Effect of day of the estrous cycle at the initiation of a timed artificial insemination protocol on reproductive responses in dairy heifers

Our objectives were to identify stages of the estrous cycle at which initiation of a timed artificial insemination (Ovsynch/TAI) protocol may reduce pregnancy rates and to monitor ovarian follicle dynamics and corpus luteum development after initiation of the Ovsynch/TAI protocol at different stages of the cycle. Cycling Holstein heifers (n = 24) were injected twice with prostaglandin F2alpha to induce estrus and were scanned by ovarian ultrasonography to determine the day of ovulation (d 0). Heifers were assigned to initiate the Ovsynch/TAI protocol at d 2 (n = 5), 5 (n = 5), 10 (n = 4), 15 (n = 5), or 18 (n = 5) of the cycle. The Ovsynch/TAI was initiated with an injection of gonadotropin-releasing hormone agonist followed 7 d later with an injection of prostaglandin F2alpha. At 36 h after injection of prostaglandin F2alpha, heifers were injected with gonadotropin-releasing hormone agonist and inseminated 16 h later. Heifers were scanned daily during the Ovsynch/TAI protocol and every other day after insemination until 16 d later. Blood samples were collected daily starting at the 1st day heifers were scanned and continued until 16 d after insemination. Initiation of the Ovsynch/TAI protocol at d 15 of the estrous cycle caused heifers to ovulate prior to insemination. A shortened return to estrus (< 16 d) was caused by ovulation failure to the second gonadotropin-releasing hormone injection, by incomplete regression of the corpus luteum, and by short life-span of the induced corpus luteum. Day of the cycle in which the Ovsynch/TAI protocol is initiated affects dynamics of follicular development, plasma progesterone profiles, and occurrence of premature ovulation. Size of the pre-ovulatory follicle was associated positively with subsequent progesterone concentrations following insemination.     

Strategies to optimize reproductive efficiency by regulation of ovarian function

Pregnancy rate to the Ovsynch protocol can be improved if cows are presynchronized (i.e., two PGF(2alpha) injections given 14 days apart and the second injection of PGF(2alpha) given 12 days prior to the first GnRH of the Ovsynch program) so that a greater proportion of cows during the Ovsynch protocol ovulate to the first GnRH injection and have a CL at PGF(2alpha) injection. Pregnancy rates were normal in anestrous cows (39.6%) if they ovulated to both injections of GnRH. Estradiol cypionate (ECP) can be used to replace GnRH to induce ovulation as a modification of the Presync-Ovsynch program (i.e., Presync-Heatsynch). Pregnancy rates after TI were 37.1+/-5.8% for Presync-Ovsynch compared to 35.1+5.0% for Presync-Heatsynch. Use of ECP to induce ovulation was an alternative to GnRH in which greater uterine tone, ease of insemination and occurrence of estrus, improved acceptance by inseminators. A GnRH agonist (Deslorelin; 750 microg) implant inserted at 48 h after injection of PGF(2alpha), as a component of the Ovsynch protocol, induced ovulation, development of a normal CL and delayed follicular growth until 24 d after implant insertion. Utilization of Deslorelin implants (450 microg and 750 microg) to induce ovulation compared to GnRH (100 microg) within the Ovsynch protocol resulted in 27 d pregnancy rates (GnRH 100 microg, 39%; Deslorelin implants 450 microg, 40% and 750 microg, 27.5%) with 12.7%, 5.0% and 9.5% embryonic losses by 41 d of pregnancy, respectively. Induction of an accessory CL with injection of hCG on day 5 after insemination improved conception rates by 7.1%. Bovine somatotrophin injected at first insemination following a Presync-Ovsynch program in cycling-lactating dairy cows increased 74 days pregnancy rates (57.1%>42.6%).     

Intravaginal progesterone devices in synchronization protocols for artificial insemination in beef heifers

Two experiments were designed to investigate the administration of intravaginal progesterone in protocols for oestrus and ovulation synchronization in beef heifers. In Experiment 1, cyclic Black Angus heifers (n = 20) received an Ovsynch protocol and were randomly assigned to receive (CIDR‐Ovsynch) or not (Ovsynch) a progesterone device between Days 0 and 7. Treatment with a controlled internal drug release (CIDR) device significantly increased the size of the dominant follicle prior to ovulation (12.8 ± 0.4 CIDR‐Ovsynch vs 11.4 ± 0.4 Ovsynch) (p < 0.02). Plasma progesterone concentrations throughout the experiment were affected by the interaction between group and day effects (p < 0.004). In Experiment 2, cyclic Polled Hereford heifers (n = 382) were randomly assigned to one of the six treatment groups (3 × 2 factorial design) to receive a CIDR, a used bovine intravaginal device (DIB), or a medroxiprogesterone acetate (MAP) sponge and GnRH analogues (lecirelin or buserelin). All heifers received oestradiol benzoate plus one of the devices on Day 0 and PGF on Day 7 pm (device withdrawal). Heifers were detected in oestrus 36 h after PGF and inseminated 8–12 h later, while the remainder received GnRH 48 h after PGF and were inseminated on Day 10 (60 h). The number of heifers detected in oestrus on Day 8 and conception rate to AI on Day 9 were higher (p < 0.01) in the used‐DIB than in the CIDR or MAP groups, while the opposite occurred with the pregnancy rate to FTAI on Day 10 (p < 0.01). There was no effect of progesterone source, GnRH analogue or their interaction on overall pregnancy rates (64.9%). Progesterone treatment of heifers during an Ovsynch protocol resulted in a larger pre‐ovulatory follicle in beef heifers. Progesterone content of intravaginal devices in synchronization protocols is important for the timing of AI, as the use of low‐progesterone devices can shorten the interval to oestrus.     

