Successful transposition of sperm freezing, in vitro fertilization and artificial insemination methodologies in the Tadjik markhor species (Capra falconeri heptneri).

The objective of the present study was to transpose sperm freezing methodology from domestic goat to the Tadjik markhor (Capra falconeri heptneri) and to address the feasibility to develop IVP and artificial insemination using such frozen semen. Semen of different adult markhor males were successfully recovered by electro-ejaculation and were then frozen using caprine methodology. Frozen semen showed good survival rates at thawing and good fertility rates were assessed in heterologous in vitro fertilization system with goat oocytes. LOPU/IVF was applied for Tadjik markhor females allowing the first successful blastocyst production in vitro. In an applied program, we also transposed successfully intrauterine AI method with frozen/thawed semen to the Tadjik markhor.

Preovulatory follicular dynamics and ovulatory events following the use of GnRH 84 h after medroxyprogesterone acetate sponge removal in postpartum buffaloes.

Herein, we evaluated the effects of Gonadotropin hormone-releasing hormone (GnRH) administration 84 h after medroxyprogesterone acetate (MAP) sponge removal on follicular growth, ovulation timing, and pregnancy per artificial insemination (AI) in cosynchronized postpartum Nili Ravi buffaloes. In this study, 58 Nili Ravi postpartum buffaloes (DIM = 103 ± 1.64) were randomly divided into two treatment groups (n = 29/treatment): GnRH-TAI-84 and TAI-84. All buffaloes were administered a MAP sponge for seven days. Upon MAP sponge removal, all the subjects received prostaglandin F2α (PGF2α), and Timed AI (TAI) was performed 84 hours after sponge removal. In the GnRH-TAI-84 group, the buffaloes received GnRH alongside insemination, whereas in the TAI-84 group, the buffaloes were inseminated without GnRH administration. Follicle diameter and blood estradiol levels were measured every 6 h from 72-108 h after MAP sponge removal. The animals were checked for pregnancy using ultrasonography 40 days after AI. Animals subjected to the GnRH-TAI-84 protocol had a higher follicular growth rate and preovulatory follicle size than those in the TAI-84 group. The follicular diameter was also larger in animals that received GnRH-TAI-84 than in those that received TAI-84 90 and 96 h after MAP sponge removal. Buffaloes in the GnRH-TAI-84 group had lower estradiol concentrations at 90, 96, 102, and 108 h than those in the TAI-84 group. Ovulation in GnRH-TAI-84 buffaloes occurred 11 h earlier than that in buffaloes from the TAI-84 group. A shorter interval between AI and ovulation in GnRH-TAI-84 buffaloes (14 h vs. 25 h) led to greater pregnancies per AI (62% vs. 17%) compared to buffaloes from the TAI-84 group.

Failure of isoxsuprine HCl to increase sheep fertility after cervical versus laparoscopic AI using chilled semen at different durations.

To improve the fertility of cervical artificial insemination (AI) in sheep, we investigated isoxsuprine HCl usage on the cervical passage during cervical AI. We also compared cervical and laparoscopic AI fertility results of using chilled semen at different durations. Semen was collected from rams and diluted as 20 × 106 or 400 × 106 spermatozoa/straw for laparoscopic and cervical AI, respectively, and chilled to 4°C within 2 h. Sheep were inseminated with chilled semen for 8 or 24 h via the laparoscopic or cervical AI method. Moreover, some of the cervical inseminated sheep were injected intramuscularly with 0.5 mg/kg of isoxsuprine HCl 15 min before AI. As a result, the use of isoxsuprine HCl did not affect cervical transit and fertility. In addition, fertility was affected by the storage duration of the semen; laparoscopic AI was more successful than cervical AI in terms of fertility; if cervical AI is performed, the duration between semen collection and AI should be less than 8 h after chilling the semen at 4°C, and if laparoscopic AI is performed, the time between semen collection and insemination can be up to 24 h after chilling the semen at 4°C. Longer storage periods should be studied.

Carboxyfullerene C60 preserves porcine sperm by enhancing antioxidant capacity and inhibiting apoptosis and harmful bacteria.

