Effect of Initial GnRH and Duration of Progesterone Insert Treatment on the Fertility of Lactating Dairy Cows

The study compared response to prostaglandin F2α (PG), synchrony of ovulation and pregnancy per AI (P/AI) in a 5‐ vs a 7‐day Ovsynch + PRID protocol and investigated whether the initial GnRH affects P/AI in lactating dairy cows. Two hundred and seventy‐six cows (500 inseminations) were assigned to one of four timed‐AI (TAI) protocols: (i) PRID‐7G; 100 μg GnRH im, and a progesterone‐releasing intravaginal device (PRID) for 7 days. At PRID removal, PG (500 μg of cloprostenol) was given im. Cows received the second GnRH treatment at 60 h after PRID removal and TAI 12 h later. (ii) PRID‐5G; as PRID‐7G except the duration of PRID, treatment was 5 days and PG was given twice (12 h apart). (iii) PRID‐7NoG; as PRID‐7G except the initial GnRH, treatment was omitted. (iv) PRID‐5NoG; as PRID‐7NoG except the duration of PRID, treatment was 5 days. Response to treatments and pregnancy status at 32 and 60 days after TAI was determined by ultrasonography. The percentage of cows ovulating before TAI was greatest in PRID‐7G (17.1%), and the percentage of cows that did not have luteal regression was greatest in PRID‐5G (9.5%). The overall P/AI at 32 and 60 days did not differ among TAI protocols. However, during resynchronization, cows subjected to the 5‐day protocols had greater (p < 0.05) P/AI (45.3% vs 33.6%) than cows subjected to the 7‐day protocols. Pregnancy loss between 32 and 60 days tended (p = 0.10) to be greater in cows that did not receive initial GnRH (14.8%) compared to those that received GnRH (8.2%). In conclusion, the PRID‐5G protocol resulted in fewer cows responding to PG, but P/AI did not differ among TAI protocols. A 5‐day protocol resulted in more P/AI in resynchronized cows, and cows that did not receive initial GnRH tended to experience more pregnancy losses.     

Effect of body condition score and reuse of progesterone‐releasing intravaginal devices on conception rate following timed artificial insemination in Nelore cows

This study had the aim of investigating the efficiency of timed artificial insemination (TAI) through the progesterone‐releasing intravaginal device (PRID), used in new condition and for the second and third times in Nelore cows. The effects of device reuse and body condition score (BCS) on the conception rate (CR) were evaluated in 1,122 multiparous Nelore cows (mean BCS of 2.7 ± 0.4), which were randomly distributed into three groups that received new (n = 330), once (n = 439) and twice used (n = 353) PRID. Among the 1,122 females that underwent TAI, 573 became pregnant, thus representing an overall CR of 51.06%. Cows with BCS between 2.75 and 4.0 had greater (p < .0001) CR (69.75%) than cows with BSC of 2.0–2.5 (32.98%). It was observed that the CR through using PRID was 60.00%, 51.71% and 41.93% for new, once and twice used PRID, respectively, with difference between all groups (p < .0001). Under tropical conditions, animals with BCS greater than 2.5 had a higher CR, and the CR decreased proportionally with the number of times that the PRID had been used.     

Resynchronization of ovulation with new and reused intravaginal progesterone‐releasing devices without previous pregnancy diagnosis in Bos taurus indicus cows subjected to timed‐artificial insemination

