To determine effects of timed artificial insemination (TAI) hormonal treatments on reproductive performance of gilts/sows and explore molecular mechanisms, gilts (TAI: 90; Control:149; Total: 239) and sows (TAI: 370; Control: 492) were utilized. Results indicated the estrus/farrowing rate and number of piglets born alive and weaned in the TAI group were greater than in the control group for both gilts and sows. To explore the molecular mechanism for TAI hormonal effects, the small RNA of the gilt endometrium at 16 and 25 of gestation were sampled and sequenced to determine potential functions of microRNA (MiRNA); 358 known and 142 novel MiRNAs were detected. With comparison of TAI and control groups, there were 54 differentially abundant MiRNAs, and functional analysis results indicated \"binding,\" \"protein/ion binding,\" and \"immune response\" were mostly enriched. In addition, representative MiRNAs were selected based on criteria including being regulated on both day 16 and 25 of gestation (ssc-miR-10a-5p, ssc-miR-345-5p, ssc-miR-370) along with reproduction-related target genes (ssc-miR-424-5p, ssc-miR-142-5p). Furthermore, target genes of selected MiRNAs were screened, and functional enrichment of those genes also indicated that the \"binding\" and \"immune response\" were mainly enriched. Results from the present study confirmed TAI-hormonal treatments improved estrous/farrowing rate and number of piglets born alive/weaned of gilts/sows and that hormonal treatment regimens leading to behavioral estrus at timed artificial insemination in gilts results in microRNA patterns in the endometrium that are more supportive of pregnancy. Results contribute valuable information for future studies of effects of TAI hormonal treatments on pig reproductive performance.

The objective of this study was to use luteinizing hormone-releasing hormone A3 (LRH-A3) and human chorionic gonadotrophin (HCG) to improve pregnancy rate of dairy cows during timed artificial insemination (TAI). In experiment 1, the TAI process (0 d, GnRH, 100 μg; 7 d, PGF2α, 0.4 mg; 56 hr, GnRH, 100 μg; 16 hr, AI) was applied to 160 dairy cows on 50th and 60th days after parturition respectively. In experiment 2, 320 postpartum dairy cows were treated with TAI (Group A), TAI + 25 μg LRH-A3 (Group B), TAI + 1,500 IU HCG 5 days after AI (Group C), and TAI + 25 μg LRH-A3 + 1,500 IU HCG 5 days after AI (Group D). In experiment 3, endometrial cells were treated with HCG. The results showed that TAI did not affect the pregnancy rate, while LRH-A3 and HCG increased the pregnancy rate of the cow. HCG of 5 IU/ml and 10 IU/ml increased the expressions of leukemia inhibitory factor but decreased those of interleukin-6, epidermal growth factor and vascular endothelial growth factor in endometrial cells. This study provided a plan for the use of LRH-A3 and HCG to increase pregnancy rate during TAI in dairy cows.

BACKGROUND: With the expansion of the donkey industry, timed artificial insemination (TAI) is becoming increasingly important in the reproductive management of jennies, however, TAI has not been widely investigated in donkeys.
OBJECTIVE: To develop efficient TAI protocols for cooled or frozen semen in jennies, based around ovulation induction with a GnRH analogue.
METHODS: Experimental exploratory study.
RESULTS: In experiment 1, the effects of different GnRH analogue (deslorelin) doses, follicle diameter (FD) at induction, repeated use of a GnRH analogue, and the influence of season on induction efficiency, as well as distribution of ovulations over time after induction were investigated. Induction efficiency was sufficient with 2.2 mg deslorelin (≥90% ovulation within 48 hours of treatment). Ovulation rate between 24 and 48 hours was highest when the FD at treatment was 31-35 mm, as compared to 25-30 mm or 36-40 mm. Repeated use of deslorelin or treatment during different seasons had no effect on induction efficiency. About 70% of ovulations occurred between 32 and 48 hours, and highest incidence of ovulation was at 36-38 hours after induction. In experiment 2, TAI using cooled semen (1 × 109 motile sperm in a 10 mL volume) was performed once at 8 hours after induction (n = 59). Pregnancy rate after TAI with cooled semen was 49.2% (29/59). In experiment 3, jennies were inseminated twice with 10 (n = 23), 5 (n = 31), 3 (n = 32), 2 (n = 82) and 1 (n = 66) straws (more than 50 × 106 motile spermatozoa in each 0.5 mL straw) of frozen semen at 34 and 42 hours after induction. The pregnancy rates were 30.4%, 35.5%, 34.4%, 29.3% and 28.8%, respectively (P > 0.05).
UNASSIGNED: In the frozen semen trial, 22.5% (68/302) jennies were excluded after failure to ovulate during the appropriate time interval. In addition, there were no control groups for the AI trials.
CONCLUSIONS: When FD reaches 31-35 mm, a donkey jenny can be inseminated once using cooled semen at 8 hours or twice using frozen semen at 34 and 42 hours after deslorelin treatment. The frozen semen TAI protocol resulted in acceptable pregnancy rates using 1 × 108 motile spermatozoa per cycle.

The purpose of this study was to determine the optimal time for ovulation induction and artificial insemination (AI) based on the relationship between estrous behavior and ovulation in jennies. Thirty-two jennies were teased by one jackass for 1 hour per day during 46 days and estrous behaviors were recorded, while the follicular development and ovulation was examined by ultrasound. Furthermore, another 31 jennies were teased by one jackass as the teasing group (group T), which were injected with Deslorelin at 2 and 4 days after the onset of estrus, and AI was performed at 8 hours after each injection. Moreover, Ultrasound was performed on the follicle development of 23 jennies as the ultrasonography group (group U). Injection with Deslorelin when the follicle diameter ≥ 30 mm, and AI was performed at 8 hours later. The results showed that mouth clapping was the specific estrous behavior of jennies and indicated the beginning of estrus. The mean time for jennies to develop dominant follicles (≥30 mm) after the onset of estrus was 3.5 ± 1.3 days, and the mean time between the onset of estrus and ovulation was 5.1 ± 1.5 days. Estrous behaviors ended 0.5 ± 1.2 days after ovulation. After AI, there were no significant differences in ovulation (96.8% vs. 91.3%) and conception rates (40.0% vs. 38.1%) between group T and U. The optimal breeding time of jennies can be determined by jackass teasing and hastening ovulation by Deslorelin injection.