This study used a porcine model to systematically investigate whether carboxyfullerene C60(CF-C60) can be used for sperm preservation. The results indicated that CF-C60 supplementation can preserve porcine sperm quality during storage at 17 °C. This effect was attributable to an improvement in the antioxidant capacity of sperm through a decrease in the reactive oxygen species (ROS) level. Additionally, CF-C60 can maintain mitochondrial function, inhibit sperm apoptosis through the ROS/Cytochrome C (Cyt C)/Caspase 3 signaling pathway, and mediate suppression of bacterial growth through the effects of ROS. Finally, the results of artificial insemination experiments indicated that insemination with CF-C60-treated sperm can increase the total number of offspring born and reduce the number of deformed piglets. Thus, CF-C60 is safe for use as a component of semen diluent for sperm storage.

The development of novel porcine sperm protective agents holds profound significance for improving fertility quality and promoting reproductive health. Excessive oxidative stress and bacterial contamination, leading to sperm apoptosis, are the 2 major factors affecting the decline of porcine sperm quality. Recently, CF-C60 has gained attention as an important nanocarbon derivative with strong antioxidant and antibacterial activity. However, the role and mechanism of CF-C60 in the preservation of mammalian sperm remain unknown. This study aimed to explore the important protective role of CF-C60 in porcine sperm.

Associations among the largest follicle, preovulatory estradiol concentrations, and predominant vaginal epithelial cells at the completion of hormonal ovarian stimulation for fixed-time artificial insemination in goats.

The objective of the present study was to investigate the association among the largest follicle (LF), preovulatory estradiol (E2), and predominant vaginal epithelial cell at the completion of hormonal ovarian stimulation for fixed-time artificial insemination (FTAI) in goats. Thirty-seven crossbred Boer does received gonadotropin-releasing hormone (GnRH) and intravaginal progesterone (P4)-releasing devices (day 0). On day 5, P4 devices were removed and does received prostaglandin F2α and equine chorionic gonadotrophin. On day 7, does received GnRH, and FTAI was undertaken. On day 7, does were divided into three groups, i.e. small-sized (3-3.9 mm; n = 5), medium-sized (4-4.9 mm; n = 8), and large-sized (≥5 mm; n = 24) according to the diameter of the ovarian LF; follicular characteristics (number and diameter) were identified, and blood samples and vaginal smears were collected. The average diameters of total antral follicles and LF and the percentage of superficial cell were greatest in large-sized LF does (p < .01). The average diameters of total antral follicle (r = .68) and LF (r = .71), number of preovulatory follicle (r = .58), and plasma E2 concentrations (r = .61) were positively correlated with the percentage of superficial cells (p < .01). The likelihood of a pregnancy outcome after the FTAI increased by 13.71 times in does with a greater average diameter of antral follicle, 14.18 times with emergence of a large preovulatory follicle, and 36.83 times with a higher percentage of vaginal superficial cells (p < .01). It was concluded that there is a relationship between the cell types of the vaginal epithelium, the diameters of the largest ovarian follicles, and the concentration of E2 in goats subjected to FTAI protocols.

Characterization of conception rate after embryo transfer in comparison with that after artificial insemination in dairy cattle.

Artificial insemination (AI) and embryo transfer (ET) are important in the reproduction of dairy cows. The conception rate after AI or ET is an essential indicator when selecting appropriate breeding methods. However, information on the environmental factors affecting ET conception rate when compared with AI is limited. We aimed to investigate environmental factors affecting ET conception rate and characterize the differences in environmental factors between AI and ET. Records of the first AI (n = 1,870,143) and ET (n = 29,922) from Holstein nulliparous, primiparous, and multiparous cows in Hokkaido, Japan, were analyzed using separate multivariable logistic regression models. For each breeding method, we grouped primiparous and multiparous cows according to milk yield at peak lactation (PY; < 25, 25-30, 30-35, ≥ 35 kg in primiparous, < 40, 40-45, 45-50, ≥ 50 kg in multiparous) and the interval from calving to first AI or ET (CFI/CFT; < 60, 60-79, 80-99, ≥ 100 d) to evaluate the effects of PY and CFI/CFT on conception rate. AI conception rate decreased with increasing PY in primiparous and multiparous cows, whereas ET conception rate did not decrease significantly. Additionally, the ET conception rate did not decrease even in primiparous and multiparous cows slightly earlier than 60 d in CFI/CFT when compared with those in CFI/CFT after 60 d, which differed from the AI conception rate. Collectively, breeding by ET leads to the avoidance of negative effects of high milk yield and calving on the conception rate, indicating that cows are fertile by ET within 60 d after calving.

