Effects of two estradiol esters (benzoate and cypionate) on the induction of synchronized ovulations in Bos indicus cows submitted to a timed artificial insemination protocol

The effects of estradiol benzoate (EB) and estradiol cypionate (EC) on induction of ovulation after a synchronized LH surge and on fertility of Bos indicus females submitted to timed AI (TAI) were evaluated. In Experiment 1, ovariectomized Nelore heifers were used to evaluate the effect of EB (n = 5) and EC (n = 5) on the circulating LH profile. The LH surge timing (19.6 and 50.5 h; P = 0.001), magnitude (20.5 and 9.4 ng/mL; P = 0.005), duration (8.6 and 16.5 h; P = 0.001), and area under the LH curve (158.6 and 339.4 ng/mL; P = 0.01) differed between the EB and EC treatments, respectively. In Experiment 2 (follicular responses; n = 60) and 3 (pregnancy per AI; P/AI; n = 953) suckled Bos indicus beef cows submitted to an estradiol/progesterone-based synchronization protocol were assigned to receive one of two treatments to induce synchronized ovulation: 1 mg of EB im 24 h after progesterone (P4) device removal or 1 mg of EC im at P4 device removal. There was no difference (P > 0.05) between EB and EC treatments on follicular responses (maximum diameter of the ovulatory follicle, 13.1 vs. 13.9 mm; interval from progesterone device removal to ovulation, 70.2 vs. 68.5 h; and ovulation rate, 77.8 vs. 82.8%, respectively). In addition, P/AI was similar (P < 0.22) between the cows treated with EB (57.5%; 277/482) and EC (61.8%; 291/471). In conclusion, despite pharmacologic differences, both esters of estradiol administered either at P4 device removal (EC) or 24 h later (EB) were effective in inducing an LH surge which resulted in synchronized ovulations and similar P/AI in suckled Bos indicus beef cows submitted to TAI.     

Effects of oestradiol and some of its esters on gonadotrophin release and ovarian follicular dynamics in CIDR-treated beef cattle

Three experiments were conducted to: (1) compare the effect of three oestradiol formulations on gonadotrophin release in ovariectomised cows; (2) compare the effects of either oestradiol-17beta (E-17beta) or oestradiol benzoate (EB), given at two doses, on the synchrony of ovarian follicular wave emergence in CIDR-treated beef cattle; and (3) determine the timing of ovulation of the dominant follicle of a synchronised follicular wave following administration of E-17beta or EB 24h after progesterone withdrawal. In Experiment 1, ovariectomised cows (n = 16) received a once-used CIDR on Day 0 (beginning of the experiment) and were allocated randomly to receive 5mg of E-17beta, EB or oestradiol valerate (EV) plus 100mg progesterone i.m. The CIDR inserts were removed on Day 7. There were effects of time, and a treatment-by-time interaction (P < 0.0001) for plasma concentrations of both oestradiol and FSH. Plasma oestradiol concentrations peaked 12h after treatment, with highest (P < 0.01) peak concentrations in cows given E-17beta; estradiol concentrations subsequently returned to baseline by 36 h in E-17beta-treated cows and by 96 h in EB- and EV-treated cows. Plasma FSH concentrations decreased by 12h after oestradiol treatment in all groups (P < 0.0001), reached a nadir at 24h, and increased by 60 h in all groups; plasma FSH reached higher (P < 0.02) concentrations in E-17beta-treated than in EB- or EV-treated cows. In Experiment 2, non-lactating Hereford cows (n = 29) received a new CIDR on Day 0 (beginning of the experiment), and were assigned randomly to receive 1 or 5mg of E-17beta or EB i.m. on Day 1. On Day 8, CIDR were removed and PGF was given. Transrectal ultrasonography was done once daily from 2 days before CIDR insertion to 2 days after CIDR removal, and then twice-daily to ovulation. Although there was no difference among groups in the interval from oestradiol treatment to follicular wave emergence (4.2 +/- 0.3 days; P = 0.5), 5mg of E-17beta resulted in the least variable interval to wave emergence (P < 0.005), compared with the other treatment groups which were not different (P = 0.1). For the interval from CIDR removal to ovulation, there were no differences among groups for either means (P = 0.5) or variances (P = 0.1). In Experiment 3, beef heifers (n = 32) received a once-used CIDR on Day 0 (beginning of the experiment) plus 100mg progesterone i.m. and were assigned randomly to receive 5mg E-17beta or 1mg EB i.m. On Day 7, CIDR were removed and all heifers received PGF. On Day 8 (24h after CIDR removal), each group was subdivided randomly to receive 1mg of either E-17beta or EB i.m. There was no effect of oestradiol formulation on interval from treatment to follicular wave emergence (4.1 +/- 0.2 days; P = 0.7) or on the median interval (76.6h; P = 0.7) or range (72-120 h; P = 0.08) from CIDR removal to ovulation. In summary, oestradiol treatments suppressed FSH in ovariectomised cows, with the duration of suppression dependent on the oestradiol formulation. Both E-17beta and EB effectively synchronised ovarian follicular wave emergence and ovulation in CIDR-treated cattle, and the interval from CIDR removal to ovulation did not differ in heifers given either E-17beta or EB 24h after CIDR removal.     