The study aimed to evaluate pregnancy per artificial insemination (P/AI) of cows subjected to synchronization and resynchronization in ovulation protocols using intravaginal progesterone‐releasing insert (P4) before pregnancy diagnosis (PD) and the relationship of PR with the diameter of preovulatory follicles (ØPOF) before TAI. Cows (n = 378) were distributed into two groups: a resynchronization group with new devices (GRN; n = 185) and resynchronization group with used devices (GRU; n = 193). On Day 0, both groups received a new P4 and estradiol benzoate (EB). On D8, P4 removal + D‐cloprostenol + eCG + estradiol cypionate (EC) was done. On d10, TAI was conducted. On d32, cows were resynchronized and divided into two groups, GRN (n = 185) and GRU (n = 193). The GRN group received a new P4 + EB, and the GRU group received a used P4 + EB. On d40, the P4 was removed + PD. The non‐pregnant cows received D‐cloprostenol + eCG + EC. US was done again on d42 to determine ØPOF before the second TAI. The P/AI of the GRN and GRU groups after synchronization were 56.2% and 57.0% (p = 0.87), respectively, and those after resynchronization were 58.0% and 37.3% (p < 0.008), respectively. The P/AI of the GRN and GRU groups observed after TAI (synchronization + resynchronization) were 81.6% and 73.1%, respectively (p = 0.047). No difference (p = 0.067) in ØPOF between the pregnant and non‐pregnant cows in the GRN was found, whereas the GRU group showed a significant difference (p = 0.003). Resynchronization protocols optimized the P/AI in both groups. New intravaginal devices resulted in greater P/AI and P/AI accumulation in resynchronization as compared with the GRU; the ØPOF was related with P/AI.     

Effect of oestrous resynchronization on the reproductive efficiency of zebu cows

The aim of this study was to develop a resynchronization strategy before the return of oestrus in cows diagnosed as not pregnant after fixed‐time artificial insemination (TAI). A total of 839 cows, approximately 45 days post‐partum, were synchronized using TAI. On day 0, intravaginal progesterone‐releasing devices were inserted and 2 mg of oestradiol benzoate was administered. Eight days later (D8), the progesterone‐releasing devices were removed and oestradiol cypionate (0.5 mg, eCG [300 IU]) and prostaglandin (7.5 mg) were administered. All cows were inseminated between 48 and 56 hr after device removal (D10). Thirty days after TAI, cows that were not diagnosed as pregnant by ultrasound were immediately resynchronized and again inseminated at a fixed time. The hormonal protocol used in the first and second rounds of TAI was the same. The pregnancy rate after the first TAI was 52%, and after the second TAI, it was 49%. The increase in the total pregnancy rate (synchronization + second oestrous synchronization) compared to a single synchronization was 23.5%. In conclusion, resynchronization of oestrus and ovulation in zebu cows that had previously undergone TAI protocols is effective in increasing the reproductive efficiency.     

ACE inhibition in goats under fixed‐time artificial insemination protocol increases the pregnancy rate and twin births

To assess the effect of the angiotensin‐converting enzyme (ACE) inhibition on the efficiency of the fixed‐time artificial insemination (TAI), 69 goats were divided randomly into two groups: enalapril (n = 35) and control (n = 34). In the experiment, all animals underwent the protocol of fixed‐time artificial insemination for 12 days. Enalapril group received enalapril maleate dissolved in saline (Enalapril, Lab Teuto Ltda) subcutaneously at the following doses: 0.2 mg/kg/day in D0‐D2; 0.3 mg/kg/day in D3‐D6 and 0.4 mg/kg/day in D7‐D11. The control group received the corresponding volume of 0.9% saline solution. We performed a single insemination 36 hr after sponge removal using frozen semen from two adult male goats with recognized fertility. The ultrasound pregnancy diagnosis was 30 days after the artificial insemination (AI). There was significant increase in pregnancy rates and twinning as well as a decrease in foetal loss in animals receiving enalapril (p < .01). The use of ACE inhibitors during the TAI protocol was shown to be a promising alternative to increase the efficiency of such reproductive biotechnology.     

Anti-Mullerian hormone and its relationship to ovulation response and fertility in timed AI Bos indicus heifers

This study evaluated the association between plasma anti-Mullerian hormone (AMH) concentration and fertility in Nelore (Bos indicus) heifers submitted to timed artificial insemination (TAI). At the onset of the synchronization protocol, heifers (n = 289) received a subcutaneous P4 ear implant (3 mg) and 2 mg of oestradiol benzoate. Eight days later, the P4 implant was removed and 0.5 mg of oestradiol cypionate, prostaglandin (0.265 mg, i.m.) and equine chorionic gonadotropin (300 UI, i.m.) was administered, and TAI was performed 48 hr after ear implant removal. Ovarian ultrasound evaluations were performed to measure number of ovarian follicles, dominant follicle size and ovulation response. Pregnancy diagnosis was performed by ultrasound 30 days after AI. Heifers with greater circulating AMH had more antral follicles, a smaller dominant follicle near timed ovulation and lower ovulation response to the timed AI protocol compared to heifers with lower circulating AMH. Although AMH and pregnancy outcome had a quadratic-shaped pattern, AMH was not significantly associated with fertility. In conclusion, heifers with lower AMH had larger follicles towards the end of the synchronization protocol and greater ovulation responses, whereas greater circulating AMH was unrelated to conception success.     

