Oocyte developmental potential and embryo production before puberty in cattle.

With the development of in vitro technologies, embryos can be produced using oocytes retrieved directly from the ovaries, i.e., regardless of ovulation. This has allowed the use of different animal categories as oocyte donors, including prepubertal cattle. The advantages of using this strategy to shorten the generation interval and accelerate genetic gain over time were soon recognized, and the first offspring generated using oocytes collected from calves were born in the early 1990s. Nevertheless, embryo production from calves and prepubertal heifers remains a challenge. The oocytes collected before puberty present low in vitro developmental potential, and the subsequent blastocyst rates are consistently lower than those from pubertal females. The acquisition of developmental competence by the oocytes occurs progressively throughout the prepubertal period, which can be subdivided into an early, intermediate, and late prepubertal (or peripubertal) phases, each characterized by different physiological and endocrine features. Therefore, embryo yield increases with age but will only achieve its maximum after puberty. The most common strategy to improve oocyte developmental potential before puberty is the use of gonadotrophic stimulation prior to oocyte retrieval. The results with superstimulation, however, vary among studies, depending on the source, dose, and length of FSH treatment, as well as the age and breed of the donors. The use of calves and prepubertal heifers as oocyte donors should also consider the possible impacts of the oocyte retrieval technique (LOPU or OPU) and the use of exogenous hormones on their subsequent fertility and productive life.

Selecting oocyte donors based on anti-Müllerian hormone (AMH) concentrations: A critical analysis of using cutoff values as exclusion criterion for an in vitro embryo production program in Gir cattle.

The aims of this study were to determine anti-müllerian hormone (AMH) cutoff values for selecting Gir (Bos taurus indicus) oocyte donors and estimate the impact of using AMH concentrations as a selection criterion. In Exp. 1, Gir heifers (n=120) were sampled for AMH analysis and submitted to ovum pick-up and in vitro embryo production (OPU-IVEP). AMH cutoff values were calculated using ROC analysis or, alternatively, by the successive exclusion of heifers with the lowest AMH values. The correlations between AMH and OPU-IVEP outcomes were significant (P<0.001), though low or moderate (r= 0.34-0.52). We estimated an improvement (P<0.05) after the use of AMH cutoff values to select donors of +15.3% for total oocyes, +19.4% for viable COC, and +23.4% for blastocysts. This selection pressure, however, led to the exclusion of 32.8%, 37.9%, and 50.0% of the initial potential donors, respectively. In Exp. 2, we analyzed data from OPU-IVEP sessions of 658 Gir donors with known genomic values for predicted transmitting ability for milk (GPTAm) and age at first calving (GPTAafc). The selection based on the number of oocytes recovered had no effect (P>0.05) on the average GPTAm nor GPTAafc values of the remaining donors. In summary, plasma AMH ≥700 pg/mL is a cutoff value that can be used to select Gir heifers with a greater potential as oocyte donors. Nevertheless, this selection leads to the exclusion of up to 50% of potential donors. Finally, exclusion of poor responders had no effect on mean genomic estimates for milk production or age at first calving in the selected subset of donors.

Comparing Bayesian models for the genetic evaluation of oocytes and embryo counts in Dairy Gir cattle.

Count traits are usually explored in livestock breeding programs, and they usually do not fit into normal distribution, requiring alternatives to adjust the phenotype to estimate accurate genetic parameters and breeding values. Alternatively, distribution such as Poisson can be used to evaluate count traits. This study aimed to compare and discuss the genetic evaluation for oocyte and embryo counts considering Gaussian (untransformed variable - LIN; transformed by logarithm - LOG; transformed by Anscombe - ANS) and Poisson (POI) distributions. The data comprised 11,343 total oocytes (TO), viable oocytes (VO), cleaved embryos (CE), and viable embryo (VE) records of ovum pick-up from 1740 Dairy Gir heifers and cows. The genetic parameters and breeding values were estimated by the MCMCglmm package of the R software. The posterior means of heritability varied from 0.40 (LIN) to 0.49 (POI) for TO, 0.39 (LIN) to 0.49 (POI) for VO, 0.30 (LOG) to 0.41 (POI) for cleaved embryos, and 0.19 (LIN) to 0.32 (POI) for viable embryos. The posterior means of repeatability varied from 0.56 (LIN) to 0.65 (POI) for TO, 0.53 (LOG) to 0.63 (POI) for VO, 0.44 (LOG) to 0.60 (POI) for CE, and 0.36 (LOG) to 0.56 (POI) for VE. Deviance information criterion and mean squared residuals indicated that POI model should be used for the genetic evaluation of embryo and oocyte count traits. Spearman's rank correlation between estimated breeding value (EBV) for embryo and oocyte count traits computed by POI, LOG, and ANS models was high (ranging from 0.77 to 0.99), indicating little reranking among the best animals. The POI model is the most adequate for genetic evaluation, resulting in more reliable EBV of oocyte and embryo count traits for Dairy Gir cattle.

