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.     

Effects of month of breeding on reproductive efficiency of Holstein cows and heifers inseminated with sex-sorted or conventional semen in a hot environment

The main objective of this study was to assess the effect of month of breeding on reproduction performance of Holstein heifers and cows inseminated with sex-sorted or conventional semen in a hot environment. Pregnancy per artificial insemination (P/AI; 64,666 services over an 8-year period) both in heifers (n?=?22,313) and cows (n?=?42,353) from a large dairy herd in northern Mexico (26°N) were evaluated with the GENMOD procedure of SAS, with respect to month of AI. Overall, P/AI with sex-sorted semen was greater (P?<?0.01) in heifers (41.6 %) than cows (17.3 %). P/AI for cows serviced with conventional semen was 10 % points higher (P?<?0.01) in January and December (31 vs. 21 %) than cows serviced with sex-sorted semen. While there was no difference in P/AI between the sex-sorted sperm and conventional semen in cows inseminated in July (16 and 18 %, respectively), P/AI plummeted for both groups of cows during the summer and fall (more severe heat stress). P/AI was not different between heifers serviced with sex-sorted or conventional semen during the hottest months of the year (July to October). However, during the coldest month of the year (January and February), P/AI was 10 percentage points greater (P?<?0.01) in heifers serviced with conventional than sex-sorted semen. It was concluded that in this hot climate cow and heifer fertility declined in the summer and fall when inseminated with conventional semen. However, the use of sex-sorted semen during summer and fall did not compromise the breeding success in heifers. Thus, this data suggest that sex-sorted semen promotes some embryonic thermoprotective mechanism, which leads to a marginal summer and fall fertility depression with this type of semen in this particular hot environment.     

Fertility of Holstein cows and heifers submitted to timed artificial insemination and receiving one or two doses (12 h apart) of semen

The objective of this retrospective study was to assess the effect of receiving a single (n = 50,285) or double (n = 4392) artificial insemination (AI), 12 h apart, within a timed artificial insemination protocol on pregnancy per AI (P/AI) in nulliparous heifers (inseminated with either sex-sorted or conventional semen) and pluriparous Holstein cows in a commercial dairy herd. Also, this study aimed to investigate the relationship between temperature-humidity index (THI) and time of the first AI and fertility. Fertility of cows receiving two AI with normothermia (THI <68) was higher (p < .05) than cows receiving a single AI (42.9% vs. 36.4%). P/AI of cows receiving two AI with severe heat stress (THI >85) was higher (p < .05) than cows receiving a single AI (21.0% vs. 12.6%). Regardless of heat stress conditions, applying the first AI in the morning increased (p < .05) P/AI in cows with double AI than in cows whose first AI occurred in the afternoon (38.4 vs. 33.3%). With moderate heat stress, and sexed-sorted semen, P/AI to timed AI was higher (65.0 vs. 51.9%; p < .05) in heifers receiving double AI than those serviced once. It was concluded that double AI, 12 h apart, enhanced fertility at timed AI than herd mates with a single AI, particularly with heat stress at breeding.     

Fertility of lactating dairy cows inseminated with sex‐sorted or conventional semen after Ovsynch,Presynch–Ovsynch and Double‐Ovsynch protocols

The objective was to compare pregnancy per artificial insemination (P/AI) with conventional (CS) or sex‐sorted semen (SS) in dairy cows subjected to one of the three timed AI protocols. Cows (n = 356) were randomly assigned to synchronization with Ovsynch (OVS), Presynch–Ovsynch (PO) or Double‐Ovsynch (DO) and inseminated on Day 77 ± 3 postpartum with either frozen‐thawed SS (n = 182) or CS (n = 184) of the same bull. More cows were cyclic at the beginning of breeding Ovsynch increased (p < 0.01) with presynchronization and it was greater for DO than PO (OVS = 78.5%, PO = 85.1%, DO = 95.6%). Overall, P/AI for SS and CS increased with presynchronization (p < 0.05) on Days 31 (OVS = 35.5%, PO = 47.1%, DO = 48.3%) and 62 (OVS = 30.1%, PO = 43.8%, DO = 43.9%). Regardless of synchronization treatments, insemination with SS reduced P/AI (p < 0.02) on Days 31 (38.1% vs. 50.6%) and 62 (34.5% vs. 45.6%) compared with CS. No interaction was observed between synchronization treatment and type of semen for P/AI, although in cows receiving CS, P/AI was numerically greatest for PO (OVS = 42.0%, PO = 59.3%, DO = 49.0%), and in cows receiving SS, it was numerically greatest for those inseminated following DO (OVS = 27.9%, PO = 35.5%, DO = 47.6%). Thus, presynchronization improved P/AI in cows inseminated with sex‐sorted or conventional semen.     