Effect of synchronization protocols on reproductive indices,progesterone profile and fertility under subtropical environmental conditions in repeat breeder Holstein cows

The objectives were to evaluate the reproductive indices and survival analysis of pregnancy outcomes in multiparous repeat breeder Holstein cows (n = 557). The cows were synchronized to ovulate by Ovsynch, new controlled internal drug release device (N‐CIDRsynch), and once‐used CIDR device (U‐CIDRsynch). The pregnancy per AI at 28 days post‐insemination (P/AI 28) in the N‐CIDRsynch group (28.75%) was significantly (COR = 1.49; p = 0.011) greater than that reported in the Ovsynch (23.46%) and U‐CIDRsynch (21.73%) groups. Furthermore, the pregnancy per AI at day 75 post‐insemination (P/AI 75) in the N‐CIDRsynch group was significantly greater than the Ovysync group (COR = 1.35; p = 0.050). The repeat breeder cows received a N‐CIDR device had a significantly higher progesterone level on day 2 and day 4 of CIDR insertion (1.38 and 1.67 ng/ml, respectively) than those received a U‐CIDR device or the control group (p = 0.012 and 0.001, respectively). The Cox regression model recorded significant associations for synchronization protocols, THI at the TAI and season of calving with the hazard of P/AI 28 and P/AI 75 (p = 0.044 and 0.046; 0.001 and 0.005; 0.003 and 0.001, respectively). Multiparous repeat breeder cows (>3) had a lower hazard ratio (HR) of P/AI 28 than that reported in the reference (2nd parity) group (HR = 0.74, p = 0.050). The repeat breeder cows inseminated at 76–80 and >80 temperature‐humidity index (THI) had significantly lower HR of P/AI 28 than those inseminated at the baseline (<70) THI value (HR = 0.73 and 0.30, p = 0.036 and 0.001, respectively). The current results indicate that the use of N‐CIDR synch protocol may achieve satisfactory pregnancy outcomes in repeat breeder cows.     

Comparison of pregnancy per AI of heifers inseminated with sex-sorted or conventional semen after oestrus detection or timed artificial insemination

The objective of the study was to compare the fertility after using sex-sorted or conventional semen either with oestrus detection (EST) or timed artificial insemination (TAI) in Holstein heifers. Holstein heifers were randomly assigned to one of the following treatments in a 2 × 2 factorial design. Heifers in the EST group were inseminated with sex-sorted (n = 114) or conventional semen (n = 100) after spontaneous or induced oestrus. Heifers in the TAI, subjected to the 5-day Cosynch+Progesterone protocol (GnRH+P4 insertion-5d-PGF_2α+P4 removal-1d-PGF_2α-2d-GnRH+TAI), were inseminated with sex-sorted (n = 113) or conventional semen (n = 88). Statistical analyses were performed using PROC GLIMMIX procedure of SAS 9.4 (SAS Institute Inc., Cary, NC). Overall P/AI was 60.7% for EST and 54.2% for TAI regardless of types of semen and 68.1% for conventional and 48.9% for sex-sorted semen regardless of insemination strategies. Fertility of heifers inseminated with either sex-sorted (53.5%; 44.2%) or conventional (69.0%; 67.0%) semen did not differ between EST and TAI respectively. Besides, the interaction between the semen type and the insemination strategy was not significant for P/AI. The embryonic loss was significantly greater with sex-sorted semen (17.1%) compared to conventional semen (1.6%). There was no sire effect with sex-sorted semen on P/AI (52.6% vs. 46.2%) and embryonic loss (16.4% vs. 18.0%). As expected, sex-sorted semen resulted in more female calves (89.8% vs. 51.6%) than conventional semen. Thus, sex-sorted semen can be used with 5-day Cosynch+Progesterone protocol to eliminate the inadequate oestrus detection and to increase female calves born in dairy heifers.     

