Expression of Progesterone Receptor in the Utero‐tubal Junction After Intra‐uterine and Deep Intra‐uterine Insemination in Sows

The aim of this study was to investigate the expression of progesterone receptor (PR) in the utero‐tubal junction (UTJ) of sows at 24 h after intra‐uterine insemination (IUI) and deep intra‐uterine insemination (DIUI) compared with conventional artificial insemination (AI) in pigs. Fifteen multiparous sows were used: AI (n = 5), IUI (n = 5) and DIUI (n = 5). The sows were inseminated with a single dose of diluted semen during the second oestrus after weaning at 6–8 h prior to ovulation (AI: 3000 × 10⁶ spermatozoa, IUI: 1000 × 10⁶ spermatozoa and DIUI: 150 × 10⁶ spermatozoa). The UTJ was collected and subject to immunohistochemical staining using avidin‐biotin immunoperoxidase technique with mouse monoclonal antibody to PR. In the oviductal part of the UTJ, the intensity of PR in the tunica muscularis and the proportion of PR‐positive cells in the surface epithelium after DIUI were lower than AI (p < 0.05). The intensity and the proportion of PR‐positive cells between AI and IUI in all compartments of the UTJ did not differ significantly (p > 0.05). When comparing between tissue compartments, prominent staining was observed in the muscular layer of the UTJ. It could be concluded that the expression of PR in the UTJ prior to fertilization after DIUI with a reduced number of spermatozoa was lower than that after AI. This might influence sperm transportation and the fertilization process.     

Reproductive efficiency in naturally serviced and artificially inseminated beef cows

A study was conducted to compare conception rates in 71 Tuli and 86 Afrikander beef cattle bred using either artificial insemination (AI) or the bull. Animals were bred using either artificial insemination or natural service at Matopos Research Station. Animals were grouped into three groups of heifers (parity 0; P0), second calvers (parity 1; P1) and mature cows (parity 2; P2) before being randomly assigned to one of the two breeding methods. A binary logistic regression was used for statistical analysis where breeding method (AI vs natural service) was the treatment factor and conception rate was the measured response while breed, parity and last calving date were non-treatment factors. No significant differences were observed in conception rates between breeds (P > 0.05). However, the method of breeding animals, parity and calving interval affected (P < 0.05) conception rates. The breeding method, parity and calving interval had a positive Kendall’s tau-b correlation coefficients to conception. More animals were pregnant when AI (77.6%) was used compared with natural mating (56.79%). Conception rates were significantly lower (P < 0.05) in C1 compared with C2 cattle. The odds ratio for breeding method and parity are positive and significant (P < 0.05). In conclusion, the study confirms that artificial inseminated animals had similar conception rates to naturally serviced animals for both Tuli and Afrikander breeds. As such, artificial insemination technology can be used to complement or substitute natural service in indigenous cattle’s of Zimbabwe.

     

Genistein Alters the Release of Oxytocin,Prostaglandins, Cortisol and LH during Insemination in Gilts

Soya products containing phytooestrogens are widely used as feed for pigs. However, limited data are available on the effects of phytooestrogen on the endocrine status of pigs. The aim of this work was to study the impact of the phytooestrogen genistein added to a soya‐free diet on the hormonal pattern in gilts during oestrus and artificial insemination (AI). Ten gilts were fed a soya‐free diet and fitted with jugular vein catheter through vena auricularis. The gilts were randomly divided into two groups (G‐ and C‐group) where the G‐group was given pure genistein, 1 mg/kg body weight (BW) twice daily, per os. Blood samples were collected before, during and after AI. Oxytocin, prostaglandin E_2, prostaglandin F_2α, 13,14‐dihydro‐15‐keto‐prostaglandin F_2α (PGFM), cortisol and LH concentrations in blood plasma were analysed. Oxytocin concentrations were almost twice as high in the G‐group as in C‐group after the AI. Prostaglandin E₂ concentrations were higher in G‐group than in C‐group during the entire sampling period. After AI, the concentrations of prostaglandin E₂ increased in G‐group but not in C‐group. Prostaglandin F_2α concentration had a pulsatile pattern, with increasing pulses after AI in G‐group. Plasma PGFM concentrations increased after AI with a small variation between the groups. Plasma cortisol concentration increased after AI in C‐group. LH decreased after AI in G‐group. Genistein stimulated elevations of plasma oxytocin and prostaglandin E₂ concentrations and a pulsative pattern in prostaglandin F_2α concentration. The possible involvement of genistein in plasma cortisol and basal LH concentrations in gilts given genistein may also be suggested.     

