Comparative evaluation of estrus synchronization protocols on reproductive performance and estrus behavior in Barbados Black Belly sheep.

Background and Aim: Estrus synchronization of ewes has been accomplished using several protocols with various degrees of success in improving reproductive efficiency and obtaining the most effective protocol used in sheep farming. This study aimed to compare the effectiveness of three treatment protocols and to record the intensity and duration of estrus signs and pregnancy rate in Barbados Black Belly (BBB) sheep. Materials and Methods: Thirty-two primipara BBB ewes aged 18-24 months were equally divided into three treatment groups. T1: the ewes were injected intramuscularly with 2 mL Lutalyse (PGF2α) (10 mg) on days 0 and 10. T2: 1 mL Fertiline (50 µg; Gonadorelin acetate) on day 0 and 2 mL lutalyse (10 mg) on day 7. T3: 1 mL fertiline (50 µg) on day 0, 2 mL lutalyse (10 mg) on day 7, and 1 mL Fertiline (50 µg) on day 9. Estrus response was assessed using naturally mating rams and ewes. Pregnancy was determined using ultrasonography between 55 and 80 days after the last hormonal injection. The following estrus signs were noted: Swollen vulva, mucus discharge, sniffing, excitement, loss of appetite, mounting, and rapid tail movement. Results: Of the expressed signs, swollen vulva was most frequent, whereas loss of appetite and mucus discharge were the least overt signs recorded. The estrus response (%), onset (%), and duration (h) in ewe synchronization of the three treatment protocols were 100%, 58.3 ± 23.4%, and 48.0 ± 18.2 h (T1), 100%, 61.7 ± 41.2%, and 45.0 ± 27.0 h (T2), and 37.5%, 32.1 ± 1.7%, and 29.2 ± 1.25 h (T3), respectively. The pregnancy rates were 87.5%, 87.5%, 37.5%, and 50.0% in T1, T2, T3, and T4, respectively. Conclusion: Prostaglandin F2α+PGF2α and GnRH+PGF2α synchronization protocols were more effective in the fertilization of BBB ewes with better expression of estrus signs compared with the GnRH+PGF2α+GnRH (OVS) protocol.

Comparison of Gonadotropin-Releasing Hormone versus Estrogen-Based Fixed-Time Artificial Insemination Protocols in Grazing Bos taurus Suckled Beef Cows.

Fixed-timed artificial insemination (FTAI) protocols for beef cattle in South America are primarily based on estradiol esters and intravaginal progesterone-releasing devices (IVPD). The objective of this study was to determine the optimal gonadotropin-releasing hormone (GnRH)-based protocol as an alternative to the use of estrogen-based protocols in grazing Bos taurus suckling beef cows. All cows received an IVPD on the day of protocol initiation and prostaglandin F2α (PG) plus equine chorionic gonadotropin (eCG) treatments at the time of IVPD removal. In Experiment 1, cows (n = 235) were randomly assigned to one of four treatments: (i) 7-day estradiol = 2 mg of estradiol benzoate (EB) at IVPD insertion on Day 9 and 1 mg of estradiol cypionate (ECP) at IVPD removal on Day 2; (ii) 7-day GnRH = 10 µg of GnRH at IVPD insertion on Day 10, IVPD removal on Day 3 and GnRH at FTAI; (iii) 7 & 7 estradiol = PG at IVPD insertion on Day 16, EB on Day 9 and ECP at IVPD removal on Day 2; (iv) 7 & 7 GnRH = PG at IVPD insertion on Day 17, GnRH on Day 10, IVPD removal on Day 3 and GnRH at FTAI. In Experiment 2, cows (n = 462) were randomly assigned to one of four treatments: (i) 6-day estradiol = EB at IVPD insertion on Day 9, IVPD removal on Day 3 and GnRH at FTAI; (ii) 7-day estradiol; (iii) 7-day GnRH; (iv) 7 & 7 GnRH. In Experiment 1, plasma progesterone concentrations and percentage of cows with a corpus luteum (CL) at IVPD removal, and pregnancy per AI (P/AI) were greater for cows subjected to GnRH-based protocols compared with cows subjected to estrogen-based protocols (p < 0.01). In Experiment 2, cows subjected to the 7 & 7 GnRH protocol had the greatest P/AI (p < 0.01). In summary, GnRH-based FTAI protocols resulted in similar or greater P/AI compared to estrogen-based FTAI protocols in grazing postpartum Bos taurus suckled beef cows. The greatest P/AI was attained with the 7 & 7 GnRH protocol.