A novel strategy for resynchronization of ovulation in Nelore cows using injectable progesterone (P4) and P4 releasing devices to perform two timed inseminations within 22 days

We aimed to evaluate the reproductive performance of Nelore lactating cows submitted to a resynchronization 12 days after timed artificial insemination (TAI) with or without a long‐acting progesterone (P4‐LA) treatment. Nelore cows were submitted to a P4/oestradiol‐based TAI protocol (D0 = insemination). On D12, cows in the control group (n = 184) received a new P4 intravaginal device (0.96 g), whereas cows in the P4‐LA group (n = 192) received the P4 device and 75 mg P4‐LA. Cows identified as non‐pregnant (n = 120) by regression of corpus luteum using colour Doppler ultrasonography on D20 had the P4 device removed and received 500ug of sodium cloprostenol, 1 mg of oestradiol cypionate and 300 IU of eCG and were re‐inseminated on D22. There was no difference (p > 0.10) in the pregnancy rate at D20, D30 and D60 after first TAI between the control (69%, 59.7% and 57%, respectively) and P4‐LA (67%, 55.7%, and 55.2%, respectively) groups. Pregnancy losses were similar between both groups (p > 0.1). For cows submitted to the second TAI, the pre‐ovulatory follicle size did not differ (p > 0.1), but the oestrous detection and pregnancy rates were greater (p < 0.05) in the P4‐LA group (92.2% [59/64] and 60.9% [39/64], respectively) than in controls (75% [42/56] and 44.6% [25/56]). The cumulative pregnancy rate after two TAIs did not differ (p > 0.1) between control (73.3% [135/184]) and P4‐LA (76% [146/192]) groups. The use of P4‐LA at 12 days after TAI potentially increases the pregnancy rates for a new early resynchronization strategy associated with the Doppler imaging for pregnancy diagnosis and results in an alternative to perform two TAIs in 22 days in beef cows.     

The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares

Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to measure and compare the post-ovulatory rise in progesterone concentration, whereas all the mares were used to compare the length of the post-ovulatory oestrous behaviour between the first and subsequent cycles of the breeding season. The persistence of the post-ovulatory oestrus was longer (p < .001) following the first ovulation of the year (median of 52 h) compared with the subsequent ovulations (median of 36 h for second and later ovulations groups; n = 38 mares). The progesterone concentration at any of the four 8 h-intervals analysed (28, 36, 76 and 84 h post-ovulation) was lower (p < .01) following the first versus the second ovulation of the year. By 36 h post-ovulation the progesterone concentration of mares at the second ovulation of the year had passed the threshold of 2 ng/ml (2.1 ± 0.33 ng/ml), whereas in the first cycle it was 1.2 ± 0.13 ng/ml. In conclusion, mares had lower progesterone concentrations in their peripheral circulation and longer persistence of oestrous behaviour following the first ovulation of the year compared with the second and subsequent ovulatory periods of the breeding season.     

Reproductive performance of immobilized cryopreserved bovine semen used for timed artificial insemination

The SpermVital^® technology comprises embedding of spermatozoa within an alginate gel to facilitate release of sperm cells over a prolonged period in utero after AI. The aim of this study was to examine whether the survival time of spermatozoa is extended when applying this immobilization technology in combination with cryopreservation. Sperm cell survival (acrosome and plasma membrane integrity) was studied in vitro for 48 hr at physiological temperature. One dose of SpermVital^® (SV) semen was compared with single doses of Biladyl^® (B) processed semen as well as double doses of B (B double). B double was obtained by adding a second B dose the following day, thereby mimicking double AI. Furthermore, reproductive performance applying single early timed AI (TAI) with SV following oestrus synchronization was studied in a field trial. Double insemination (TAI on two consecutive days) with B semen served as control. Number of acrosome‐intact live sperm cells decreased over time in vitro for all treatments (p < .05). There was no difference between SV sperm cell survival and B double after 24 hr (p > .05). However, after 48 hr, SV sperm cell survival was higher than B double (p < .05). Moreover, multivariate analysis showed that the outcome of single early TAI with SV was not significantly different from B double (p > .05). Likelihood of pregnancy and calving in the heifer group was higher than in the cow group (p < .05). These results imply that spermatozoa immobilized in alginate gel have prolonged survival.     