Pregnancy and offspring sex ratio following insemination with SexedULTRA and conventional semen in cows in a commercial beef operation

Pregnancy rate per AI (PR/AI) and breeding season pregnancy rates between insemination with sexed semen (SS; at 18 hr after the onset of oestrus) and conventional semen (CS; at 12 hr after the onset of oestrus,) and offspring gender ratio between two groups were compared. Angus cross cows (n = 686, during 2019 and 2020 breeding seasons) were oestrus-synchronized using Select-Synch + CIDR protocol and were observed thrice daily for oestrus until 72 hr after PGF2α administration. Cows expressed oestrus (n = 513) were inseminated with either SS (n = 246; SexedULTRA 4M™; y chromosome-bearing sperm) or CS (n = 267). Cows (n = 173) that failed to express oestrus at 72 hr after PGF2α received 100 μg of GnRH and CS insemination concomitantly. Two weeks later, cows were penned with natural service sires (bull:cow ratio 1:25) for 45 days. Pregnancy was diagnosed 30 days after bull removal. Calves' gender was determined at birth. For cows that expressed oestrus, PR/AI did not differ (p > .1) between SS (65.0%) and CS (66.7%) groups. The overall PR/AI differed (p < .05) between SS (65.0%) and CS (56.4%) groups. The natural service PR differed (p < .001) but breeding season PR (p > .05) did not differ between SS vs. CS groups. Bull:heifer gender ratio following AI was 88:12 and 52:48 for SS and CS groups, respectively, with an overall 66:34 ratio. Bull:heifer gender ratio for the two breeding seasons was 79:21 and 52:48 for SS and CS groups, respectively, with an overall 62:38 ratio. In conclusion, the fertility of SS insemination at 18 hr after onset of oestrus was 97% of CS insemination at 12 hr after onset of oestrus. Though breeding season pregnancy did not differ between SS and groups, preferred calf gender was 25 percentage points greater for SS over CS application. The gender accuracy was 88%.     

Fertility in Single‐ovulating and Superovulated Dairy Heifers after Insemination with Low Dose Sex‐sorted Sperm

The present study was performed to test fertility in single‐ovulating and superovulated dairy heifers after insemination with low dose sex‐sorted sperm under field conditions. Some parameters, including the dosage, deposition site and timing, were assessed with the pregnancy rates after artificial insemination (AI). Moreover, the use of oestrus synchronization in combination with sorted sperm was evaluated. Besides that, we also improved the embryo production efficiency in superovulated dairy heifers by optimizing the timing of inseminations and repartitioning the sexed sperm dosage among multiple inseminations. The conception rate (52.8%) in heifers after low dose (2 × 10⁶) insemination with sorted sperm deep into the uterine horn did not differ (p > 0.05) from that (59.6%) of conventional AI (1 × 10⁷ non‐sorted sperm) and that of deep insemination with low dose non‐sorted sperm (57.7%). There was also no difference (p > 0.05) between conception rates after single (51.7%) and double (53.8%) deep insemination with sorted semen. Heifers inseminated with sorted sperm at synchronous oestrus had a lower pregnancy rate (48.1%) than heifers at spontaneous oestrus (53.6%), but this did not reach statistical difference (p > 0.05). The average number of transferable embryos collected in vivo from heifers inseminated with sorted sperm (4.81 ± 2.04) did not differ (p > 0.05) from that obtained from heifers after insemination with non‐sorted sperm (5.36 ± 2.74). Thus, we concluded that the pregnancy rate after deep intra‐uterine insemination with low dose sorted sperm was similar to that of non‐sorted sperm, which was either also deposited at a low dose deep intra‐uterine or into the uterine body. Sychronization of oestrus can be beneficial in combination with sorted sperm to optimize the organization and management of dairy herds. The results from superovulated heifers demonstrated that our insemination regime can be used to obtain a comparable embryo production efficiency with sorted sperm than with non‐sorted sperm.     