Regulation of pulsatile LH secretion by ovarian steroids in the heifer

Two experiments were conducted to evaluate relationships among luteinizing hormone (LH), estradiol-17 beta (E2) and progesterone secretion during the preovulatory period in the heifer after prostaglandin F2 alpha (PGF2 alpha)-induced regression of the corpus luteum. A second objective was to elucidate the effects of E2 in regulating LH secretion. In Exp. 1, LH, E2 and progesterone concentrations were determined in serial samples collected during the preovulatory period after PGF2 alpha-induced luteal regression in five Red Angus X Hereford heifers. Progesterone declined to 1 ng/ml by 12 h after the second injection of PGF2 alpha. Frequency of LH pulses increased linearly (P less than .01), whereas no change in amplitude of LH pulses was detected before the preovulatory LH surge. This resulted in a linear increase (P less than .01) in mean LH concentrations. Estradiol also increased in a linear manner (P less than .01), and the rise in E2 was parallel to the increase in mean LH concentrations. In Exp. 2, 12 Angus X Hereford heifers were ovariectomized and administered either 13.5- or 27-cm silastic implants containing E2 at ovariectomy. Four heifers served as nonimplanted controls. Thirty-one days after ovariectomy all heifers were bled at 12-min intervals for 6 h. Frequency of LH pulses declined linearly (P less than .03) while mean LH (P less than .09) and pulse amplitude (P less than .01) increased linearly as E2 dose increased. These results indicate that a reduction in progesterone increases the frequency of LH pulses during the follicular phase of the estrous cycle in cattle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of inducing luteal regression before a modified controlled internal drug-releasing device treatment on control of follicular development

At the initiation of most controlled internal drug-releasing (CIDR) device protocols, GnRH has been used to induce ovulation and reset follicular waves; however, its ability to initiate a new follicular wave is variable and dependent on stage of the estrous cycle. The objectives of the current studies were to determine 1) if inducing luteal regression before the injection of GnRH at time of insertion of a CIDR resulted in increased control of follicular development, and 2) if removing endogenous progesterone by inducing luteal regression before insertion of the CIDR decreased variation in LH pulse frequency. In Exp. 1 and 2, Angus-cross cycling beef heifers (n = 22 and 38, respectively) were allotted to 1 of 2 treatments: 1) heifers received an injection of PGF(2α) on d -3, an injection of GnRH and insertion of a CIDR on d 0, and a PGF(2α) injection and CIDR removal on d 6 (PG-CIDR) or 2) an injection of GnRH and insertion of a CIDR on d 0 and on d 7 an injection of PGF(2α) and removal of CIDR (Select Synch + CIDR). In Exp. 3, Angus-cross beef heifers (n = 15) were assigned to 1 of 3 treatments: 1) PG-CIDR; 2) PGF(2α) on d -3, GnRH on d 0, and PGF(2α) on d 6 (PG-No CIDR); or 3) Select Synch + CIDR. Follicular development and ovulatory response were determined by transrectal ultrasonography. Across all experiments, more (P = 0.02) heifers treated with PG before GnRH initiated a new follicular wave after the injection of GnRH compared with Select Synch + CIDR-treated heifers. In Exp. 1, after CIDR removal, interval to estrus did not differ (P = 0.18) between treatments; however, the variance for the interval to estrus was reduced (P < 0.01) in PG-CIDR heifers compared with Select Synch + CIDR heifers. In Exp. 3, there was a tendency (P = 0.09) for LH pulse frequency to be greater among PG-CIDR and PG-No CIDR compared with the Select Synch + CIDR, but area under the curve, mean LH concentrations, and mean amplitude did not differ (P > 0.76). In summary, induction of luteal regression before an injection of GnRH increased the percentage of heifers initiating a new follicular wave. Removal of endogenous progesterone tended to increase LH pulse frequency, and the modified treatment increased the synchrony of estrus after CIDR removal.     

