Gyr Cows Research Articles

Association between BMP4 gene polymorphism and in vitro embryo production traits in Gyr cows

Association between BMP4 gene polymorphism and in vitro embryo production traits in Gyr cows

Association between BMP4 gene polymorphism and in vitro embryo production traits in Gyr cows

Association between BMP4 gene polymorphism and in vitro embryo production traits in Gyr cows

Modelos para avaliação genética da produção de leite em múltiplas lactações

O objetivo deste trabalho foi avaliar componentes de covariância e valores genéticos para produção de leite acumulada em até 305 dias, a partir dos dados das três primeiras lactações de vacas Gir. Foram analisados dados de 14.659 lactações, de 9.079 vacas, por meio dos modelos de repetibilidade, multicaracterístico (Mult) e de regressão aleatória com variância residual homogênea (MRAHo) ou heterogênea (MRAHe). A produção de leite foi considerada como característica distinta em cada lactação, no modelo Mult. Polinômios lineares foram utilizados nos modelos de regressão aleatória para ajuste das trajetórias médias e dos efeitos genético aditivo e de ambiente permanente individuais, de acordo com a ordem de parto. As médias a posteriori da herdabilidade foram semelhantes entre os diferentes modelos e lactações, e variaram entre 0,24 e 0,29. Os modelos Mult e MRAHe ajustaram-se melhor aos dados, uma vez que observou-se heterogeneidade de variâncias genéticas e residuais entre lactações. As correlações genéticas da produção acumulada de leite em até 305 dias nas três primeiras lactações foram próximas de 1,0; portanto, a seleção de reprodutores já pode ser feita a partir dos resultados da primeira lactação. Modelos de regressão aleatória com variâncias genéticas e residuais heterogêneas permitem modelar adequadamente as covariâncias das produções de leite acumuladas em múltiplas lactações e obter valores genéticos para seleção de reprodutores com base nos dados já da primeira lactação.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: III. Placental adaptations and placentome gene expression

This study investigated the influence of day of gestation (DG) and feeding regimens (FR) on the expression of genes responsible for placenta development, nutrient transfer, and angiogenic factors in Holstein × Gyr cows. Forty pregnant multiparous Holstein × Gyr cows with an average initial body weight of 482±10.8kg and an initial age of 5±0.8 yr were allocated to 1 of 2 FR: ad libitum (AL; n=20) or maintenance level (ML; n=20). Maintenance level was considered to be 1.15% of body weight (dry matter basis) and met 100% of the net energy requirements and AL provided 190% of the total net energy requirements. Cows were slaughtered at 4 DG: 139, 199, 241, and 268d. After the cows were slaughtered, the placenta and uterus were separated and weighed. Caruncles and cotyledons were individually separated, counted, and weighed. Placenta expressed as kilograms and grams per kilogram of empty body weight (EBW) was heavier in ML- than in AL-fed cows at 268d of gestation. Placenta expressed as kilograms and grams per kilogram of EBW was the lightest at 139d of gestation, and the greatest mass was observed at 268d in ML-fed cows. However, in AL-fed cows, the heaviest placenta expressed as grams per kilogram of EBW was observed from 199d of gestation. Placentomes expressed as grams per kilogram of EBW were heavier in ML-fed cows during gestation, and the number of placentomes was greater in ML-fed cows at 268d of gestation. We observed that IGFR1 and IGFR2 were involved in placenta adaptations when ML was provided, as their expression in placentome cells was greater in ML-fed cows at 268d of gestation. The genes responsible for angiogenesis were also greater in ML-fed cows: VEGFA, GUCY1B3, HIFA, FGF2, and NOS3 were altered by FR and DG interaction and they were greater in ML-fed cows at 268d of gestation. In addition, VEGFB and ANGPT2 did not show interactions between FR and DG, but they were greater in ML-fed cows. Thus, we suggest that the placenta from an ML-fed cow develops adaptations to the reduced nutrient supply by altering its structure and gene expression, thereby developing mechanisms for potential increased nutrient transfer efficiency to the fetus.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: II. Maternal and fetal visceral organ mass

