d-G groups, whereas 7d-0, 8d-0 and 9d-0 groups did not receive GnRH at AI. Estrus was detected using tail-chalk between Day -2 and Day 0. Pregnancy per AI (P/AI) was evaluated by ultrasound 30 d after AI. Effects were considered significant when P ≤ 0.05, whereas a tendency was designated when P ≤ 0.10 and P > 0.05. In heifers (Exp. 1.1), incidence of estrus was similar regardless of protocol length (7 or 8 d). There was no independent treatment effect on P/AI or interaction between protocol length and GnRH at AI for P/AI (7d-0: 46.9, 7d-G: 51.4, 8d-0: 47.7, and 8d-G: 43.6%). Heifers in estrus had greater P/AI, and GnRH had no additional effect. More cows (Exp. 1.2) from the 8 d protocol were in estrus than cows submitted to the 7 d protocol. Additionally, despite no interaction between protocol length and GnRH on P/AI (7d-0: 55.9, 7d-G: 60.9, 8d-0: 56.2, and 8d-G: 60.8%), GnRH at AI increased P/AI. There was no interaction between estrus and GnRH, but cows displaying estrus had greater P/AI. Cows not expressing estrus tended (P = 0.06) to have greater P/AI when receiving GnRH. In Exp. 2, more 9 d cows were in estrus than 7 d cows. Protocol length did not affect P/AI but tended (P = 0.08) to interact with GnRH (7d-G had greater P/AI [57.9%] than 7d-0 [47.6%], but 9d-0 [54.6%] and 9d-G [55.4%] were not different from other groups). Moreover, GnRH increased P/AI only for the 7 d protocol. No interaction between estrus and GnRH was detected but estrus improved P/AI, and GnRH tended (P = 0.09) to improve P/AI of cows in estrus. In conclusion, despite longer protocols being more conducive to expression of estrus, there were no detectable effects of protocol length on P/AI. In addition, GnRH at FTAI may improve fertility in cows, particularly when cows are treated with shorter protocols.

Interactions of circulating estradiol and progesterone on changes in endometrial area and pituitary responsiveness to GnRH†.

Changes in circulating progesterone (P4) and estradiol (E2) during proestrus produce dynamic changes in endometrial function and pituitary release of gonadotropins. Independent and combined effects of P4 and E2 on endometrium and pituitary were evaluated. In a preliminary study, an exogenous hormone model of proestrus was created by removal of corpus luteum and follicles ≥5 mm followed by gradual removal of intravaginal P4 implants during 18 h and treatment with increasing doses of estradiol benzoate during 48 h to mimic proestrus using high E2 (n = 9) or low E2 (n = 9). Decreased P4, increased E2, and increased endometrial area (EA) simulated proestrus in high-E2 cows and this was used subsequently. The main experiment used a 2 × 2 factorial design with: high E2 and low P4 (n = 11); high E2 and high P4 (n = 11); low E2 and high P4 (n = 11); low E2 and low P4 (n = 10). At 48 h, gonadotropin-releasing hormone (GnRH)-induced luteinizing hormone (LH) and follicle stimulating hormone (FSH) release was determined. Variables were analyzed using PROCMIXED of Statistical Analysis System. The EA increased dramatically during 48 h only in high-E2 and low-P4 cows. For FSH, high-E2 cows had greater area under the curve (AUC) and FSH peak after GnRH than low E2, with mild negative effects of high P4. For LH, concentration at peak and AUC were 2-fold greater in high E2 compared to low-E2 groups, with low P4 also 2-fold greater than high-P4 groups. Thus, maximal changes in uterus and pituitary during proestrus depend on both low P4 and high E2, but different physiologic responses are regulated differently by E2 and P4. Changes in endometrium depend on low P4 and high E2, whereas GnRH-induced FSH secretion primarily depends on high E2, and GnRH-induced LH secretion is independently increased by high E2 or reduced by high P4.