Effects of supplementation of grazing Nellore cows with ß-carotene and vitamins A + D3 + E + biotin on follicle diameter, oestrus, establishment of pregnancy, and foetal morphometry.

The objectives of this experiment were to evaluate the effects of supplementation of Nellore (Bos indicus) cows with ß-carotene + vitamins A + D3 + E + biotin on body condition score (BCS), oestrus, pregnancy, and foetal morphometry. Lactating cows (n = 497) from two herds were balanced for BCS and calving period [early calving (EC); late calving (LC)] and were assigned randomly to: Control (n = 251)-supplementation with a mineral supplement; and SUP (n = 246)-supplementation with the mineral supplement fed to control + ß-carotene (150 mg/day) + vitamin A (40,000 IU/day) + vitamin D3 (5000 IU/day) + vitamin E (300 mg/day) + biotin (20 mg/day). Cows were supplemented from Days -30 to 30 (Day 0 = timed artificial insemination; TAI). Pregnancy was diagnosed 30 days after TAI and foetal crown-rump distance and thoracic diameter were measured at 30 and 77 days of gestation. Cows in the SUP treatment were more likely to have BCS ≥3.0 on Day 0 (63.0 ± 3.1 vs. 60.2 ± 3.1; p < .01) and were more likely to gain BCS from Days -30 to 30 (57.7 ± 3.3 vs. 44.1 ± 3.3%; p < .01). Fewer LC cows in the SUP treatment were detected in oestrus at the time of the first TAI (Control: LC: 75.4 ± 4.4 vs. SUP: LC: 64.0 ± 5.2 vs. Control: EC: 65.3 ± 4.0 vs. SUP: EC: 71.8 ± 3.7; p = .04). There was a tendency for the SUP treatment to increase pregnancy to the first TAI (64.2 ± 3.0 vs. 56.6 ± 3.1%; p = .08). A greater percentage of SUP cows was detected in oestrus at the time of the second TAI (70.1 ± 5.0 vs. 52.3 ± 4.8%; p = .01). The SUP treatment increased pregnancy to the second TAI among LC cows (SUP: LC: 75.9 ± 8.0% vs. Control: LC: 50.0 ± 8.3% vs. Control: EC: 52.0 ± 5.9% vs. SUP: EC: 41.4 ± 6.5%; p = .02). The SUP treatment increased foetal size (crown-rump; p = .04 and thoracic diameter; p < .01) at 30 days of gestation and, despite decreasing crow-rump length at 77 days after the first TAI among EC cows (p < .01), it increased the thoracic diameter at 77 days after the first TAI independent of calving season. Our results support that pregnancy establishment and foetal growth can be improved when grazing Nellore cows are supplemented with ß-carotene and vitamins A + D3 + E + biotin.

Alterations in the rate of fertility, egg viability, and hatch parameters of adult geese exposed to different breeding methods.

In a 10-wk study, alterations in the rate of fertility, egg viability, and hatch parameters of adult geese exposed to different breeding methods were investigated. Twenty-four matured geese (4.0 ±â€…0.45 average weight) were randomly divided into three groups (TNM-natural mating group, TIM-artificial insemination group, TNI-natural mating and insemination group) of two replicates with four geese per replicate in a completely randomized design. Fresh semen collected from six ganders (5.2 ±â€…0.69 average weight) was pooled and used to inseminate the geese in TIM and TN1 at 0.2Ml at insemination times. The geese in TNM and TNI were allowed to mate naturally. Insemination and mating was done at 3 d interval and eggs from each treatment were collected daily. Incubation of eggs was done weekly, candling and transfer to hatcher were done on day 27 and goslings hatched out on day 30. Fertility, early embryo mortality (EEM), mid embryo mortality (MEM), late embryo mortality (LEM), hatch of fertile eggs (HOF), and hatch of set eggs (HOS) were obtained and analyzed using descriptive statistics and ANOVA and means separated using least significant difference test. Geese in TNI had significantly higher fertility (93.33 ±â€…10.97%) than TNM (59.67 ±â€…31.29%) and TIM (83.60 ±â€…17.14%). The EEM was higher in TIM than in the two other groups while the HOF and HOS were higher in TNM and TNI than in TIM. This study suggests that in comparison with TIM, higher fertility, hatchability, and lower embryo mortality can be obtained when geese are inseminated and naturally mated simultaneously.