Comparison of the effect of estradiol benzoate plus progesterone and GnRH on the follicular wave emergence and subsequent follicular development in CIDR-treated, lactating dairy cows with follicular cysts

This study examined the effect of estradiol benzoate (EB) plus progesterone (P4) as compared with GnRH on follicular wave emergence and follicular development, and synchrony of ovulation and pregnancy rates following a second injection of GnRH in a controlled internal drug release (CIDR)-based timed AI (TAI) protocol in lactating dairy cows with follicular cysts. Lactating dairy cows diagnosed with follicular cysts received a CIDR device, with an injection of 2mg EB plus 50mg P4 (EB+P4 group) or with an injection of 100 microg GnRH (GnRH group) at the beginning of the experiment (day 0). Thereafter, all received PGF(2alpha) at the time of CIDR removal on day 7, GnRH on day 9, and TAI 16 h later. Follicular wave emergence occurred within 7 days in 12/15 EB plus P4-treated and 14/15 GnRH-treated cows (P>0.05). The interval to wave emergence was longer in the EB+P4 group (4.8+/-0.4 days) than in the GnRH group (2.0+/-0.2 days). The mean diameters of preovulatory follicles and the proportion of cows with preovulatory follicles greater than 12 mm on day 9 did not differ between groups (P>0.05). The proportion of cows with synchronized ovulations by 40 h after the GnRH injection on day 11 and pregnancy rates to TAI did not differ between the EB+P4 (13/15 and 36.7%) and the GnRH (14/15 and 53.3%) groups, respectively. Results suggest that a single treatment with EB plus P4 as compared with GnRH simultaneously with CIDR insertion in lactating dairy cows with follicular cysts will result in relatively asynchronous emergence of a new follicular wave, but subsequently similar sizes of preovulatory follicles and synchronous ovulation, resulting in similar pregnancy rates to TAI.     

Importance of estrus on pregnancy per insemination in suckled Bos indicus cows submitted to estradiol/progesterone-based timed insemination protocols

The objective was to evaluate the effect of estrus occurrence (based on removal of tail-head marks) on ovarian responses and pregnancy per AI (P/AI; 30 d after AI) in suckled Bos indicus beef cows submitted to timed AI (TAI) protocols. Cows received an intravaginal device containing 1.0 g progesterone, and 2.0 mg estradiol benzoate im; 8 d later, the intravaginal device was removed, and they were given PGF_2α (0.25 mg of cloprostenol sodium) and 300 IU of eCG, with TAI 48 to 52 h later. In Experiment 1, cows were assigned to receive one of three treatments: 1 mg of estradiol cypionate (ECP) im at progesterone (P4) device removal (N = 178); 10 μg of GnRH im at TAI (N = 190); or both treatments (N = 172). In cows given estradiol (ECP or ECP + GnRH), more displayed estrus (P = 0.002) and became pregnant (P < 0.0001) compared with those receiving only GnRH. In Experiment 2, the effect of the occurrence of estrus on ovarian responses was evaluated in cows (N = 53) synchronized using ECP at device removal. Cows that displayed estrus had a greater diameter of the largest follicle (LF) at device removal (P < 0.0001), a greater diameter at TAI (P < 0.0001), a greater ovulation rate (P = 0.02), a larger CL (P = 0.02), and a greater P4 concentration (P < 0.0001) than cows that did not display estrus. In Experiment 3, the effect of GnRH treatment on P/AI at TAI was evaluated in cows that received ECP at device removal, and either displayed, or did not display, estrus (N = 726). There was no estrus by GnRH interaction (P = 0.22); the P/AI was greater (P < 0.0001) in cows that displayed estrus (61.9%) than cows that did not display estrus (41.4%). However, GnRH did not improve (P = 0.81) P/AI (GnRH = 53.7% vs. no GnRH = 52.6%). In conclusion, exogenous estradiol at device removal increased both the proportion of suckled Bos indicus cows that displayed estrus and P/AI. Cows that displayed estrus had better ovarian responses (i.e., larger follicles at TAI, a greater ovulation rate, larger CL, and greater P4 concentrations) following an estradiol/P4-based synchronization protocol. Although occurrence of estrus improved pregnancy outcomes, GnRH at TAI did not improve P/AI in suckled Bos indicus cows treated with ECP, regardless of estrus occurrence.     

Effects of estradiol cypionate (ECP) on ovarian follicular dynamics,synchrony of ovulation,and fertility in CIDR-based,fixed-time AI programs in beef heifers

Estradiol cypionate (ECP) was used in beef heifers receiving a controlled internal drug release (CIDR; insertion = Day 0) device for fixed-time AI (FTAI) in four experiments. In Experiment 1, heifers (n = 24) received 1mg ECP or 1mg ECP plus 50mg commercial progesterone (CP) preparation i.m. on Day 0. Eight or 9 days later, CIDR were removed, PGF was administered and heifers were allocated to receive 0.5mg ECP i.m. concurrently (ECP0) or 24h later (ECP24). There was no effect of treatment (P = 0.6) on mean (+/-S.E.M.) day of follicular wave emergence (3.9+/-0.4 days). Interval from CIDR removal to ovulation was affected (P<0.05) only by duration of CIDR treatment (88.3+/-3.8h versus 76.4+/-4.1h; 8 days versus 9 days, respectively). In Experiment 2, 58 heifers received 100mg progesterone and either 5mg estradiol-17beta or 1mg ECP i.m. (E-17beta and ECP groups, respectively) on Day 0. Seven (E-17beta group) or 9 days (ECP group) later, CIDR were removed, PGF was administered and heifers received ECP (as in Experiment 1) or 1mg EB 24h after CIDR removal, with FTAI 58-60h after CIDR removal. Follicular wave emergence was later (P<0.02) and more variable (P<0.002) in heifers given ECP than in those given E-17beta (4.1+/-0.4 days versus 3.3+/-0.1 days), but pregnancy rate was unaffected (overall, 69%; P = 0.2). In Experiment 3, 30 heifers received a CIDR device and 5mg E-17beta, with or without 100mg progesterone (P) i.m. on Day 0. On Day 7, CIDR were removed and heifers received ECP as described in Experiment 1 or no estradiol (Control). Intervals from CIDR removal to ovulation were shorter (P<0.05) in ECP0 (81.6+/-5.0h) and ECP24 (86.4+/-3.5h) groups than in the Control group (98.4+/-5.6h). In Experiment 4, heifers (n = 300) received a CIDR device, E-17beta, P, and PGF (as in Experiment 3) and after CIDR removal were allocated to three groups (as in Experiment 2), with FTAI 54-56h (ECP0) or 56-58h (ECP24 and EB24) after CIDR removal. Pregnancy rate did not differ among groups (overall, 63.6%, P = 0.96). In summary, although 1mg ECP (with or without progesterone) was less efficacious than 5mg E-17beta plus 100mg progesterone for synchronizing follicular wave emergence, 0.5mg ECP (at CIDR removal or 24h later) induced a synchronous ovulation with an acceptable pregnancy rate to fixed-time AI.     