Transrectal ultrasonic diagnosis of ovarian follicular cysts in goats and treatment with GnRH

Cystic ovarian disease is an important cause of reproductive failure. The objective of this study was to evaluate transrectal ultrasonography as a diagnostic tool and gonadotropin-releasing hormone (GnRH) as a therapeutic approach for ovarian follicular cysts in goats. Goats were considered to have a follicular cyst(s) if a non-echoic structure >10 mm in diameter was detected in the absence of corpora lutea (CL) in three ultrasonic examinations performed at 5-day intervals. After diagnosis (Day 0), goats with ovarian follicular cysts (n = 5) were treated with a single bolus injection of 10.5 microg synthetic GnRH followed by administration of 125 microg prostaglandin F2alpha (PGF2alpha) 10 days later. Five blood samples were collected at 5-day intervals for determination of progesterone and estradiol-17beta. For detection of LH surge, blood samples were collected every 2 h. Ovulation rate was determined and pregnancy was confirmed by transrectal ultrasonography. The results showed that transrectal ultrasonography is reliable for diagnosis of ovarian follicular cysts and the mean diameter of the follicular cysts was 12.6 +/- 0.4 mm. Plasma concentrations of progesterone and estradiol-17beta at the time of diagnosis of follicular cysts (Day 0) were 0.7 +/- 0.2 ng/ml and 12.7 +/- 0.9 pg/ml, respectively. The concentration of progesterone increased to 4.0 +/- 0.5 ng/ml 10 days after administration of GnRH indicating luteinization of the ovarian follicular cysts concomitant with a decrease in the concentration of estradiol-17beta (3.5 +/- 0.4 pg/ml). Administration of GnRH to cystic goats resulted in a surge of LH within 2 h of treatment. The interval from PGF2alpha injection to the preovulatory LH surge was 62.8 +/- 1.4 h. All goats exhibited estrus 55.2 +/- 2.3 h after PGF2alpha injection and four goats out of the five ovulated. The ovulation rate was 1.5 +/- 0.3. In conclusion, results of this study suggest that transrectal ultrasonography is a reliable tool for diagnosis of ovarian follicular cysts. In addition, GnRH can be used to effectively treat ovarian follicular cysts in goats with 80% success rate.     

Comparison of three approaches for synchronization of ovulation for timed artificial insemination in Bos indicus-influenced cattle managed on the Texas gulf coast

Our objectives were to compare the relative efficacies of three protocols designed to synchronize ovulation for timed artificial insemination (AI) of predominantly Brahman-influenced cows and heifers. In Exp. 1, 273 Brahman x Hereford (F1) cows at three locations were stratified by BW, body condition score (BCS), age, and days postpartum and assigned randomly to three treatments: 1) Syncro-Mate-B (SMB), 2) norgestomet-prostaglandin (NP), and 3) Ovsynch. The management goal required that cows have a minimum BCS of 5 and be at least 36 d postpartum (PP) at treatment onset. However, final results included 23 cows (8.4%) whose BCS fell below 5. In Exp. 2, 286 pubertal beef heifers were stratified by BW and BCS and allocated randomly to the three treatments. Heifers were predominantly Brahman crossbred (n = 265; Brahman x Hereford, F1; Santa Cruz) or purebred Brahman-influenced (Santa Gertrudis) with a smaller number (n = 21) of Hereford heifers also included. For both experiments, SMB treatment consisted of a 9-d norgestomet ear implant plus an estradiol valerate/norgestomet injection on d 0. Norgestomet-prostaglandin-treated females were implanted with a SMB implant without the estradiol valerate/norgestomet injection at the time of implant insertion and received 25 mg prostaglandin F2alpha (PGF) i.m. 2 d before implant removal. Ovsynch consisted of 100 microg GnRH i.m. on d 1, 25 mg PGF i.m. on d 8, and a second GnRH injection on d 10. Beginning on d 9, calves were removed for 48 h in Exp. 1. Cattle in SMB and NP groups in both experiments were timed-inseminated 48 to 54 h after implant removal and at 12 to 24 h after the second GnRH injection (Ovsynch). Timed AI conception rates did not differ between the SMB (45.1%) and Ovsynch (42.4%) groups; however, conception rate in the NP group tended (P < 0.12) to be lower overall than in the other groups due to a reduced (P < 0.05) conception rate in cows that were < 60 d PP at treatment onset. Conversely, timed-AI conception was greatest (P < 0.056) in NP (54.7%) compared with SMB (40.4%) and Ovsynch (39.1%) for heifers in Exp. 2. We conclude that in mature, suckled beef cows with Brahman genetic influence, SMB and Ovsynch perform similarly when cow eligibility relies primarily on BCS and minimum days PP. The NP treatment results in lower conception in cows < 60 d PP compared with SMB and Ovsynch. However, in nulliparous Brahman-influenced heifers that are confirmed to be pubertal, NP may be superior to the other two treatments for timed AI.     