This study investigated the influence of day of gestation (DG) and feeding regimens (FR) on maternal and fetal visceral organ mass in Holstein × Gyr cows. Forty-four pregnant multiparous Holstein × Gyr cows with an average initial body weight of 480±10.1 kg and an average initial age of 5±0.5 yr were allocated to 1 of 2 FR: ad libitum (AL; n=20) or maintenance level (ML; n=24). Maintenance level was considered to be 1.15% of body weight (dry matter basis) and met 100% of the energy requirements; AL provided 190% of the total net energy requirements. Cows were individually fed a corn silage and concentrate-based diet composed of 93% roughage and 7% concentrate (dry matter basis) as a total mixed ration twice daily. Pregnant cows were slaughtered at 4 DG: 139 (n=11), 199 (n=11), 241 (n=11), and 268 (n=11) d, which was followed by necropsy. Mass of heart, liver, and gastrointestinal tract was heavier in AL- than in ML-fed cows. Mammary gland mass was heavier in AL- than in ML-fed cows, and the heaviest mass was observed at 268 d of gestation. Feeding regimen did not influence fetal body weight in this study. The majority of the visceral organ masses were similar in fetuses from cows fed AL or ML. These data indicate that maternal feed restriction does not affect the development of most fetal organs or fetal development; however, some maternal organs are affected by the FR provided. Moreover, the negative effect on mammary gland mass caused by ML feeding will probably not affect the subsequent lactation because the crude protein concentration in the mammary gland increased with ML feeding. However, we suggest that the AL diet in pregnant dry cows should be provided with caution because the amount of fat in the mammary gland increased at 268 d of gestation.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: I. Apparent total-tract digestibility, nitrogen balance, and fat deposition

This study investigated how feeding regimen (FR) alters apparent total-tract digestibility, performance, N balance, excretion of purine derivatives, and fat deposition in Holstein × Gyr cows at different days of gestation (DG). Forty-four pregnant multiparous Holstein × Gyr cows with an average initial body weight of 480±10.1 kg and an initial age of 5±0.5 yr old were allocated to 1 of 2 FR: ad libitum (AL; n=20) and maintenance level (ML; n=24). Maintenance level was considered to be 1.15% of body weight on a dry matter (DM) basis and met 100% of the energy requirements, whereas AL provided 190% of total net energy requirements. Data for hot and cold carcass dressing, fat deposition, average daily gain, empty body gain, and average daily gain without the gravid uterus were analyzed as a 4×2 factorial design. Intake, apparent total-tract digestibility, N balance, urinary concentration of urea, and purine derivatives data were analyzed as repeated measurements taken over the 28-d period (122, 150, 178, 206, 234, and 262 d of gestation). Cows were individually fed a corn silage-concentrate based diet composed of 93% roughage and 7% concentrate (DM basis) as a total mixed ration. Pregnant cows were slaughtered on 4 different DG: 139 (n=11), 199 (n=11), 241 (n=11), and 268 d (n=11). Overall, DM intake decreased as DG increased. This decrease observed in DM intake may be associated with the reduction in ruminal volume caused by the rapid increase in fetal size during late gestation. We observed an interaction for DM and organic matter apparent total-tract digestibility between FR and DG; at 150, 178, and 206 d of gestation, ML-fed cows had greater DM and organic matter apparent total-tract digestibility values than AL-fed cows. Rib fat thickness, mesentery, and kidney, pelvic, and heart fat were greater in AL-fed than in ML-fed cows at all DG, with the exception of rib fat thickness on d 139. Ad libitum-fed cows excreted more N in their feces and urine compared with ML-fed cows. Pregnant cows that were fed at maintenance had greater digestibility during some DG, excreted less N in feces and less N and urea in urine, and deposited less fat in the body. We therefore recommend ML (1.15% of body weight with 93% of roughage) as a FR for pregnant dry cows; however, during the last month of gestation, AL seems to be the most appropriate FR to avoid loss of body weight.