Comparison of the efficacy of altrenogest versus transdermal progestin patches on estrus synchronization and ovulation in mares.

Background and Aim: The adhesive progestin patch was investigated for estrus synchronization in mares because this method is convenient and safe in other species. This study aimed to compare the efficacy of a transdermal progestin patch versus oral altrenogest on estrus synchronization, preovulatory follicle development, and ovulation. Materials and Methods: Twenty-four broodmares were randomly divided into two groups. In Group 1, mares (n = 12) were fed 0.044 mg/kg altrenogest daily for 14 days. Group 2 (n = 12) mares were treated with adhesive transdermal progestin patches for 14 days. Mares were observed for estrus daily, from day 3 until day 7, after hormone withdrawal. Follicular development was determined by ultrasonography. The preovulatory follicle size and ovulation time after hormone treatments and ovulation rate were determined for both groups. Plasma progesterone levels were measured during the experimental periods and 2 days after hormone termination in both groups. Results: The results revealed that the transdermal progestin patch efficiently controlled follicular growth and estrus synchronization in mares. The percentage of mares exhibiting estrus was similar in the altrenogest (100.00%) and transdermal patch (91.67%) groups. Ovulation rates were equivalent with either altrenogest or progestin patch protocols (91.67% [11/12] vs. 83.33% [10/12]). In addition, the preovulatory follicle size was similar in mares treated with altrenogest and progestin patches. Conclusion: This study revealed that the transdermal progestin patch provides effective estrus synchronization and ovulation, similar to altrenogest treatment. However, the transdermal patch was more convenient with a shorter predictable ovulation time after estrus synchronization and should be considered as an alternative method for mares.

Development and evaluation of an injectable slow-release progesterone formulation for estrus synchronization in ewes out of the breeding season.

This study was aimed at developing a type of slow-release progesterone micro-particles useable in a single intramuscular injection for estrus synchronization in non-breeding season ewes. A total of 66 ewes were randomly assigned into four groups: CIDR (n = 16): exposed to intravaginal CIDR for 12 days, and three experimental groups, i.e., T100 (n = 16), T150 (n = 17) and T200 (n = 17), receiving a single intramuscular injection of 100, 150 and 200 mg slow-release progesterone, respectively. Blood sampling was performed on all ewes at five different times, and the ELISA method measured progesterone levels. No significant differences were observed in progesterone levels among the groups in each sampling time. More than 90% of ewes in the CIDR, T100 and T150 groups and all those in T200 showed estrus behaviour, and the rate was not significantly different between groups. The difference in the mean interval from progesterone treatment to estrus was also insignificant. The parturition rate declined by increasing the dose of injected progesterone; although it was similar in CIDR and T100 groups, it decreased significantly in T150 and T200 . Since our injectable progesterone formulation was successful in the induction and synchronization of estrus in ewes out of the breeding season, it can be applied as an alternative to the conventional progesterone containing intravaginal devices.

Effect of cloprostenol on luteolysis and comparison of different estrus synchronization protocols in jennies.

Estrus synchronization is necessary for intensive donkey farming. Studies on estrus synchronization in jennies are, however, scarce. We aimed to investigate the susceptibility of the donkey corpus luteum to cloprostenol and design a successful estrus synchronization protocol. Firstly, the effects of different cloprostenol doses and the timing effect of cloprostenol treatment on estrous cycle was investigated. The time from treatment to luteolysis, the ovulation interval, pre-ovulatory diameter, and ovulation rates were compared between groups. Secondly, to identify the best protocol, eight estrus synchronization protocols from three categories were examined. In the first category, jennies in groups A (n = 55) and B (n = 30) received a progesterone releasing intra-vaginal device (JVID®) and cloprostenol treatment. In the second category (group C to F), jennies were pretreated with deslorelin, and then treated with JVID and cloprostenol, including groups C (n = 50), D (n = 50), E (n = 70), and F (n = 65). In the third category, jennies were treated with deslorelin and cloprostenol, including groups G (n = 40) and H (n = 40). Comparisons were made among groups regarding the degree of synchronization, ovulation, and pregnancy rates. Treatment with 0.4 mg cloprostenol on the third day following ovulation minimized the length of the luteal phase and estrous cycle. Synchronization rate varied from 60.0% to 88.6% among groups and was highest in group E. Pregnancy rates did not differ among the eight protocols. In conclusion, cloprostenol effectively induced luteolysis in jennies and a treatment protocol combining deslorelin, cloprostenol, and JVID is efficient for estrus synchronization in donkeys.