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.     

Detection of APAF1 mutation in Holstein cows and mummified foetuses in Japanese dairy herds

Some of the highest genetic merit sires have been shown to harbour recessive mutations affecting fertility, which may spread rapidly in the population through AI. These disorders may result in abortion and decline in pregnancy per insemination in cows. This study was carried out on 240 Holstein‐Friesian cows and 15 mummified foetuses. Blood and tissue samples were collected from the cows and mummified foetuses, respectively, for DNA extraction. Allele‐specific PCR was designed for the detection of the cows and foetuses carrying the nonsense mutation (C/T) in apoptosis peptide activating factor 1 gene (APAF1). The mutant allele frequency of the APAF1 in carrier cows and mummified foetuses was calculated. Milk samples were taken from the carrier and non‐carrier cows for progesterone assay. The allele‐specific PCR reaction efficiently distinguished the C/T mutation in APAF1. Of 240 cows, seven cows (2.9%) were diagnosed to carry one copy of the mutant allele of APAF1. However, the carrier frequency was 33.3% in mummified foetuses (five of 15). The mutant allele frequency was 0.02 and 0.17 in the cows and mummified foetuses, respectively. Concentrations of progesterone did not differ between cows with APAF1 mutation and non‐carrier cows during 45 days post‐insemination. This study provided allele‐specific PCR for the detection of APAF1 mutation in cows. Moreover, it reports the carrier and mutant allele frequencies of APAF1 in dairy cows and mummified foetuses in Japan.     

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.     

Evaluation of vaginal discharge with the Metricheck device and the relationship to reproductive performance in postpartum dairy cows

Vaginal mucus during estrus was examined with the Metricheck device and the relationship to the reproduction of high‐yielding dairy cows was studied. The study was conducted in 99 dairy herds located in Western Germany and 1348 Holstein‐Friesian heifers and cows showing spontaneous estrus were examined. Independent of the Metricheck result, the animals were inspected by professional insemination technicians and those suitable for insemination (n = 989) were bred by artificial insemination (AI). Reproductive performance was characterized by non‐return rate at 90 days (NRR90). The discharge of the animals predominantly had a clear appearance (70%) and a stringy consistency (80%). Animals with clear vaginal discharge had higher NRR90 (56%; n = 697) than animals with abnormal (turbid, mucopurulent, purulent, sanguineous) vaginal secretion (48%, n = 292; P < 0.05). NRR90 was lower in animals with short calving to AI interval (< 70 days; 39%) than with medium (70–130 days; 54%) or long (> 130 days; 62%) intervals (P < 0.05). NRR90 decreased by 12% from the lowest (< 15 kg) to the highest (> 45 kg) milk yield class. In conclusion, the use of the Metricheck device integrated into the insemination procedure is recommended to identify dairy cows suffering severely from uterine disease.     

Low Dose Insemination in the Sow – A Review

Artificial insemination (AI) in pigs has been established for about four decades but ejaculates are still used insufficiently. Higher demand of semen for AI and new techniques that involve low sperm concentration require the optimization of insemination protocols. Based on the knowledge of the physiology of sperm transportation and events in the female genital tract prior to fertilization, new strategies are under development to minimize sperm losses. One goal is to deposit the semen into the uterine horn rather than into the proximal cervix. It was shown that the minimal number of spermatozoa necessary for surgical AI at the utero‐tubal junction (UTJ) were at least 1 × 10⁶ diluted in 0.5 ml of a special extender. Artificial insemination into the distal part of the uterine horn required about 1 × 10⁷ million sperm in 20 ml of extender. Meanwhile, first insemination devices for non‐surgical intra‐uterine AI are commercially available. Using similar sperm concentrations as for surgical AI, non‐surgical uterine insemination did not differ significantly from control inseminations in terms of pregnancy rate and litter size. With respect to the fertilizing capacities of their ejaculates, boars have to be selected more strictly for sperm quality parameters as most of the compensatory effects of sperm cells disappear in maximally extended semen samples.     

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.     

Neospora caninum Infection Does Not Affect the Fertility of Dairy Cows in Herds with High Incidence of Neospora‐associated Abortions

This present study, was designed to establish whether Neospora caninum infection affects the fertility of high‐producing dairy cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection in aborted foetuses. Assessment was made of 7518 artificial inseminations (AI) (applied to parous cows) performed in three herds. Of these inseminations, 2540 (33.8%) resulted in pregnancy; 34% of which corresponded to seronegative cows (2226 of 6556 AI performed in seronegative cows) and 32.6% to seropositive animals (314 of 962). Abortion occurred in 97 (30.1%) of the 314 pregnancies recorded in dams seropositive for N. caninum, while only 93 (4.2%) of the 2226 pregnancies in seronegative animals ended in abortion. Logistic regression analysis indicated no significant effects of N. caninum antibody titre, days in milk and milk production at insemination on fertility. Factors found to affect fertility were herd, season of insemination, lactation and insemination number, semen‐providing bull and AI technician. Our results indicate that N. caninum infection does not affect the fertility of high‐producing dairy cows.     