Follicular wave emergence, luteal function and synchrony of ovulation following GnRH or estradiol benzoate in a CIDR-treated, lactating Holstein cows

This study evaluated the effect of GnRH or estradiol benzoate (EB) on follicular wave emergence and progesterone concentrations, and following a second injection of GnRH, synchrony of ovulation, and pregnancy rates in a controlled internal drug release (CIDR)-based timed AI (TAI) protocol in lactating Holstein cows. Cows received a CIDR device without hormone (controls), with an injection of 100 microg GnRH or with an injection of 4 mg EB. Thereafter, all received PGF(2 alpha) at the time of CIDR removal on Day 7, GnRH on Day 9, and TAI 16 h later. Follicular wave emergence occurred within 7 days in 19/20 GnRH-treated, 14/20 EB-treated and 5/20 control cows (P < 0.05). The interval to wave emergence was the shorter and less variable (P < 0.01) in the GnRH group (2.9 +/- 0.2 days) than in the EB (4.7 +/- 0.5 days) or control (4.8 +/- 1.0 days) groups. Serum progesterone concentrations from Days 4 to 7 were higher (P < 0.01) in the GnRH-treated cows that ovulated than in those that did not ovulate, or in control and EB-treated cows. The diameters of dominant follicle on Day 7 differed among groups (P < 0.01), and the diameters of the preovulatory follicle on Day 9 were larger (P < 0.01) in the control and GnRH groups than in the EB group. The proportion of cows with synchronized ovulations did not differ among groups, but pregnancy rate to TAI was higher (P < 0.05) in the GnRH group (65%; 13/20) than in the control (30%; 6/20) or EB (35%; 7/20) groups. Results suggest that GnRH treatment of CIDR-treated lactating Holstein cows will result in synchronous follicular wave emergence, large preovulatory follicles and synchronous ovulation, resulting in an acceptable pregnancy rates to TAI.     

The use of PGF2α as ovulatory stimulus for timed artificial insemination in cattle

The objective of this study was to evaluate the effect of a PGF2α-analogue (PGF) on ovulation and pregnancy rates after timed artificial insemination (TAI) in cattle. In experiment 1, crossbred dual-purpose heifers, in a crossover design (3 × 3), were given an intravaginal progesterone-releasing insert (controlled internal drug release [CIDR]) plus 1 mg estradiol benzoate (EB) intramuscularly (im) and 250 μg of a PGF-analogue im on Day 0. The CIDR inserts were removed 5 days after follicular wave emergence, and the heifers were randomly divided into three treatment groups to receive the following treatments: (1) 1 mg of EB im (EB group, n = 13); (2) 500 μg of PGF im (PG group, n = 13); or (3) saline (control group, n = 13), 24 hours after CIDR removal. Ovulation occurred earlier in EB (69.81 ± 3.23 hours) and PG groups (73.09 ± 3.23 hours) compared with control (83.07 ± 4.6 hours; P = 0.01) after CIDR removal. In experiment 2, pubertal beef heifers (n = 444), 12 to 14 months of age were used. On Day 0, the heifers were given a CIDR insert plus 2 mg EB im. On Day 9, the CIDR was removed and the heifers were given 500 μg of PGF im. Heifers were randomly assigned into one of three treatment groups: (1) 1 mg of EB (EB group; n = 145); (2) 500 μg of PGF (PG group; n = 149), both 24 hours after CIDR removal; or (3) 600 μg of estradiol cypionate (ECP group; n = 150) at CIDR removal. Timed artificial insemination occurred 48 hours after CIDR removal in the ECP group and 54 hours in the PG and EB groups. The percentage of heifers ovulating was higher in the PG group compared with the other groups (P = 0.08). However, the pregnancy rates did not differ among groups (47.6%, 45%, and 46.6%, for EB, PG, and ECP, respectively; P = 0.9). In experiment 3, 224 lactating beef cows, 40 to 50 days postpartum with 2.5 to 3.5 of body condition score were treated similarly as described in experiment 2, except for the ECP group, which was excluded. The treatments were as follows: 1 mg EB (EB group; n = 117) or 500 μg PGF (PG group; n = 107), 24 hours after CIDR removal. The calves were temporarily separated from their dams from Days 9 to 11. No difference was detected on the pregnancy rate between the EB and PG groups (58.1% vs. 47.6%, respectively; P = 0.11). Taken together, the combined results suggested that PGF2α could be successfully used to induce and synchronize ovulation in cattle undergoing TAI, with similar pregnancy rates when compared with other ovulatory stimuli (ECP and EB).     

Timed artificial insemination plus heat I: effect of estrus expression scores on pregnancy of cows subjected to progesterone–estradiol-based protocols

Expression of estrus near timed artificial insemination (TAI) is associated with greater fertility, and estrus detection could improve TAI fertility or direct TAI management, although accurate estrus detection can be difficult and time-consuming using traditional methods. The aim of this study is to evaluate influence of estrus on pregnancy (artificial insemination pregnancy rates (P/AI)) and to validate an alternative method to classify estrus/heat expression using tail chalking (HEATSC) in postpartum Bos indicus cows subjected to TAI in progesterone–estrogen-based protocols. In experiment 1 (Exp. 1), cows (5491) were subjected to visual observation of estrus after progesterone device removal, before TAI, and P/AI was evaluated according to estrus and body condition score (BCS). Cows received a progesterone device and 2 mg estradiol benzoate (EB). After 8 days, the device was removed and 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotrophin was given. Later, animals in Exp. 1 received 1 mg EB and TAI 44 to 48 h. In the Exp. 2 – 3830 cows using similar protocol, received different ovulation inducers: 1 mg EB (n=1624) or 1 mg estradiol cypionate (EC; n=2206) on day 8 (D8). Cows were then marked with chalk, and HEATSC evaluated at TAI on D10 (HEATSC1 – no chalk removal=no estrus expression; HEATSC2 – partial chalk removal=low estrus expression; HEATSC3 – near complete/complete chalk removal=high estrus expression). In Exp. 1, cows showing estrus presented greater P/AI (48.4% v. 40.2%, P<0.05). In Exp. 2, P/AI (HEATSC1 – 40.0%; HEATSC2 – 49.7%; HEATSC3 – 60.9%; P<0.001), and larger follicle timed artificial insemination (LFTAI) (<0.001) varied according to HEATSC. There was no difference in P/AI (P=0.41) or LFTAI (P=0.33) according to ovulation inducer. Cows with greater BCS showed greater P/AI in both experiments (P<0.05). Estrus presence and greater HEATSC improved P/AI, and EC v. EB used promoted differential estrus manifestation (cows showing HEATSC2 and HEATSC3: 79.5% with EB v. 69.98% with EC use, P<0.001), however, with similar P/AI. The use of HEATSC in B. indicus cows subjected to TAI is useful to identify cows with greater estrus expression and consequently improved pregnancy rates in TAI, allowing the cows with low HEATSC to be targeted for additional treatments aimed at improving P/AI.     