Impact of heat stress on genetic evaluation of oocyte and embryo production in Gir dairy cattle.

Identifying and selecting genotypes tolerant to heat stress might improve reproductive traits in dairy cattle, including oocyte and embryo production. The temperature-humidity index (THI) was used, via random regression models, to investigate the impact of heat stress on genetic parameters and breeding values of oocyte and embryo production in Gir dairy cattle. We evaluated records of total oocytes (TO), viable oocytes (VO), cleaved embryos (CE), and viable embryos (VE) from dairy Gir donors. Twenty-four models were tested, considering age at ovum pick-up (AOPU) and THI means as a regressor in the genetic evaluation. We computed THI in eight periods, from 0 to 112 days before ovum pick-up, which were adjusted by different orders of Legendre polynomials (second, third, and fourth). The best-fit model according to Akaike's information criterion (AIC) and Model Posterior Probabilities (MPP) considered Legendre polynomials of third order and THI means of 112 days for TO, fourth order and 56 days for VO, second order and 28 days for CE, and second order and 42 days for VE, respectively. The heritability (h2) estimates across AOPU and THI scales ranged from 0.34 to 0.62 for TO, 0.31 to 0.58 for VO, 0.26 to 0.39 for CE, and 0.15 to 0.26 for VE, respectively. The fraction of the phenotypic variance explained by the permanent environment in different AOPU and THI scales ranged from 0.03 to 0.25 for TO, 0.05 to 0.26 for VO, 0.09 to 0.36 for CE, and 0.15 to 0.27 for VE, respectively. Spearman's rank correlation between the estimated breeding values in different AOPU and THI scale from the top 5% sires and females ranged from 0.18 to 0.90 for TO, 0.31 to 0.95 for VO, 0.14 to 0.85 for CE, and 0.47 to 0.94 for VE, respectively. The h2 estimates for all evaluated traits varied from moderate to high magnitude across AOPU and THI scales, indicating that genetic selection can result in rapid genetic progress for the evaluated traits. There was a reranking among the best animals in different AOPU and THI. It is possible to select dairy Gir cattle tolerant to heat stress to improve oocyte and embryo production.

Outstanding Gir oocyte donors: How does individual factor affect in vitro embryo production efficiency?

The oocyte donor plays a pivotal role in bovine in vitro embryo production (IVP) success. The individual factor affects blastocyst/oocyte ratio and determine the existence of outstanding performing animals. The aim of this study was to assess the extent of individual factor effect to IVP efficiency, in a population of Gir oocyte donors. Extreme (high or low IVP efficiency based on blastocyst/oocyte ratio) animals were selected out of a population of 250 oocyte donors (1,734 observations) to form high (>0.48, n = 40), average (0.17-0.48, n = 168), and low (<0.17, n = 42) efficiency donor groups. Cumulus-oocyte complex indicators (total number, IVF-grade number, and IVF-grade/total ratio) were lower (p < 0.05) in high efficiency donors. The number of blastocysts per OPU was analyzed for highest performing bull, and an increase (p < 0.05) in high efficiency donors and a decrease (p < 0.05) in low efficiency donors were noticed, compared to average efficiency donors. The number of pregnancies obtained per OPU was affected (p = 0.017) by donor's efficiency (low: 0.60 ± 0.09 $$ 0.60\pm 0.09 $$ , average: 1.17 ± 0.07 $$ 1.17\pm 0.07 $$ , high: 2.57 ± 0.26 $$ 2.57\pm 0.26 $$ ), being 4.3-fold higher in high than in low efficiency donors. We conclude that producing embryos from high efficiency blastocyst/oocyte ratio donors increases blastocyst and pregnancy numbers by OPU, being an important indicator for donor selection in IVP programs.

Imputation accuracy for genomic selection using embryo biopsy samples in Gir.