Improved synchrony of estrus and ovulation with the addition of GnRH to a melengestrol acetate-prostaglandin F2alpha synchronization treatment in beef heifers

The objective of this experiment was to determine the effect of a GnRH injection within a melengestrol acetate (MGA)-PGF2alpha (PGF) estrus synchronization protocol on follicular dynamics and synchronization of estrus. Pubertal crossbred beef heifers (n = 34) were randomly assigned to one of two treatments. Both treatment groups were fed MGA (0.5 mg x hd(-1) x d(-1)) for 14 d and injected (i.m.) with PGF (25 mg of Lutalyse) 19 d after MGA withdrawal. Melengestrol acetate was delivered in a feed supplement of 1.8 kg x hd(-1) x d(-1). Seventeen heifers received an injection of GnRH (100 microg Cystorelin) 12 d after MGA withdrawal and 7 d before PGF. The control group (n = 17) received only MGA-PGF. Estrus was detected four times/d for 7 d beginning on the day PGF was injected. Transrectal ultrasonography was performed daily on eight heifers from each treatment to monitor ovarian activity and characterize changes in follicular dynamics after MGA withdrawal and until ovulation after PGF. Each of the GnRH-treated heifers either ovulated or had a luteinized dominant follicle following GnRH and subsequently initiated a new follicular wave (8/8, 100%). All GnRH-treated heifers (17/17, 100%) and 94% of controls (16/17) exhibited estrus after PGF. Estrus was exhibited over a 132-h period (12 to 144 h) for control heifers compared with 60 h (48 to 108 h) for GnRH-treated heifers. The peak synchronized period for both treatments was between 48 and 72 h after PGF, during which time 76% (13/17) of the GnRH-treated heifers exhibited estrus compared with 63% (10/16) for controls. Seventy-one percent (12/17) of the GnRH-treated heifers exhibited estrus from 48 to 60 h after PGF, compared with 38% (6/16) for controls (P < 0.05). In summary, injection of GnRH within a 14- to 19-d MGA-PGF protocol increased the synchrony of estrus during the synchronized period and concentrated the period of detected estrus. This protocol may offer potential for the fixed-time insemination of replacement beef heifers.     

Inhibitory effects of CIDR-based ovulation-synchronization protocols on uterine PGF2alpha secretion at the following luteal phase in early postpartum non-cycling beef cows

We investigated whether CIDR-based ovulation-synchronization protocols inhibit secretion of prostaglandin (PG) F2alpha from the uterus in the following luteal phase in non-cycling beef cows. Ten early (a month) postpartum non-cycling Japanese Black beef cows were treated with (1) Ovsynch (GnRH analogue on Day 0, PGF2alpha analogue on Day 7, and GnRH analogue on Day 9; n=3), (2) Ovsynch+CIDR (Ovsynch protocol plus a CIDR for 7 days from Day 0; n=4), or (3) estradiol benzoate (EB) Ovsynch+CIDR (EB on Day 0 in lieu of the first GnRH treatment followed by the Ovsynch+CIDR protocol; n=3). An oxytocin challenge was administered on Day 24 to examine uterine PGF2alpha secretion. Plasma concentrations of 13,14-dihydro-15-keto- PGF2alpha were lower at 30-120 min after oxytocin administration in the Ovsynch+CIDR group and 75 min after administration in the EB Ovsynch+CIDR group than in the Ovsynch group (P<0.05). Plasma progesterone concentrations were higher from Days 1 to 7 in the Ovsynch+CIDR group and from Days 1 to 5 in the EB Ovsynch+CIDR group than in the Ovsynch group (P<0.05). The progesterone concentrations were higher on Days 27 and 29 in both CIDR-treated groups than in the Ovsynch group (P<0.05). In conclusion, in non-cycling beef cows, CIDR-based ovulation-synchronization protocols inhibit uterine PGF2alpha secretion in the following luteal phase and prevent premature luteolysis as is seen with the Ovsynch protocol.     

Effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given GnRH

The effects of plasma progesterone concentrations on LH release and ovulation in beef cattle given 100 microg of GnRH im were determined in three experiments. In Experiment 1, heifers were given GnRH 3, 6 or 9 days after ovulation; 8/9, 5/9 and 2/9 ovulated (P<0.02). Mean plasma concentrations of progesterone were lowest (P<0.01) and of LH were highest (P<0.03) in heifers treated 3 days after ovulation. In Experiment 2, heifers received no treatment (Control) or one or two previously used CIDR inserts (Low-P4 and High-P4 groups, respectively) on Day 4 (estrus=Day 0). On Day 5, the Low-P4 group received prostaglandin F(2alpha) (PGF) twice, 12 h apart and on Day 6, all heifers received GnRH. Compared to heifers in the Control and Low-P4 groups, heifers in the High-P4 group had higher (P<0.01) plasma progesterone concentrations on Day 6 (3.0+/-0.3, 3.0+/-0.3 and 5.7+/-0.4 ng/ml, respectively; mean+/-S.E.M.) and a lower (P<0.01) incidence of GnRH-induced ovulation (10/10, 9/10 and 3/10). In Experiment 3, 4-6 days after ovulation, 20 beef heifers and 20 suckled beef cows were given a once-used CIDR, the two largest follicles were ablated, and the cattle were allocated to receive either PGF (repeated 12h later) or no additional treatment (Low-P4 and High-P4, respectively). All cattle received GnRH 6-8 days after follicular ablation. There was no difference between heifers and cows for ovulatory response (77.7 and 78.9%, P<0.9) or the GnRH-induced LH surge (P<0.3). However, the Low-P4 group had a higher (P<0.01) ovulatory response (94.7% versus 61.1%) and a greater LH surge of longer duration (P<0.001). In conclusion, although high plasma progesterone concentrations reduced both GnRH-induced increases in plasma LH concentrations and ovulatory responses in beef cattle, the hypothesis that heifers were more sensitive than cows to the suppressive effects of progesterone was not supported.     

JSAR Innovative Technology Award. Development of ovulation synchronization and fixed time artificial insemination in dairy cows

Recently, reproductive management has become more difficult as a result of increased herd size. Problems with missing estrous signs and decrease in conception rate by artificial insemination (AI) performed at wrong timing have caused low AI conception rates. In 1995, ovulation synchronization and fixed-time AI (Ovsynch/TAI) was developed in the USA as a new reproductive technology, which was accepted as an useful reproductive management tool in many countries. However, no information on the use of Ovsynch/TAI was available in Japan. It was, therefore, warranted to show the ovulation rate and conception rate after Ovsynch/TAI using gonadotropin releasing hormone analogue (GnRH-A, fertirelin acetate) and prostaglandin F2alpha (PGF2alpha)-THAM, both were commercially available in this country. The conception rate after Ovsynch/TAI has been known to vary among different herds and individuals. Investigation and analysis of factors affecting the conception rate was also warranted to improve the conception rate. A series of experiments were carried out to establish Ovsynch/TAI using domestically produced GnRH-A and PGF2alpha and to study factors affecting conception rate after Ovsynch protocol. Ovsynch using 100 microg GnRH-A and 25 mg PGF2alpha were observed using ultrasonography. As a result, a high synchronization rate of ovulation at 16 to 20 h after the second GnRH injection was confirmed. The conception rate after Ovsynch/TAI was compared in 87 cows with the conception rate after AI at estrus induced by PGF2alpha (139 cows). Conception rate after Ovsynch/TAI was higher than the figure after AI at induced estrus (59.1% vs 20.9%, P<0.05). The dose of GnRH-A was also studied and a practical dose of GnRH-A was found to be 50 microg per cow. To clarify some factors affecting the conception rate after Ovsynch/TAI, 1,558 cows were investigated for the state of their ovaries, days after calving, parity, season, ovarian cyclicity postpartum and nutritional state at the day of Ovsynch. The overall conception rate after Ovsynch/TAI was 51.5%. Fifty-six cows (3.6%) showed estrus at 6 to 7 d after the first injection of GnRH-A. The conception rate after Ovsynch/TAI was low in cows that were 40 to 60 d postpartum, those in their 5th lactation or more, those bred in July to August, and those recovering ovarian cyclicity later than 56 d postpartum. The conception rate after Ovsynch/TAI was high in cows in which body condition score (BCS) was 3.75 at dry period and 3.0 at the day of Ovsynch. In conclusion, Ovsynch/TAI is an effective tool for the reproductive management of dairy cows. A steady and sufficient conception rate after Ovsynch/TAI could be expected by taking the factors affecting the conception rate into the consideration.     

Ovsynch plus CIDR protocol for timed embryo transfer in suckled postpartum Japanese Black beef cows