Fertility in beef cattle given a new or previously used CIDR insert and estradiol, with or without progesterone

The objective was to compare pregnancy rates following fixed-time AI (FTAI) in beef cattle given a new or previously used CIDR insert and injections of estradiol, with or without progesterone, to synchronize follicular wave emergence. In Experiment 1, heifers (n=616) received a new or once-used CIDR insert for 9 days and were given 1mg estradiol cypionate (ECP), with or without 100 mg of a commercial progesterone preparation (CP4), at CIDR insertion. Heifers were treated with PGF at CIDR removal and 0.5 mg ECP i.m. 24h later, with FTAI 55 to 60 h after CIDR removal. Pregnancy rate was not affected by either the number of CIDR uses (P=0.59; 48.3% versus 46.2% for new versus once-used CIDRs, respectively) or the addition of progesterone (P=0.42; 45.6% versus 48.8% for ECP+CP4 and ECP, respectively). In Experiment 2 (replicated at two locations), heifers (n=56) and lactating beef cows (n=307) received a once- or twice-used CIDR and an i.m. injection of 1mg estradiol benzoate (EB), with or without 100 mg progesterone, at CIDR insertion. Cattle received PGF in the ischiorectal fossa at CIDR removal (Day 7) and 1mg EB i.m. 24h later, with FTAI 52 to 56 h after CIDR removal. Pregnancy rate was affected by location (P<0.002; 46.0% versus 61.1% for Locations A and B, respectively), parity (P<0.04; 67.9% versus 53.1% in heifers and cows, respectively), and numbers of times the CIDR had been used (P<0.03; 62.4% versus 48.4% for once- and twice-used CIDRs, respectively). However, the addition of progesterone to the injection of EB at CIDR insertion did not affect pregnancy rate (P=0.6). In Experiment 3, heifers (n=187) received one new, one once-used, one twice-used or two twice-used CIDRs for 7 days and 2 mg EB plus 50 mg of CP4 at the time of CIDR insertion. Heifers were treated with PGF at CIDR removal and 1mg EB i.m. 24 h later, with FTAI 52-56 h after CIDR removal. Pregnancy rate was not affected by treatments (P=0.28, 57.5, 63.8, 47.9, 47.9% for one new, one once-used, one twice-used, or two twice-used CIDRs, respectively). In summary, pregnancy rate to FTAI did not differ between cattle synchronized with a new or once-used CIDR, but pregnancy rate was lower in cattle synchronized with a twice-used CIDR; however, the insertion of two twice-used CIDRs did not affect pregnancy rates. The addition of an injection of progesterone to the estradiol treatment at CIDR insertion did not enhance pregnancy rate to FTAI.     

Length of progesterone exposure needed to resolve large follicle anovular condition in dairy cows

Hypothalamic unresponsiveness to an estradiol surge appears to be an underlying cause of large follicle anovular condition (follicular cysts), but progesterone exposure for 7 days resolves this condition. In this study, dairy cows with induced (Experiment 1) or naturally occurring (Experiment 2) follicular cysts were treated for different times with progesterone. In Experiment 1, 16 of 26 cows (62%) were induced into anovulation by causing a GnRH/LH surge when no ovulatory follicle was on the ovary. Anovular cows (n = 16) were assigned to one of four treatment groups ( 0, 1, 3, or 7 days of progesterone treatment) using an intravaginal, progesterone-releasing implant (CIDR). All anovular cows had low circulating progesterone concentrations before controlled internal drug releasing (CIDR) and greater concentrations that reached steady state (1.3 +/- 0.1 ng/mL progesterone) by 3 h after CIDR insertion. Circulating progesterone decreased to basal concentrations by 4 h after CIDR removal. Cows were treated with 5mg estradiol benzoate (EB) 12 h after CIDR removal. None (n = 4) of the control cows (0 day) had an LH surge after EB. All of the 3 days (5/5) and 7 days (4/4) CIDR-treated cows had an LH surge following EB, but only one of the 1 day (1/3) CIDR-treated cows. Magnitude of the LH peak was similar in the 3 and 7 days cows. All cows treated for 7 days ovulated (4/4), whereas, ovulation occurred in only 3/5, 1/3, and 0/4 of the cows treated for 3, 1, and 0 day, respectively. The two cows in the 3 days group that did not ovulate had a normal LH surge, but these two cows had a smaller maximal follicle size than cows that ovulated. In Experiment 2, naturally anovular lactating dairy cows (24 of 248) were identified using weekly ultrasonography. All anovular cows grew follicles to >12 mm, with 54% (13 of 24) having follicles larger than ovular size (15-24 mm) and 33% (8 of 24) having follicles that would be considered cystic (>25 mm). Anovular cows were randomly assigned to CIDR treatment for 0, 1, or 3 days. All (7/7) of 3 days, 33% (3/9) of 1 day, and 25% (2/8) of control (0 day) cows ovulated by 1 week after CIDR removal. Thus, 3 days but not 1 day of progesterone exposure appears to be sufficient to reinitiate estradiol responsiveness of the hypothalamus.     