The aim of this study was to establish a platform for genomic selection of in vitro-fertilized (IVF) Gir embryos. Multiple displacement amplification (MDA)-based embryo biopsy samples were genotyped, and genomic estimated breeding values (GEBV) for milk yield (305MY) were calculated. The concordance of GEBV and accuracy between embryo biopsies and the respective liveborn were assessed. Imputation was performed using two panels (Z-Chip and Bovine HD, Illumina) based on a database of 73,110 lactating cow's database and pedigree files from 147,131 animals. Biopsied embryos had similar pregnancy rates (39% vs 40%), pregnancy loss rates (18% vs 20%), and pregnancy length compared to Control embryos. After genotyping, low call rate means were detected for biopsy samples compared to the respective calf samples (0.80 vs 0.98). Imputation presented 0.83 (Z-Chip) and 0.96 (HD) accuracy (CORRanim). Embryo GEBV accuracy levels were higher in BovineHD imputation (0.82) than Z-Chip imputation (0.55) or no imputation (0.62), and the correlation between embryo/calf pairs' accuracy was 0.85 for BovineHD imputation, 0.11 for Z-Chip imputation, and 0.02 for no imputation. GEVB estimates correlation between embryo/calf pairs was 0.87 for BovineHD imputation, 0.80 for Z-Chip imputation, and 0.41 before imputation. The call rate of embryo samples did not affect the correlation between embryo/calf pairs for accuracy and GEBV before and after BovineHD imputation. Embryos obtained on the same farm presented GEBV 305MY differences of up to 800 kg, emphasizing the expected impact of embryo genomic selection for the Gir breed.

Likelihood of pregnancy after the transfer of embryos derived from follicle aspiration and in vitro embryo production sessions with different relative efficiencies.

The aim of the present study was to evaluate the likelihood of pregnancy of in vitro-produced (IVP) embryos from batches with distinct relative efficiencies. Data were retrospectively analyzed from 605 transvaginal ultrasonic-guided follicle aspiration sessions (OPU) followed by in vitro embryo production (IVEP) and 2456 fresh embryo transfers (ET), performed between 2008 and 2012 in individuals of the Gir (dairy Bos indicus) breed. The OPU and IVEP were performed using standard procedures by a single group of technicians at the same laboratory facility. Records were stratified into quartiles (I to IV) according to the total of cumulus-oocytes complexes (COC) produced per donor, or in percentile ranges (0%-25%, 26%-50%, 51%-75%, and 76%-100%) for endpoints related to COC quality or efficiency of embryo production. Pregnancy per embryo transfer (P/ET) was compared among quartiles or ranges using the chi-squared test. Donors producing a greater number of total COC (quartile I) also had more viable and grade I COC, and a greater number of embryos than donors ranked in quartiles II, III or IV, respectively (P < 0.0001). Nevertheless, P/ET did not differ (P > 0.05) among embryos produced by donors ranked in Quartiles I to IV. Similarly, there was no difference (P > 0.05) in P/ET for embryos derived from OPU sessions with a relatively greater or lesser percentage of viable or Grade I COC. Cleavage and blastocyst rates within each IVEP batch had no effect (P > 0.05) on P/ET. In conclusion, data suggest that there is no relationship among oocyte yield after OPU, or efficiency of IVEP, and the likelihood of pregnancy after ET of fresh IVP embryos.

Protected fatty acid supplementation during estrus synchronization treatment on reproductive parameters of dairy goats.

This study evaluated the effect of the protected fatty acid inclusion during estrus synchronization on reproductive parameters. Goats (n = 32) received progestagen sponges for 6 days and 200 IU equine chorionic gonadotropin and 30 µg d-cloprostenol were given on Day 5. No difference was found among control (C), 1% protected fatty acid inclusion (C + 1%) or 4% protected fatty acid inclusion (C + 4%) groups, respectively, in estrus (100.0, 100.0 or 90.9%), estrus duration (31.6 ± 12.3; 43.2 ± 12.9 or 40.8 ± 14.1 h), animals ovulating (100.0, 90.0 or 100.0%) or ovulation rate (1.3 ± 0.5; 1.1 ± 0.3 or 1.2 ± 0.4). The interval from sponge removal to ovulation and from estrus to ovulation, respectively, were shorter for C + 4% (45.2 ± 8.0 h; 18.3 ± 11.0 h) compared with C (56.3 ± 12.6 h; 30.6 ± 10.5 h) or C + 1% (57.7 ± 8.7 h; 30.3 ± 11.1 h). The average ovulatory follicle diameter was smaller for C + 4% (6.2 ± 0.7 mm) than C (7.5 ± 0.8 mm), but similar to C + 1% (7.0 ± 1.5 mm). Insulin, insulin-like growth factor 1, glucose and progesterone concentrations were similar among groups. The inclusion of protected fatty acid during synchronization treatment promoted no benefits on ovulation rate, but 4% anticipated the ovulation time.