Progesterone-based timed AI protocols for Bos indicus cattle I: Evaluation of ovarian function.

Three experiments evaluated ovarian dynamics and circulating progesterone (P4) during P4-based protocols initiated with GnRH, estradiol benzoate (EB), or no additional treatment in Nelore (Bos indicus) cattle. In Exp 1 (n = 59 cows), a 5-d P4-only protocol (P-5d; D0: P4 implant alone (1g); D5: P4 removal, 0.5 mg estradiol cypionate [EC], 0.526 mg cloprostenol [PGF], and 300 IU equine chorionic gonadotropin [eCG]; D7: 8.4 µg buserelin acetate [GnRH]) was compared to a 9d protocol initiated with EB (EB-9d; D0: 2 mg EB + P4; D9: P4 removal + EC + PGF + eCG), and to a 7d GnRH protocol (G-7d; D0: 16.8 µg GnRH + P4; D6: PGF + eCG; D7: P4 removal + PGF; D9: GnRH). Exp 2 (n = 55 cows) compared G-7d and EB-7d protocols (similar to EB-9d, but D9 treatments were done on D7). Exp 3 (n = 64 heifers) compared EB-7d, G-7d, and P-5d protocols. For all experiments, daily ovarian ultrasonography was done from D0 until 4d after implant withdrawal and blood samples were collected at D0 and first PGF. Follicle dynamics were determined for each individual animal, analyzed within individual experiments, and afterwards combined to determine overall effects of treatments. The protocol that began with GnRH, G-7d, had greater ovulation rate after D0 with subsequently greater number of CL and circulating P4 at time of PGF (52.8%, 1.0 ± 0.1 CL, 4.0 ± 0.4 ng/mL) than for EB protocols (12.1%, 0.4 ± 0.05 CL, 2.0 ± 0.2 ng/mL), or P-5d (2.5%, 0.6 ± 0.09 CL, 2.6 ± 0.3 ng/mL). The G-7d and EB protocols had synchronized follicle wave emergence in 92.1% of animals but with distinct patterns. For the G-7d group, wave emergence occurred earlier in ovulating than non-ovulating animals (1.4 ± 0.2 d vs 2.5 ± 0.4 d). By comparison, most animals in EB-7d or EB-9d (80.3%) displayed atresia of the dominant follicle, followed by wave emergence 2-3 d after EB treatment. In contrast, P-5d protocol synchronized wave emergence in only 30.0% of cows. Nevertheless, no differences among treatments were detected for ovulation at end of the protocol (85.7%). In conclusion, the P-5d protocol did not synchronize follicle wave emergence but produced similar final ovulation, whereas, GnRH and EB protocols had follicle dynamics synchronized by distinct mechanisms that produced differences in CL number and P4 at the time of PGF treatment but similar final ovulation. Based on ovarian function, each of these synchronization methods are promising for use in FTAI, although fertility still needs to be evaluated.

Progesterone-based timed AI protocols for Bos indicus cattle II: Reproductive outcomes of either EB or GnRH-type protocol, using or not GnRH at AI.