The use of GnRH or estradiol to facilitate fixed-time insemination in an MGA-based synchronization regimen in beef cattle

Two experiments were conducted to compare pregnancy rates when GnRH or estradiol were given to synchronize ovarian follicular wave emergence and ovulation in an MGA-based estrus synchronization program. Crossbred beef cattle were fed melengestrol acetate (MGA, 0.5 mg per day) for 7 days (designated days 0-6, without regard to stage of the estrous cycle) and given cloprostenol (PGF; 500 microg intramuscular (im)) on day 7. In Experiment 1, lactating beef cows (n=140) and pubertal heifers (n=40) were randomly allocated to three groups to receive 100 microg gonadorelin (GnRH), 5 mg estradiol-17beta and 100 mg progesterone (E+P) in canola oil or no treatment (control) on day 0. All cattle were observed for estrus every 12 h from 36 to 96 h after PGF. Cattle in the GnRH group that were detected in estrus 36 or 48 h after PGF were inseminated 12 h later; the remainder were given 100 microg GnRH im 72 h after PGF and concurrently inseminated. Cattle in the E+P group were randomly assigned to receive either 0.5 or 1.0 mg estradiol benzoate (EB) in 2 ml canola oil im 24 h after PGF and were inseminated 30 h later. Cattle in the control group were inseminated 12 h after the first detection of estrus; if not in estrus by 72 h after PGF, they were given 100 microg GnRH im and concurrently inseminated. In the absence of significant differences, all data for heifers and for cows were combined and the 0.5 and 1.0 mg EB groups were combined into a single estradiol group. Estrus rates were 57.6, 57.4 and 60.0% for the GnRH, E+P and control groups, respectively (P=0.95). The mean (+/-S.D.) interval from PGF treatment to estrus was shorter (P<0.001) and less variable (P<0.001) in the E+P group (49.0+/-6.1 h) than in either the GnRH (64.2+/-15.9 h) or control (66.3+/-13.3 h) groups. Overall pregnancy rates were higher (P<0.005) in the GnRH (57.6%) and E+P (55.7%) groups than in the control group (30.0%) as were pregnancy rates to fixed-time AI (47.5, 55.7 and 28.3%, respectively). In Experiment 2, 122 crossbred beef heifers were given either 100 microg GnRH or 2 mg EB and 50 mg progesterone in oil on day 0 and subsequently received either 100 microg GnRH 36 h after PGF and inseminated 14 h later or 1 mg EB im 24 h after PGF and inseminated 28 h later in a 2 x 2 factorial design. Pregnancy rates were not significantly different among groups (41.9, 32.2, 33.3 and 36.7% in GnRH/GnRH, GnRH/EB, EB/GnRH and EB/EB groups, respectively). In conclusion, GnRH or estradiol given to synchronize ovarian follicular wave emergence and ovulation in an MGA-based synchronization regimen resulted in acceptable pregnancy rates to fixed-time insemination.     

Timing of insemination and fertility in dairy and beef cattle receiving timed artificial insemination using sex-sorted sperm

The objective was to evaluate the effects of timing of insemination and type of semen in cattle subjected to timed artificial insemination (TAI). In Experiment 1, 420 cyclic Jersey heifers were bred at either 54 or 60 h after P4-device removal, using either sex-sorted (2.1 × 10⁶ sperm/straw) or non-sorted sperm (20 × 10⁶ sperm/straw) from three sires (2 × 2 factorial design). There was an interaction (P = 0.06) between time of AI and type of semen on pregnancy per AI (P/AI, at 30 to 42 d after TAI); it was greater when sex-sorted sperm (P < 0.01) was used at 60 h (31.4%; 32/102) than at 54 h (16.2%; 17/105). In contrast, altering the timing of AI did not affect conception results with non-sorted sperm (54 h = 50.5%; 51/101 versus 60 h = 51.8%; 58/112; P = 0.95). There was an effect of sire (P < 0.01) on P/AI, but no interaction between sire and time of AI (P = 0.88). In Experiment 2, 389 suckled Bos indicus beef cows were enrolled in the same treatment groups used in Experiment 1. Sex-sorted sperm resulted in lower P/AI (41.8%; 82/196; P = 0.05) than non-sorted sperm (51.8%; 100/193). In addition, there was a tendency for greater P/AI (P = 0.11) when TAI was performed 60 h (50.8%; 99/195) versus 54 h (42.8%; 83/194) after removing the progestin implant. In Experiment 3, 339 suckled B. indicus cows were randomly assigned to receive TAI with sex-sorted sperm at 36, 48, or 60 h after P4 device removal. Ultrasonographic examinations were performed twice daily in all cows to confirm ovulation. On average, ovulation occured 71.8 ± 7.8 h after P4 removal, and greater P/AI was achieved when insemination was performed closer to ovulation. The P/AI was greatest (37.9%) for TAI performed between 0 and 12 h before ovulation, whereas P/AI was significantly less for TAI performed between 12.1 and 24 h (19.4%) or >24 h (5.8%) before ovulation. In conclusion, sex-sorted sperm resulted in a lesser P/AI than non-sorted sperm following TAI. However, improvements in P/AI with delayed time of AI were possible (Experiments 1 and 3), and seemed achievable when breeding at 60 h following progestin implant removal, compared to the standard 54 h normally used in TAI protocols.     