Invasion of Chicken Anemia Virus in Specific-Pathogen-Free Chicken Flocks and Its Successful Elimination from the Colony.

A specific-pathogen-free (SPF) chicken colony was maintained with successive groups a month apart in age. The absence of specific pathogens, including chicken anemia virus (CAV), was confirmed through periodic serological tests for each group. However, some groups became CAV seropositive. The procedures of removing seropositive and the adjacent seronegative chickens followed with chemically disinfecting the housing did not halt CAV outbreaks. The full genome sequence of the CAV strain that appeared was closely related to low-virulence isolates in China. The outbreaks of CAV decreased with an increase in the seropositive chicken population, indicating that the progeny is protected from CAV infection by maternal anti-CAV antibodies. The persistence of CAV in erythroid and lymphoid tissues or reproductive tissues from CAV seropositive chickens was examined in chickens of various ages using polymerase chain reaction (PCR). Since a low persistence of CAV was observed in the colony, we isolated eggs from CAV seropositive hens through artificial insemination using semen collected from roosters and confirmed as CAV-free by PCR. Fertilized eggs were transferred to a new SPF facility and used for generating CAV-free progeny. To date, chickens reared in the new facility have been CAV-free for longer than two years. Redirection of eggs from seropositive hens was an effective means of eliminating CAV from chickens.

Application of antioxidants in extender on bull sperm cryopreservation to reduce the male effect in dairy fertility.

Because male and female effects on fertility must be considered, it may be difficult to achieve accurate and repeatable fertility predictions using only sperm characteristics given differences in breed, health, and season. Improving sperm quality after cryopreservation may be a method to reduce the male effect on the fertility outcome. This study was conducted using 2 different Certified Semen Service approved extenders, one containing plant-derived antioxidants, to assess cryopreserved sperm quality and determine pregnancy per artificial insemination (P/AI) in a commercial dairy farm. Beef bull ejaculates (n = 24 ejaculates, n = 4 bulls) were split and frozen in egg-yolk-TRIS (EYT) or GameteGuard-FB cryopreservation extenders. Frozen-thawed sperm was evaluated for sperm quality and used for insemination in dairy cows (n = 4856). There was an improvement in total and progressive motility, acrosome integrity, and reduction of DNA fragmentation at 0 and 3 h after thawing in sperm frozen in GameteGuard-FB when compared with the EYT extender. An overall 12.7% improvement in pregnancy per AI (P/AI) was observed when GameteGuard-FB (40.8%) was used for cryopreservation when compared with EYT (36.2%). More importantly, GameteGuard-FB frozen semen increased P/AI for first service by 19.5% (35.9% EYT and 42.9% GameteGuard-FB) and 45.0% was also observed at third service (31.3% EYT and 45.4% GameteGuard-FB). The use of GameteGuard-FB resulted in an average 40% P/AI among bulls. Taken together, the increase in cryopreserved sperm quality using GameteGuard-FB can be used to minimize the male effects on pregnancy outcomes in commercial dairy farms.

Pregnancy risk in beef and dairy cows after supplementing semen with transforming growth factor beta-1 at the time of artificial insemination.

