Factors influencing upfront single-and multiple-ovulation incidence, progesterone, and luteolysis before a timed insemination resynchronization protocol.

Our objectives were to determine relationships among factors influencing responses to the first GnRH injection in a timed artificial insemination (TAI) protocol and subsequent fertility after altering timing of the second GnRH injection and AI relative to PGF(2alpha) injection. Replacement heifers (n = 86) and 613 lactating cows previously inseminated were diagnosed not pregnant to form 77 breeding clusters spanning 36 mo. At not-pregnant diagnosis (d 0), females received 100 mug of GnRH, and then 7 d later, they received 25 mg of PGF(2alpha). Females in 2 treatments received GnRH 48 h (G48) after PGF(2alpha) injection and TAI at the time of the second GnRH injection (G48 + TAI48) or 24 h later (G48 + TAI72). Females in the third treatment received GnRH 72 h after PGF(2alpha) when inseminated (G72 + TAI72). Neither timing of GnRH nor time of AI altered TAI pregnancy rates (average of 20.4%). Ovaries of females in 65 clusters were scanned on d 0 (first GnRH injection) and 7 d later (PGF(2alpha) injection). Ovarian structures were mapped and ovulation in response to the first GnRH injection was evaluated on d 7. When estrus was detected before scheduled TAI, females were inseminated; otherwise, TAI conception of remaining females was based on timing of GnRH and AI in 3 treatments. On d 7, 1 or more new corpora lutea (CL) were detected in 43% of females and their pregnancy rate was subsequently greater (28 vs. 18%) than those not ovulating. Follicle diameters on d 0 did not differ between females that did (11.9 +/- 0.3 mm) and did not (11.8 +/- 0.4 mm) subsequently ovulate in response to GnRH. Follicle diameter and number of follicles >/=5 mm increased with increasing lactation number, but decreased with increasing number of CL. Diameter of follicles in which more than 1 follicle ovulated decreased linearly from that in which only 1 follicle ovulated. Incidence of ovulation increased with increasing lactation number and total number of follicles >/=5 mm, but decreased with increasing number of CL. Incidence of multiple ovulations (15%) was greater in females having more follicles >/=5 mm and in those in early diestrus. Multiple ovulation did not occur in heifers, but was decreased in cows having more than 1 CL. In cows having more than 1 CL, luteal regression was reduced by 5.6 percentage units compared with those having 1 CL. In a TAI protocol, pregnancy rate was greater for females in early diestrus compared with females in other stages of the cycle, in those that ovulated after the first GnRH injection, in those having luteolysis, and in those inseminated during nonsummer months.

Ovarian traits after gonadotropin-releasing hormone-induced ovulation and subsequent delay of induced luteolysis in an Ovsynch protocol.

Our objective was to determine whether delaying the PGF2alpha injection by 24 or 48 h after the first GnRH injection in an Ovsynch protocol (from a standard 7 d) altered ovarian characteristics in lactating dairy cows. Beginning 9 d after removal of a progesterone-releasing controlled internal drug release (CIDR) insert and injection of PGF2alpha (d 6.4 of the estrous cycle), 36 Holsteins (average body weight = 707 +/- 12 kg and body condition score = 2.3 +/- 0.1) were administered 100 microg of GnRH (81 +/- 2 d in milk) and assigned randomly to receive a treatment injection of PGF2alpha 7, 8, or 9 d later. Timed artificial insemination was performed at 48 h after PGF2alpha at which time a second injection of GnRH was administered. Ovarian structures were mapped by ultrasonography on d 0 (first GnRH injection); on d 2 to determine responses to the first GnRH injection; at PGF2alpha injection; and daily thereafter through 72 h after PGF2alpha to monitor ovulation of preovulatory follicles. Blood was collected on d 0, 2, at PGF2alpha injection, and at 24 and 48 h after PGF2alpha to monitor serum changes in estradiol-17beta (E2-17beta) and progesterone (P4). Based on serum P4 and ovarian exams, 2 cows were eliminated because of anestrus and their failure to ovulate a follicle in response to the first GnRH injection. Two other cows in which luteolysis failed to occur after PGF2alpha treatment also were eliminated. Final numbers of cows per treatment were: 7 d (n = 13), 8 d (n = 9), and 9 d (n = 10). Twenty-nine of 32 cows ovulated (90.6%) in response to the first GnRH injection. Of those cows not ovulating in response to the first GnRH injection, 2 had 1 original corpus luteum and 1 had 2 original corpora lutea. Despite a 24- or 48-h delay between first GnRH and PGF2alpha injections, the diameter (mm) and volume (mm3) of the ovulatory follicle did not differ among treatments: 14.3 +/- 0.6 and 1,526 +/- 62 at 7 d; 14.1 +/- 0.8 and 1,479 +/- 97 at 8 d; and 15.3 +/- 0.9 and 1,490 +/- 69 at 9 d. In all 32 cows, at least 1 follicle ovulated after treatment, but ovulation rates did not differ: 1.2 +/- 0.1, 1.1 +/- 0.1, and 1.3 +/- 0.2, respectively, for the 7-, 8-, and 9-d treatments. Serum concentrations of E2-17beta did not differ among treatments. Four cows in the 7-d treatment were inseminated 24 h late and were excluded before assessing conception rates, which were 5/9 (55.6%), 5/9 (55.6%), and 1/10 (10%), respectively. We conclude that delaying PGF2alpha injection by 24 h had no effect on outcomes.