Ovarian follicular development in Holstein cows following synchronisation of oestrus with oestradiol benzoate and an intravaginal progesterone releasing insert for 5-9 days and duration of the oestrous cycle and concentrations of progesterone following ovulation

The aim of this study was to determine if the duration of treatment with an intravaginal progesterone releasing insert (IVP(4)) after treatment with oestradiol benzoate (ODB) at the time of insertion and 24 h after removal would affect selected variables including: size of ovarian follicles at the time of removal of inserts, diameter of ovulatory follicles, plasma concentrations of progesterone following ovulation, and duration of the following oestrous cycle. Characteristics of oestrus at a synchronised and spontaneous oestrus were also monitored. Non-lactating Holstein cows were synchronised with an IVP(4) for 5 (n = 10), 7 (n = 10), 8 (n = 9) or 9 (n = 9) days together with injections of ODB at device insertion (2 mg) and 24 h after removal (1 mg). Ultrasonography showed no significant effect of treatment on the day of emergence of preovulatory follicles relative to the day of removal of inserts (overall mean = -4.22 +/- 0.58; P = 0.15) for cows that ovulated within 120 h insert removal (n = 36). Treatment with ODB and an IVP(4) for 5 days reduced the diameter of preovulatory follicles at the time of removal of inserts and for the following 2 days compared to cows treated for 7-9 days (mean difference 2.56 +/- 1.15 mm; P = 0.033) but did not reduce the diameter of the ovulatory follicle (P = 0.21). Day of emergence relative to removal of inserts was associated with the diameter of the ovulatory follicle (R2 = 0.69; P < 0.001). Concentrations of progesterone and the diameter of the corpus luteum following ovulation were not affected by treatment (P > 0.20), but were affected by the diameter of the ovulatory follicle (P < 0.01). Diameter of the ovulatory follicle did not affect interoestrous and interovulatory intervals (P > 0.40). We conclude that treatment with an IVP(4) for 5 compared to 7-9 days with ODB administered at device insertion, and 24 h after removal reduced the diameter of preovulatory follicles at the time of removal of the insert but did not reduce the diameter of the ovulatory follicle or concentrations of progesterone in plasma. Emergence of preovulatory follicles closer to the time of removal of inserts reduced the diameter of the ovulatory follicle when oestrus was induced with ODB. Ovulation of smaller follicles reduced concentrations of progesterone in plasma following ovulation but did not affect oestrous cycle duration.     

Follicular growth, estrus and pregnancy after fixed-time insemination in beef cows treated with intravaginal progesterone inserts and estradiol benzoate

An experiment was performed to compare the effects of 3 short-term treatments with progesterone and estradiol benzoate (EB) on follicular growth, synchrony of estrus and pregnancy rate after fixed-time insemination in lactating postpartum beef cows. In Treatment 1 (n = 46), each cow received a progesterone-containing intravaginal insert for 7 d with injection of EB (2 mg, i.m.) at the time of device insertion. In Treatment 2 (n = 46), the insert was used for only 5 d with injection of EB (2 mg, i.m.) at the time of insertion. Cows in Treatment 3 (n = 47) received an insert for 5 d with no EB at the time of insertion. Each cow in the 3 groups received PGF2 alpha (25 mg, i.m.) at the time of insert removal, followed by EB (1 mg, i.m.) 30 h later. The cows were then inseminated 28 to 30 h after treatment with EB (58 to 60 h after insert removal). Treatment with 2 mg EB terminated the growth of the largest ovarian follicle (> 5 mm in diameter) at device insertion in 16/16 and 14/15 cows in Treatments 1 and 2, respectively. Estrus was detected within an 8-h target period (48 to 56 h after insert removal) in 93, 87 and 81% of cows in Treatments 1, 2 and 3, respectively (P > 0.05). Pregnancy rates at 39 d post insemination were 60, 50 and 51% for Treatments 1, 2 and 3, respectively (P > 0.05). The pregnancy rates did not differ between cows that were anovulatory or those that had ovulated before the initiation of treatments (54%), or among cows that were 28 to 40, 41 to 60 or > 60 days post partum at insemination (43, 59 and 54%, respectively). Treatment with progesterone inserts for 5 or 7 d, PGF2 alpha at the time of insert removal and 1 mg EB 30 h later induced the high degree of synchrony of estrus and ovulation necessary for fixed-time insemination.     

Synchronization of ovulation in beef cows (Bos indicus) using GnRH, PGF2alpha and estradiol benzoate

The objective of this study was to evaluate protocols for synchronizing ovulation in beef cattle. In Experiment 1, Nelore cows (Bos indicus) at random stages of the estrous cycle were assigned to 1 of the following treatments: Group GP controls (nonlactating, n=7) received GnRH agonist (Day 0) and PGF2alpha (Day 7); while Groups GPG (nonlactating, n=8) and GPG-L (lactating, n=9) cows were given GnRH (Day 0), PGF2alpha (Day 7) and GnRH again (Day 8, 30 h after PGF2alpha). A new follicular wave was observed 1.79+/-0.34 d after GnRH in 19/24 cows. After PGF2alpha, ovulation occurred in 19/24 cows (6/7 GP, 6/8 GPG, 7/9 GPG-L). Most cows (83.3%) exhibited a dominant follicle just before PGF2alpha, and 17/19 ovulatory follicles were from a new follicular wave. There was a more precise synchrony of ovulation (within 12 h) in cows that received a second dose of GnRH (GPG and GPG-L) than controls (GP, ovulation within 48 h; P<0.01). In Experiment 2, lactating Nelore cows with a visible corpus luteum (CL) by ultrasonography were allocated to 2 treatments: Group GPE (n=10) received GnRH agonist (Day 0), PGF2alpha (Day 7) and estradiol benzoate (EB; Day 8, 24 h after PGF2alpha); while Group EPE (n=11), received EB (Day 0), PGF2alpha (Day 9) and EB (Day 10, 24 h after PGF2alpha). Emergence of a new follicular wave was observed 1.6+/-0.31 d after GnRH (Group GPE). After EB injection (Day 8) ovulation was observed at 45.38+/-2.03 h in 7/10 cows within 12 h. In Group EPE the emergence of a new follicular wave was observed later (4.36+/-0.31 d) than in Group GEP (1.6+/-0.31 d; P<0.001). After the second EB injection (Day 10) ovulation was observed at 44.16+/-2.21 h within 12 (7/11 cows) or 18 h (8/11 cows). All 3 treatments were effective in synchronizing ovulation in beef cows. However, GPE and, particularly, EPE treatments offer a promising alternative to the GPG protocol in timed artificial insemination of beef cattle, due to the low cost of EB compared with GnRH agonists.     