Our objective was to determine if the addition of a concentrated human recombinant transforming growth factor beta-1 (TGF) to bovine semen at the time of AI would result in increased risk of pregnancy in beef and dairy cows. Suckled beef cows (n = 1,132) in 11 herds across 2 states and lactating dairy cows (n = 2,208) in one organic-certified herd were enrolled. Beef cows received fixed-time AI (FTAI) following a 7 d CO-Synch + controlled internal drug release estrous synchronization protocol. Dairy cows were inseminated following observation of natural estrus expression. Cows received either no treatment as a control (CON) or 10 ng of TGF in 10 µL added through the cut-end of a thawed straw of semen immediately prior to AI. At the time of FTAI of beef cows, the mean ±â€…SD age was 5.0 ±â€…2.4 yr, BCS was 5.3 ±â€…0.7, and days postpartum was 78.2 ±â€…15.5 d. The overall pregnancy risk (PR) in beef cows was 55.2% to AI and 90.5% season-long. PR in beef cows was not affected (P = 0.27) by the addition of TGF (53.1% vs. 58.1%). Furthermore, there was no difference (P = 0.88) for season-long PR in beef cows that received TGF (91.2% vs. 91.5%). At the time of insemination of dairy cows, the mean ±â€…SD lactation was 3.0 ±â€…1.3 lactations, BCS was 2.9 ±â€…0.3, days in milk was 115.6 ±â€…56.6 d, and cows had received 2.4 ±â€…1.5 inseminations/cow. The overall pregnancy risk to AI in dairy cows was 23.1%. PR to AI for dairy cows was not affected (P = 0.32) by addition of TGF (22.0% vs. 23.8%). In conclusion, PR to AI was not affected by addition of TGF to thawed semen immediately prior to AI in beef or dairy cows.

Seminal plasma is the fluid portion of the ejaculate that is routinely removed or significantly diluted when preparing semen for artificial insemination. Seminal plasma has been shown to elicit changes to the tissues of the uterus at the time of insemination that improves pregnancy outcomes in rodents and swine. Here, we supplemented the molecule of seminal plasma, transforming growth factor beta-1, to semen at the time of artificial insemination in an attempt to improve pregnancy rates in beef and dairy cattle. In total, 3,340 cows were inseminated; half received no treatment, and the other half received a supplementation of transforming growth factor beta-1. We found that supplementing transforming growth factor beta-1 did not improve the pregnancy rate in beef or dairy cattle. We conclude that the pregnancy rate was not affected by the supplementation of transforming growth factor beta-1 to semen at the time of insemination. Future studies should consider the effects of transforming growth factor beta-1 on other pregnancy outcomes, such as calving rate, birth weight, and postnatal growth.

Influence of early progesterone removal on follicular development, expression of estrus, and pregnancy rates in presynchronized postpartum beef cows.

The objective of this study was to evaluate the impact of early progesterone removal on pregnancy rates to fixed-time artificial insemination (FTAI) in presynchronized beef cows. Postpartum beef cows (n = 882) were randomly assigned to 1 of 2 treatments: 1) 7&7 Synch: cows received a controlled internal drug release insert (CIDR) and a 25-mg injection of prostaglandin F2α on day 0, 100 µg of GnRH on day 7, a second injection of prostaglandin F2α (PG2) at CIDR removal on day 14, and a second injection of GnRH at FTAI 60-66 h after PG2 (day 17); 2) 7&6 Synch: cows received the same treatment as 7&7 Synch; however, CIDR removal occurred in conjunction with PG2 on day 13, while FTAI remained at 60-66 h after CIDR removal (day 16). Ovarian ultrasonography was performed to determine follicle diameter at PG2 and FTAI in a subset of cows (n = 40). Cows exposed to the 7&7 Synch tended to have larger follicle diameter at PG2 compared with 7&6 Synch cows (P = 0.09); however, there were no differences in follicle diameter at FTAI. No differences were determined between treatments for the expression of estrus (7&7 Synch: 61.6 ± 5.30; 7&6 Synch: 54.1 ± 5.45; P = 0.31) or pregnancy rates to FTAI (7&7 Synch: 60.8 ± 3.83; 7&6 Synch: 57.0 ± 3.84; P = 0.42). In conclusion, early removal of progesterone did not impact pregnancy rates in presynchronized beef cows.

Effects of a simple method for determining the time of insemination and different methods on artificial insemination in common bottlenose dolphins (Tursiops truncatus).