The Use of a Brief Synchronization Treatment after Weaning, Combined with Superovulation, Has Moderate Effects on the Gene Expression of Surviving Pig Blastocysts.

The combination of estrus synchronization and superovulation (SS) treatments causes alterations in ovarian and endometrial gene expression patterns, resulting in abnormal follicle and oocyte growth, fertilization, and embryo development. However, the impact of combined SS treatments on the transcriptome of the surviving embryos remains unidentified. In this study, we examined gene expression changes in day 6 blastocysts that survived a brief regimen of synchronization treatment combined with superovulation. The sows were included in one of three groups: SS7 group (n = 6), sows were administered Altrenogest (ALT) 7 days from the day of weaning and superovulated with eCG 24 h after the end of ALT treatment and hCG at the onset of estrus; SO group (n = 6), ALT nontreated sows were superovulated with eCG 24 h postweaning and hCG at the onset of estrus; control group (n = 6), weaned sows displaying natural estrus. Six days after insemination, the sows underwent a surgical intervention for embryo collection. Transcriptome analysis was performed on blastocyst-stage embryos with good morphology. Differentially expressed genes (DEGs) between groups were detected using one-way ANOVA with an un-adjusted p-value < 0.05 and a fold change 1.5. The effect of SO treatment on the number of altered pathways and DEGs within each pathway was minimal. Only four pathways were disrupted comprising only a total of four altered transcripts, which were not related to reproductive functions or embryonic development. On the other hand, the surviving blastocysts subjected to SS7 treatments exhibited moderate gene expression changes in terms of DEGs and fold changes, with seven pathways disrupted containing a total of 10 transcripts affected. In this case, the up-regulation of certain pathways, such as the metabolic pathway, with two up-regulated genes associated with reproductive functions, namely RDH10 and SPTLC2, may suggest suboptimal embryo quality, while the down-regulation of others, such as the glutathione metabolism pathway, with down-regulated genes related to cellular detoxification of reactive oxygen species, namely GSTK1 and GSTO1, could depress the embryos' response to oxidative stress, thereby impairing subsequent embryo development. The gene expression changes observed in the present study in SS7 embryos, along with previous reports indicating SS7 can negatively affect fertilization, embryo production, and reproductive tract gene expression, make its use in embryo transfer programs unrecommendable.

Role of aquaporin 3 in reproductive performance of dairy goats after repeated estrus synchronization stimulation.

This study explored the specific molecular mechanisms through which repeated estrus synchronization (ES) treatments reduce the reproductive performance of dairy goats. Ninety-six goats (n = 24/group) were randomly assigned to two groups receiving ES treatments thrice every fortnight (3-equine chorionic gonadotropin [eCG] and 3-follicle stimulating hormone [FSH] groups) and two groups receiving one ES treatment (1-eCG and 1-FSH groups). ES treatments of 1- and 3-eCG goats were performed via the intravaginal insertion of a controlled internal drug release (CIDR) device containing 300 mg progesterone (P4), followed by 300 IU eCG injections 48 h before CIDR withdrawal. The 1- and 3-FSH goats received CIDR for 10 days, followed by 50 IU FSH and 100 µg PGF2α within 12 h of CIDR withdrawal. Ovaries of three goats in estrus from both groups were harvested for analysis. Subsequently, all the goats in estrus were artificially inseminated twice. Consequently, 3-eCG and 3-FSH goats showed a considerably reduced estrus rate and litter size than 1-eCG and 1-FSH goats. AQP3 mRNA and protein expression were significantly higher in the 3-eCG and 3-FSH groups than in the 1-eCG and 1-FSH groups. AQP3 overexpression led to cell apoptosis and decreased steroid hormone secretion ability of ovarian granulosa cells. Moreover, it resulted in a decrease in maturation and cleavage rates after parthenogenetic activation and in vitro fertilization, respectively. AQP3 gene was involved in reducing the reproductive performance of repeated ES-treated dairy goats. These findings provide a theoretical foundation for the effective use of reproductive hormones in breeding techniques for livestock.

Evaluation of Corpus Luteum and Plasma Progesterone the Day before Embryo Transfer as an Index for Recipient Selection in Dairy Cows.

