Modifying the double-Ovsynch protocol to include human chorionic gonadotropin to synchronize ovulation in dairy cattle.

The objectives were to determine whether rates of conception, ovulation, presynchronization, or follicle and CL characteristics were altered after modifying the Double-Ovsynch (DO) protocol to include hCG compared with the DO protocol. Primiparous and multiparous lactating dairy cows (N = 183), and nulliparous dairy heifers (N = 51) were used. Cows were blocked by parity and heifers were stratified by age and breed before being randomly assigned to one of two treatments. All females received either 100 μg GnRH or 2000 IU hCG im, at initiation of the Pre-Ovsynch (PO) portion of the DO protocol (PO: GnRH/hCG, 7 days PGF(2α) and 3 days GnRH). After 7 days, females started the Breeding-Ovsynch portion of the DO protocol (Breeding-Ovsynch: GnRH, 7 days, PGF(2α), 48 or 56 h and GnRH 16 hours timed artificial insemination with sex-sorted semen). Transrectal ultrasonography and blood samples were used to assess ovarian structures, ovulation, pregnancy diagnosis, and concentration of progesterone in plasma. Conception rates were similar in females treated with GnRH or hCG in cows (32.2 and 25.0%) and in heifers (30.8 and 36.0%). Ovulation rates in cows at the onset of PO were increased with hCG compared to GnRH (77.2 vs. 62.2%, P < 0.05). Concentrations of progesterone 7 days post-hCG or GnRH were greater in cows treated with hCG compared with GnRH (least significant mean ± SEM; 4.3 ± 0.3 and 3.0 ± 0.3 ng/mL, P < 0.01), but did not differ in heifers (4.5 ± 0.9 and 2.9 ± 0.9 ng/mL). More cows ovulated within 7 days post-hCG and a greater proportion of these cows tended to have failed luteal regression by Day 3 post-PGF(2α) compared with cows that had ovulated to GnRH (29.6 vs. 16.1%, P ≤ 0.10). The overall percentage of females which were synchronized to PO did not differ between GnRH- or hCG-treated cows (61.5% and 52.2%) and heifers (42.3% and 40.0%). In conclusion, no overall improvement in fertility was achieved by replacing the first injection of GnRH in the DO protocol with hCG.

Binversie JA ，Pfeiffer KE ，Larson JE 《-》

Effect of presynchronization with human chorionic gonadotropin or gonadotropin-releasing hormone 7 days before resynchronization of ovulation on fertility in lactating dairy cows.

Our objectives were to (1) compare the effect on pregnancies per artificial insemination (P/AI) of presynchronization of the estrous cycle with human chorionic gonadotropin (hCG) 7d before resynchronization of ovulation (Resynch) initiated 25 d after timed artificial insemination (TAI) and compare the presynchronization treatment with the Double-Ovsynch (DO) protocol, and (2) evaluate whether hCG for presynchronization could be replaced with GnRH. In experiment 1, lactating Holstein cows were blocked by parity and were randomly assigned to receive (1) Resynch-25 (D25), the Resynch protocol (GnRH-7 d-PGF(2 α)-56 h-GnRH-16 h-TAI) initiated 25 d after TAI (n=418); (2) HGPG, presynchronization with hCG (2,000 IU of Chorulon) 7d before D25 (n=450); and (3) DO (Pre-Resynch, GnRH-7 d-PGF(2 α)-72 h-GnRH; Breeding-Resynch, GnRH-7 d-PGF(2 α)-56 h-GnRH-16 h-TAI) initiated 22d after TAI (n=405). At 29 d after TAI, cows in the HGPG (37.3%) and DO (35.8%) groups had more P/AI than did cows in the D25 group (28.0%), and cows in the HGPG and DO groups continued to have more P/AI than did cows in the D25 group at 53 d after TAI. Presynchronization with hCG induced ovulation in 76% of the cows, which increased the percentage of HGPG cows with a corpus luteum at the initiation of Resynch compared with cows in the D25 group. In experiment 2, the D25 (n=368) and HGPG (n=338) treatments described in experiment 1 were compared in addition to a third treatment (GGPG; n=351), in which the hCG injection 18 d after TAI was replaced with a GnRH injection (200 µg of gonadorelin). At 32 d after TAI, cows in the HGPG group had more P/AI than did cows in the D25 group (33.7 vs. 25.5%), whereas cows in the GGPG group had intermediate P/AI (31.6%). At 53 d after TAI, P/AI tended to be greater for cows in the HGPG group than for those in the D25 group, whereas P/AI for cows in the GGPG group did not differ from that for cows in the D25 group. Treatment with hCG and GnRH 18d after TAI induced ovulation in 58.8 and 48.2% of cows, respectively, but did not increase the percentage of cows with a corpus luteum at the initiation of Resynch. More cows in the HGPG and GGPG groups had their estrous cycles synchronized after the resynchronization protocols compared with cows in the D25 group. We conclude that presynchronization with hCG increased fertility by increasing synchronization to the Resynch protocol, whereas presynchronization with GnRH improved synchronization to the Resynch protocol but did not improve fertility when compared with no presynchronization or presynchronization with hCG.

Giordano JO ，Wiltbank MC ，Guenther JN ，Ares MS ，Lopes G Jr ，Herlihy MM ，Fricke PM ... -  《-》

Prostaglandin F2α and gonadotropin-releasing hormone administration improve progesterone status, luteal number, and proportion of ovular and anovular dairy cows with corpora lutea before a timed artificial insemination program.

