Proteomic profile of seminal plasma from Pêga donkeys (Equus asinus) with high sperm motility and vigor: Implications for assisted reproduction.

Donkeys of the Pêga breed (Equus asinus) have been used for two centuries to produce breeding stock and create hybrids for labor and transport in southeast Brazil, and for exporting meat and milk to other countries. Furthermore, they are used in competitions, as they are docile and easy to handle. However, assisted reproduction success rates for frozen donkey semen are remarkably low, with no standardized method for cryopreserving sperm after removal of seminal plasma. This work aims to reveal the biological involvement of seminal plasma proteins from Pêga donkeys in aiding the development of assisted reproduction. This study was carried out with 14 ejaculates collected every eight days, throughout the breeding season, from three healthy fertile Pêga donkeys, with an average age of four years. After confirming the high freezability of fresh semen by evaluating quality parameters, the seminal plasma was separated by centrifugation and an aliquot from each collection was microfiltered and frozen. A label-free technique followed by LC-MS/MS analysis applied to pools of seminal plasma samples from each animal revealed 522 proteins in the proteomic profile, of which 49.8 % (260 proteins) are related to cellular energy transformation, and many proteins involved in reproduction (76), spermatogenesis (38), fertilization (29), among other biological process. By comparison with literature, Pêga donkeys share many proteins with donkeys of Dezhou breed that present great potential as fertility biomarkers. Our results showed proteins positively related to fertilization for different breeds of donkeys around the world, helping to enhance the assisted reproduction of Pêga donkeys.

Calving date as a potential predictor for the probability of approval in the first breeding soundness evaluation of Nellore bulls.

Beef production systems primarily use natural service (NS) for breeding. However, a significant number of bulls used for NS are subfertile, limiting the profitability of the cow-calf operations. Therefore, producers should select bulls based on breeding soundness evolutions (BSE) to ensure higher pregnancy rates. Several factors can affect the bull ability to pass a BSE. We hypothesize that calving date would be a factor that affects the bull probability of approval at the first BSE. For this purpose, a multivariate logistic regression in a dataset of 14,737 BSEs from young Nellore bulls was used. Correlations between calving date, biometrics, and semen traits were evaluated using Pearson's correlation coefficient. Our results demonstrated that the calving date affected the probability of approval at the first BSE (p < .05). Indeed, the variable that added more information to our model was the calving date, far more than the age group of the bulls according to Akaike's information criterion. Hence, bulls born on day 0 of the calving season have 1.26 more chances to be approved at the first BSE than bulls born 21 days later. This result highlights the importance of getting the dams of future bulls pregnant as soon as possible in the breeding season. In addition, the calving season should be no longer than 47 days to achieve 80% BSE approval in 20-22 months old Nellore bulls. The strongest correlation was with SC, which decreased as the calving date increased. Therefore, the calving date may be used as a predictor of the outcome of the first BSE in young bulls. In that way, the calving date can help seedstock producers to maximize efficiency in making crucial management decisions during the breeding and calving season including nutrition, reproductive, and culling.

Dietary L-arginine supplementation during early gestation of gilts affects conceptuses development.

l-arginine supplementation of sows has led to improvement of reproductive performance, but the mechanisms responsible for the positive effects of arginine during gestation on conceptuses survival and development are still poorly understood. Thus, we aimed to evaluate effects of 1.0% l-arginine supplementation (ARG) on phenotypic traits of commercial gilts, embryos and fetuses, concentration of gilts' blood metabolites, expression of developmental and cellular apoptosis genes in conceptuses of 25 and 35 days. At 25 days, IGF1 gene was more expressed in embryos from ARG than in embryos from control gilts (CON) (P = 0.05). At this same gestational age, ARG embryos tended to be heavier compared to CON (P = 0.07) and ARG gilts showed a trend to have a greater arginine concentration in blood plasma (P = 0.06). However, at 35 days of gestation, arginine concentration in blood plasma of ARG gilts tended to be lower compared to CON (P = 0.06) and ARG fetuses showed smaller cephalic-caudal length (P = 0.05). These results indicate that duration of supplementation is determinant for arginine effects, not only on the females performance but also on the conceptuses, since supplementation upregulated IGF1 expression at 25 days, in addition to the reduction of cephalic-caudal length of 35-day fetuses.

Soybean grain is a suitable replacement with soybean meal in multiple supplements for Nellore heifers grazing tropical pastures.

