Effects of ascorbic acid on in vitro culture of bovine preantral follicles

The objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine their effects on growth activation and viability of early-stage follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed (control) and the other fragments were cultured in minimum essential medium (MEM) supplemented or not with various doses of ascorbic acid. Ovarian tissue was processed for histology, transmission electron microscopy (TEM) and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Compared with control fragments, the percentage of primordial follicles was reduced (p < 0.05) and the percentage of growing follicles had increased (p < 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (p < 0.05), but not when cultures were supplemented with 25, 50 or 100 μg/ml of ascorbic acid. Ultrastructural and immunohistochemical analysis of 8 day cultured ovarian cortical fragments, however, showed the integrity and viability of follicles only when fragments were cultured in presence of 50 μg/ml of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg/ml not only stimulates the activation of 8 day in vitro cultured cattle primordial follicles and subsequent growth of activated follicles, but also safeguards the viability of these early-stage follicles. © 2012 Copyright Cambridge University Press.

Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle.

Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the 'P-36 protocol' for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT(2) receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus-oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.

Índice de seleção bioeconômico para fêmeas de corte da raça nelore

The reproductive efficiency of Nellore females was described on the basis of sexual precocity, staying productive in the herd (NP), maternal productivity (PM) and estimated cost for maintenance (CM). The combination of these traits resulted in the bioeconomic maternal revenue index (RMat) ensures reproductive efficiency. The index estimates the return in kilograms of live weight produced per cow per year. In addition, the composition of calf weight produced was considered, adding to the PM the information on the scores of conformation, muscling and early finishing at weaning, in order to input the calf biotype. The females considered precocious had their age at first calving before 30 months. The NP was expressed by the number of calving till 53 months of age. The CM was calculated according to the estimated feed intake of dry matter. The estimated average RMat was 62.02±24.12 kg/ cow/year. Estimates of additive genetic and residual variances for RMat, using restricted maximum likelihood under a single trait animal model was equal to 195.35 and 242.96, respectively. The heritability estimated was 0.45±0.02, indicating that RMat is inheritable and can be applied in the breeding program to improve reproductive efficiency. The NP was the main component of variation for RMat. Sires selected based on RMat tended to have more efficient daughters.

Análise Genética de Características Reprodutivas Na Raça Brahman

Aiming to establish selection criteria for the Brahman cattle in Brazil, the objective of this paper was to estimate genetic and environmental parameters for birth weight (BW), gestation length (GL) and scrotal circumference at 365 (SC365), 455 (SC455) and 550 (SC 550) days of age, and relative efficiency of indirect versus direct selection for these traits. Data were obtained from 15 851 animals belonging to 16 herds in six states, enrolled in a genetic breeding program. (Co)variance and heritability coefficients and genetic and environmental correlations between traits were estimated by restricted maximum likelihood (REML) under a multi trait animal model. The model included as fixed effects the contemporary group and the age of cow at calving as a covariate (linear and quadratic), and as random the direct genetic, the permanent environmental and residual effects. For BW and GL it was also considered the maternal genetic effect as random. Estimates of direct heritabilities were 0.36, 0.29, 0.55, 0.43 and 0.40 for BW, GL, SC365, SC455 and SC550, respectively. Genetic correlations between BW and GL, GL and SC365, GL and SC455, GL and SC550, SC365 and SC455, SC365 and SC550 and SC455 and SC550, were respectively 0.06, 0.13, 0.20, 0.13, 0.96, 0.98 and 0.99. The heritabilities estimates for all traits indicated these show enough additive genetic variability to respond favorably to selection. For sexual precocity, the best option would be SC365, due to the high value of heritability and to indirect selection at this age being more efficient than direct selection for PE455.