Soybean in different forms of processing in the feeding of crossbred cows on brachiaria grass pastures

The objective was to evaluate intake and digestibility of nutrients, as well as milk production and composition of the milk from F1 Holstein × Gyr cows kept on pasture, supplemented with sugarcane and concentrate (28% CP). Five cows with 150±14 lactation days and average milk production of 7.1±2.1 kg/day were distributed in a 5 × 5 Latin square design. The treatments were: soybean meal-based concentrate; soy bean-based concentrate; ground soybean-based concentrate; roasted soybean-based concentrate; and ground, roasted soybean-based concentrate. Dry matter (DM), organic matter, crude protein (CP), neutral detergent fiber (NDF), total carbohydrates and total digestible nutrients intakes were not affected by the diet, but ether extract intake was higher for the animals fed soy bean-based diets than those fed soybean meal. The digestibility of DM, NDF and CP did not differ. The corrected milk yield differed between treatments; animals on the treatment with soy bean-based concentrate had the lowest production in relation to the others, which did not differ from each other. There was no difference between treatments for milk composition. Thus, soybean meal can be replaced by ground soy beans or roasted soy beans (ground or whole) in diets for low-yield cows reared on Brachiaria decumbens pastures with no harm to milkproduction or composition. Therefore, the use of these alternative raw materials is recommended whenever their inclusion represents lower feeding costs.

Random regression analysis of test-day milk yields in the first and second lactations of Brazilian Gyr cows.

The objective of the present study was to estimate the genetic parameters for test-day milk yields (TDMY) in the first and second lactations using random regression models (RRM) in order to contribute to the application of these models in genetic evaluation of milk yield in Gyr cattle. A total of 53,328 TDMY records from 7118 lactations of 5853 Gyr cows were analyzed. The model included the direct additive, permanent environmental, and residual random effects. In addition, contemporary group and linear and quadratic effects of the age of cows at calving were included as fixed effects. A random regression model fitting fourth-order Legendre polynomials for additive genetic and permanent environmental effects, with five classes of residual variance, was applied. In the first lactation, the heritabilities increased from early lactation (0.26) until TDMY3 (0.38), followed by a decrease until the end of lactation. In the second lactation, the estimates increased from the first (0.29) to the fifth test day (0.36), with a slight decrease thereafter, and again increased on the last two test days (0.34 and 0.41). There were positive and high genetic correlations estimated between first-lactation TDMY and the remaining TDMY of the two lactations. The moderate heritability estimates, as well as the high genetic correlations between half the first-lactation TDMY and all TDMY of the two lactations, suggest that the selection based only on first lactation TDMY is the best selection strategy to increase milk production across first and second lactations of Gyr cows.

Efeito da idade de aquisição de fêmeas de reposição na rentabilidade da atividade leiteira

Analisou-se, por meio de simulação, o efeito da idade de aquisição de fêmeas para reposição na rentabilidade de um sistema de produção de leite com vacas F1 Holandês x Gir, no período de 01 de maio de 2011 a 30 de abril de 2012. Durante o período chuvoso, na época do verão, as vacas foram mantidas em pastos e, durante o período da seca, foram confinadas com o uso da cana-de-açúcar in natura enriquecida com ureia e sulfato de amônio. A referência zootécnica foi do rebanho da Fazenda Experimental de Felixlândia (FEFX) da Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG), localizada no município de Felixlândia, MG. O inventário, bem como as despesas, as receitas e os demais dados, foram cadastrados no software CUSTO BOVINO LEITE 1.0, visando obter a análise de rentabilidade. Tal software contempla as metodologias de custo total de produção (clásssica), que envolve o custo fixo e variável, e custo operacional. A aquisição da fêmea de reposição, aos 10 ou aos 30 meses de idade, é rentável para o sistema de produção de leite analisado. Como alternativa mercadológica, a aquisição da fêmea de reposição com 10 meses de idade foi a pior opção, devido ao alto valor de aquisição e devido a necessidade da aquisição de maior quantidade de fêmeas, em função da taxa de mortalidade.

Phenotypic correlations between ovum pick-up in vitro production traits and pregnancy rates in Zebu cows.