In this study, we examined the usefulness of a simpler and more feasible method for determining the optimal timing of artificial insemination and the conditions for its success in six female common bottlenose dolphins. Pregnancy was successfully achieved in five dolphins by performing intrauterine insemination, using chilled semen stored for less than 3 days or frozen semen within 24 hr of exhibiting a peak serum estradiol (E2) level of 100 pg/mL or higher or on the day with a serum E2 level of approximately 100 pg/mL, measured with a simple measuring device. We concluded that the determining the optimal timing of intrauterine insemination by measuring serum E2 levels is a simpler and more useful method compared with the conventional approach.

Vaginitis with purulent vaginal discharge caused by artificial insemination using frozen Histophilus somni-contaminated semen.

In April 2020, two cows in Japan, developed reproductive disorders accompanied by vaginitis with purulent discharge within 3 days of artificial insemination (AI) with the same lot of frozen semen. Histophilus somni was isolated from the vaginal swabs of both cows as well as from the same lot of frozen semen used for the AI. This incident marks the first reported case of H. somni infection in cattle through AI. The major outer membrane protein gene sequences and pulsed-field gel electrophoresis profiles of the isolates were identical. Moreover, we investigated the antimicrobial activity of 12 frozen semen straws against an H. somni isolate using a disk diffusion test. These straws were sourced from five AI centers and included the same lot of semen used for the AI. Although the composition of semen diluents from individual AI centers is not publicly available, both the same lot of frozen semen used in the AI and other lots produced by the same manufacturer showed lower antimicrobial activity than semen from other manufacturers. These results strongly suggest that the two vaginitis were caused by AI using H. somni-contaminated frozen semen because of insufficient antimicrobial activity to inhibit bacterial growth. The minimum inhibitory concentrations of the six antimicrobials recommended for addition to frozen semen in isolates were below the recommended concentrations, suggesting that proper addition could have prevented this incident. This highlights the importance of conducting periodical checks on the antibacterial activity of frozen semen to prevent the transmission of pathogens via AI.

Equine chorionic gonadotropin (eCG) treatment in heifers: Double ovulation, twinning rate, and pregnancy losses in twin pregnancies.

This study aimed to evaluate the effects of different doses of equine chorionic gonadotropin (eCG; 200 and 300 IU) administered at the end of a fixed-time artificial insemination (FTAI) treatment protocol on ovulation, pregnancy, and twin rates in Bos taurus beef heifers. In addition, pregnancy losses in heifers with singleton and twin pregnancies were determined. A total of 2382 Angus heifers treated with a 6-day estradiol/progesterone-based protocol for FTAI (J-Synch protocol) were randomly allocated to two experimental groups to receive 200 or 300 IU of eCG administered intramuscularly at the time of intravaginal progesterone device removal; FTAI was performed from 60 to 72 h after device removal. The pregnancy rate did not differ (P = 0.89) between the 200 and 300 IU eCG groups. The number of corpus luteum induced by both eCG doses was determined by ultrasonographic examination 14 days after insemination and those treated with 300 IU of eCG had a greater double ovulation rate (P < 0.05). In addition, 300 IU eCG treated heifers had a higher twinning rate on day 30 of gestation (P < 0.05) and parturition (P < 0.05). Pregnancy losses from 30 days of gestation to calving did not differ between heifers treated with 200 and 300 IU of eCG (P = 0.70). However, regardless of the experimental group, heifers bearing twins had greater pregnancy losses than heifers with singletons (P < 0.05). In conclusion, reducing the dose of eCG from 300 to 200 IU under FTAI treatment protocol decreases double ovulation and twinning rates, maintaining a similar pregnancy rate in heifers. Nulliparous cows carrying two fetuses suffer greater pregnancy losses than cows with singletons.

The male effect associated with prostaglandins and reproductive outcomes in photo-stimulated Saanen goats during the non-breeding season.