Crescimento, comportamento ingestivo e desempenho reprodutivo de novilhas mestiças holandês x zebu, em pastejo, submetidas à suplementação protéica durante a época das águas

The objective of this study was to evaluate the effect of supplemental protein during the rainy season on body growth, ingestive behavior and reproductive performance of crossbred Gir x Holstein dairy heifers, with mean age of 21 ± 4.1 months and average initial body weight of 296.7 ± 57.3 kg, distributed in a completely randomized design with four treatments and six replicates per treatment. Heifers remained on Brachiaria brizantha cv. Marandu pasture, managed according to a rotational grazing system and, as treatment, it was supplemented with protein concentrate (34%, 43%, 56% and 66% BW) and level of supply of 0.30% body weight (BW). The animals were weighed and the mean weight of the lot was used for the calculation of adjustment of the amount of concentrate offered. Heifers that reached 330.0 kg of live weight were artificially inseminated and, 45 days after insemination, pregnancy diagnosis was performed for the calculation of pregnancy rate and age at conception. No significant difference of protein levels was observed on grazing, eating behavior, productive performance and age at conception of dairy heifers on pasture. The average daily weight gain of BW and age at conception were, on average, 0.907 kg/animal and 24.03 months, respectively.

Glicerina bruta na dieta de bovinos de corte confinados: Efeito sobre o hemograma

The aim of this study was to evaluate the hemogram in cattle fed crude glycerin. 30 Nellore steers, were used for blood sampling, which were done each 28 days. The animals were confined for 103 days and were fed with five diets containing 0, 7.5, 15, 22.5 and 30% crude glycerin based on dry matter, formulated in a forage:concentrate ratio of 30:70. Corn silage was used as forage and the concentrate was composed by corn grain, soybean hulls, sunflower meal, glycerin, limestone, dicalcium phosphate and salt. The experimental design was a randomized block with five treatments. Orthogonal contrasts were used to determine the linear, quadratic, cubic effects and gliceryn treatments × the control one. There was no treatment effect (P>0,05) on blood concentrations of erythrocytes, hematocrit, basophils, eosinophils, band neutrophils and lymphocytes, but were linearly effect on the concentrations of platelets, hemoglobin and monocytes (P=0.009, P=0.001 and P=0.043, respectivelly), and cubic effect on the concentrations of total leukocytes and segmented neutrophils (P=0.008 and P=0.004, respectivelly). Diets for beef cattle, with forage:concentrate ratio of 30:70 added with crude glycerin promote changes in the erytrhogram and leucogram of these animals.

Genotype×production environment interaction for weaning weight in Angus populations of Brazil and Uruguay

The objective of this study was to define production environments by grouping different environmental factors and, consequently, to assess genotype by production environment interactions on weaning weight (WW) in the Angus populations of Brazil and Uruguay. Climatic conditions were represented by monthly temperature means (°C), minimum and maximum temperatures in winter and summer respectively and accumulated rainfall (mm/year). Mode in month of birth and weaning, and calf weight (kg) and age (days) at weaning were used as indicators of management conditions of 33 and 161 herds in 13 and 34 regions in Uruguay and Brazil, respectively. Two approaches were developed: (a) a bi-character analysis of extreme sub-datasets within each environmental factor (bottom and top 33% of regions), (b) three different production environments (including farms from both countries) were defined in a cluster analysis using standardized environmental factors. To identify the variables that influenced the cluster formation, a discriminant analysis was previously carried out. Management (month, age and weight at weaning) and climatic factors (accumulated rainfalls and winter and summer temperatures) were the most important factors in the clustering of farms. Bi or trivariate analyses were performed to estimate heritability and genetic correlations for WW in extreme sub-datasets within environmental factor or between clusters, using MTDFREML software. Heritability estimates of WW in the first approach ranged from 0.27 to 0.54, and genetic correlations between top and bottom sub-datasets within environmental factors, from -0.29 to 0.70. In the cluster approach, heritabilities were 0.58±0.04 for cluster 1, 0.31±0.01 for Cluster 2 and 0.40±0.02 for Cluster 3. Genetic correlations were 0.27±0.08, 0.32±0.09 and 0.33±0.09, between clusters 1 and 2, 1 and 3, and 2 and 3, respectively. Both approaches suggest the existence of genotype x environment interaction for weaning weight in Angus breed of Brazil and Uruguay. © 2012 Elsevier B.V.