Fertility of Angus cross beef heifers after GnRH treatment on day 23 and timing of insemination in 14‐day CIDR protocol

This study compared artificial insemination pregnancy rate (AI‐PR) between 14‐day CIDR‐GnRH‐PGF2α‐GnRH and CIDR‐PGF2α‐GnRH synchronization protocol with two fixed AI times (56 or 72 hr after PGF2α). On day 0, heifers (n = 1311) from nine locations assigned body condition score (BCS: 1, emaciated; 9, obese), reproductive tract score (RTS: 1, immature, acyclic; 5, mature, cyclic) and temperament score (0, calm; and 1, excited) and fitted with a controlled internal drug release (CIDR, 1.38 g of progesterone) insert for 14 days. Within herd, heifers were randomly assigned either to no‐GnRH group (n = 635) or to GnRH group (n = 676), and heifers in GnRH group received 100 μg of GnRH (gonadorelin hydrochloride, IM) on day 23. All heifers received 25 mg of PGF2α (dinoprost, IM) on day 30 and oestrous detection aids at the same time. Heifers were observed for oestrus thrice daily until AI. Within GnRH groups, heifers were randomly assigned to either AI‐56 or AI‐72 groups. Heifers in AI‐56 group (n = 667) were inseminated at 56 hr (day 32 PM), and heifers in AI‐72 group (n = 644) were inseminated at 72 hr (day 33 AM) after PGF2α administration. All heifers were given 100 μg of GnRH concurrently at the time AI. Controlling for BCS (p < .05), RTS (p < .05), oestrous expression (p < .001), temperament (p < .001) and GnRH treatment by time of insemination (p < .001), the AI‐PR differed between GnRH treatment [GnRH (Yes – 60.9% (412/676) vs. No – 55.1% (350/635); p < .05)] and insemination time [AI‐56 – 54.6% (364/667) vs. AI‐72 – 61.8% (398/644); (p < .01)] groups. The GnRH treatment by AI time interaction influenced AI‐PR (GnRH56 – 61.0% (208/341); GnRH72 – 60.9% (204/335); No‐GnRH56 – 47.9% (156/326); No‐GnRH72 – 62.8% (194/309); p < .001). In conclusion, 14‐day CIDR synchronization protocol for FTAI required inclusion of GnRH on day 23 if inseminations were to be performed at 56 hr after PGF2α in order to achieve greater AI‐PR.     

Comparative efficiency of novel laparoscopic and routine vaginal inseminations with cryopreserved semen in rabbits

Laparoscopic artificial insemination technique (LAI) is described to overcome reduced fertility problems in sheep artificial insemination (AI) programmes with frozen semen. Later on, this technology was modified for endangered non-domestic cats to deposit low quality or reduced number of sperm cells hardly obtained by electro-ejaculation into the oviduct. This technique by passes the complex structure of cervix and efficiently transfers the sperm cells to the point of fertilization. In recent years, rabbits are becoming popular transgenic animal models producing various therapeutic and commercial products, as well as being experimental animals for disease models. The worldwide transportation of frozen semen and re-establishment of transgenic lines using AI technology has become a common practice. Therefore, this study was designed to describe a laparoscopic intrauterine insemination technique, which might assist in conceiving the animals with limited number of sperm cells. The female rabbits were laparoscopically (n = 22) or vaginally (n = 13) inseminated with frozen–thawed semen samples containing approximately 10 × 10⁶ motile sperm. The laparoscopic insemination technique provided higher pregnancy rate (45.5%) than vaginal insemination technique (7.7%) (p < .05). In conclusion, the described laparoscopic AI might be a new alternative technique, thus enabling limited or low-quality frozen sperm samples to establish pregnancy in rabbits.     