The aim of these experiments was to study ovarian dynamics and fertility of Bos indicus beef cattle submitted to 7-d progesterone (P4)-based fixed-time AI (FTAI) protocols using different hormonal treatments. In Exp. 1, 2 yr old Nelore heifers (n = 973) were randomly assigned to one of four treatments: EB-0 (estradiol benzoate, EB on D0 and no GnRH at AI), EB-G (EB on D0 and GnRH at AI), G-0 (GnRH on D0 and no GnRH at AI), or G-G (GnRH on D0 and at AI). On D0, heifers received an intravaginal P4 implant (0.5 g) for 7 d and EB (1.5 mg) or GnRH (16.8 µg). On D7, the P4 implant was withdrawn and heifers received cloprostenol (PGF; 0.5 mg) and estradiol cypionate (EC, 0.5 mg). Heifers in G groups also received PGF and eCG (200 IU) on D6, whereas EB heifers received eCG on D7. At FTAI on D9, only EB-G and G-G groups received GnRH (8.4 µg). In Exp. 2, Nelore cows (n = 804) received the same treatments (EB-0, EB-G, G-0, or G-G) using a 1.0 g P4 implant, 2.0 mg EB, and 300 IU eCG. Effects were considered significant when P ≤ 0.05. After treatment on D0, G had more ovulations than EB in heifers (60.3 [287/476] vs. 12.7% [63/497]) and cows (73.7 [83/112] vs. 24.4% [28/113]). Luteolysis after D0 was greater in EB than G in heifers (39.2 [159/406] vs. 20.0% [77/385]) and cows (25.5 [14/55] vs. 1.6% [1/64]). Heifers in G had larger follicles (mm) than EB on D7 (10.3 ± 0.2 vs. 9.2 ± 0.2) and at AI (11.9 ± 0.2 vs. 11.3 ± 0.2). Cows had larger follicles in G than EB on D7 (11.0 ± 0.3 vs. 9.9 ± 0.3) but not at AI. More estrus was observed in G than EB for heifers (80.3 [382/476] vs. 69.6% [346/497]) and cows (67.6 [270/400] vs. 56.2% [227/404]). There was no interaction between D0 and D9 treatments on pregnancy per AI (P/AI) in heifers (EB-0: 56.7 [139/245], EB-G: 53.6 [135/252], G-0: 52.6 [127/241], and G-G: 57.5% [135/235]). However, cows from EB-G had greater P/AI than EB-0 (69.5 [142/204] vs. 60.2% [120/200]), whereas P/AI for G-0 (62.7% [127/203]) was similar to G-G (60.9% [120/197]). In heifers, there was no interaction of GnRH at AI with estrus, however, cows that did not display estrus had greater P/AI if they received GnRH at AI (GnRH = 59.1 [91/154] vs. No GnRH = 48.2% [78/162]). Thus, protocols initiated with EB or GnRH for Bos indicus heifers and cows had differing ovarian dynamics but similar overall fertility, enabling their use in reproductive management programs. Treatment with GnRH at time of AI increased fertility in some instances in Bos indicus cows but not in heifers.

Oxytocin-induced prostaglandin F2-alpha release is low in early bovine pregnancy but increases during the second month of pregnancy†.

Circulating prostaglandin F2α metabolite (PGFM) after an oxytocin challenge was evaluated throughout the first 2 months of pregnancy in lactating Holstein cows. On day 11, 18, and 25 after artificial insemination (AI), and on days 32, 39, 46, 53, and 60 of pregnancy, cows were challenged with 50 IU oxytocin, i.m. Blood was collected before (0 min), 30, 60, 90, and 120 min after oxytocin for plasma PGFM concentrations. Ultrasound evaluations were performed for pregnancy diagnosis on day 32-60 post-AI. Nonpregnant (NP) cows on day 18 were designated by a lack of interferon-stimulated genes in peripheral blood leukocytes and Pregnant (P) based on day 32 ultrasound. On day 11, P and NP were similar with low PGFM and no effect of oxytocin on PGFM. On day 18, oxytocin increased PGFM (3-fold) in NP with little change in P cows. Comparing only P cows from day 11 to 60, basal circulating PGFM increased as pregnancy progressed, with day 11 and 18, lower than all days from day 25 to 60 of pregnancy. Oxytocin-induced PGFM in P cows on day 25 was greater than P cows on day 18 (2.9-fold). However, oxytocin-induced PGFM was lower on day 25 compared to day 53 and 60, with intermediate values on day 32, 39, and 46 of pregnancy. Thus, the corpus luteum (CL) of early pregnancy (day 11, 18) is maintained by suppression of PGF, as reflected by suppressed PGFM in this study. However, during the second month of pregnancy, uterine PGF secretion was not suppressed since basal PGFM and oxytocin-induced PGFM secretion were elevated. Apparently, mechanisms other than suppression of oxytocin receptors maintain CL after day 25 of pregnancy.