The research aimed to evaluate the effect of replacing soybean meal with soybean grain on the nutritional parameters and productivity of heifers grazing on Urochloa decumbens in the rainy-dry transition period. Forty crossbred heifers with the initial age and weight of 18 months and 292 ± 6.1 kg, respectively, were used. The experimental design was a completely randomized design, with five treatments and eight replications. The evaluated treatments were as follows: SM0.5-supply of 0.5 kg/animal/day of soybean meal supplement; SG0.5-supply of 0.5 kg/animal/day of soybean grain supplement; SM1.0-supply of 1.0 kg/animal/day of soybean meal supplement; SG1.0-supply of 1.0 kg/animal/day of soybean grain supplement; MM-only mineral mix ad libitum. The supplement was isoprotein with 350 g of crude protein/kg of dry matter. Supplementation improved the performance of the heifers and this fact can be verified by the higher average daily gain (ADG) and final BW (fBW) of the supplemented animals (P < 0.10). The two supplemented treatments with 1 kg/day demonstrated similar performance (P > 0.10), the same happens for the two treatments receiving 0.5 kg/day (P > 0.10). However, animals receiving 1 kg/day of supplementation had an ADG and final BW higher than animals receiving 0.5 kg/day (P < 0.10). Supplementation (P < 0.10) affected the intake of dry matter (DM), organic matter, crude protein, ether extract, non-fiber carbohydrates, total digestible nutrients, and neutral detergent fiber corrected for ash and protein (apNDF). Supplementation improved DM digestibility and all constituents of the diet (P < 0.10), except for apNDF (P > 0.10). In summary, it is concluded that multiple supplementations improve the performance of grazing heifers in the rainy-dry transition period and the total replacement of soybean meal by soybean grain does not alter the performance of the animals.

Sêmen sexado através de citometria de fluxo e centrifugação por gradiente de concentração / Semen sexed using flow cytometry and concentration gradient centrifugation / Semen sexado a través de citometría de flujo y centrifugación por gradiente de concentración

Resumo O objetivo deste trabalho foi apresentar uma revisão completa e atualizada sobre a origem do sêmen sexuado, os requisitos e as técnicas de sexagem espermáticas. Nas últimas décadas, desenvolveram-se várias tecnologias na área da reprodução animal. O sêmen sexado é uma biotecnologia recente que ainda se encontra em fase de estudo e aperfeiçoamento em diversas etapas, devido a que o processo de sexagem causa estresse ao espermatozoide, e o deixa mais sensível ao processo de armazenamento; isto interfere diretamente na fertilidade. Esta tecnologia, que chegou ao Brasil em 2004, vem ganhando adeptos entre produtores que desejam produzir animais mais homogêneos para o sacrifício ou para a produção de leite. O uso do sêmen sexado tem sido difundido em muitos rebanhos do mundo, já que se apresenta como uma técnica com um direcionamento maior na seleção do sexo. Muitas técnicas têm sido utilizadas com a finalidade de buscar uma maior separação de espermatozoides portadores de cromossomas X e Y, com mínima agressão espermática. Desta forma, a utilização da sexagem espermática tem obtido resultados na aplicação de biotecnologias da reprodução assistida, e com isso tem elevado o lucro no melhoramento genético e tem otimizado a produtividade no sexo escolhido. Entretanto, ainda existem muitas limitações, principalmente referentes al tempo de exposição, o índice de aproveitamento e os resultados de índices de gestação em condições de campo. Por outro lado, a dose de sêmen sexado pode custar de duas a oito vezes o valor de uma convencional; da mesma forma, o seu uso avança no Brasil.

Abstract This study aimed to present a complete and updated review on the origin of sexed semen, as well as on the requirements and techniques of sperm sexing. In the last decades, several technologies were developed in the field of animal reproduction. Sexed semen is a recent biotechnology that is still under study and refinement in various stages, because the sexing process causes stress to the sperm, leaving it more sensitive to the storage process; this directly interferes with fertility. This technology, which arrived in Brazil in 2004, has been gaining support among producers who want to produce more homogeneous animals for slaughter or for dairy production. The use of sexed semen has been disseminated in many herds of the world, since it is a technique with better results in the selection of sex. Many techniques have been used in order to seek a greater separation of sperms carrying X and Y chromosomes, with minimal sperm aggression. In this way, the use of sperm sexing has obtained results in the application of assisted reproductive biotechnologies, which has increased the gain in genetic improvement and has optimized productivity in the chosen sex. In the meantime, there are still many limitations, mainly concerning exposure time, exploitation rate, and the results of pregnancy rates in field conditions. On the other hand, the dose of sexed semen can cost two to eight times the value of conventional semen; nevertheless, its use progresses in Brazil.