Effect of delaying the time of insemination with sex-sorted semen on pregnancy rate in Holstein heifers

The objective of the study was to evaluate the interval from onset of oestrus to time of artificial insemination (AI) to obtain the optimum pregnancy rate with sex-sorted semen in Holstein heifers. Heifers in oestrus were detected and inseminated only by using heat–rumination neck collar comprised electronic identification tag at the age of 13–14 months. Heifers (n = 283) were randomly assigned to one of three groups according to the timing of insemination at 12–16 hr (G1, n = 97), at 16.1–20 hr (G2, n = 94) and at 20.1–24 hr (G3, n = 92) after reaching the activity threshold. The mean duration of oestrus was 18.6 ± 0.1 hr, and mean peak activity was found at 7.5 ± 0.1 hr after activity threshold. The mean interval from activity threshold to ovulation was 29.4 ± 0.4 hr. The overall pregnancy per AI (P/AI) was 53.0% at 29–35 days and 50.9% at 60–66 days after AI. There was a significant reduction between G1 (13.8 ± 1.4 hr) and G3 (7.9 ± 1.4 hr) related to the intervals from AI to ovulation time. Sex-sorted semen resulted in significantly higher P/AI at 29–35 days when heifers inseminated in G3 (60.9%) after oestrus than those inseminated in G1 (49.5%) and G2 (48.9%). In terms of fertility, when the temperature–humidity index (THI) was below the threshold value (THI ≤65) at the time of AI, there was a tendency (≤65; 57.2% vs. > 65; 47.1%) for high pregnancy rate. There was no effect of sire on P/AI. In addition, the interaction of the technician with the time of AI was found significant, and three-way interaction of technician, sire and time of AI was tended to be significant on pregnancy rate. Thus, in addition to delaying the time of insemination (between 20.1 and 24 hr) after oestrous detection, THI and experienced technician were also found to be critical factors in increasing fertility with the use of sex-sorted semen in Holstein heifers.     

The effect of select seminal plasma proteins on endometrial mRNA cytokine expression in mares susceptible to persistent mating‐induced endometritis

In the horse, breeding induces a transient endometrial inflammation. A subset of mares are unable to resolve this inflammation, and they are considered susceptible to persistent mating‐induced endometritis PMIE Select seminal plasma proteins cysteine‐rich secretory protein‐3 (CRISP‐3) and lactoferrin have been shown to affect the innate immune response to sperm in vitro. The objective of this study was to determine whether the addition of CRISP‐3 and lactoferrin at the time of insemination had an effect on the mRNA expression of endometrial cytokines in susceptible mares after breeding. Six mares classified as susceptible to PMIE were inseminated during four consecutive oestrous cycles with treatments in randomized order of: 1 mg/ml CRISP‐3, 150 μg/ml lactoferrin, seminal plasma (positive control) or lactated Ringer's solution (LRS; negative control) to a total volume of 10 ml combined with 1 × 10⁹ spermatozoa pooled from two stallions. Six hours after treatment, an endometrial biopsy was obtained for qPCR analysis of selected genes associated with inflammation (pro‐inflammatory cytokines interleukin (IL)‐1β, IL‐8, tumour necrosis factor (TNF)‐α, interferon (INF)‐γ, anti‐inflammatory cytokines IL‐1RN and IL‐10, and inflammatory‐modulating cytokine IL‐6). Seminal plasma treatment increased the mRNA expression of IL‐1β (p = .019) and IL‐8 (p = .0068), while suppressing the mRNA expression of TNF (p = .0013). Lactoferrin also suppressed the mRNA expression of TNF (p = .0013). In conclusion, exogenous lactoferrin may be considered as one modulator of the complex series of events resulting in the poorly regulated pro‐inflammatory response seen in susceptible mares.     

LRH-A3 and HCG increase pregnancy rate during timed artificial insemination in dairy cows

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.     