Babesia bovis infection in cattle in the southwestern Brazilian Amazon

The present study provides the first epidemiological data on infection with Babesia bovis in cattle raised in the southwestern Brazilian Amazon. Blood clot samples were filtered through nylon cloth before being submitted to DNA extraction. PCR and nested-PCR were applied to assess the frequency of infection with B. bovis in calves with ages from 4 to 12 months bred in 4 microregions each in the states of Rondônia and Acre. After the DNA was extracted from the samples, the infection in cattle was investigated by amplification of the rap1 gene from B. bovis. The DNA amplification results revealed a frequency of infection with B. bovis of 95.1% (272/286) in the samples from Rondônia and 96.1% (195/203) in those from Acre. The high frequency of B. bovis infection in the animals with ages from 4 to 12 months indicates a situation of enzootic stability in the regions studied. The infection rates are comparable to those detected by immunodiagnostic techniques in other endemic regions of Brazil. © 2012 Elsevier GmbH.

Supplementation with the histone deacetylase inhibitor trichostatin A during in vitro culture of bovine embryos

Summary Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA. © Cambridge University Press 2011.

Phenotypic plasticity of composite beef cattle performance using reaction norms model with unknown covariate

The objective of the present study was to determine the presence of genotype by environment interaction (G × E) and to characterize the phenotypic plasticity of birth weight (BW), weaning weight (WW), postweaning weight gain (PWG) and yearling scrotal circumference (SC) in composite beef cattle using the reaction norms model with unknown covariate. The animals were born between 1995 and 2008 on 33 farms located throughout all Brazilian biomes between latitude -7 and -31, longitude -40 and -63. The contemporary group was chosen as the environmental descriptor, that is, the environmental covariate of the reaction norms. In general, higher estimates of direct heritability were observed in extreme favorable environments. The mean of direct heritability across the environmental gradient ranged from 0.05 to 0.51, 0.09 to 0.43, 0.01 to 0.43 and from 0.12 to 0.26 for BW, WW, PWG and SC, respectively. The variation in direct heritability observed indicates a different response to selection according to the environment in which the animals of the population are evaluated. The correlation between the level and slope of the reaction norm for BW and PWG was high, indicating that animals with higher average breeding values responded better to improvement in environmental conditions, a fact characterizing a scale of G × E. Low correlation between the intercept and slope was obtained for WW and SC, implying re-ranking of animals in different environments. Genetic variation exists in the sensitivity of animals to the environment, a fact that permits the selection of more plastic or robust genotypes in the population studied. Thus, the G × E is an important factor that should be considered in the genetic evaluation of the present population of composite beef cattle. © The Animal Consortium 2012.

Short communication: Acute but transient increase in serum insulin reduces messenger RNA expression of hepatic enzymes associated with progesterone catabolism in dairy cows

The objective of this experiment was to evaluate the effects of glucose infusion on serum concentrations of glucose, insulin, and progesterone (P4), as well as mRNA expression of hepatic CYP2C19 and CYP3A4 in nonlactating, ovariectomized cows in adequate nutritional status. Eight Gir × Holstein cows were maintained on a low-quality Brachiaria brizantha pasture with reduced forage availability, but they individually received, on average, 3. kg/cow daily (as fed) of a corn-based concentrate from d -28 to 0 of the experiment. All cows had an intravaginal P4-releasing device inserted on d -14, which remained in cows until the end of the experiment (d 1). On d 0, cows were randomly assigned to receive, in a crossover design containing 2 periods of 24. h each (d 0 and 1), (1) an intravenous glucose infusion (GLUC; 0.5. g of glucose/kg of BW, over a 3-h period) or (2) an intravenous saline infusion (SAL; 0.9%, over a 3-h period). Cows were fasted for 12. h before infusions, and they remained fasted during infusion and sample collections. Blood samples were collected at 0, 3, and 6. h relative to the beginning of infusions. Liver biopsies were performed concurrently with blood collections at 0 and 3. h. After the last blood collection of period 1, cows received concentrate and returned to pasture. Cows gained BW (16.5 ± 3.6. kg) and BCS (0.08 ± 0.06) from d -28 to 0. Cows receiving GLUC had greater serum glucose and insulin concentrations at 3. h compared with SAL cohorts. No treatment effects were detected for serum P4 concentrations, although mRNA expression of CYP2C19 and CYP3A4 after the infusion period was reduced for cows in the GLUC treatment compared with their cohorts in the SAL treatment. In conclusion, hepatic CYP3A4 and CYP2C19 mRNA expression can be promptly modulated by glucose infusion followed by acute increases in circulating insulin, which provides novel insight into the physiological mechanisms associating nutrition and reproductive function in dairy cows. © 2013 American Dairy Science Association.