The aim of the present study was to evaluate the effects of synchronization method, season, parity, corpus luteum (CL) size, and progesterone (P4) levels on the pregnancy rate after bovine embryo transfer (ET). Among 165 recipient candidates who received 1 of 2s estrus synchronization treatments, 96 heifers and 43 cows were selected through rectal examination and used as recipients. The day before ET, the CL size and plasma P4 concentration were evaluated. The CL sizes and plasma P4 levels were not different between the selected and unselected candidates, and the pregnancy rates with the two synchronization methods were not different. However, the pregnancy rates were higher in heifers than in lactating cows, and also higher after ET performed from September to February than from March to August (p < 0.05). The recipients with a CL larger than 1.5 cm showed statistically higher pregnancy rates, and although there was no statistical significance, the pregnancy rate was higher when the plasma P4 levels were between 2.0 and 4.0 ng/mL. Exposure to a stressful environment and repeated manipulations can reduce the success rate of ET, and recipient selection with an optimal CL size and P4 level can increase the success rate of ET.

Administration of GnRH at day 20 of AI and embryonic losses in synchronized crossbred buffalo.

Introduction: Following the application of different artificial insemination and synchronization protocols, the synchronized buffaloes had a higher incidence of early embryonic or fetal death, thus impairing reproductive performance. Therefore, there is a need to devise a synchronizing program that can improve conception, allow early pregnancy diagnosis, reduce early embryonic losses, and provide an early clue for pregnancy diagnosis and establishment. The present study aimed (1) to determine the effect of administration of different GnRH doses at day 20 of artificial insemination (AI) on the reproductive performance of buffaloes and (2) to observe the influence of synchronization protocol, parity, age, milk production, and body condition score (BCS) on early embryonic loss and progesterone (P4) levels in crossbred buffaloes. Methods: Crossbred buffaloes (n = 101) were synchronized using the GPGMH protocol. At day 20 of AI, the synchronized buffaloes were randomly divided into four groups and administrated GnRH doses (0, 100, 150, and 200 µg). The buffaloes were monitored for ovarian dynamics, P4 level, non-return rate, embryonic/fetal losses, and pregnancy rates. The previously synchronized buffaloes were also classified for synchronization protocol (with or without GnRH), parity (nulli- or multiparous), milk production (high or low), BCS (low, medium, or good) or age (>3 or < 3 years) groups for observing the embryonic loss and P4 level variations. Results: The results indicated no difference (P > 0.05) in CL size, P4 level, pregnancy rate and embryo/fetal losses across the treatment groups at different observation periods. There was a high (P < 0.05) incidence of early embryonic mortality in aged, multiparous, low BCS and low milk-producing buffaloes treated without GnRH. Conclusion: The data suggest that GnRH 200 µg at day 20 of AI improves embryo survival and pregnancy maintenance in crossbred buffaloes.

Efficiency evaluation of two estrus synchronization protocols in estrus response and conception rate of dairy cows in the Dalocha district, Ethiopia.

The study aimed to determine the effectiveness of single and double-shot estrus synchronization protocols on conception and estrus response rates in dairy cows. Among 195 sampled female animals, only 174 cows and heifers met a standard of the protocols. Animals were prepared for hormone injections based on their breed, parity number, and body condition score (BCS). Among 174 sampled animals, 120 were indigenous and the rest 54 were crossbreds, and 143 cows and 31 heifers were selected for single alfagladin C (Cloprostenol) injections. Whereas, 16 of them were given double alfagladin C (Cloprostenol) injections when they did not showed estrus prior to single injection. All data was collected and analyzed by SPSS version 20 software. Through observation, inspection and rectal palpation, 90.8% of 174 synchronized animals showed estrus response with a single dose, whereas 16 of them (100%) showed estrus response with double injections. Overall, 48.3% of cows and heifers became pregnant. The conception rate of cows/heifers varied significantly through each protocol, with 50% of animals conceived by double injection treatment and 48.1% of cows conceived through single shot treatment. Additionally, there was a substantial difference in estrus response across breeds, BCS, and parity number. Furthermore, the rates of conception in local and crossbred cows were 40.8 and 64.8%, respectively. In terms of body condition, a high conception rate (58.6%) was observed in the good body condition score. The conception rate and estrus response were significantly different in parity number. In general, the double protocol outperform than the single protocol in terms of estrus and conception rate. Accordingly, stakeholders or artificial insemination technicians might apply double protocols after a single injection to obtain remarkable results. Nevertheless, strict follow-up is required, and more resources at the farmer's management level are required.

