Sheep with ovarian androgen excess have fibrosis and follicular arrest with increased mRNA abundance for steroidogenic enzymes and gonadotropin receptors.

An androgen excess ovarian micro-environment may limit follicle progression in sheep. Two populations of ewes with divergent follicular fluid androstenedione (A4) were identified in a flock in Jordan: High A4; (A4) ≥ 30 ng/mL, (N = 12) or Control A4 (Control); A4 ≤ 15 ng/mL; (N = 12). We hypothesized High A4 ewes would have increased steroidogenic enzyme mRNA abundance, inflammation, and follicular arrest. Messenger RNA abundance for steroidogenic enzymes StAR, CYP17A1, CYP11A1, and HSD3B1 were increased in theca cells while CYP17A1, CYP19A1, and HSD3B1 were increased in granulosa cells in High A4 ewes compared to Control. Gonadotropin receptor mRNA expression for LHCGR was increased in theca and FSHR in granulosa in High A4 ewes. Messenger RNA expression of FOS when reduced, increases expression of CYP17A1 which was observed in High A4 granulosa cells compared to Control. Furthermore, High A4 ewes had greater numbers of primordial follicles (P < 0.001) and fewer developing follicles compared to Control before, and after 7 d of culture, indicating follicular arrest was not alleviated by cortex culture. Increased fibrosis in the ovarian cortex was detected in High A4 ewes relative to Control (P < 0.001) suggesting increased inflammation and altered extracellular matrix deposition. Thus, this High A4 ewes population has similar characteristics to High A4 cows and women with polycystic ovary syndrome suggesting that naturally occurring androgen excess occurs in multiple species and may be a causative factor in follicular arrest and subsequent female sub- or infertility.

Excess androgen (androstenedione; A4) in ewes can result in ovarian follicular arrest and fibrosis contributing to anovulation in sheep. We have identified a naturally occurring ovarian A4 excess in a sheep population with similar characteristics to High A4 cows, both of which are similar to that in women with polycystic ovary syndrome indicating that several mammalian species experience naturally occurring androgen excess resulting in infertility or follicle arrest. Somatic cells, theca and granulosa, surrounding the egg in High A4 ewes had increased expression of steroidogenic enzymes, similar to that seen in High A4 cows, permitting more ovarian cells to manufacture androgens, which may be the cause of androgen excess. Thus, naturally occurring androgen-excess in domestic livestock females can be utilized as models to research the causes of androgen excess and determine the mechanisms that result in follicular arrest and sub- or infertility.

Effect of two prostaglandin injections on days 5 and 6 in a timed AI protocol after estrus expression on pregnancy outcomes in dairy cows during cold or hot seasons of the year.

The objective of this study was to test whether prostaglandin (PG) injection on day 30 postpartum (pp) and detection of estrus can affect the efficacy of injecting PG on days 5 and 6 in the timed artificial insemination (TAI) protocol on pregnancy rate in a large dairy herd in hot or cold seasons. Out of 2235 cows, 1998 received an injection of PG at 30 ± 3  d pp and estrus was observed. Cows that displayed estrus during the estrous observation period after PG injection were classified as estrus (E), while those that did not show estrus were classified as nonestrus (NE). Cows in each group were assigned to two treatments: CO-72 (control treatment) (ECO-72 and NECO-72) (day 44 GnRH, day 51 PGF 2 α , day 54 GnRH + TAI) or PG-PG (EPG-PG and NEPG-PG) (day 44 GnRH, day 49 PGF 2 α , day 50 PGF 2 α , day 52 GnRH + TAI). Pregnancy was diagnosed on days 33 and 47 after artificial insemination (AI). The proportion of cows in estrus on the day of TAI was higher ( P ≤ 0.05 ) for cows that received two PG than for cows that received one PG. Pregnancies per AI ( P /AI) on days 33 and 47 for cows inseminated during and after a voluntary waiting period in the NEPG-PG treatment had higher rates than for cows in the EPG-PG, ECO-72 and NECO-72 treatments. Moreover, P /AI were significantly ( P ≤ 0.05 ) affected by parity. Primiparous had higher P /AI (37.0 %) than multiparous cows (31.6 %). Cows inseminated in cold months had higher P /AI and reduced PL (35.6 % and 20.8 %) than cows inseminated in hot months (29.1 % and 30.6 %, respectively). In conclusion, treatments with PG on days 5 and 6 after the first GnRH injection increased P /AI. Estrus detection before the beginning of TAI protocol did not affect fertility. To maximize P /AI cows exhibiting heat at any time during the synchronization protocol should be inseminated.

Treatment with hCG 4 or 6 days after TAI to improve pregnancy outcomes in repeat-breeding dairy cows.

A study was conducted to evaluate the effect of human chorionic gonadotropin (hCG) administration 4 or 6 days after timed AI (TAI) on P4 concentration and pregnancy outcomes in repeat breeding dairy cows. All cows were treated by Cosynch protocol before being assigned into 3 groups: CON (n=139): did not receive hormonal treatments; D4 (n=136): received 1500 IU hCG 4 days post TAI; and D6 (n=131): received 1500 IU hCG 6 days post TAI. Plasma P4 concentration was evaluated on Day of hCG and 12 days post TAI. Pregnancies per AI (P/AI) on Day 45 were greater for D6 (38.9%) than CON (30.9%) groups. Pregnancy losses (PL) were lower for D6 (15.0%) and D4 (26.7%) than CON (37.7%) groups. Treatment by season revealed a greater P/AI and lower PL for D6 (30.3% and 18.2%) and D4 (33.3% and 31.0%) than CON (19.2% and 58.3%) groups during summer, while P/AI was greater for D6 (57.1%) than D4 (30.6%) and CON (37.9%) groups during winter. Treatment by parity indicated a greater P/AI for D4 (46.4%) and D6 (31.6%) than CON (15.8%) groups within primiparous cows. Pregnancy losses were lower for D6 (15.1%) than CON (36.5%) groups within multiparous cows while D4 tended to be lower (13.3%) than CON (50.0%) groups. Plasma P4 concentration on Day 12 post TAI was higher for treated than for CON groups. Applying hCG 6 days after TAI was beneficial in improving P/AI either in summer or winter as a result of reducing PL and increasing P4 concentration.