Resistance against gastrointestinal nematodes in Crioulo Lageano and crossbred Angus cattle in southern Brazil

Gastrointestinal nematode (GIN) infection is a major cause of production losses in cattle. This study was carried out to evaluate the natural resistance against nematode infection in Crioulo Lageano and crossbred Angus male calves. Crioulo Lageano is a local cattle breed in the state of Santa Catarina, in southern Brazil. Ten weaned calves of each breed were grazed together on pasture and naturally infected with nematodes between July 2009 and December 2010. Once every 28 days, we collected fecal and blood samples for parasitological and immunological tests, as well as recording body weights. After 19 samplings, all animals were slaughtered for quantification and identification of GINs. We found that the animals had been infected with the following nematode species, in decreasing order by the mean number of specimens: Trichostrongylus axei, Cooperia punctata, Ostertagia ostertagi, Haemonchus placei, Oesophagostomum radiatum, and Trichuris spp. There were no significant differences between the Crioulo Lageano and crossbred Angus groups in terms of worm burden or nematode fecal egg count, nor in terms of the mean levels of immunoglobulin (G and A) against C. punctata and H. placei antigens, except in IgA mean level in abomasal mucus against H. placei adult worms that was significantly higher in crossbred Angus cattle (p<0.05). At the end of the study, the crossbred Angus cattle were heavier than were the Crioulo Lageano cattle (mean live weight, 507.35 and 390.3. kg, respectively). Comparative parasitological and immunological evaluation revealed no difference between two breeds in terms of their natural resistance against GINs. © 2012 Elsevier B.V.

Effects of vaccination against reproductive diseases on reproductive performance of lactating dairy cows submitted to AI

Four experiments evaluated the effects of vaccination against bovine herpesvirus-1 (BoHV-1), bovine viral diarrhea virus (BVDV), and Leptospira spp. on reproductive performance of lactating dairy cows without (experiments 1, 2, and 3) or with previous vaccination against these diseases (experiment 4). Cows were assigned to a fixed-time AI protocol (FTAI; d -11 to 0) in all experiments, as well as AI 12. h upon estrus detection in experiment 3. Pregnancy status was determined with transrectal ultrasonography on d 30 and 71 (d 60 for experiment 3) after AI. Pregnancy loss was considered in cows pregnant on d 30 but non-pregnant on the subsequent evaluation. In experiment 1, 853 cows received (VAC) or not (CON) vaccination against BoHV-1, BVDV, and Leptospira spp. at the beginning of the FTAI (d -11) and 30. d after AI. Pregnancy loss was reduced (P=0.03) in VAC cows compared with CON. In experiment 2, 287 cows received VAC or CON 30. d prior to (d -41) and at the beginning (d -11) of the FTAI. Pregnancy rates on d 30 and 71 were greater (P≤0.03) in VAC cows compared with CON. In experiment 3, 1680 cows with more than 28. d in milk were randomly assigned to receive VAC or CON with doses administered 14. d apart, and inseminated within 15-135. d after the second dose. Pregnancy rates on d 30 and 60 were greater (P≤0.02) in VAC cows compared with CON. In experiment 4, 820 cows received (REVAC) or not (CON) revaccination against BoHV-1, BVDV, and Leptospira spp. at the beginning of the FTAI protocol (d -11). Pregnancy rates and loss were similar (P≥0.54) between treatments. Hence, vaccinating naïve cows against BoHV-1, BVDV, and Leptospira spp. improved reproductive efficiency in dairy production systems, particularly when both doses were administered prior to AI. © 2013 Elsevier B.V.