Effect of sexed semen,puberty and breeding ages on fertility of Holstein dairy heifers treated with double Ovsynch protocol

The objective of this study was to evaluate the fertility of sexed semen compared with conventional semen with regard to the puberty and breeding ages of Holstein dairy heifers subjected to double Ovsynch protocol with fixed time of artificial insemination. A total of 468 Holstein heifers were divided into two groups. The first group was 122 dairy heifers inseminated via conventional semen, while the second group was 346 heifers inseminated with sexed semen. The puberty and breeding ages of heifers were determined from the farm records. Estrus was synchronized using the double Ovsynch protocol. Numbers were estimated for pregnancy at 40 and 60 days post insemination, embryonic loss, and abortion. The results revealed that the heifers inseminated with sexed semen had a significantly lower first-service pregnancy rate (51.45%) than those inseminated with conventional semen (61.47%). Heifers achieving puberty before 350 days old had a higher pregnancy rate. Embryonic losses and abortion rates did not differ between the two types of semen. Holstein heifers subjected to Ovsynch protocol with sexed semen had an acceptable first-insemination pregnancy rate. Even the applications of sexed semen reduce the reproductive fertility and pregnancy rate in Holstein heifers.

     

Timed artificial insemination in crossbred mares: Reproductive efficiency and costs

Timed artificial insemination (TAI) has boosted the use of conventional artificial insemination (CAI) by employing hormonal protocols to synchronize oestrus and ovulation. This study aimed to evaluate the efficiency of a hormonal protocol for TAI in mares, based on a combination of progesterone releasing intravaginal device (PRID), prostaglandin (PGF_2α) and human chorionic gonadotropin (hCG); and compare financial costs between CAI and TAI. Twenty-one mares were divided into two groups: CAI group (CAIG; n = 6 mares; 17 oestrous cycles) and TAI group (TAIG; n = 15 mares; 15 oestrous cycles). The CAIG was subjected to CAI, involving follicular dynamics and uterine oedema monitoring with ultrasound examinations (US), and administration of hCG (1,600 IU) when the dominant follicle (DF) diameter's ≥35 mm + uterine oedema + cervix opening. The AI was performed with fresh semen (500 × 10⁶ cells), and embryo was recovered on day 8 (D8) after ovulation. In TAI, mares received 1.9 g PRID on D0. On D10, PRID was removed and 6.71 mg dinoprost tromethamine was administered. Ovulation was induced on D14 (1,600 IU of hCG) regardless of the DF diameter's, and AI was performed with fresh semen (500 × 10⁶ cells). On D30 after AI, pregnancy was confirmed by US. The pregnancy rate was 80.0% in TAIG and 82.3% in CAIG (p > .05). The TAI protocol resulted in 65% reduction in professional transport costs, and 40% reduction in material costs. The TAI was as efficient as CAI, provided reduction in costs and handlings, and is recommended in mares.     

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.     