Profiles of prostaglandin F2α metabolite in dairy cattle during luteal regression and pregnancy: implications for corpus luteum maintenance†.

Mechanisms of bovine corpus luteum (CL) maintenance during the second month of pregnancy have not been adequately investigated, despite significant reproductive losses. In the first month, interferon-tau is believed to suppress oxytocin-stimulated prostaglandin F2α (PGF) production, yet there are conflicting reports of circulating PGF metabolite (PGFM). In this study, characterization of PGFM and P4 occurred through continuous bihourly blood sampling in cows undergoing CL regression (day 18-21, n = 5), and during the first (day 18-21, n = 5) and second month (day 47-61; n = 16) of pregnancy. Cattle in the second month were assigned to control (n = 8) or oxytocin treatment (n = 8; three pulses to mimic luteolysis) to evaluate if oxytocin receptors were active. All cows but one (which had elevated PGFM prior to oxytocin treatment) maintained the pregnancy. Basal PGFM concentrations were low (11.6 ± 0.7 pg/mL) in the first month but increased 2.54-fold in the second month. Few (0.26 ± 0.12 pulses/day) PGFM pulses with low peak concentrations (28.8 ± 3.1 pg/mL) were observed during the first month of pregnancy, similar to cows not undergoing regression. However, in the second month, frequency (1.10 ± 0.26 pulses/day) and peak concentration (67.2 ± 5.0 pg/mL) of PGFM pulses increased, displaying similar frequency but lower peak PGFM than seen in regression (1.44 ± 0.14 pulses/day; 134.5 ± 18.9 pg/mL). Oxytocin treatment increased likelihood of PGFM pulses post-treatment and increased peak concentration (89.7 ± 10.1 pg/mL) in cows during the second month. Thus, cows have more PGFM pulses during second than first month of pregnancy, possibly induced by endogenous oxytocin, indicating suppression of PGF production is an important mechanism for CL maintenance during first but not second month of pregnancy.

Effect of different chorionic gonadotropins on final growth of the dominant follicle in Bos indicus cows.

The aim of this study was to evaluate the effect of eCG or hCG on the final growth of the dominant follicle in Nelore (Bos indicus) cows submitted to fixed-time AI (FTAI). Eighty-four lactating cows with body condition score (BCS) of 2.9 (range 1-5) were used. At a random day of the estrous cycle (D0) cows received 2 mg estradiol benzoate and a reused intravaginal progesterone device (1.9 g). At D8, when the device was removed, 0.5 mg cloprostenol and 1 mg estradiol cypionate was given i.m., and cows were randomly assigned to receive on D8 one of the following treatments: Control (no treatment; n = 17), eCG (300 IU i.m.; n = 17), hCG 300 (300 IU i.m.; n = 18), hCG 200 IM (200 IU i.m.; n = 16) and hCG 200 SC (200 IU s.c.; n = 16). On the same day and 2 days later, cows were subjected to ovarian ultrasonography to evaluate the diameter of the largest follicle and to calculate follicular growth rate (D8 to D10). No differences were observed for the diameter of the largest follicle on D8 (P = 0.3) or D10 (P = 0.4) among treatments. However, the growth rate of the dominant follicle between D8 and D10 was greater for the groups eCG and hCG 300 and there were no differences between the other treatments (Control = 1.1 mm/day; eCG = 1.8 mm/day; hCG 300 = 1.8 mm/day; hCG 200 IM = 1.3 mm/day; hCG 200 SC = 1.3 mm/day; P = 0.02). In addition, more cows from the Group hCG 300 presented premature ovulation (44.4%) than cows from Control (5.8%), eCG (0%), or hCG 200 IM (12.5%), but did not differ from Group hCG 200 SC (18.7%). Regardless of treatment, the size of the largest follicle on D8 was different between cows that presented premature ovulation vs. cows that did not ovulate prematurely (11.3 mm vs. 9.9 mm, respectively; P = 0.01). Treatment with different hCG doses on D8 of a FTAI protocol failed to produce similar effects compared to eCG in terms of final follicular growth support and greater ovulatory follicle size. In addition, the groups hCG 300 and hCG 200 SC induced premature ovulation in a greater portion of cows. Thus, a single administration of hCG on D8 does not appear to be a reliable alternative to eCG treatment in FTAI protocols.