Analysis of the risk factors related to the immune humoral anti-Anaplasma marginale in dairy cattle

Anaplasma marginale is the most prevalent pathogen of cattle transmitted by ticks in the world. This study aimed to evaluate the risk factors for anaplasmosis in dairy cattle. Fifty dairy cattle from the herd of Empresa de Pesquisa Agropecuaria do Estado do Rio de Janeiro were selected by proportional stratified sampling. The risk factors evaluated were: physiological state, race pattern, number of lactations, milk production, infestation by Rhipicephalus microplus and animal density. Antibody activity against A. marginale was determined using the indirect enzyme-linked immunosorbent assay. The percentual values of seroprevalence for A. marginale were submitted to X2 test, and the level of minimum significance, to keep a factor in the model of logistic regression, was fixated in 5%. It was observed that pregnancy and lactation influenced significantly (p<0.05) in the seropositivity of the animals. Bos indicus animals had 5.21 times more chances of being seropositive than B. taurus animals. Primiparous female had 88% more chances of being seropositive than pluriparous female. Animals with high milk production were 63% more positive than low production animals. When infested by ticks the animals had 39% more chance of being seropositive to A. marginale. Bos indicus animals presented 5.21 times more chance of being seropositive than B. taurus animals. Primiparous females presented 88% more chance of being seropositive than the pluriparous ones. High milk production animals were 63% more positive than the low production ones. When infested by ticks the animals had 39% more chance of being seropositive to A. marginale. High density grazing provided for the animals 3.2 times more chances of being seropositive than low density ones. The herd was classified as erratic to A. marginale, even being placed in a steady enzootic area.

Evaluation of the hypothalamus-pituitary axis response to exogenous GnRH, estradiol benzoate, And LH during the postpartum period in Nellore cows

The objective was to evaluate when the LH reserve was re-established in postpartum Nellore (Bos indicus) cows by evaluating the response of the hypothalamic-pituitary axis responsiveness to exogenous GnRH or estradiol benzoate (EB). Additionally, we tested the influence of dietary supplementation (SUPL) and calf removal (CR) on the duration of postpartum anestrus. Ninety multiparous lactating Nellore cows were randomly assigned to eight groups. The EB and GnRH groups received 1.0 mg EB (N = 7), and 50 μg lecireline (N = 16), respectively. Additional cows were given the same hormones, and subjected to either nutritional supplementation (EB-SUPL, N = 9; GnRH-SUPL, N = 16), or calf removal at 72 hours after calving (EB-CR, N = 4; GnRH-CR, N = 13). The remaining two groups were the LH (12.5 mg, N = 14) and control groups (saline, N = 11). Hormones were administered weekly from 7 (±5) days postpartum to first ovulation (detection of a CL during a weekly ultrasonographic examination). Blood samples were collected just before and 2 hours (GnRH, LH, and control groups) or 18 hours (EB groups) after hormone or saline (control) administration. Ovulation occurred as early as 15 days postpartum in the GnRH group. The mean ± SEM intervals (days) from calving to first ovulation were EB, 87.7 ± 4.2; EB-CR, 20.3 ± 1.2; EB-SUPL, 60.3 ± 3.2; GnRH, 40.4 ± 2.1; GnRH-CR, 21.0 ± 1.1; GnRH-SUPL, 26.4 ± 1.1; LH, 35.6 ± 1.1; and control, 60.9 ± 2.1. We concluded that there was sufficient LH in the pituitary gland (of Nellore cows) from the second week postpartum to induce ovulation in response to exogenous GnRH. Additionally, calf removal and nutritional supplementation reduced, by 2 to 4 weeks, the interval from calving to an LH increase and ovulation induced by GnRH or EB. © 2013.