The use of estradiol and/or GnRH in a two-dose PGF protocol for breeding management of beef heifers

The objective was to determine reproductive performance following AI in beef heifers given estradiol to synchronize ovarian follicular wave emergence and estradiol or GnRH to synchronize ovulation in a two-dose PGF-based protocol. In Experiment 1, 561 cycling (confirmed by ultrasonography), Angus heifers received 500 microg cloprostenol, i.m. (PGF) twice, 14 days apart (days 0 and 14) and were equally allocated to four groups in a 2 x 2 factorial design. On Day 7, heifers received either 2 mg estradiol benzoate (EB) and 50 mg progesterone (P), i.m. in oil (EBP group) or no treatment (NT group). Half the heifers in each group received 1mg EB, i.m. in oil on Day 15 (24h after the second PGF treatment) with TAI 28 h later (52 h after PGF), and the other half received 100 microg GnRH, i.m. on Day 17 (72 h after PGF) concurrent with TAI. All heifers were observed for estrus twice daily from days 13 to 17; those detected in estrus more than 16 h before scheduled TAI were inseminated 4-16 h later and considered nonpregnant to TAI. Overall pregnancy rate (approximately 35 days after AI) was higher in heifers that received EBP than those that did not (61.6% versus 48.2%, respectively; P < 0.002); but was lower in heifers that received EB after PGF than those that received GnRH (50.0% versus 59.8%; P < 0.02). Although estrus was detected prior to TAI in 77 of 279 heifers (27.6%) treated with EBP (presumably due to induced luteolysis), they were inseminated and 53.2% became pregnant. Overall pregnancy rates were 51.4, 68.3, 45.0, and 55.0% in the NT/GnRH, EBP/GnRH, NT/EB, and EBP/EB groups, respectively (P < 0.05). In Experiment 2, 401 cycling, Angus heifers were used. The design was identical to Experiment 1, except that 1.5mg estradiol-17beta (E-17beta) plus 50mg progesterone (E-17betaP) and 1mg E-17beta were used in lieu of EBP and EB, respectively. All heifers receiving E-17beta 24h after the second injection of PGF (NT/E-17beta and E-17betaP/E-17beta) were TAI 28 h later without estrus detection, i.e. 52 h after PGF. Heifers in the other two groups received 100 microg GnRH, i.m. 72 h after PGF and were concurrently TAI; heifers in these two groups that were detected in estrus prior to this time were inseminated 4-12h later and considered nonpregnant to TAI. Estrus rate during the first 72 h after the second PGF treatment was higher (P < 0.05) in the E-17betaP/GnRH group (45.0%; n = 100) than in the NT/GnRH group (16.0%; n = 100), but conception rate following estrus detection and AI was not different (mean, 57.4%; P = 0.50). Overall pregnancy rate was not significantly different among groups (mean, 46.9%; P = 0.32). In summary, the use of EB or E-17beta to synchronize follicular wave emergence and estradiol or GnRH to synchronize ovulation in a two-dose, PGF-based protocol resulted in acceptable fertility to TAI. However, when 2mg EB was used to synchronize follicular wave emergence, early estrus occurred in approximately 28% of heifers, necessitating additional estrus detection. A combination of estrus detection and timed-AI in a two-dose PGF protocol resulted in highly acceptable pregnancy rates.     

Effects of equine chorionic gonadotropin and type of ovulatory stimulus in a timed-AI protocol on reproductive responses in dairy cows

The objectives were to evaluate the effects of equine chorionic gonadotropin (eCG) supplementation (with or without eCG) and type of ovulatory stimulus (GnRH or ECP) on ovarian follicular dynamics, luteal function, and pregnancies per AI (P/AI) in Holstein cows receiving timed artificial insemination (TAI). On Day 0, 742 cows in a total of 782 breedings, received 2 mg of estradiol benzoate (EB) and one intravaginal progesterone (P4) insert (CIDR). On Day 8, the CIDR was removed, and all cows were given PGF2α and assigned to one of four treatments in a 2 × 2 factorial arrangement: (1) CG: GnRH 48 h later; (2) CE: ECP; (3) EG: eCG + GnRH 48 h later; (4) EE: eCG + ECP. There were significant interactions for eCG × ovulatory stimulus and eCG × BCS. Cows in the CG group were less likely (28.9% vs. 33.8%; P < 0.05) to become pregnant compared with those in the EG group (odds ratio [OR] = 0.28). There were no differences in P/AI between CE and EE cows (30.9% vs. 29.1%; OR = 0.85; P = 0.56), respectively. Thinner cows not receiving eCG had lower P/AI than thinner cows receiving eCG (15.2% vs. 38.0%; OR = 0.20; P < 0.01). Treatment with eCG tended to increase serum progestesterone concentrations during the diestrus following synchronized ovulation (P < 0.10). However, the treatment used to induce ovulation did not affect CL volume or serum progesterone concentrations. In conclusion, both ECP and GnRH yielded comparable P/AI. However, eCG treatment at CIDR removal increased pregnancy rate in cows induced to ovulate with GnRH and in cows with lower BCS.     

Pregnancy rates to timed artificial insemination in dairy cows treated with gonadotropin-releasing hormone or porcine luteinizing hormone

We compared the effects of porcine luteinizing hormone (pLH) versus gonadotropin-releasing hormone (GnRH) on ovulatory response and pregnancy rate after timed artificial insemination (TAI) in 605 lactating dairy cows. Cows (mean ± SEM: 2.4 ± 0.08 lactations, 109.0 ± 2.5 d in milk, and 2.8 ± 0.02 body condition score) at three locations were assigned to receive, in a 2 × 2 factorial design, either 100 μg GnRH or 25 mg pLH im on Day 0, 500 μg cloprostenol (PGF) on Day 7, and GnRH or pLH on Day 9, with TAI 14 to 18 h later. Ultrasonographic examinations were performed in a subset of cows on Days 0, 7, 10, and 11 to determine ovulations, presence of corpus luteum, and follicle diameter and in all cows 32 d after TAI for pregnancy determination. In 35 cows, plasma progesterone concentrations were determined 0, 3, 4, 5, 6, 7, and 12 d after ovulation. The proportion of noncyclic cows and cows with ovarian cysts on Day 0 were 12% and 6%, respectively. Ovulatory response to first treatment was 62% versus 44% for pLH and GnRH and 78% versus 50% for noncyclic and cyclic cows (P < 0.01). Location, ovulatory response to first pLH or GnRH, cyclic status, presence of an ovarian cyst, and preovulatory follicle size did not affect pregnancy rate. Plasma progesterone concentrations after TAI did not differ among treatments. Pregnancy rate to TAI was greater (P < 0.01) in the GnRH/PGF/pLH group (42%) than in the other three groups (28%, 30%, and 26% for GnRH/PGF/GnRH, pLH/PGF/GnRH, and pLH/PGF/pLH, respectively). Although only 3% of cows given pLH in lieu of GnRH on Day 9 lost their embryo versus 7% in those subjected to a conventional TAI using two GnRH treatments, the difference was not statistically significant. In summary, pLH treatment on Day 0 increased ovulatory response but not pregnancy rate. Cows treated with GnRH/PGF/pLH had the highest pregnancy rate to TAI, but progesterone concentrations after TAI were not increased. In addition, preovulatory follicle diameter did not affect pregnancy rate.     