We compared synchronization and pregnancy rates, and the increase in blood progesterone concentrations during luteal development, between (1) Ovsynch plus an intravaginal controlled internal drug release (CIDR) device protocol followed by timed embryo transfer (timed ET), and (2) a conventional estrus synchronization method using PGF(2 alpha) and ET in suckled postpartum Japanese Black beef cows. Cows in the PGF group (n=18) received a PGF(2 alpha) analogue when a CL was first palpated per rectum at 10-d intervals after 1 to 2 month postpartum. Cows (n=11), which showed estrus (Day 0) within 5 d of the PGF(2 alpha), and had a CL on Day 7, received ET. Cows in the Ovsynch+CIDR group (n=19) underwent the Ovsynch protocol plus a CIDR for 7 d (GnRH analogue and CIDR on Day-9, PGF(2alpha) analogue with CIDR removal on Day-2, and GnRH analogue on Day 0), with ET on Day 7. The ovulation synchronization (100%) and embryo transfer (100%) rates in the Ovsynch+CIDR group were greater (P<0.01) than the estrus synchronization (66.7%) and the embryo transfer (61.1%) rates in the PGF group. The postpartum interval at ET in the Ovsynch+CIDR group (62.5 +/- 2.5 d) was shorter (P<0.01) than in the PGF group (74.9 +/- 3.9 d). The pregnancy rate in the Ovsynch+CIDR group (57.9%) did not differ significantly from that in the PGF group (50.0%). Plasma progesterone concentrations were not significantly different in the two groups on Days 0, 1, 2, 5, 7, 14 and 21. In summary, higher synchronization and transfer rates, and shorter postpartum interval to ET, can be achieved with timed ET following the Ovsynch plus CIDR protocol than after estrus with the single PGF(2 alpha) treatment followed by ET in suckled postpartum recipient beef cows. Pregnancy rates were similar. Also, the increase in blood progesterone concentrations during luteal development following ovulation synchronized by the Ovsynch plus CIDR protocol was similar to that after estrus induced by the PGF(2 alpha) treatment.     

Preventive effects of CIDR-based protocols on premature ovulation before timed-AI in Ovsynch in cycling beef cows

Ovsynch is a program developed to synchronize ovulation for timed breeding. In this paper, the authors investigate whether controlled internal drug release (CIDR)-based protocols prevent premature ovulation before timed-artificial insemination (AI) when Ovsynch is started a few days before luteolysis in cycling beef cows. Nine beef cows at 16 days after oestrus were treated with (1) Ovsynch, i.e. gonadotropin releasing hormone (GnRH) analogue on day 0, prostaglandin (PG) F(2alpha) analogue on day 7 and GnRH analogue on day 9 with timed-AI on day 10, (n=3); (2) Ovsynch+CIDR (Ovsynch protocol plus a CIDR for 7 days from day 0, n=3), or (3) oestradiol benzoate (OB)+CIDR+GnRH (OB on day 0 in lieu of the first GnRH treatment, followed by the Ovsynch+CIDR protocol, n=3). In the Ovsynch group (1) plasma progesterone concentrations fell below 0.5 ng/mL earlier (day 5) than in both CIDR-treated groups (2) and (3), where this occurred on day 8. Plasma oestradiol-17beta concentrations peaked on day 8 in the Ovsynch group and on day 9 in both CIDR-treated groups. The dominant follicle ovulated on day 10 in the Ovsynch group and on day 11 in both CIDR-treated groups. Thus, both CIDR-based protocols prevented premature ovulation before timed-AI in Ovsynch when the protocol was started a few days before luteolysis. This reflects the fact that progesterone levels remained high until the beef cattle were treated with PGF(2alpha).     

Melengestrol acetate blocks the preovulatory surge of luteinizing hormone,the expression of behavioral estrus,and ovulation in beef heifers

We tested the hypothesis that melengestrol acetate (MGA), an orally active progestin, blocks estrus and the preovulatory surge of luteinizing hormone (LH) in beef heifers. Cycling yearling Angus heifers were divided randomly into two groups: MGA-treated (n = 6) and control (n = 5). All heifers received injections of prostaglandin F2alpha (PGF) on d -25, -11, and 0 to synchronize estrus. Following the last PGF injection on d 0, heifers were fed either 0.5 mg MGA in a carrier or the MGA carrier each day for 8 d. At 4-h intervals on d 1 through 6, all heifers were observed for expression of estrous behavior, and blood samples were collected and assayed for LH. Daily blood samples were collected at 0800 on d 1 through 10 and assayed for circulating progesterone concentrations. All control heifers exhibited estrus and a preovulatory surge of LH. In each case, this was followed by increases in circulating concentrations of progesterone indicative of ovulation and normal luteal function. In contrast, none of the MGA-treated heifers exhibited estrus, LH surges, or evidence of ovulation. The results of this experiment show that MGA prevents ovulation in cattle by inhibiting the preovulatory surge of LH.     