The efficiency of combining oestrous induction via a light program (16 h of light and 8 h of darkness for 60 days, ending on Day 0 - D0) with cloprostenol administration, followed by the male effect or not, was tested in acyclic Saanen goats during the non-breeding season (June/2019 to January/2020). Initially, all animals (males and females) were submitted to the described light program; 60 days after its ending (D60), the females were divided into two groups, with (G1; n = 67) or without (G2; n = 61) a male effect from D60 to D75 after the light program. At D75, both groups received two cloprostenol doses (120 µg; intramuscular) 7.5 days apart (D75 and D82.5). Artificial insemination was performed at a specific time according to the oestrous onset (approximately 68.4 ± 1.2 h between the second cloprostenol dose and IA). Ultrasound scans were performed at different intervals to evaluate follicular dynamics and confirm pregnancy. At the first cloprostenol dose (D75), the proportion of does with at least a corpus luteum (CL), which indicates resumed cyclicity, was greater in G1 than in G2 (85.2% vs. 48.8%; p < .05), although no difference was found at the second dose (p > .05). The adjusted pregnancy rates (number of pregnant goats/oestrous goats) differed between G1 and G2 (21.7% vs. 42.0%; p < .05). G1 also showed a higher frequency of functional CL (based on blood flow and morphology) compared to G2 (96.9% vs. 66.7%; p < .05) at D116. A male effect using photo-stimulated bucks after the first cloprostenol dose increased the number of does presenting CL after buck removal, and no impairment in the pregnancy rates of multiparous does was found.

Establishment of the uterine microbiome following artificial insemination in virgin heifers.

Introduction: The concept of a sterile uterus was challenged by recent studies that have described the microbiome of the virgin and pregnant uterus for species including humans and cattle. We designed two studies that tested whether the microbiome is introduced into the uterus when the virgin heifer is first inseminated and whether the origin of the microbiome is the vagina/cervix. Methods: The uterine microbiome was measured immediately before and after an artificial insemination (AI; Study 1; n = 7 AI and n = 6 control) and 14 d after insemination (Study 2; n = 12 AI and n = 12 control) in AI and non-AI (control) Holstein heifers. A third study (Study 3; n = 5 Holstein heifers) that included additional negative controls was subsequently conducted to support the presence of a unique microbiome within the uterus despite the low microbial biomass and regardless of insemination. Traditional bacteriological culture was performed in addition to 16S rRNA gene sequencing on the same samples to determine whether there were viable organisms in addition to those detected based on DNA sequencing (16S rRNA gene sequence). Results and discussion: Inseminating a heifer did not lead to a large change in the microbiome when assessed by traditional methods of bacterial culture or metataxonomic (16S rRNA gene) sequencing (results of Studies 1 and 2). Very few bacteria were cultured from the body or horn of the uterus regardless of whether an AI was or was not (negative control) performed. The cultured bacterial genera (e.g., Bacillus, Corynebacterium, Cutibacterium, Micrococcus, Staphylococcus, and Streptococcus) were typical of those found in the soil, environment, skin, mucous membranes, and urogenital tract of animals. Metataxonomic sequencing of 16S rRNA gene generated a large number of amplicon sequence variants (ASV), but these larger datasets that were based on DNA sequencing did not consistently demonstrate an effect of AI on the abundance of ASVs across all uterine locations compared with the external surface of the tract (e.g., perimetrium; positive control samples for environment contamination during slaughter and collection). Major genera identified by 16S rRNA gene sequencing overlapped with those identified with bacterial culture and included Cutibacterium, Staphylococcus, and Streptococcus.

Acceptance and rejection of "morally challenging" behaviour in online sperm donation communities: narrative interviews with recipients and donors.