Reduction in size of the ovulatory follicle reduces subsequent luteal size and pregnancy rate

We hypothesized that reducing the size of the ovulatory follicle using aspiration and GnRH would reduce the size of the resulting CL, reduce circulating progesterone concentrations, and alter conception rates. Lactating dairy cows (n=52) had synchronized ovulation and AI by treating with GnRH and PGF2alpha as follows: Day -9, GnRH (100 microg); Day -2, PGF2alpha (25 mg); Day 0, GnRH (100 microg); Day 1, AI. Treated cows (aspirated group; n=29) had all follicles > 4 mm in diameter aspirated on Days -5 or -6 in order to start a new follicular wave. Control cows (nonaspirated group: n=23) had no follicle aspiration. The size of follicles and CL were monitored by ultrasonography. The synchronized ovulation rate (ovulation rate to second GnRH injection: 42/52=80.8%) and double ovulation rate of synchronized cows (6/42=14.3%) did not differ (P > 0.05) between groups. Aspiration reduced the size of the ovulatory follicle (P < 0.0001; 11.5 +/- 0.2 vs 14.5 +/- 0.4 mm), and serum estradiol concentrations at second GnRH treatment (P < 0.0002; 2.5 +/- 0.4 vs 5.7 +/- 0.6 pg/mL). The volume of CL was less (P < 0.05) for aspirated than nonaspirated cows on Day 7 (2,862 +/- 228 vs 5,363 +/- 342 mm3) or Day 14 (4,652 +/- 283 vs 6,526 +/- 373 mm3). Similarly, serum progesterone concentrations were less on Day 7 (P < 0.05) and Day 14 (P < 0.10) for aspirated cows. Pregnancy rate per AI for synchronized cows was lower (P < 0.05) for aspirated (3/21=14.3%) than nonaspirated (10/21=47.6%) cows. In conclusion, ovulation of smaller follicles produced lowered fertility possibly because development of smaller CL decreased circulating progesterone concentrations.     

Induction of ovarian follicular wave emergence and ovulation in progestin-based timed artificial insemination protocols for Bos indicus cattle

The present study aimed to evaluate the efficacy of different inducers of new follicular wave emergence (FWE) and ovulation in fixed-time artificial insemination (FTAI) synchronization protocols using norgestomet ear implants (NORG) in Bos indicus cattle. In Experiment 1, the synchronization of FWE was evaluated when two different estradiol esters in different doses [2mg estradiol benzoate (EB), 2.5mg EV or 5mg estradiol valerate (EV)] were administered with NORG implant insertion in B. indicus cattle (estrous cyclic heifers and cows with suckling calves; n=10 per treatment). After estradiol treatment, ovarian ultrasonic exams were performed once daily to detect the interval between treatment and FWE. There were significant treatment-by-animal category interaction (P=0.05) on the interval from the estradiol treatment to FWE. An earlier (P<0.0001) and less variable (P=0.02) interval from estradiol treatment to FWE was observed in heifers treated with EB (2.5±0.2; mean±SE) than in those treated with 2.5mg EV (4.2±0.3) or 5mg EV (6.1±0.6). Cows treated with 5mg EV (4.0±0.5) had longer (P=0.05) interval than cows receiving EB (2.5±0.2), however, there was an intermediate interval in those cows treated with 2.5mg EV (3.1±0.4). In Experiment 2, the number of uses of the NORG implant (new; n=305 or previously used once; n=314) and three different ovulation induction hormones [0.5mg estradiol cypionate (EC) at implant removal (n=205), 1mg EB given 24h after implant removal (n=219), or 100μg gonadorelin (GnRH) given at FTAI (n=195)] were evaluated in Nelore heifers (2×3 factorial design). Similar pregnancy per AI (P/AI; 30 days after FTAI; P>0.05) were achieved using each of the three ovulation induction hormones (EB=40.6%; EC=48.3%, or GnRH=48.7%) and with a new (47.2%) or once-used NORG implant (44.3%). In Experiment 3, the effect of different ovulation induction hormones for FTAI [1mg EC at NORG implant removal (n=228), 10μg buserelin acetate at FTAI (GnRH; n=212) or both treatments (EC+GnRH; n=215)] on P/AI was evaluated in suckled beef cows treated with a once-used NORG implant and EB to synchronize the FWE. Similar P/AI (P=0.71) were obtained using GnRH (50.9%), EC (51.8%) or both treatments (54.9%) as ovulation induction hormones. Therefore, both doses of EV (2.5 or 5.0mg) with NORG implant delayed and increased the variation of the day of new FWE compared with EB in B. indicus cattle. These effects were more pronounced in B. indicus heifers than cows. Synchronization protocols for FTAI with either a new or once-used NORG implant with EB at insertion to induce a new FWE and either the use of EB, EC or GnRH as ovulation induction hormones may be successful in B. indicus heifers. Also, when a once-used NORG implant was used, either the administration of EC, GnRH or both as ovulation inducers resulted in similar P/AI in suckled B. indicus cows, showing no additive effect of the combination of both ovulation induction hormones.