Response to hormonal treatment and conception rates of Sahiwal cows subjected to fixed time artificial insemination in pastoral dairy systems.

This study aimed at determining factors influencing response of Sahiwal cows/heifers to fixed time artificial insemination protocol in pastoral systems in Kenya. Available cows/heifers were inspected for conformity to Sahiwal breed characteristics, parity, body condition score, and subsequently rectal palpation to determine pregnancy status, ovarian structures, and estimated ovarian diameter. Consequently, these animals were injected with 100 µg of gonadotrophin-releasing hormone. On days 7 and 9, only responsive cows/heifers were injected with 500 µg of cloprostenol and 100 µg of gonadorelin Acetate, respectively. On day 10, animals were inseminated and separated from bulls for 45 days and pregnancy diagnosis done after 90 days. Analysis of variance was performed to determine the effects of production system, parity, and ovarian structures on ovary diameters pre- and post-hormonal treatment. Logistic regression was used fitting a logit function to account for the binomial distribution of conception. Overall, 56.2%, 23.1%, and 20.7% of the animals had follicles (F), corpus luteum (CL), and corpus albicans (CA), respectively, at day 0, and 16.6%, 68.6%, and 14.8%, respectively, at day 7. Human and environmental factors had no influence on conception. Among the animal factors, only the ovarian structures at day 7 had a significant effect on conception. Ovaries with CL at this time were about 6 times significantly more likely to conceive than those with F. For higher conception rates, animals with ovaries with CL should be recruited into the FTAI program as they are significantly more likely to conceive than those with other ovarian structures.

Dynamic changes in gene expression during follicle development and the efficacy of four timed AI protocols in non-suckling female yaks (Bos grunniens).

The objectives of the study were to investigate changes in the mRNA expression levels of five genes during antral follicle development and to assess the efficacy of four timed-artificial insemination (TAI) protocols in female yaks (Bos grunniens). RT-qPCR analysis revealed that expression levels were greater for follicle-stimulating hormone receptor and bone morphogenic protein 15 in the small follicle, luteinizing hormone receptor, and kit ligand in the large follicle, and growth differentiation factor 9 in the medium follicle (p < 0.05). Non-suckling yaks were treated as a 7-d CIDR, and PGF2α + eCG at CIDR withdrawal and TAI with frozen yak semen at 56-58 h after PGF2α (PPe-7d); either a 7-d CIDR (PPG-7d) or a 5-d CIDR (PPG-5d), and PGF2α at CIDR withdrawal and TAI + GnRH at 70-72 h after PGF2α; and GnRH treatment on Day 0, followed by PGF2α on Day 7 and TAI + GnRH on Day 9 (GPG-7d). The results showed that the pregnancy rate (P/AI) was greater in PPG-5d than in GPG-7d (p < 0.05), but the P/AI was not different among the other TAI protocols. In conclusion, the expression levels of these genes in follicles are dynamically changed during antral follicle development in yaks. The PPG-5d protocol achieved a greater P/AI.

Influence of repeated estrus synchronization treatment on hormone secretion, growth, and development of dairy goats.

In large-scale intensive farms, dairy goats often undergo frequent estrus synchronization (ES) treatment, which may result in a decline in reproductive performance; however, the underlying mechanism remains unclear. The present study aimed to investigate the effect of pregnant mare serum gonadotropin (PMSG) and progesterone (P4)-mediated ES treatment on fertility in dairy goats, while also identifying key metabolic and endocrine mechanisms that influence reproductive performance in does subjected to repeated ES treatment. Forty-eight Saanen does were randomly assigned to two groups (24 goats each) that received ES treatments either thrice fortnightly (3-PMSG) or once (1-PMSG) simultaneously with the third ES treatment of the 3-PMSG group during the breeding season. ES treatment was performed via the intravaginal insertion of a controlled internal drug release (CIDR) device impregnated with 300 mg P4, followed by 300 IU PMSG injections 48 h before CIDR withdrawal. Blood was collected to detect the level of hormones and blood biochemical indices. Additionally, estrus rate, fecundity rate, body weight, size, and lactation performance were measured. The results showed that repeated ES treatment markedly decreased the estrus rate and fecundity rate of goats. Among the does in all groups, there was no substantial difference in follicle stimulating hormone, luteinising hormone, gonadotropin-releasing hormone, melatonin, growth hormone, PMSG, total cholesterol, total protein, and glucose levels, as well as the body weight, body size, and lactation performance. Repeated ES treatment elevated estrogen (E2) levels 36, 48, and 72 h post-CIDR removal; increased P4 upon CIDR insertion; and raised PMSG antibody levels 24, 48, and 72 h post-CIDR removal. The results suggest that elevated anti-PMSG levels are the primary reason for the decline in ES efficiency, and that high E2 and P4 levels at some time points also impair reproductive performance. These findings provide novel insights into the metabolic effects of repeated PMSG stimulation in goats, guiding future reproductive hormone use in breeding practices.

