Comparison of GnRH vs. estradiol benzoate plus GnRH to initiate a progesterone-based timed-artificial insemination resynchronization protocol in lactating dairy cows.

The present study compared 2 strategies to initiate a progesterone (P4)-based timed-artificial insemination (TAI) protocol for lactating dairy cows: only GnRH or estradiol benzoate (EB) plus GnRH. Lactating Holstein cows (n = 487; 184 primiparous and 303 multiparous) from 2 commercial dairy herds were used for their second or greater services postpartum. Weekly nonpregnant cows at pregnancy diagnosis 32 d after a previous AI were randomly assigned to 1 of 2 experimental groups that differed only in the strategy to initiate (d 0) the TAI protocol. On d 0, every cow received a 2.0 g P4 implant, and in Group EB+GnRH, cows were treated with 2.0 mg i.m. of EB and 16.8 µg i.m. of buserelin acetate (GnRH), while in Group GnRH, cows received only 16.8 µg i.m of GnRH. Seven d later (d 7), 0.530 mg i.m. of cloprostenol sodium (PGF) was administered in all cows, followed by a second dose on d 8, concomitant with 1.0 mg i.m. of estradiol cypionate (EC) and P4 implant withdrawal. The TAI was performed on d 10 (48 h after P4 device withdrawal) in both experimental groups. Only conventional Holstein semen was used throughout the study. The percentage of cows with corpus luteum (CL) on d 0 (73%) and overall ovulation rate after d 0 (54%) did not differ between groups. The CL regression between d 0 and the first PGF treatment was greater in EB+GnRH than GnRH group (42 vs. 31%). Consequently, the proportion of cows with CL at PGF was greater when only GnRH was used on d 0 compared with EB plus GnRH (86 vs. 82%), and the mean number of CL at PGF was greater (1.23 vs. 1.11). The expression of estrus near TAI was greater in GnRH group (84 vs. 77%), and cows showing estrus had greater (44 vs. 10%) pregnancy per AI (P/AI) on d 32 for both treatments. There was no effect of the presence of CL on d 0 or at PGF, nor of ovulation after d 0 or CL regression between d 0 and d 7 on fertility. However, fertility was critically impaired when cows did not have CL at both times, d 0 and at PGF treatment. There was no interaction between treatment and other variables, and the P/AI was similar in cows receiving EB plus GnRH or only GnRH on d 0 (37.8 vs. 36.6%). In summary, although there was no detectable difference in P/AI between treatments, this study demonstrated potential negative physiological outcomes caused by EB treatment on d 0 (greater incidence of luteolysis after d 0 and fewer cows with CL at PGF treatment). In conclusion, there was no benefit of adding EB at the initiation of a P4-based TAI protocol on fertility compared with using GnRH alone, despite differences in ovarian dynamics and expression of estrus.

Effect of progesterone from corpus luteum, intravaginal implant, or both on luteinizing hormone release, ovulatory response, and subsequent luteal development after gonadotropin-releasing hormone treatment in cows.

This study aimed to determine the effect of circulating progesterone (P4) concentrations produced by a corpus luteum (CL) or released by an intravaginal P4 implant (IPI) on GnRH-induced LH release, ovulatory response, and subsequent CL development, after treatment with 100 µg of gonadorelin acetate (GnRH challenge). Nonlactating multiparous Holstein cows were synchronized and GnRH was used to induce ovulation (d -7). Over 4 replicates, cows that ovulated (n = 87) were randomly assigned to a 2 × 2 factorial arrangement (presence or absence of CL and insertion or not of an IPI at GnRH challenge), creating 4 groups: CL_IPI, CL_NoIPI, NoCL_IPI, and NoCL_NoIPI. On d -1.5, NoCL_IPI and NoCL_NoIPI received 2 doses of 0.53 mg of cloprostenol sodium (PGF2α), 24 h apart to regress CL. On d 0, cows were treated with 100 µg of GnRH and, simultaneously, cows from IPI groups received a 2-g IPI maintained for the next 14 d. Diameter of dominant follicle, ovulatory response, and subsequent CL volume were assessed by ultrasonography on d -1.5, 0, 2, 7, and 14. Blood samples were collected on d -1.5, 0, 1, 2, 3, 5, 7, and 14 for analysis of circulating P4 and at 0, 1, 2, 4, and 6 h after GnRH challenge for analysis of circulating LH. In a subset of cows (n = 34), the development of the new CL was evaluated daily, from d 5 to 14. The presence of CL at the time of GnRH challenge affected the LH peak and ovulatory response (CL: 5.3 ng/mL and 58.1%; NoCL: 13.2 ng/mL and 95.5%, respectively). However, despite producing a rapid increase in circulating P4, IPI insertion did not affect LH concentration or ovulation. Regardless of group, ovulatory response was positively correlated with LH peak and negatively correlated with circulating P4 on d 0. Moreover, new CL development and function were negatively affected by the presence of CL and by the IPI insertion. In summary, circulating P4 produced by a CL exerted a suppressive effect on GnRH-induced LH release and subsequent ovulation of a 7-d-old dominant follicle, whereas the IPI insertion at the time of GnRH had no effect on LH concentration or ovulation. Finally, elevated circulating P4, either from CL or exogenously released by the IPI, compromised the development and function of the new CL, inducing short cycles in cows without CL at the time of GnRH treatment.