Effect of progesterone (P(4)) intravaginal device (CIDR) to reduce embryonic loss and to synchronize return to oestrus of previously timed inseminated lactating dairy cows.

A study on postpartum dairy cows was done to evaluate the effect of using progesterone intravaginal device (CIDR) to synchronize return to oestrus of previously timed inseminated (TAI) cows and to evaluate embryo survival and pregnancy rate in the return to oestrus cows. All cows were subjected to TAI program. On Day 14 after AI, cows were assigned randomly into two groups: treated group (CTAI; n=126) and control group (TAI; n=172). Every cow in the CTAI group received CIDR device for 7 days. Cows were observed for oestrus after CIDR removal in the first period (Days 21-28). Cows, which did not show oestrus, were retreated with CIDR on Day 28 for 7 days and they were observed for oestrus after CIDR removal in the second period (Days 35-42). Pregnancy was diagnosed on Days 28, 45 and 90 after AI. In the first period, more cows in the CTAI group showed oestrus within 2.4 days after CIDR removal compared to cows within 4 days in the control. In the second period, more cows in the CTAI group showed oestrus within 4.5 days compared to cows within 3.7 days in the control. Pregnancy rates on Day 28 were not different between both groups. Pregnancy rates on Days 45 and 90 were higher for cows in the CTAI (42.1+/-5.3%, P=0.028 and 38.9+/-5.2%, P=0.020) compared to that in the TAI group (35.5+/-3.9% and 31.9+/-3.8%). Moreover, pregnancy rates were lower for multiparous cows (34.2+/-4.1, P=0.017 and 30.2+/-3.9%, P=0.008) compared to primiparous cows (42.7+/-5.2% and 39.9+/-5.1%) on Days 45 and 90, respectively. Pregnancy losses between Days 28-45 and 45-90 tended to be lower for the CTAI group and primiparous cows compared to the TAI group and multiparous cows. Pregnancy rates of first (P=0.020), second, third and up to fourth AI were higher (P=0.001) for the CTAI group (38.9+/-0.05%, 78.6+/-0.05%, 92.1+/-0.04%, 93.6+/-3.71%) compared to that for the TAI group (31.9+/-0.03%, 54.1+/-0.04%, 68.0+/-0.03%, 81.4+/-2.75%, respectively). Days open for pregnant cows were similar in both groups, but lower (P=0.025) for primiparous cows (81.6+/-4 day) compared to multiparous cows (93.8+/-3 day). Number of services for pregnant cows was lower (P=0.002) for the CTAI group (1.70+/-0.10) compared to that for the TAI group (2.14+/-0.09), whereas it was similar for primiparous and multiparous cows. Results indicate that the CIDR device improved synchronization to return to oestrus and increased pregnancy rate to first, second, third and fourth AI by reducing embryonic and fetal losses.

Comparison of an oestrus synchronisation protocol with oestradiol benzoate and PGF2alpha and insemination at detected oestrus to a timed insemination protocol (Ovsynch) on reproductive performance of lactating dairy cows.

A total of 226 out of 245 postpartum lactating dairy cows in a commercial dairy farm were allocated to two groups of oestrous synchronisation protocols in order to evaluate reproductive performance. One group was treated with oestradiol benzoate (ODB) and PGF2alpha on day 10 of the oestrous cycle with insemination at the detected oestrus, the second group underwent the Ovsynch (OVS) protocol (GnRH + PGF2alpha + GnRH) with timed AI. Pregnancy was diagnosed by ultrasonography on day 28 after AI and confirmed by rectal palpation on day 45. A higher (P < 0.001) proportion of cows in OVS (100%) were inseminated within (19.2 +/- 3.8 h) following the second GnRH injection than those of cows in EPE (ODB + PGF2alpha + ODB) (70.6%) inseminated at the detected oestrus within (35.6 +/- 5.2 h) following the second ODB injection. Pregnancy rates for the first AI at day 28 (64.0 +/- 4.6, 62.4 +/- 5.5%) and at day 45 post-insemination (40.4 +/- 4.7, 40.0 +/- 5.6%) for OVS and EPE cows respectively, did not differ between the two treatments, whereas, the overall pregnancy rates tended to be higher (P < 0.08) for the OVS (85.1 +/- 3.8%) cows than the EPE cows (74.1 +/- 4.5%). No differences were observed in pregnancy rates for first AI and overall up to fourth AI between primiparous (34.7 +/- 5.8 and 85.3 +/- 4.7%) and multiparous cows (43.5 +/- 4.5 and 77.4 +/- 3.6%). Days open for pregnant cows tended to be lower (P < 0.08) for OVS (76.2 +/- 3) than for EPE cows (84.7 +/- 4), while days open were higher (P < 0.05) in primiparous cows (85.3 +/- 4) than in multiparous cows (75.6 +/- 3). The results indicate that pregnancy rates for first AI were similar, but overall pregnancy rates up to the fourth AI tended to be higher for OVS than EPE cows, while days open was tended to be lower for OVS than EPE cows.