Oestrus detection techniques and insemination strategies in Bos indicus heifers synchronised with norgestomet-oestradiol

SUMMARY Oestrus was synchronised in 57 Bos indicus heifers using norgestometoestradiol and pregnant mare serum gonadotrophin. Oestrus was detected by observations made at six-hourly intervals, using oestrogen-treated and chin-ball harnessed steers, heatmount detectors, tail-paint and visual observation. Heifers were inseminated once at either a fixed time of 49.2 ± 0.4 h (mean ± SE; n = 29) after implant removal or 12.6 ± 1.5 h (n = 28) after oestrus was detected. The mean (± SE) time to the onset of oestrus was 47.1 ± 1.9 h, while 90% of heifers recorded in oestrus were detected within 66 h of implant removal. Heatmount detectors were significantly more efficient at detecting oestrus than chin-ball harnessed steers, tail paint or visual observation (P < 0.001). A higher pregnancy rate was obtained in heifers inseminated after oestrus detection compared with heifers inseminated at a fixed-time (57.1 vs 34.5%; P = 0.043) and a higher pregnancy rate was obtained in heifers classified as easy to inseminate compared with heifers classified as difficult to inseminate (57.8 vs 0%, P < 0.001). We conclude that heatmount detectors are an efficient means of detecting oestrus in synchronised B indicus heifers and that pregnancy rates can be increased when insemination follows oestrus detection compared with a fixed-time insemination regimen.     

Artificial insemination of Holstein heifers with sex-sorted semen during the hot season in a subtropical region

Our aim was to investigate insemination techniques in order to improve pregnancy rates of artificial insemination (AI) using sex-sorted semen (sexed AI) in cattle in tropical and subtropical (T/ST) regions. In T/ST regions, the pregnancy rates by sexed AI are reportedly the lowest in the hottest months of the year, with less than 15% in cows and 35–40% in heifers (PMID 24048822). We compared sexed AI by depositing the semen into the uterine body (UB-AI, n = 12) versus the unilateral uterine horn (UUH-AI, n = 14) of pre-ovulation heifers. The ovary and follicle were assessed by rectal ultrasound before AI. After insemination, pregnancy was determined by ultrasound at approximately 40 days and approximately 70 days. In the present study, we demonstrated that high pregnancy rates (>70%) by sexed AI in the hottest season in a subtropical region such as Taiwan can be achieved when heifers with pre-ovulation follicles are used. The overall pregnancy rates were 54% higher in the UUH-AI (71%) group than in the UB-AI (42%) group (P = 0.06), examined on approximately 40 days post-sexed AI. Surprisingly, however, the pregnancy outcome appeared to be higher in the hot season (62%) than in the cool season (46%) although this difference was not statistically significant. Based on the present study, we recommend that cattle breeders perform UUH-AI using sex-sorted semen for heifers with pre-ovulation follicles in order to achieve satisfactory pregnancy outcome in the hot seasons in T/ST regions.     

Artificial insemination of Bos indicus heifers: The effects of body weight, condition score, ovarian cyclic status and insemination regimen on pregnancy rate

SUMMARY Effects of body weight, condition score, ovarian cyclic status and insemination regimen on pregnancy rates were investigated in 164 Bos indicus heifers synchronised with norgestomet-oestradiol and pregnant mare serum gonadotrophin (PMSG). Oestrus detection techniques were also compared. Heifers were inseminated at either a fixed time (group 1, n = 83) of 48.0 ± 0.2 h (mean ± SEM) after implant removal or at 8.9 ± 0.5 h after oestrus was detected (group 2, n = 81). Group 2 heifers that were not detected in oestrus by 72 h after implant removal were inseminated at that time. Oestrus was detected for the purpose of insemination using heatmount detectors. Tail-paint and oestrogen treated, chin-ball harnessed steers were used to compare the efficiency of oestrus detection. The probability of ovarian cyclicity increased with increasing body weight and condition score (P < 0.001). A higher proportion of heifers that were acyclic at the commencement of treatment, compared with cyclic heifers, were detected in oestrus at the time of insemination in the fixed-time inseminated group (P <0.01). Analysis of covariance revealed that intervals from implant removal to oestrus were influenced by ovarian cyclic status (P < 0.01) and insemination group (P < 0.05). A higher pregnancy rate (%± SEM) was obtained in acyclic compared with cyclic heifers in the group 1 heifers (50.0 ± 10 vs 28.1 ± 6; P = 0.055) but not among the group 2 heifers (45.8 ± 10 vs 49.1 ± 7; P = 0.787). The probability of pregnancy was found to be associated negatively with body weight (P = 0.01) while a higher pregnancy rate was obtained in the group 2 compared with group 1 heifers (48.2%vs 34.9%; P = 0.093). The efficiency of oestrus detection was highest using heatmount detectors compared with tail-paint and chin-ball harnessed steers (90.7%vs 37.0% and 23.5%, respectively; P < 0.0001). We conclude that pregnancy rates can be increased in extensive environments when insemination follows oestrus detection using heatmount detectors compared with a fixed-time insemination. The fertility of heifers inseminated at a fixed time is influenced by ovarian cyclic status due to its influence on oestrus-to-insemination intervals.     