Relationships among total mixed ration nutritional components and reproductive performance in high-producing dairy herds.

The main objective of the present study was to determine whether composition of total mixed ration influences reproductive performance in high-producing commercial dairy farms. Dairy producers and nutritional consultants from 48 dairy farms located in Wisconsin agreed to provide reproductive data and dietary information on high milk production pens during the main breeding period for the previous 12 mo. Dietary components (percentage of dry matter) were crude protein (CP), rumen degradable (RDP) and undegradable (RUP) protein, neutral detergent fiber (NDF), nonfiber carbohydrates (NFC), starch, and fat. Reproductive data were service rate (SR), overall pregnancy per artificial insemination (P/AI) and P/AI at the first service, 21-d pregnancy rate (PR), days open, and percentage of cows pregnant by 150 d in milk (PREG150). Participating herds had lactating Holstein cows (range = 143 to 2,717) housed in freestall facilities. Statistical analyses were performed with CORR and GLIMMIX of SAS (SAS Institute Inc.). Daily average milk production of herds was 38.9 ± 0.60 kg/d (30.0 to 50.4 kg/d). Overall SR was 58.5% (39-73) and P/AI was 36.1% (22-49). Overall 21-d PR was 20.3% (10-42%). Correlation between SR and PR was 0.59, whereas correlation of overall P/AI and P/AI at first service with PR were both 0.72. Similarly, for PREG150, correlation with overall P/AI (0.63) and P/AI at first service (0.66) were greater than with SR (0.48). There was large variation in diet composition, with CP varying from 16.0 to 18.7%, NDF from 24.9 to 35.1%, NFC from 31.7 to 46.6%, starch from 20.1 to 30.8%, and fat from 3.1 to 6.7%. Overall, there were no detectable associations of CP, RDP, and RUP with reproductive measures. The strongest relationship was a decrease in reproductive performance with increasing dietary NFC including overall P/AI (-0.48), P/AI at first service (-0.51), and PREG150 (-0.33). Starch also had a negative relationship with P/AI at first service (-0.35). Conversely, greater NDF was positively associated with P/AI at first service (0.34). Fat content was also positively associated with P/AI at first service (0.34). When NFC was divided in tertiles (<40, 40 to 42.2, and >42.2% NFC), the highest tertile had lower overall P/AI (39 vs. 36 vs. 31%), P/AI at first service (43 vs. 40 vs. 33%), and PREG150 (54 vs. 53 vs. 47%). In conclusion, farms with greater dietary NFC may have compromised reproductive performance. Correspondingly, herds with greater NDF content may achieve high milk production with potentially positive associations with reproduction. Other relationships of dietary components on reproduction were not as obvious in this herd-level analysis.

What are the factors associated with pregnancy loss after timed-artificial insemination in Bos indicus cattle?

Pregnancy loss (PL) has important impacts on the profitability of livestock production systems, although it is not widely reported, particularly in Bos indicus cattle. The present study retrospectively evaluated PL after timed-artificial insemination (TAI) in Bos indicus (Nelore) beef cows corresponding to several factors, such as parity, body condition score (BCS), presence of corpus luteum (CL) at the beginning of TAI protocols, expression of estrus, and hormonal manipulations during the TAI protocol. Data from two experiments performed during three breeding seasons (BS) were reanalyzed. Both experiments evaluated adding GnRH treatment at TAI in a 7-d estradiol (E2) plus progesterone (P4)-based protocol, with intravaginal P4 implant removal on Day 7 combined with treatment with 0.5 or 1.0 mg E2 cypionate and 300 IU eCG, and TAI on Day 9. In addition, during BS 2 and 3 (Exp 2), cows were randomized to receive or not a PGF treatment on Day 0 (beginning of the TAI protocol). In all BS, presence of CL and BCS were evaluated at the beginning of TAI protocols, follicle size and expression of estrus were evaluated at TAI. The PL was assessed between the first pregnancy diagnosis (∼35d) and parturition. There were no effects of hormonal manipulations within TAI protocols of different BS on PL. There was no interaction between GnRH treatment at TAI and the other variables within BS, and there was no main effect of GnRH treatment on PL (without = 10.1% [102/1007] vs. with = 10.4% [114/1100]). The addition of PGF on Day 0 had no effect on PL (11.5% [102/886] vs. 10.5% [89/850]), as well as EC dose to induce final ovulation (10.8% [89/827] vs. 11.2% [102/909] for 0.5 and 1.0 mg, respectively). Primiparous had greater PL than multiparous cows (14.0% [77/550] vs. 8.9% [139/1557]), and cows not expressing estrus near TAI had greater PL than those expressing estrus (13.5% [57/422] vs. 9.7% [156/1617]). There was no interaction between follicle size at TAI and GnRH treatment on PL. However, probability of PL decreased linearly as follicle size at TAI increased. There were no effects of service number (first TAI or resynchronization), BCS, or presence of CL on D0 on PL. In addition, PL was not affected by sire within any of the BS. In conclusion, some factors that are known to impact pregnancy per AI also influenced PL, such as parity and expression of estrus, although, other aspects such as BCS, number of services, and presence of CL on D0 did not affect PL. Moreover, commonly implemented treatments to increase fertility (e.g., PGF on Day 0 and increasing EC dose to 1.0 mg) did not affect PL. Finally, the GnRH treatment at TAI had no effect on PL and did not interact with any of the variables, an important result, since GnRH at TAI also increases fertility in Bos indicus beef cows.