Resumen El objetivo de este trabajo fue presentar una revisión completa y actualizada sobre el origen del semen sexado, los requisitos y las técnicas de sexaje espermáticas. En las últimas décadas, se desarrollaron varias tecnologías en el área de la reproducción animal. El semen sexado es una biotecnología reciente que aún se encuentra en fase de estudio y perfeccionamiento en diversas etapas, debido a que el proceso de sexaje causa estrés al espermatozoide, y lo deja más sensible al proceso de almacenamiento; esto interfiere directamente en la fertilidad. Esta tecnología, que llegó a Brasil en 2004, viene ganando adeptos entre productores que desean producir animales más homogéneos para el sacrificio o para la producción lechera. El uso del semen sexado ha sido difundido en muchos rebaños del mundo, ya que se presenta como una técnica con un direccionamiento mayor en la selección del sexo. Muchas técnicas han sido utilizadas con el fin de buscar una mayor separación de espermatozoides portadores de cromosomas X y Y, con mínima agresión espermática. De esta forma, la utilización del sexaje espermático ha obtenido resultados en la aplicación de biotecnologías de la reproducción asistida, y con eso ha elevado la ganancia en el mejoramiento genético y ha optimizado la productividad en el sexo escogido. Entretanto, aún existen muchas limitaciones, principalmente referentes al tiempo de exposición, la taza de aprovechamiento y los resultados de tazas de gesta ción en condiciones de campo. Por otro lado, la dosis de semen sexado puede costar de dos a ocho veces el valor de una convencional; así mismo, su uso avanza en Brasil.

Estimation of daily milk yield of Nellore cows grazing tropical pastures.

Beef cows' milk yield is typically determined by measuring milk yield once daily and then doubling this value to estimate daily production. However, it is not known whether this is accurate. Thus, we aimed to determine the association between morning and afternoon milk yield in grazing Nellore cows. Eighty Nellore cows were used, with initial weight of 516.0 ± 1.0 kg. The experiment was a completely randomized factorial scheme, with 20 replications and four treatments (i.e., + or - pre-partum supplementation in combination with + or - post-partum supplementation): PRMM-1 kg of supplement/cow/day for 90 days pre-partum; MMPS-1 kg of supplement/cow/day for 90 days post-partum; PRPS-1 kg of supplement/cow/day for 90 days pre-partum and 90 days post-partum; and MM-only mineral mix ad libitum during pre- and post-partum. Milk was sampled on days 45, 135, and 225 post-partum (early, middle, and late lactation, respectively). No effects were observed of pre- and post-partum supplementation on milk yield (P > 0.05). The afternoon/morning proportion of 0.45 in the early third of lactation was higher than other stages, which had a proportion of 0.41 (P < 0.05). Post-partum supplementation increased milk protein in the morning and afternoon milking (P < 0.05). There was also no effect of pre- and post-partum supplementation on afternoon-morning proportion other milk components (P > 0.05). We conclude that estimating daily milk production of grazing beef cattle by multiplying a once daily milking amount times two is not accurate. Under the conditions of this study, proportion of total daily production represented by the ratio of afternoon/morning milking was 0.45 in early lactation (first third) and 0.41 in mid- and late lactation.

Can scrotal circumference-based selection discard bulls with good productive and reproductive potential?

Nonlinear mixed models were used to describe longitudinal scrotal circumference (SC) measurements of Nellore bulls. Models comparisons were based on Akaike's information criterion, Bayesian information criterion, error sum of squares, adjusted R2 and percentage of convergence. Sequentially, the best model was used to compare the SC growth curve in bulls divergently classified according to SC at 18-21 months of age. For this, bulls were classified into five groups: SC < 28cm; 28cm ≤ SC < 30cm, 30cm ≤ SC < 32cm, 32cm ≤ SC < 34cm and SC ≥ 34cm. Michaelis-Menten model showed the best fit according to the mentioned criteria. In this model, ß1 is the asymptotic SC value and ß2 represents the time to half-final growth and may be related to sexual precocity. Parameters of the individual estimated growth curves were used to create a new dataset to evaluate the effect of the classification, farms, and year of birth on ß1 and ß2 parameters. Bulls of the largest SC group presented a larger predicted SC along all analyzed periods; nevertheless, smaller SC group showed predicted SC similar to intermediate SC groups (28cm ≤ SC < 32cm), around 1200 days of age. In this context, bulls classified as improper for reproduction at 18-21 months old can reach a similar condition to those considered as good condition. In terms of classification at 18-21 months, asymptotic SC was similar among groups, farms and years; however, ß2 differed among groups indicating that differences in growth curves are related to sexual precocity. In summary, it seems that selection based on SC at too early ages may lead to discard bulls with suitable reproductive potential.