Induction of ovulation with GnRH and PGF2alpha in lactating Bos taurus x Bos indicus cows

Induction of ovulation for timed artificial insemination (TAI) with the Ovsynch protocol was evaluated in 49 anoestrous and lactating Bos taurus x Bos indicus cows. Palpation per rectum and transrectal ultrasonography were used on Days -30, -20, -10 and 0 (start of treatment) to confirm anoestrus but with the presence of follicles > or = 10 mm, and every other day during treatment to determine ovarian activity. Cows were randomly assigned to: (1) Ovsynch (n = 24; Day 0, 200 microg GnRH; Day 7, 150 microg PGF2alpha; Day 9, 200 microg GnRH + TAI 16 to 20 h later) and (2) control (n = 25; no treatment). Rates of ovulation for the first GnRH injection, detection of a corpus luteum (CL) at PGF2alpha injection, pregnancy and induction of cyclicity were greater (P < 0.05) with Ovsynch. There was no effect of body condition score (P > 0.05). In conclusion, the Ovsynch protocol was not effective in obtaining acceptable pregnancy rate for TAI, but it was effective for induction of cyclicity in anoestrous and lactating Bos taurus x Bos indicus cows under tropical conditions.     

Ovarian responses to intramuscular or intravenous administration of prostaglandin F2 alpha in control and FSH-treated beef heifers

Sixty Simmental crossbred heifers 18 to 20 mo of age were detected in estrus and assigned at random to a 2 x 2 factorial design study with 30 controls, and 30 given FSH. Half of each group was given prostaglandin F2 alpha (PGF) i.v. and the rest i.m. Injections of follicle-stimulating hormone were started on d 7 to 14 of an estrous cycle and continued for 5 d or until ovariectomy; PGF was administered either i.v. or i.m. at 48 (25 mg) and 60 (15 mg) h after the initial FSH injection. Control females received a similar PGF treatment on a day between d 9 and 15 of the estrous cycle. Blood samples were collected from all animals immediately before PGF administration and every 12 h thereafter until ovariectomy. Within each of the four experimental subgroups, ovariectomies were performed at either 24, 48 or 72 h (five/time group) after initial PGF injection. Ovarian and corpus luteum (CL) weights were recorded as well as number and size of follicles and number of ovulations. Regression of the CL was slower (P less than .05) after administration of PGF i.v. than i.m. (CL weight was 2.6 vs 3.3 +/- .2 g for i.m. and i.v. groups, respectively). Exogenous FSH increased estradiol-17 beta (E2) concentrations, and FSH-treated heifers had more (P less than .05) early ovulations than control heifers did. Ovulations in FSH-treated heifers had begun to occur by 24 h after i.v. and i.m. PGF injection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characteristics of estrus before and after first insemination and fertility of heifers after synchronized estrus using GnRH,PGF2alpha,and progesterone

Our objectives were to determine fertility of heifers after synchronization of estrus using PGF2alpha, preceded by progesterone (P4), GnRH, or both, and to examine the variability of estrual characteristics in heifers before first and second AI. Dairy (n = 247) and beef (n = 193) heifers were assigned randomly to each of three treatments: 1) 50 microg of GnRH (injected i.m.) administered on d -7 followed by 25 mg of PGF2alpha (i.m.) on d -1 (GnRH + PGF; modified Select Synch protocol); 2) placement of an intravaginal progesterone (P4)-releasing insert on d -7, PGF2alpha on d -1, and insert removal on d 0 (P4+PGF); and 3) 50 microg of GnRH plus a P4 insert on d -7, followed by 25 mg of PGF2alpha on d -1, and insert removal on d 0 (P4+GnRH+PGF). Characteristics of estrus were examined before first AI and before the next eligible AI (18 to 26 d later), including duration of estrus, number of standing events, and total and individual duration of standing events. In addition, all heifers were checked visually at least twice daily for estrus. Blood samples were collected on d -7, -1, and 0 for determination of P4, and pregnancy status was diagnosed by ultrasonography 27 to 34 d after AI. Rates of detected estrus were less (P < 0.05) in dairy than in beef heifers, and greater (P < 0.05) in heifers treated with P4. Pattern of conception and pregnancy rates among treatments differed between beef and dairy heifers (treatment x group interaction; P < 0.05). In dairy heifers, conception and pregnancy rates were greatest with P4+PGF, followed by P4+GnRH+PGF and GnRH+PGF, respectively. The opposite was observed among treatments in beef heifers. Administration of P4 without the preceding injection of GnRH produced the lowest pregnancy rates in beefheifers. Ofthe quantified sexual behavioral characteristics during the synchronized estrus, the number of standing events and total duration of standing events were greater (P < 0.01) than those observed during the next eligible estrus before second AI, whereas duration of estrus was unaffected.     

Preovulatory changes in follicular prostaglandins and their role in ovulation in cattle.