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.     

Evaluation of three hormonal protocols for anovulatory lactating cows under regulations restricting the use of estrogenic compounds

When European Union regulations restricted the use of estrogenic compounds in food‐producing animals, refined hormonal protocols were no longer applicable for anovulatory cows. However, Ovsynch and its adaptations are routinely and uniformly applied to all cows regardless of ovarian function. To evaluate their efficacy on anovulatory cows, 143, 147 and 144 anovulatory cows received Ovsynch, Presynch and G6G protocols, respectively. In comparison, 150 cyclic cows were bred without using a synchronized protocol. Results showed that cows in the Presynch group had luteolysis responding to the last prostaglandin F_2α (PGF_2α) injection greater than the Ovsynch group. The serous progesterone levels at the first gonadotropin‐releasing hormone of Ovsych and the last PGF_2α injection was greater in the G6G group than the other two hormonal treatment groups. Concentrations of Ca²⁺ and total protein in cervical mucus in all three hormone‐treated groups before artificial insemination (AI) were significantly different from the controls. The G6G group obtained a greater pregnancy rate compared with Ovsynch and Presynch, but significantly less than the controls. For open cows in the Ovsynch group, estrus rate within 24 days after the first AI was significantly less than the controls. In conclusion, the G6G treatment resulted to better reproductive performance in anovulatory cows.     

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 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.     

Characterization of the LH peak after short and long fixed‐time artificial insemination protocols in sheep raised in the tropics

The aim of this study was to evaluate the peak in luteinizing hormone (LH) and the pregnancy rate of sheep (Texel × Santa Inês) in the tropics using short‐ (6 days) and long‐term (12 days) progesterone protocols followed by artificial insemination (AI) both in and out of the breeding season. Experiment 1 was conducted within (IN) the breeding season (autumn, n = 36), and experiment 2 was conducted outside (OUT) of the breeding season (spring, n = 43). In each experiment, the sheep were divided into two groups (6 or 12 days) according to the duration of treatment with a single‐use progesterone release vaginal device (CIDR^®, Pfizer, São Paulo, SP, Brazil), and blood samples were collected from 10 animals per group every 4 hr to measure the LH and progesterone concentrations. In the spring, the characteristics of the LH peak did not differ between groups; but in the autumn, there were differences between groups at the beginning (G‐6 IN: 36.44 ± 5.46 hr; G‐12 IN: 26.57 ± 4.99 hr) and end of the LH peak (G‐6 IN: 46.22 ± 7.51 hr; G‐12 IN: 34.86 ± 8.86 hr). The results showed alterations in the LH peak during the breeding season only in the sheep undergoing the short‐term protocol.     

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%.     

Factors Affecting the First Service Conception Rate of Cows in Smallholder Dairy Farms in Bangladesh

The successful outcome of an insemination is a combination of both male and female fertility‐linked factors. We investigated the first service conception rate of cows at artificial insemination (AI) in the smallholder dairy farms in Bangladesh. Frozen straws were prepared from ejaculates of Bos indicus (n = 7) and Bos indicus × Bos taurus (n = 7) AI bulls. Fertility was determined from 6101 first services in cows that were performed by 18 technicians in four regions between April 2004 and March 2005. Pregnancy was diagnosed by rectal palpation between 60 and 90 days post‐insemination. The Asian version of Artificial Insemination Database Application (AIDA ASIA) was used for bulls‐, cows‐ and AI‐related data recording, and later retrieved for analysis. The mean ± SD number of inseminations performed from individual bulls and their conception rates were 436.0 ± 21.6 and 50.7 ± 1.9%, respectively. Logistic regression demonstrated body condition scores (BCS), heat detection signs, months of AI and their interactions had greatest effects (odds ratios: 1.24–16.65, p < 0.04–0.001) on first service conception rate in cows. Fertility differed (p < 0.02–0.001) between the regions, previous calving months, months of AI, BCS, parity and heat detection signs of cows. Inseminations based on mounting activity (n = 2352), genital discharge (n = 3263) and restlessness and/or other signs (n = 486) yielded a conception rate of 53.6%, 48.8% and 50.1%, respectively (p < 0.05). Conception rate between technicians ranged between 43.4% and 58.6% (p < 0.05). The days interval from calving to first service (overall mean ± SD = 153.4 ± 80.6) had relationship (p < 0.001) with BCS, months of previous calving and parity of the cows. Fertility at AI in smallholder farms can be improved by training farmers on nutrition and reproductive management of the cows.     