Nellore cows and their calves during the lactation period: performance, intake, milk composition, and total apparent digestibility.

An experiment was conducted to evaluate intake and nutrient digestibility, performance, milk composition, and microbial efficiency of Nellore cows and their calves during lactation. Fifteen Nellore cows were fed corn silage and concentrate (85:15). After calving, calves were kept with their dams for 2 months then separated and housed in individual pens adjacent to their dams. Cows were milked every 15 days over 7 months to estimate milk intake of the calves. During lactation, nutrient intake decreased (P < 0.01) for cows, while nutrient digestibility increased (P < 0.001). Starting 2 months post-calving, calves were offered concentrate (5 g/kg body weight (BW)) and roughage. An increase in DM digestibility was observed for calves in the third month of age, probably due to an associative effect between milk, concentrate, and roughage, which increased DM digestibility without increasing DM intake. Average daily gain (ADG) of the calves increased (P < 0.001) during lactation period. Microbial efficiency (g mCP/kg total digestible nutrients (TDN)) did not change (P > 0.05) in either cows or their calves during lactation period. However, nutrient digestibility decreased for calves during this period. Nellore cows produced milk with average of 4.58 % lactose, 5.61 % fat, and with milk protein increasing from 3.6 to 4.0 % as lactation progressed.

Effect of phase-feeding crude protein on performance and carcass characteristics of crossbred beef bulls: an application to reduce nitrogen compounds in beef cattle diets.

Beef cattle (24) with an average initial body weight of 417 ± 54 kg were assigned in a 2 × 2 factorial design with six repetitions. The factors studied were two levels of CP: 11 (low CP) or 13 % (high CP) in the initial feedlot phase (days 1 - 36) and in the final phase (days 37 - 72). After 36 days, half of the bulls fed with each level of CP were selected to reverse the CP level. No interaction was observed (P > 0.05) between the level of protein in the initial and final phases on the intake of the nutrients evaluated. There was no effect (P > 0.05) of the protein levels in the initial and final phases on intake of dry matter, organic matter, CP, non-fiber carbohydrates, and total digestible nutrients. No differences were observed among treatments (P > 0.05) for average daily gain (X = 1.99 kg/day) and carcass traits. We conclude that it is possible to feed Holstein-Zebu crossed bulls with an average daily gain of approximately 2 kg using a fixed level of 11 % CP during the entire feedlot period, and this diet is economically viable and environmentally sound.

Performance, growth, and maturity of Nellore bulls.

The objectives of this study were to evaluate the dry matter intake (DMI), digestibility, average daily gain (ADG), microbial efficiency, empty body weight (EBW) gain, and body composition of Nellore bulls. Additionally, Nellore bull maturity was estimated, and the prediction equation for DMI, suggested by the Brazilian nutrient requirements system (BR CORTE; Azevêdo et al. 2010), was evaluated. Thirty-three Nellore bulls, with a mean initial weight of 259 ± 25 kg and age of 14 ± 1 months, were used in this study. Five animals were slaughtered at the beginning of the experiment (control group), and the remaining 28 were divided into 4 groups, each slaughtered at 42-day intervals. Their diet was composed of corn silage and concentrate (55:45). The power model was used to estimate muscle tissue, bone tissue, crude protein (CP), mineral matter (MM), and water present in the empty body, while the exponential model was used to estimate adipose tissue and ether extract (EE) present in the empty body. When expressed in kilograms per day, differences were observed (P < 0.05) only for the intake of EE and neutral detergent fiber as a function of feedlot time periods. Although there was a difference in relation to nutrient intake, it did not affect (P > 0.05) digestibility, with the exception of EE digestibility. The equation suggested by BR CORTE correctly estimates the DMI of Nellore bulls. ADG was not affected (P > 0.05) by time spent in the feedlot. No differences were observed (P > 0.05) for microbial efficiency; a mean value of 142 g microbial crude protein/kg total digestible nutrients was achieved. The muscle and bone tissues, CP, MM, and water present in the empty body increased as the animal grew, although at a lower rate. The adipose tissue and EE present in the empty body increased their deposition rate when the animal reached its mature weight. Maturity is defined as when an animal reaches 22 % EE in the empty body, which corresponds to 456 kg of EBW in Nellore bulls. Therefore, this study can conclude that the feedlot time period does not affect DMI, nutrient intake, ADG, or microbial efficiency. The equation proposed by BR CORTE (Azevêdo et al. 2010) correctly estimates the DMI of Nellore bulls, which reach maturity when an EBW of 456 kg is attained.