Indomethacin (INDO, n = 5) or vehicle (CONTROL, n = 4) was injected into superovulated heifers at 48 and 60 h following a luteolytic cloprostenol injection (0 h). One heifer from each group was ovariectomized (OVX) at 48, 56, 64 and 72 h. The fifth heifer of the INDO group was OVX at 80 h. Blood samples were collected at 0 h, every 2 h between 37 and 47 h, and at the time of each OVX to monitor plasma progesterone (P4) and luteinizing hormone (LH) concentrations. Following each OVX, the number and size of follicles were recorded and the incidence of ovulation determined. Follicular fluid (FF) was aspirated from follicles greater than or equal to 8 mm to determine the concentration of prostaglandins E2 (PGE2) and F2 alpha (PGF2 alpha). The highest PG concentrations were measured in both groups at 24-25 h following the preovulatory LH surge and the PGF2 alpha concentration at this time was significantly greater (p less than 0.01) in the CONTROL group compared to the INDO group. By 35-36 h after the LH surge, 75% (25/34) of the CONTROL follicles had ovulated, whereas there were no ovulations (0/50) on either ovary of the INDO treated heifer. These preliminary results suggest that the preovulatory rise of PGs in FF, particularly PGF2 alpha, is essential for ovulation and that suppression of this rise with indomethacin will inhibit ovulation in heifers.     

Development of an estrus synchronization protocol for beef cattle with short-term feeding of melengestrol acetate: 7-11 synch

An estrus synchronization protocol (7-11 Synch) was developed to synchronize the first follicular wave and timing of ovulation in postpartum beef cows. In Exp. 1, follicular development and timing of ovulation in response to the following protocol were evaluated. Beef heifers (n = 12) and cows (n = 6), at random stages of the estrous cycle, were fed melengestrol acetate (MGA; .5 mg x animal(-1) x d(-1)) for 7 d and injected with PGF2alpha (PG; 25 mg) on the last day of MGA. A second injection of PG was administered 11 d after cessation of MGA. After the second injection of PG, estrus was synchronized in 6/12 heifers and 3/6 cows. The interval to estrus in heifers and cows was 54 and 64 h, respectively (P > .10). All animals exhibiting estrus ovulated first-wave follicles. Animals that failed to respond to the second injection of PG were in estrus later than 6 d after cessation of MGA and had corpora lutea that were unresponsive to the injection of PG. Based on the variation in interval to estrus following the first PG injection on the last day of MGA feeding in Exp. 1, an injection of GnRH (100 microg) was added to the protocol 4 d after the cessation of MGA to ensure ovulation or luteinization of dominant follicles and synchronization of first-wave follicular development. This revised protocol was termed "7-11 Synch." In Exp. 2, two estrus synchronization protocols were compared. Multiparous beef cows were stratified by breed and postpartum interval and randomly assigned to the 7-11 Synch (n = 44) or Select Synch protocols (GnRH injection followed by PG injection 7 d later; n = 45). Timing of estrus after the last PG injection (0 h) ranged from 42 to 102 h in the 7-11 Synch group and -30 to 114 h in the Select Synch group. Eight cows (18%) in the Select Synch group exhibited estrus 30 h before to 18 h after PG. Synchronized estrus peaked between 42 and 66 h after the last PG injection, and a maximum number of cows were in estrus at 54 h for both treatment groups. Synchrony of estrus from 42 to 66 h was greater (P < .05) in 7-11 Synch (91%: 41/44) than in Select Synch cows (69%: 31/45). Artificial insemination pregnancy rate from 42 to 66 h was greater (P < .05) in the 7-11 Synch group (66%: 29/44) than in the Select Synch group (40%: 18/45). In summary, the 7-11 Synch protocol improved synchrony of estrus without reducing fertility. This protocol has potential future application for fixed-time AI in beef cattle production systems.     

Effect of exogenous progesterone administration on luteal sensitivity to PGF during the early development of the corpus luteum in mares and cows

The objective of this study was to determine the effect of exogenous progesterone administration at ovulation and during the early development of the CL, on its future sensitivity to a single administration of PGF2a in mares and cows. Horse Retrospective reproductive data from an equine clinic in the UK during three breeding seasons were used. Mares were divided into: control group, cycles with single ovulations; double ovulation group cycles with asynchronous double ovulations; and PRID group: cycles with single ovulations and treatment with intravaginal progesterone device (CIDR) immediately after the ovulation. All mares were treated with d‐cloprostenol (PGF) at either: (i) 88 hr; (ii) 96 hr; (iii) 104 hr; or (iv) 112 hr after the last ovulation. Cattle A total of nine non‐lactating Holstein cows were used. All cows were administered PGF14 d apart and allocated to one of two groups control group GnRH was administered 56 hr after the second PGF administration. CIDR group CIDR was inserted at the same time of GnRH administration. All cows were administered PGF at 120 hr post‐ovulation. The complete luteolysis rate of mares with double ovulation (66.7%) and those treated with exogenous progesterone (68.4%) was significantly higher than the rate of mares with single ovulation (35.6%) at 104 hr. In the cow, however, the treatment with CIDR did not increase the luteolytic response in cows treated at 120 hr post‐ovulation. In conclusion, the degree of complete luteolysis can be influenced by increasing the concentration of progesterone during the early luteal development in mares.