Evaluation of human chorionic gonadotropin as a replacement for gonadotropin-releasing hormone in ovulation-synchronization protocols before fixed timed artificial insemination in beef cattle

Two experiments were conducted during 2 yr to evaluate differences in ovulation potential and fertility in response to GnRH or hCG. In Exp. 1, 46 beef cows were given 100 microg of GnRH or 500, 1,000, 2,000, or 3,000 IU of hCG. Ovulation incidence was not different between GnRH and any of the hCG doses, indicating that ovulatory capacity of at least 500 IU of hCG was equivalent to GnRH. In Exp. 2, beef cows (n = 676) at 6 locations were assigned randomly to a 2 x 3 factorial arrangement of treatments. Main effects were: 1) pre-timed AI (TAI) treatment (GnRH or hCG) and 2) post-TAI treatment (saline, GnRH, or hCG) to initiate resynchronization of ovulation in previously inseminated cattle. Blood samples were collected (d -21 and -10) to determine progesterone concentrations and assess cyclicity. Cattle were treated with a progesterone insert on d -10 and with 100 microg of GnRH or 1,000 IU of hCG. A PGF(2alpha) injection was given at insert removal on d -3. Cows were inseminated 62 h (d 0) after insert removal. On d 26 after first TAI, cows of unknown pregnancy status were treated with saline, GnRH, or hCG to initiate a CO-Synch protocol. Pregnancy was diagnosed 33 d after first TAI to determine pregnancies per AI (P/AI). Nonpregnant cows at 6 locations in yr 1 and 1 location in yr 2 were given PGF(2alpha) and inseminated 56 h later, concurrent with a GnRH injection. Five weeks later, pregnancy diagnosis was conducted to determine pregnancy loss after first TAI and pregnancy outcome of the second TAI. Injection of pre-TAI hCG reduced (P < 0.001) P/AI compared with GnRH, with a greater reduction in cycling cows. Post-TAI treatments had no negative effect on P/AI resulting from the first TAI. Serum progesterone was greater (P = 0.06) 7 d after pre-TAI hCG than after GnRH and greater (P < 0.05) after post-TAI hCG on d 26 compared with saline 7 d after treatment in association with greater frequency of multiple corpora lutea. Compared with saline, injections of post-TAI GnRH and hCG did not increase second insemination P/AI, and inconsistent results were detected among locations. Use of hCG in lieu of GnRH is contraindicated in a CO-Synch + progesterone insert protocol. Compared with a breeding season having only 1 TAI and longer exposure to cleanup bulls, total breeding season pregnancy rate was reduced by one-third, subsequent calving distribution was altered, and 50% more AI-sired calves were obtained by applying 2 TAI during the breeding season.     

Effect of environmental factors on estrus synchronization and artificial insemination success in farmers flock in sheep under semi-arid tropical region

Environmental cues in the different seasons might influence the estrus and lambing percentages. The present study was conducted to assess the effect of environmental factors on estrus synchronization (ES) and artificial insemination success in sheep . During the period from 2012 to 2017, a total of 1,197 ewes of Kheri and Malpura breeds of 34 farms originating from 10 villages were synchronized for estrus and then fixed-time artificial insemination (FTAI) was done. Oestrous synchronization was done by intravaginal progesterone sponges and eCG protocol. Fixed-time cervical insemination was performed 48 and 56 hr after sponge removal in ewes exhibiting estrus, using liquid chilled semen containing 100 million sperm per dose of Patanwadi/Malpura rams. Mean sunshine hours, maximum and minimum ambient temperatures, temperature amplitude, mean relative humidity (RH), temperature-humidity index (THI), mean temperature during sponge-in to sponge-out—12-day period, sponge-out and next day, AI day and AI to next 15 days have been calculated. The estrus response and lambing percentage were higher (p < .05) when ES and FTAI were done during hot-humid with rainfall season. Successful (p < .05) ES occur when the minimum temperature was higher in cold-humid season, and the sunshine hour was higher in hot-humid with scanty rainfall season during sponge-in to sponge-out period. The success of the lambing percentage was higher (p < .05) when maximum temperature, minimum temperature, mean temperature and THI was higher on the day of AI and the next 15-day period, irrespective of the season. The success of FTAI indicates the adaptability of the local breed to tolerate the harsh climate of the hot semi-arid region without much hindrance in lambing percentages.     