Effects of temperament on reproductive performance of Bos taurus heifers enrolled in the 7-day CO-Synch + controlled internal drug release protocol.

Cattle temperament significantly impacts production traits such as reproduction. The objective of this study was to assess the effects of temperament on pregnancy rates to fixed-timed artificial insemination (TAI) in Bos taurus beef heifers. A total of 297 Angus influenced heifers from 3 different locations were evaluated for temperament based on chute score and exit velocity on the first day of the estrus synchronization protocol (d-9) and classified by temperament type based on temperament score (calm ≤ 3 < excitable). Pregnancy status was determined by transrectal ultrasonography approximately 40 d after TAI. Hair from the tail switch was collected at d-9 and at d0 (TAI) for cumulative cortisol evaluation. A subset of 43 heifers from location 3 had blood samples collected at all handling events and evaluated for cortisol concentrations. Overall, 71% of heifers were classified as calm whereas 29% as excitable. Pregnancy rates to TAI were reduced (P = 0.042) in excitable heifers compared to calm heifers (36% vs. 55%, respectively). Mean concentration of cortisol in the hair was reduced over time (P < 0.001) from d-9 (3.5 ± 0.3 pg/mg) to d0 (1.74 ± 0.3 pg/mg) in all heifers, regardless of temperament. In addition, excitable heifers had increased circulating concentrations of cortisol when compared to calm heifers in all handling events (P = 0.015). More interestingly, despite of temperament, concentrations of circulating cortisol were reduced as the protocol events progressed, with greater cortisol concentrations on d-9, intermediate on d-3, and lesser on d0 (P = 0.031). Therefore, the present study demonstrates that heifer temperament has negative effects in pregnancy rates to TAI programs. However, according to blood cortisol concentrations, no chronic stress response was detected due to cattle handling for the TAI protocol. In fact, blood cortisol concentration was reduced between the initiation and completion of the protocol, indicating that heifers were acclimated to handing. These results support the adoption of acclimation protocols and proper cattle handling as a strategy to increase fertility of heifers that are exposed to TAI.

Current status of assisted reproductive technologies in buffaloes.

Buffaloes are raised by small farm holders primarily as source of draft power owing to its resistance to hot climate, disease, and stress conditions. Over the years, transformation of these animals from draft to dairy was deliberately carried out through genetic improvement program leading to the development of buffalo-based enterprises. Buffalo production is now getting more attention and interest from buffalo raisers due to its socioeconomic impact as well as its contribution to propelling the livestock industry in many developing countries. Reproduction of buffaloes, however, is confronted with huge challenge and concern as being generally less efficient to reproduce compared with cattle due to both intrinsic and extrinsic factors such as poor estrus manifestation, silent heat, marked seasonal infertility, postpartum anestrus, long calving interval, delayed puberty, inherently low number of primordial follicles in their ovaries, high incidence of atresia, and apoptosis. Assisted reproductive technologies (ARTs) are major interventions for the efficient utilization of follicle reserve in buffaloes. The present review focuses on estrus and ovulation synchronization for fixed time artificial insemination, in vitro embryo production, intracytoplasmic sperm injection, cryopreservation of oocytes and embryos, somatic cell nuclear transfer, the factors affecting utilization in various ARTs, and future perspectives in buffaloes.

The effect of progesterone length in timed AI in ewes.