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.     

Influence of Temperament Score and Handling Facility on Stress,Reproductive Hormone Concentrations,and Fixed Time AI Pregnancy Rates in Beef Heifers

The objectives were (i) to evaluate the effect of temperament, determined by modified 2‐point chute exit and gait score, on artificial insemination (AI) pregnancy rates in beef heifers following fixed time AI and (ii) to determine the effect of temperament on cortisol, substance‐P, prolactin and progesterone at initiation of synchronization and at the time of AI. Angus beef heifers (n = 967) at eight locations were included in this study. At the initiation of synchronization (Day 0 = initiation of synchronization), all heifers received a body condition score (BCS), and temperament score (0 = calm; slow exit and walk or 1 = excitable; fast exit or jump or trot or run). Blood samples were collected from a sub‐population of heifers (n = 86) at both synchronization initiation and the time of AI to determine the differences in serum progesterone, cortisol, prolactin and substance‐P concentrations between temperament groups. Heifers were synchronized with 5‐day CO‐Synch+ controlled internal drug release (CIDR) protocol and were inseminated at 56 h after CIDR removal. Heifers were examined for pregnancy by ultrasound 70 days after AI to determine AI pregnancy. Controlling for synchronization treatment (p = 0.03), facility design (p = 0.05), and cattle handling facility design by temperament score interaction (p = 0.02), the AI pregnancy differed between heifers with excitable and calm temperament (51.9% vs 60.3%; p = 0.01). The alley‐way with acute bends and turns, and long straight alley‐way had lower AI pregnancy rate than did the semicircular alley‐way (53.5%, 56.3% and 67.0% respectively; p = 0.05). The serum hormone concentrations differed significantly between different types of cattle handling facility (p < 0.05). The cattle handling facility design by temperament group interactions significantly influenced progesterone (p = 0.01), cortisol (p = 0.01), prolactin (p = 0.02) and substance‐P (p = 0.04) both at the initiation of synchronization and at the time of AI. Inter‐ and intra‐rater agreement for temperament scoring were moderate and good (Kappa = 0.596 ± 0.07 and 0.797 ± 0.11) respectively. The predictive value for calm and pregnant to AI was 0.87, and excited and non‐pregnant to AI was 0.76. In conclusion, the modified 2‐point temperament scoring method can be used to identify heifers with excitable temperament. Heifers with excitable temperament had lower AI pregnancy. Further, cattle handling facility design influenced the temperament and AI pregnancy.     

Methods to reduce or eliminate detection of estrus in a melengestrol acetate-PGF2alpha protocol for synchronization of estrus in beef heifers