Introduction: Online sperm donation allows those hoping to conceive a baby ("recipients") to meet prospective sperm donors online, via "connection" websites or social media. These sites offer some advantages to clinical donation (including lower costs and greater choice over donation arrangements) but previous research has suggested that these sites may also pose risks and challenges to those who use them. Therefore, the aim of this exploratory research was to better understand online sperm donation communities and the experiences of both recipients and donors, particularly with respect to situations that could be "morally challenging" or involve harm. Methods: Three prolific donors and five recipients were interviewed using an in-depth narrative approach. Carol Gilligan's Listening Guide was employed to analyse the data. Results: The findings demonstrated that the donors sought to find ways to maintain autonomy in their donating practices and were concerned about the character and parenting abilities of recipients, as well as the potential for recipients to make unwarranted complaints. The recipients were concerned about their safety and finding a donor they could trust, discussing issues relating to donor "dishonesty", online abuse, and a lack of support from connection sites and related authorities. Both donors and recipients identified "morally challenging" behaviour relating to donor anonymity (donor use of fake online profiles or aliases) and the sexual motivations or (mis)conduct of some donors. The participants each discussed the ways in which they managed perceived risks. Discussion: The degree to which the participants voiced their acceptance or rejection of challenging behaviour in online sperm donation communities varied across and within participants, highlighting the complexity of the way in which people interact in this environment. Further research is required to understand how this form of sperm donation can be as safe and supportive as possible, while also respecting the importance to donors and recipients of autonomy and choice when making donation arrangements.

Inseminate without hormonal treatment in dairy sheep farms: exploring the consequences on the sustainability of several contrasted production systems.

Alternatives to hormonal treatments (HTs) in dairy sheep reproduction management are being explored in response to increasing societal concerns regarding animal welfare and food safety. However, hormone-free reproduction implies higher variability in flock performances and additional constraints for timely synchronised artificial insemination (AI) in the flock, impacting the diffusion of genetic progress. The use of the male effect, a well-known practice to induce synchronised oestrus, combined with precision tools (e.g., heat detector), is a plausible way to implement AI without HT in dairy sheep farms. To date, the consequences of such alternative reproduction management on the whole farm sustainability remain unknown. To anticipate these potential impacts, a multiagent model (REPRIN'OV) was used to simulate dairy sheep farms' sustainability indicators (biotechnical, economic, environmental and workload). A reproduction management scenario, including the use of the male effect followed by AI on the adult ewes (HFAI), was simulated and compared to the current reproduction management of four case study farms (Early_conv, Late_conv, Early_org and Late_org). They were selected to represent the different agricultural models (Conventional or Organic) and reproduction seasons (Early - during spring, out of ewes' natural reproduction season - or Late -from early summer to the end of autumn) of the Roquefort Basin's farms in Southern France. Simulation results showed that the HFAI scenario had different consequences depending on the farm's production system type. A negative effect on most key sustainability indicators of the Conv farms was observed, as a significant reduction in the fertility rate, in the proportion of young ewes born from AI (-54% in both farms; P < 0.05) and in the flock's milk production were observed; while the workload and greenhouse gas (GHG) emissions were increased compared to the initial scenario. In the Org farms, HFAI had neutral to positive effects on most indicators as the fertility, milk production of the flock, workload during milking and GHG emissions were barely affected by this scenario, while an increase in the proportion of young ewes born from AI was observed (+39% and + 43% in each farm, respectively; P < 0.05), allowing a better farm gross margin. Still, the workload during lambing was increased in Early_org (+18%; P < 0.05), as Early farms, tended to be more negatively impacted by HFAI than Late ones. Overall, our simulation approach provides interesting elements to exchange with stakeholders on how to progress towards a socially acceptable reproduction management system, for the dairy sheep sector.

Selection and direct biomarkers of reproductive capacity of breeding boars.

Efficient management of pig reproduction is paramount for the sustainability and productivity of the global pork industry. Modern artificial insemination (AI) breeding programs have greatly benefited from the integration of advanced selection methods and biomarkers to enhance the reproductive performance of boars. While traditional selection methods have relied soley on boar phenotype, such as growth rate and conformation, modern pig breeding has shifted more and more toward molecular and genetic tools, which are still complemented by phenotypic traits. These methods encompass genomics, transcriptomics, and proteomics. Biomarkers serve as critical indicators of boar reproductive capacity. They can help to identify individuals with superior fertility and aid in the early identification of potential fertility issues, allowing for proactive management strategies. This review summarizes current knowledge of various biomarkers associated with semen quality, sperm function, and overall reproductive fitness in boars. Furthermore, we explore advanced technologies and their potential applications in uncovering novel selection methods and biomarkers for predicting boar fertility. A comprehensive understanding of selection criteria and biomarkers governing boar reproductive capacity is essential for developing effective breeding programs to enhance swine reproductive performance.