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.     

Predictors of beef calf temperament at weaning and its impact on temperament at breeding and reproductive performance

Two experiments were conducted to determine (i) factors influencing calf temperament at weaning, (ii) association between heifer–calf temperament at weaning and temperament at breeding and (iii) effect of heifer–calf temperament on pregnancy rate per artificial insemination (P/AI). In experiment 1, beef cows and their calves (n = 285) from three farms were used. Sire docility estimated progeny difference (EPD) score, birth type (normal or assisted), calf gender, calf behaviour (during 1st 4 weeks) and calf health status (until weaning) were recorded. Cows and calves were assigned a temperament score (0—calm; 1—excitable), and all cows were given a body condition score (BCS, 1–9; 1—emaciated; 9—obese) at weaning. Calf's illness (p < .05), low sire docility EPD score (p < .05), altered gait (p < .05), altered resting behaviour (p < .01), reduced/no play behaviour (p < .05) and cow excitable temperament (p < .001) increased calf excitable temperament at weaning. In experiment 2, replacement heifer–calves (n = 758) from 12 farms were assigned a temperament score at weaning and later at breeding. Blood from 40 calves at weaning and 31 heifers at initiation of synchronization (same animals) was collected by coccygeal venipuncture for determination of circulating cortisol and substance P concentrations. Heifers were assigned a BCS and reproductive tract score (RTS, 1–5; 1—immature, acyclic; 5—mature, cyclic), synchronized for fixed time AI, observed for oestrus and were artificially inseminated. Cortisol concentrations were increased in excitable heifer–calves compared to calm heifer–calves at weaning (p < .05), and substance P was increased in excitable compared to calm females both at weaning and breeding (p < .05). Low sire EPD docility score (p < .01), heifer–calf excitable temperament at weaning increased excitable temperament at breeding (p < .01). Controlling for BCS categories (p < .01), oestrous expression (p < .0001) and temperament at breeding by oestrous expression (p < .05), the calf's excitable temperament at weaning (p < .001) reduced P/AI (Calm, 62.7 (244/389) vs. Excitable, 53.4% (197/369); p < .01). In conclusion, selection of docile cows and sires with greater docility EPD score should be given consideration to reduce calf excitement. Temperament in beef female can be detected earlier in their life and could be used as a tool in the selection process and to improve their performances.     

Effects of Hysteroscopic and Uterine Body Insemination on the Presence of Selected Immune Cells in the Equine Endometrium

The effects of standard uterine body and hysteroscopic insemination on endometrial health were investigated. For this purpose, 33 mares were assigned to five different protocols: control (no insemination; n = 7), sham AI (sham uterine body insemination; n = 6), sham HysAI (sham hysteroscopic insemination; n = 7), standard AI (standard uterine body insemination, 300 × 10⁶ progressively motile sperms (PMS); n = 7) and HysAI (hysteroscopic insemination, 100 × 10⁶ PMS; n = 6). Sampling included uterine swabbing for microbiological examination, cytology for determination of polymorphonuclear neutrophils (PMNs) in the uterus, and endometrial biopsy collection for histology and characterization of endometrial immune cells on day 18 after ovulation (B1) as well as 8–10 hours (B2, day 20) and 72 hours after insemination (B3, day 23). Microbial contamination increased throughout the experiment in the sham insemination groups. Significant effects (P < .05) over time were detected for PMNs (cytology: sham HysAI, standard AI, and HysAI; histology: standard AI and HysAI), macrophages (immunohistochemistry: standard AI and HysAI) and T cells (immunohistochemistry: standard AI), showing an increase at B2 and a subsequent decrease toward baseline levels at B3. At B2, significant differences (P < .05) existed for PMNs (mean ± SEM) between control (1.3 ± 1.9%) and sham AI (2.2 ± 2.7%) versus standard AI (12.2 ± 4.7%) and for macrophages between control (4.1 ± 3.5%) and sham AI (2.5 ± 1.3%) versus standard AI (25.4 ± 15.8%). Thus, the cellular immune response of the endometrium depends on sperm deposition in the uterus and does not differ between hysteroscopic and standard uterine body insemination.