This study aimed to evaluate the reproductive performance of ewes submitted to timed AI protocol with 7 (D7) or 9 (D9) days of progesterone. A total of 220 crossbred ewes (Doper × Santa Ines) were distributed in the treatments according to body weight (52.67 ± 11.76 kg), body condition score (2.5 ± 0.8; scale of 1-5), and the female category (multiparous, nulliparous and primiparous). Ewes received an intravaginal device of 0.3 g of progesterone (CIDR® Easy breed) on day 0, keeping in ewes for 7 or 9 days, which were the experimental treatments. At the time of P4 withdrawal, ewes received 300 IU of eCG (Novormon®) and 6.70 mg of dinoprost tromethamine (Lutalyse®). Insemination by laparoscopy was performed between 46 and 59 h after device removal, lasting from 1 to 5 min/ewe, which did not differ between protocols. The estrus detection rate was higher in the 9-day protocol compared to the 7-day protocol (82% and 65%, respectively, P = 0.0096). However, the 7-day protocol tended to increase pregnancy rate on the TAI compared to the 9-day protocol (45% and 33%, respectively, P = 0.09). The plasma concentration of progesterone at device removal was higher in the 7-day protocol than in the 9-day protocol (2.35 and 1.22 ng/mL, respectively, P = 0.04). We conclude that reducing the length of the estrus synchronization protocol to 7 days is recommended, which improved the reproductive response in ewes.

Reproductive Hormones Mediate Intestinal Microbiota Shifts during Estrus Synchronization in Grazing Simmental Cows.

To study shifts in the intestinal microbiota during estrus synchronization in ruminants, we characterized the intestinal microbiota in grazing Simmental cows and the possible mechanism that mediates this shift. Fourteen postpartum Simmental beef cows were synchronized beginning on day 0 (D0) with a controlled internal release device (CIDR), and cloprostenol was injected on D9 when the CIDR was withdrawn. Synchronization ended with timed artificial insemination on D12. Serum and rectal samples harvested on D0, D9, and D12 were analyzed to assess the reproductive hormones and microbiota. Reproductive hormones in the serum of the host were measured using enzyme-linked immunosorbent assay. The microbiota was characterized using 16S rRNA sequencing of the V3−V4 hypervariable region, alpha diversity and beta diversity analyses (principal coordinate analysis, PCoA), cladogram of the linear discriminant analysis effect size (LEfSe) analysis, and microbiota function analysis. Levels of the reproductive hormones, except gonadotropin-releasing hormone (p > 0.05), shifted among D0, D9, and D12 (p < 0.05). Decreased community diversity (Chao1 and ACE) was observed on D12 compared with D0 (p < 0.05). The beta diversity (PCoA) of the microbiota shifted markedly among D0, D9, and D12 (p < 0.05). The LEfSe analysis revealed shifts in the intestinal microbiota communities among D0, D9, and D12 (p < 0.05 and LDA cutoff >3.0). The KEGG pathway analysis showed that carbohydrate metabolism, genetic information and processing, the excretory system, cellular processes and signaling, immune system diseases, and the metabolism were altered (p < 0.05). Reproductive hormones (especially estradiol) were correlated with the alpha diversity indices, beta diversity indices, and an abundance of biomarkers of the shifting intestinal microbiota (p < 0.05). In conclusion, the structure, composition, and function of the intestinal microbiota were shifted during estrus synchronization in a grazing Simmental cow model, and these shifts were mediated by reproductive hormones.

Impact of vitamin AD3E treatment before application of controlled intra-vaginal drug releaser on clinical and hematologic findings, ovarian hormones, and calving rates in she-camels (Camelus dromedarius).

Background: The scope of using controlled intra-vaginal drug releasers (CIDR) in the camel industry was small relative to that of cow, sheep, and goat. Aim: The purpose of this study was to determine the effects of intramuscular injection of AD3E (25 ml/400 kg) as a premedication protocol before CIDR treatment on ovarian hormones profile and calving rate in dromedary she-camels (Camelus dromedarius) through measuring concentrations of progesterone (P4) and estradiol (E2) before, during, and after CIDR use. Methods: The study was conducted on she-camels (C. dromedarius) (n = 24) aged between 6 and 14 years. The animals were assigned into two equal groups: CIDR treated group (CIDRgr) and AD3E plus CIDR treated group (CIDR+ gr). Results: Except for hemoglobin, the clinical and hematological findings showed no remarkable change either within each independent group or between CIDRgr and CIDR+ gr. Serum P4 and E2 concentrations differed significantly between the first 7 days (during CIDR use) and days from 9 to 15 (after CIDR use) in dromedary camels. Serum concentrations of P4 and E2 showed significant elevation at CIDR+ gr at day 0 compared with CIDRgr. This significant increase in serum of P4 and E2 at CIDR+ gr compared to those of CIDRgr was also observed either during CIDR use (days 3, 5, and 7) or after CIDR use (days 9, 11, 13, and 15). During CIDR use, serum concentrations of P4 were negatively correlated with those of E2 either in CIDRgr or CIDR+ gr, however, they were positively correlated after CIDR removal either in CIDRgror CIDR+ gr. Regarding the calving rate, it was highest in CIDR+ gr whereas CIDRgr showed the lowest one. Conclusion: It was concluded that the AD3E pretreatment protocol of she-camels before insertion of CIDR could be used successfully during the breeding season to improve fertility in she-camels whereas the serum ovarian hormones profile and calving rates were clearly changed due to pretreatment of she-camels with AD3E before CIDR insertion.