Three experiments were conducted to evaluate methods to decrease or eliminate the detection of estrus inherent to a melengestrol acetate (MGA)-PGF2alpha (PGF) protocol for synchronization of estrus in heifers. In each experiment, all heifers received 0.5 mg of MGA x animal(-1) x d(-1) for 14 d (d -32 to -19) and PGF (25 mg, i.m.; d 0, 0 h) 19 d after the last feeding of MGA (MGA-PGF protocol). In Exp. 1, heifers (n = 709) were assigned to each of the following protocols: 1) the MGA-PGF protocol with AI 6 to 12 h after detection of estrus (estrus AI; MGA-PGF); 2) MGA-PGF plus 100 microg, i.m. of GnRH on d -7 (1x GnRH) and estrus AI; or 3) MGA-PGF, GnRH on d -7, and GnRH (100 microg, i.m.) at 48 h after PGF, coincident with insemination (2x GnRH-TB48). In Exp. 2, heifers (n = 559) received the MGA-PGF protocol and were inseminated by either estrus AI or fixed-time AI (TAI) at 60 h, coincident with an injection of GnRH (GnRH-TB60). In Exp. 3, all heifers (n = 460) received the MGA-PGF protocol and were inseminated by estrus AI when detected up to 73 h. Heifers not observed in estrus by 73 h received TAI between 76 and 80 h. Half the heifers inseminated by TAI received no further treatment (TB80), and the remaining half was injected with GnRH at insemination (GnRH-TB80). Variance associated with the interval to estrus and the proportion in estrus from d 0 to 5 was similar for 1x GnRH and MGA-PGF treatments in Exp. 1. Pregnancy rate (d 0 to 5) did not differ for the MGA-PGF and 1x GnRH treatments (62.5 and 60.4%, respectively), and both were greater (P < 0.05) than TAI pregnancy rate in the 2x GnRH-TB48 treatment (42.3%). In Exp. 2, the peak estrous response occurred 60 h after PGF. Pregnancy rate during the synchrony period was greater (P < 0.05) for the MGA-PGF (255/401; 63.6%) than the GnRH-TB60 (74/158; 46.6%) treatment. In Exp. 3, 75.7% of heifers (348/460) were detected in estrus by 73 h and were inseminated, with a conception rate of 74.4%. Pregnancy rates after TAI did not differ between TB80 and GnRH-TB80 (14/56 = 25% and 19/ 56 = 33.9%, respectively). Total pregnancy rate was 63.5% for heifers inseminated after detected estrus and by TAI. Collectively, these data indicate that the exclusive use of TAI for heifers treated with the MGA-PGF protocol resulted in lower pregnancy rates than when AI was performed after detection of estrus. However, estrus AI for 3 d and TAI at the end of d 3 could result in pregnancy rates similar to those achieved after a 5-d period of detecting estrus.     

Timing of Ovulation and Fertility of Heifers After Synchronization of Oestrus with GnRH and Prostaglandin F2α

Synchronization of oestrus and/or ovulation can reduce workload in heifer reproductive management. The objective of this study was to compare two protocols to synchronize oestrus and/or ovulation using GnRH and prostaglandin F_2α (PGF_2α) in dairy heifers concerning their effect on follicular dynamics and reproductive performance. Four trials were carried out. In trial 1, 282 heifers were treated with GnRH and PGF_2α 7 days apart (GP protocol). One group was inseminated on detection of oestrus (IDO 1), and the other group received two timed artificial inseminations (AI) 48 and 72 h after PGF_2α administration (TAI 1). In trial 2, 98 heifers were synchronized with the same GP protocol. Heifers in IDO 2 were treated as in IDO 1, heifers in TAI 2 received two TAI 48 and 78 h after PGF_2α administration. In trial 3, heifers in IDO 3 (n = 71) were again treated as in IDO 1. Heifers in TAI 3 (n = 166) received a second dose of GnRH 48 h after PGF_2α (GPG protocol) and TAI together with this treatment and 24 h later. Trial 4 compared the timing of ovulation after the GP and the GPG protocol, using a subgroup of the heifers from trials 1 to 3. The ovaries of the heifers were scanned via ultrasound at 48, 56, 72, 80, 96 and 104 h after PGF_2α administration. Timing of ovulation and size of the ovulatory follicles were compared between the two groups. In trials 1 to 3, conception rates to first service were between 49 and 66%. They did not differ significantly between IDO and TAI groups within or between trials. Pregnancy rates per synchronization were numerically higher in the TAI groups, but the difference was not significant. Conception rates to breeding on spontaneous oestrus in heifers returning to oestrus were higher than that after synchronized oestrus. In trial 4, more heifers ovulated before the end of the observation period in GPG than in GP (96.5% vs 74.7%; p < 0.001). Overall, ovulatory follicles were smaller in GPG (13.1 ± 1.9 mm vs 14.3 ± 1.9 mm; p < 0.001).     