Improved fertility following a gonadotropin-releasing hormone treatment on day 2 of an estradiol and progesterone-based timed-artificial insemination protocol in lactating dairy cows.

The present study evaluated the addition of gonadotropin-releasing hormone (GnRH) concomitant or 2 d after the beginning of protocols initiated with estradiol benzoate (EB). A total of 459 multiparous and 371 primiparous lactating Holstein cows were enrolled in the study. Weekly cohorts of cows were randomly assigned to 1 of 3 experimental groups that differed in the strategy to initiate the timed AI (TAI protocol. On d 0, all cows received a 1.55-g progesterone (P4) implant. Additionally, cows in the EBd0 group received 2 mg of EB i.m.; cows in the EBd0-GnRHd0 group were treated simultaneously on d 0 with 2 mg of EB plus 100 µg of gonadorelin diacetate tetrahydrate (GnRH) i.m.; and cows in the EBd0-GnRHd2 group received 2 mg of EB on d 0 and 100 µg of GnRH 48 h later (d 2). The remaining treatments in the protocol were similar among groups and included 0.53 mg (i.m.) of cloprostenol sodium (PGF2α) on d 7, followed by a second PGF2α treatment on d 9 (at the time of P4 implant withdrawal) and 1 mg of estradiol cypionate i.m. Then, TAI was performed on d 11 (48 h after P4 removal) in all experimental groups. We detected an effect of treatment on pregnancy per AI (P/AI) on d 30, in which cows from the EBd0-GnRHd2 group demonstrated greater fertility than EBd0 cows, whereas cows in the EBd0-GnRHd0 group did not differ among EBd0 and EBd0-GnRHd0 (40.5 vs. 30.4 vs. 34.4%, respectively). In summary, GnRH treatment at the beginning of an estradiol and P4-based TAI protocol increased fertility only when GnRH was given on d 2. Moreover, a more pronounced positive effect of this strategy was observed in particular classes of cows: multiparous cows, cows with greater milk production, and those receiving the first service.

Accessory corpus luteum regression during pregnancy I: timing, physiology, and P4 profiles.

Inappropriate corpus luteum (CL) regression can produce pregnancy loss. An experimental model was utilized to investigate regression of accessory CL during pregnancy in dairy cows. Cows were bred (day 0) and treated with gonadotrophin-releasing hormone 6 days later to form accessory CL. Transrectal ultrasound (every other days) and blood samples for progesterone (P4; daily) were performed until day 56 of pregnancy. On day 28, 13 cows were confirmed pregnant, and accessory CL were found contralateral (n = 9) or ipsilateral (n = 4) to previous ovulation. On day 18, CL biopsy was performed to analyze mRNA expression for interferon-stimulated genes (ISGs). Luteolysis occurred more frequently in cows that had contralateral accessory CL (88.9% (8/9)) than in cows with ipsilateral accessory CL (0% (0/4)). Luteolysis of contralateral accessory CL occurred either earlier (days 19-23; 2/8) or later (days 48-53; 6/8) in pregnancy and occurred rapidly (24 h), based on daily P4. After onset of earlier or later accessory CL regression, circulating P4 decreased by 41.2%. There was no difference in luteal tissue mRNA expression for ISGs on day 18 between accessory and original CL and between CL that subsequently regressed or did not regress. On day 56, an oxytocin challenge dramatically increased prostaglandin F2α metabolite (PGFM) in all cows but produced no pregnancy losses, although cows with previous accessory CL regression had greater PGFM. In summary, ipsilateral accessory CL did not regress during pregnancy, whereas most contralateral CL regressed by 63 days of pregnancy, providing evidence for local mechanisms in regression of accessory CL and protection of CL during pregnancy.

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.

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.