A Short-Term Altrenogest Treatment Post-weaning Followed by Superovulation Reduces Pregnancy Rates and Embryo Production Efficiency in Multiparous Sows.

Although embryo transfer (ET) is a biotechnology ready for the swine industry, there are factors to be solved, the availability of embryo donors as one. Multiparous sows as donors ought to be considered since weaning is a natural and efficient method for estrus synchronization. In addition, superovulation treatments at weaning are effective in increasing the efficiency of donor embryo production. However, ET programs typically require more donors than those available from a single weaning, imposing grouping several weanings to establish a batch for ET. Since short-term administration of Altrenogest is effective in delaying estrus after weaning without effects on ovulation and embryo development, we investigated how Altrenogest combined with superovulation would affect reproductive parameters and embryo quality and quantity of weaned multiparous donor sows. The sows were administered Altrenogest from the day of weaning for 14 (SS-14 group; N = 26), 7 (SS-7 group; N = 31) and 4 (SS-4 group; N = 32) days. The sows were superovulated with eCG 24 h after the last administration of Altrenogest and with hCG at the onset of estrus. Sows not treated with Altrenogest that were superovulated with eCG 24 h post-weaning and hCG at the onset of estrus (SC group; N = 37) and sows with natural estrus after weaning (C group; N = 34) were used as control groups. The percentage of sows showing estrus within 10 days was not affected by the treatment, but the interval from Altrenogest withdrawal to estrus was longer (P < 0.05) in the SS groups than the interval from weaning to estrus in the controls. SS treatments increased (P < 0.05) the percentage of sows with ovarian cysts and the development of polycystic ovaries. The pregnancy and the fertilization rates, and the overall embryo production efficiency were also negatively affected by the SS treatments (P < 0.05). Interestingly, almost 70% of the structures classified as unfertilized oocytes or degenerated embryos in sows from the SS groups were immature oocytes. In conclusion, although superovulation of weaned sows was highly efficient, short-term administration of Altrenogest in combination with superovulation had negative effects on most of the reproductive parameters assessed, particularly affecting the overall efficiency of pregnancy and embryo production.

Study of sexual behaviours with different types of estrus synchronization protocols in Boer goats.

There is still a lack of information on estrus synchronization in goats. Understanding the estrus synchronization protocols and the subsequent effects is important to improve the efficiency of assisted reproductive technologies (ARTs) and subsequently would improve the breeding procedures. This study will help in determining the most suitable estrus synchronization protocol and understand better the effect on the sexual behaviour and hormonal effects in goats. A total of 127 Boer does were used and divided into three groups with different duration of CIDR insertion intravaginally either for 14 (two groups) or 9 days (one group). Approximately 0.5 ml Estrumate® (PG) was administered intramuscularly to all groups at CIDR removal, and only groups PMSG14 and PMSG9 were administered with 200IU of Pregnant Mare Serum Gonadotropin (PMSG) intramuscularly. Estrus signs were observed at 4 h intervals and blood samples were collected for progesterone and luteinizing hormone determination. The percentage of does in estrus within 24 to 72 h post CIDR removal was significantly higher (P<0.05) in groups with PMSG compared to the group without the PMSG. The numbers of does display estrus signs within 24 to 28 h post CIDR removal were significantly higher (P<0.05) in group shorter period (9 days) compared to groups with 14 days CIDR. The P4 concentrations at 24 hours post CIDR removals and LH concentration was not significantly different (P>0.05) in all groups. The time of the LH peak in the group without the PMSG was significantly delayed (P<0.05) when compared to group 9 days CIDR and administered with PMSG. It is recommended to use the treatment for 9 days CIDR since the estrous cycle can be shortened.