The Post‐Cervical Insemination does not Impair the Reproductive Performance of Primiparous Sows

The study evaluated the reproductive performance of primiparous sows submitted to post‐cervical insemination (PCAI) compared with cervical artificial insemination (CAI). Difficulty with catheter introduction, the occurrence of bleeding or semen backflow during insemination, and volume and sperm cell backflow up to 60 min after insemination were also evaluated. Sows were homogenously distributed, according to body weight loss in lactation, lactation length, weaned piglets, weaning‐to‐oestrus interval and total born in previous farrowing, in two treatments: PCAI (n = 165) with 1.5 × 10⁹ sperm cells in 45 ml (2.4 ± 0.04 doses per sow) and CAI (n = 165) with 3 × 10⁹ sperm cells in 90 ml (2.5 ± 0.04 doses per sow). During PCAI, sows were inseminated in the absence of boars. Transabdominal real‐time ultrasonography was performed at oestrus onset, immediately before the first insemination and at 24 h after last insemination. There was no difference (P > 0.05) between treatments in farrowing rate (91.5% × 89.1%) and litter size (12.5 × 11.9 piglets born, respectively for PCAI and CAI sows). Successful passage of the intrauterine catheter in all the inseminations was possible in 86.8% (165/190) of sows initially allocated to PCAI treatment. Difficulty of introducing the catheter in at least one insemination did not affect the reproductive performance of PCAI sows (P > 0.05). Bleeding during insemination did not affect (P > 0.05) the farrowing rate in both treatments, but litter size was reduced in CAI and PCAI sows (P ≤ 0.06). Percentage of spermatozoa present in backflow within 1 h after insemination was greater in CAI than PCAI sows (P < 0.01). More than 85% of primiparous sows can be successfully post‐cervical inseminated with doses containing 1.5 × 10⁹ sperm cells in the absence of the boar during insemination without impairing the reproductive performance.     

Use of two cloprostenol administrations 11.5 days apart efficiently synchronizes oestrus in photostimulated multiparous dairy goats in the non-breeding season

This study assessed the efficiency of synchronous oestrous induction by light programme followed by two doses of cloprostenol in acyclic Saanen goats of different parity orders. Primiparous (n = 22) and multiparous (n = 33) goats were subjected to 16 hr of light and 8 hr of darkness for 60 days (D0-D60), starting 10 days after the winter solstice. All goats received 120 µg cloprostenol doses on D130 (morning) and D141.5 (afternoon) (11.5 days apart). Oestrus behaviour, ovarian follicular dynamics and serum progesterone (P4) analyses were recorded from D0 to D174 at different intervals. Animals in oestrus after D141.5 were randomly assigned into two groups: assisted natural mating (NM) or artificial insemination (AI; 10–24 hr after oestrus onset with frozen-thawed semen). From D57 to D120, 89.0% of goats presented large follicles (5–8 mm) and P4 concentrations were subluteal from D0 to D120. More multiparous compared to primiparous goats (54.5%, 18/33 vs. 18.2%, 4/22) exhibited oestrus after both injections. More primiparous compared to multiparous goats (54.5%, 12/22 vs. 12.1%, 4/33) did not exhibit oestrus after any injection. A total of 35 goats (64%) were in oestrus after the second prostaglandin injection and were subjected to NM or AI. The conception rate was similar among primiparous (70.0%, 7/10) and multiparous (68.0%, 17/25) goats but the pregnancy rate differed, being 31.8% (7/22) and 51.5% (17/33), respectively. No interaction was found between parity order and P4 concentrations in does that became pregnant or not. Thus, the association between light programme (60 days, starting at the beginning of winter) and two cloprostenol administrations 11.5 days apart (starting 70 days after the end of the light treatment) resulted in sufficient synchronous oestrous response in multiparous acyclic Saanen goats to reach satisfactory fertility levels after both NM and AI.