Interventions after artificial insemination: conception rates, pregnancy survival, and ovarian responses to gonadotropin-releasing hormone, human chorionic gonadotropin, and progesterone.

We hypothesized that increasing concentrations of progesterone (P4) after artificial insemination would increase fertility. Our objective was to assess changes in ovarian structures, incidence of ovulation, and change in serum P4 in response to GnRH, human chorionic gonadotropin (hCG), or exogenous P4 (controlled internal drug release; CIDR insert) treatment beginning 4 to 9 d after artificial insemination (d 0) and again 7 d later (experiment 1). Blood was collected from 753 cows in 3 herds on d 0 and 7. Ovaries of 162 cows were scanned and mapped to confirm the presence of a corpus luteum (CL), and cows were assigned randomly to serve as controls (n = 41) or to receive a CIDR insert for 7 d (n = 41), 100 microg of GnRH (n = 40), or 3,300 IU of hCG (n = 40). More cows were induced to ovulate in response to GnRH (60%) and hCG (78%) compared with controls (2.4%). Compared with controls, cows treated with GnRH or hCG had more induced CL (d 7) and more total CL (d 7), but serum P4 was increased only in response to hCG. Largest follicle diameters on d 7 were less after GnRH and hCG, but total follicular volume on d 7 was reduced by GnRH, hCG, and CIDR, compared with that of controls. Volume of the original luteal structures was increased by hCG but tended to be reduced by CIDR and GnRH compared with luteal volume in controls. Total CL volume was increased by hCG, but reduced by CIDR, compared with CL volume of controls. Conception rates and pregnancy survival were assessed in response to the same treatments described in experiment 1: controls (n = 708), CIDR (n = 711), GnRH (n = 719), and hCG (n = 714). Tendencies for interactions of treatment x herd and treatment x lactation group were detected, but no 3-way interactions were found. Treatment with hCG increased conception rates in second-lactation cows. The CIDR tended to increase, and hCG increased, conception rates in 2 herds, whereas the CIDR decreased conception rates in 1 herd. Pregnancy survival was reduced by GnRH compared with that in controls. We concluded that GnRH and hCG effectively induced ovulation, and increased number of CL, but only increased serum P4 in hCG-treated cows. Further, treatment with the CIDR or hCG increased conception rates but only in some herds.

Pregnancy rates in lactating dairy cows after presynchronization of estrous cycles and variations of the Ovsynch protocol.

Our objectives were to determine pregnancy rates after altering times of the second GnRH injection, insemination, or both in a combined Presynch + Ovsynch protocol, to accommodate once-daily lockup of dairy cows. Lactating dairy cows (n = 665) from 2 dairy herds in northeastern Kansas were studied. Cows ranged from 24 to 44 d in milk (DIM) at the start of the Pre-synch protocol, which consisted of 2 injections of PGF(2alpha) 14 d apart, with the second injection given 12 d before initiating the Ovsynch protocol. Cows were blocked by lactation number and assigned randomly to 3 treatments consisting of variations of the Ovsynch protocol. Cows in 2 treatments received injections of GnRH 7 d before and 48 h (G48) after the PGF(2alpha) injection. Timed AI (TAI) was conducted at the time of the second GnRH injection (G48 + TAI48) or 24 h later (G48 + TAI72). Cows in the third treatment received the injections of GnRH 7 d before and at 72 h after PGF(2alpha) and were inseminated at the time of the second GnRH injection (G72 + TAI72). Pregnancy was diagnosed weekly by palpation per rectum of uterine contents on d 40 or 41 after TAI. Pregnancy rates differed between herds, but they were consistently greater for G72 + TAI72 than for G48 + TAI48 and G72 + TAI72. Subsequent calving rates were consistent with differences in initial TAI pregnancy rates. Pregnancy loss was least for cows on the G72 + TAI72 treatment. Body condition scores (BCS) ranged from 1.0 to 4.0 when assessed on Monday of the breeding week. An interaction of BCS and herd was detected in which cows in herd 1 having poorer BCS (<2.25) had greater pregnancy rates than cows of greater BCS (>/=2.25), whereas the reverse was true in herd 2 in which overall pregnancy rates were greater. We concluded that inseminating at 48 or 72 h after PGF(2alpha), when GnRH was administered at 48 h after PGF(2alpha), produced fewer pregnancies than inseminating and injecting GnRH at 72 h after PGF(2alpha) for cows whose estrous cycles were synchronized before initiating this variant of the Ovsynch protocol.