Relationship of the conception rate and the side (left or right) of preovulatory follicle location at artificial insemination in dairy heifers

In this study, we examined the locational effect (left or right ovary) of the preovulatory follicle (PF) on fertility in dairy heifers. In total, 1,111 artificial inseminations (AI) were analyzed. At AI, PF locations were examined using rectal palpation, and heifers were divided into two groups on their PF locations: (i) the PF located in the left ovary (L‐PF); and (ii) the PF located in the right ovary (R‐PF). Pregnancy was diagnosed by rectal palpation 60 days after AI. The conception rate was 50.7% in all heifers. Conception rate was significantly higher in the L‐PF (60.1%) than in the R‐PF (46.2%). The conception rate was significantly lower by sexed semen (48.6%) than conventional semen (59.1%). Conception rates divided by the semen type (sexed: n = 896, conventional: n = 215) were significantly higher in the L‐PF than in the R‐PF for both semen types (sexed; L‐PF vs. R‐PF: 57.3% vs. 44.4%, conventional; L‐PF vs. R‐PF: 72.3% vs. 53.3%). In addition, season, age, AI number, and the number of re‐inseminations at the same estrus did not affect conception rates. In summary, PF development in the left ovary was associated with increased conception rates in dairy heifers.     

Fertility in Dairy Cows After Artificial Insemination Using Sex‐Sorted Sperm or Conventional Semen

The aim of this study was to compare pregnancy per artificial insemination (P/AI) after timed AI with sex‐sorted sperm (SS) or conventional semen (CS) in lactating dairy cows. Cyclic cows (n = 302) were synchronized by Ovsynch and randomly assigned into two groups at the time of AI. Cows with a follicle size between 12 and 18 mm and clear vaginal discharge at the time of AI were inseminated with either frozen‐thawed SS (n = 148) or CS (n = 154) of the same bull. A shallow uterine insemination was performed into the uterine horn ipsilateral to the side of probable impending ovulation. Pregnancy per AI on Day 31 tended (p = 0.09) to be less for SS (31.8%) than CS (40.9%). Similarly, P/AI on Day 62 was less (p = 0.01) for cows inseminated with SS (25.7%) compared with CS (39.0%). The increased difference in fertility between treatments from Days 31 to 62 was caused by the greater (p = 0.02) pregnancy loss for cows receiving SS (19.2%) than CS (4.8%). Cow parity (p = 0.02) and season (p < 0.01) when AI was performed were additional factors affecting fertility. Primiparous cows had greater P/AI than multiparous cows both on Day 31 (41.7% vs 25.0% in SS and 53.0% vs 31.8% in CS groups) and on Day 62 (33.3% vs 20.5% in SS and 48.5% vs 31.8% in CS groups). During the hot season of the year, P/AI on Day 31 was reduced (p = 0.01) in the SS group (19.6%) when compared with the rates during the cool season (38.1%). In conclusion, sex‐sorted sperm produced lower fertility results compared to conventional semen even after using some selection criteria to select most fertile cows.     

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.     

A new device to inseminate cows at the base of the uterine horns

A new device (Chapingo device) to deposit semen at the base of the uterine horns of cattle was developed at Universidad Autonoma Chapingo, Mexico. Nine Holstein heifers were inseminated by transvaginal laparoscopy, using a laparoscope for cattle and the Chapingo device. A dose of sexed semen (2.1 × 10⁶ spermatozoa) was deposited at the base of the uterine horn ipsilateral to the ovary where the preovulatory follicle was identified. Insemination was achieved in all the heifers, taking on average 13.7 ± 3.1 min per animal. In all cases, it was possible to see both ovaries, the base of the uterine horns and the oviducts. After the procedure, none of the heifers showed any type of complications such as haemorrhage, adhesions or trauma. On days 21 and 22 after insemination, four of the nine heifers (44.4%) returned into oestrus; on day 30 after insemination, one heifer was found to be pregnant by ultrasound. The results show the feasibility of generating pregnancies by transvaginal laparoscopy in heifers inseminated with sexed semen.