Equine chorionic gonadotropin increases fertility of grazing dairy cows that receive fixed-time artificial insemination in the early but not later postpartum period.

This study evaluated effects of equine chorionic gonadotropin (eCG) on fertility of 679 crossbred (Bos taurus x Bos indicus) lactating grazing cows synchronized for fixed-time AI (FTAI). At a random day of the estrous cycle cows received an intravaginal progesterone (P4) implant, 2 mg estradiol benzoate (EB) and 100 µg gonadorelin (D0-AM). On D7-AM, cows received 0.5 mg sodium cloprostenol and were randomly assigned into two treatments: eCG (n = 340; 400 IU eCG on D7), or Control (n = 339; no eCG). On D8-PM, P4 implants were removed and cows received 0.5 mg sodium cloprostenol and 1 mg EB. Insemination was performed on D10-AM. Pregnancy was diagnosed 30 and 60 d after AI. Treatment with eCG tended to increase pregnancy per AI (P/AI) compared to Control at 30 (37.8 vs. 30.2%; P = 0.06) and 60 (31.9 vs. 25.1%; P = 0.08) d. Pregnancy loss and twinning did not differ between groups. Treatment with eCG increased (P < 0.05) P/AI at 30 (39.0 vs. 25.2%) and 60 (32.8 vs. 21.3%) d for cows inseminated at ≤ 70 d in milk (DIM) but had no effect in cows receiving AI after 70 DIM. Thus, eCG on D7 of a FTAI protocol increased fertility of crossbred dairy cows inseminated in the early postpartum period.

In vivo embryo production in cows superovulated 1 or 2 days after ovum pick-up.

The present study evaluated superovulatory responses and in vivo embryo production in cows treated with FSH starting 1 or 2 days after ovum pick-up (OPU). Thirty-three non-lactating Nelore cows were subjected to aspiration of all follicles ≥3mm for OPU. After OPU, cows were randomly divided into two groups in which the follicle superstimulatory treatments with FSH started 1 or 2 days after OPU (Groups D1 and D2, respectively). Data are presented as the least squares mean±s.e.m. The number of follicles ≥3mm before OPU was similar between groups (~34); however, cows in Group D2 had more follicles ≥3mm on the first day of FSH (15.2±2.3 vs 7.6±1.7; P=0.04) and a higher ratio of the number of follicles at first FSH/number of follicles before OPU (0.41±0.04 vs 0.24±0.02; P=0.01). In addition, Group D2 cows had a greater superovulatory response than did cows in Group D1 (18.9±2.8 vs 9.1±1.9 corpora lutea, respectively; P<0.03). However, there was no difference in the total number of recovered ova and embryos from cows in Groups D2 and D1 (5.1±1.4 vs 4.9±1.3, respectively; P>0.10). Nevertheless Group D2 cows had more freezable embryos than Group D1 cows (3.2±1.1 vs 1.3±0.5, respectively; P<0.05). Cows from Group D2 had a much higher proportion (P<0.001) of follicles ≥8mm compared with follicles ≥6mm and <8mm at the time of the last treatment with FSH. In conclusion, to obtain a greater production of viable embryos in superovulated cows after OPU, it is recommended to wait at least 2 days before starting FSH treatment.