The objective of this research was to increase the proportion of cows with at least 1 functional corpus luteum (CL) and elevated progesterone at the onset of the timed artificial insemination (TAI) protocol. Postpartum Holstein cows in one herd were stratified by lactation number at calving (September 2009 through August 2010) and assigned randomly to 2 treatments: 1) Presynch-10 (n=105): two 25-mg injections of PGF(2α) (PG) 14 d apart (Presynch); and 2) PG-3-G (n=105): one 25-mg injection of PG 3 d before 100-μg GnRH (Pre-GnRH) injection, with the PG injection administered at the same time as the second PG injection in the Presynch-10 treatment. Cows were enrolled in a TAI protocol [Ovsynch; injection of GnRH 7 d before (GnRH-1) and 56 h after (GnRH-2) PG injection with AI 16 to 18 h after GnRH-2] 10 d after the second or only PG injection. Blood samples for progesterone or estradiol analyses were collected on median days in milk (DIM): 36, 39, 50, 53 (Pre-GnRH), 60 (GnRH-1), 67 (PG), 69 (GnRH-2), and 70 (TAI). Ovarian structures were measured by ultrasonography on median DIM 53, 60, 67, 69, and 6 d post-TAI to determine follicle diameters, ovulation response to GnRH, or both. Although progesterone concentration did not differ between treatments before Pre-GnRH injection, the proportion of cows with at least 1 CL tended to be greater for PG-3-G than Presynch-10 cows, and more PG-3-G cows ovulated after Pre-GnRH injection than ovulated spontaneously in Presynch-10. Further, the diameter of follicles that ovulated tended to be smaller in PG-3-G than in Presynch-10 cows after Pre-GnRH injection. At GnRH-1, the proportion of cows with progesterone ≥1 ng/mL, the number of CL per cow, and the proportion of cows with at least 1 CL were greater for PG-3-G than Presynch-10. Neither follicle diameter nor percentage of cows ovulating after GnRH-1 differed between treatments. At PG injection during the week of TAI, progesterone concentration and the proportion of cows with progesterone ≥1 ng/mL tended to be greater for PG-3-G than Presynch-10, and PG-3-G had more CL per cow than Presynch-10. No ovarian characteristics differed between treatments after GnRH-2, including progesterone concentration, number of CL per cow, and total luteal volume 7 d after GnRH-2. Many of the previous ovarian traits were improved in both ovular and anovular cows after PG-3-G compared with Presynch-10. Pregnancies per AI at d 32 and 60 were only numerically greater for PG-3-G than for Presynch-10 cows, largely because of differences detected during months without heat stress. We concluded that the PG-3-G treatment increased ovulation rate and luteal function 7 d before the onset of Ovsynch, resulting in improved follicular synchrony and predisposing potentially greater pregnancies per AI in lactating dairy cows.

Stevenson JS ，Pulley SL ，Mellieon HI Jr 《-》

Double-Ovsynch in high-producing dairy cows: effects on progesterone concentrations and ovulation to GnRH treatments.

Previous studies reported increased fertility using Ovsynch for presynchronization before Ovsynch (Double-Ovsynch), as compared with presynchronization with two prostaglandin F(2α) (PGF(2α)) treatments before Ovsynch (Presynch-Ovsynch). This study compared ovarian follicular dynamics and hormone concentrations during Double-Ovsynch versus Presynch-Ovsynch. Lactating Holstein cows (N = 193) were assigned to one of two treatment groups: (1) Presynch (N = 93), two injections of PGF(2α) 14 days apart, followed by the Ovsynch-timed AI protocol 12 days later; and (2) Double-Ovsynch (N = 100), one injection of GnRH, PGF(2α) 7 days later, and GnRH 3 days later, followed by the Ovsynch-timed AI protocol 7 days later. All cows received the same Ovsynch-timed AI protocol: GnRH (G1) at 68 ± 3 days in milk (mean ± SEM), PGF(2α) 7 days later, and GnRH (G2) 56 hours after PGF(2α). Ultrasonographic evaluations of the ovaries and blood sampling were performed at G1, PGF(2α), G2, and 6 days after the G2 injection of the Ovsynch-timed AI protocol. Double-Ovsynch decreased the percentage of cows with low circulating progesterone (P4) concentrations (<0.50 ng/mL) at G1 (12.0% vs. 30.1%; P = 0.003) and increased the percentage of cows with medium P4 concentrations (0.50 > P4 ≤ 3.0 ng/mL) at G1 (80.0% vs. 57.0%; P < 0.01), and with CL at G1 (94.0% vs. 67.8%; P < 0.01). Double-Ovsynch also increased the percentage of cows with high P4 (>3.0 ng/mL) at PGF(2α) (88.0% vs. 76.3%; P = 0.04) and tended to increase average circulating P4 at PGF(2α) (3.52 ± 0.17 ng/mL vs. 3.09 ± 0.21 ng/mL; P = 0.11). Double-Ovsynch also tended to increase percentage of cows ovulating to G1 (80.0% vs. 69.9%; P = 0.11) and G2 (98.0% vs. 93.5%; P = 0.08). Thus, presynchronization of cows with Double-Ovsynch induced ovulation in noncycling cows and appeared to increase most aspects of synchronization during the Ovsynch protocol.

Ayres H ，Ferreira RM ，Cunha AP ，Araújo RR ，Wiltbank MC ... -  《-》

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.

Stevenson JS ，Portaluppi MA ，Tenhouse DE 《-》