The growth of the Gyr breed in Brazil in terms of genetic gain for milk, along with conditions for market, has led to the use of ovum pick-up in vitro production (OPU-IVP) as a leader in biotechnology for the multiplication of genetic material. The aim of this study was to investigate phenotypic correlations between OPU-IVP-linked characteristics and pregnancy rates registered in an embryo transfer program using Gyr cows as oocyte donors. Data collected from 211 OPU sessions and 298 embryo transfers during the years 2012 and 2013 were analyzed and statistical analysis was performed. Estimates of simple Pearson correlations were calculated for NVcoc and PVcoc (number and proportion of viable cumulus-oocyte complexes, respectively); NcleavD4 and PcleavD4 (number and proportion of cleaved embryos on day 4 of culture, respectively); NTembD7 and PTembD7 (number and proportion of transferable embryos on day 7 of culture, respectively); NPrD30 and PPrD30 (number and proportion of pregnancies 30 days after transfer, respectively); and NPrD60 and PPrD60 (number and proportion of pregnancies 60 days after transfer, respectively). Moderate to moderately high correlations were found for all numerical characteristics, suggesting these as the most suitable parameters for selection of oocyte donors in Gyr programs. NVcoc is proposed as a selection trait due to positive correlations with percentage traits and pregnancy rates 30 and 60 days after transfer.

202 ASSOCIATION BETWEEN BMP4 GENE POLYMORPHISM AND OVUM PICK-UP IN VITRO PRODUCTION TRAITS IN BRAZILIAN Gyr COWS

In Bos taurus taurus, previous studies have reported an association between single nucleotide polymorphism (SNP) in BMP4 gene and the blastocyst rate in cows evaluated for in vitro embryo production efficacy. The aim of this study was to evaluate the degree of association between a polymorphism in the BMP4 gene and characteristics related to in vitro embryo production as well as pregnancies thereof in Bos taurus indicus cattle (Gyr breed). Data from 212 ovum pick-up in vitro (OPU-IPV) sessions were collected of 50 Gyr cows in the Rio de Janeiro state, Brazil. The OPU-IVP procedures were performed and associated with marker SNP. The OPU-IVP traits were number and proportion of viable cumulus-oocyte complexes, number and proportion of cleaved embryos at Day 4 of culture, number and ratio of transferable embryos at Day 7 of culture, number and proportion of pregnancies at Day 30 after transfer, and number and proportion of pregnancies at Day 60 after transfer. DNA was extracted from hair follicles. BMP4 polymorphism, the segment of the BMP4 gene (exon 2, SNP rs109778173) was amplified by PCR with a specific primer. PCR-restriction fragment length polymorphism (RFLP) was used for genotyping. HinfI restriction enzyme that recognises and cleaves the sequence 5′ G ? ANTC 3′ was used. The samples were submitted to capillary electrophoresis Fragment Analyzer™ (Advanced Analytical Technologies Inc., Ames, IA) for allelic discrimination by size and polymorphism identification. Genotypic frequency, allele frequency, Hardy–Weinberg equilibrium probability, gene homozygosity, gene heterozygosity, effective allele number, and polymorphism information content were statistically analysed in the software PowerMarker v.3.25 (North Carolina State University, Raleigh, NC, USA). The association between DNA marker and the OPU-IVP traits and pregnancy rates was make by analysis of variance of repeated data PROC GLIMMIX of SAS 2009 (SAS Institute Inc., Cary, NC, USA). The domain of the protein coding of BMP4 gene is located in exon 2. A fragment of the sequence of DNA in exon 2 was amplified, where the mutation SNP rs109778173 was identified (G > T) in Gyr cows using PCR-RFLP and capillary electrophoresis methods. Three genotypes were identified (GG, TT, and GT) with genotypic frequencies of 0.64, 0.32, and 0.04 respectively; allelic frequency and Hardy–Weinberg equilibrium. The polymorphism was significantly associated with the number of viable oocyte complexes, ratio of viable oocyte complex, and ratio of pregnancies at 30 days (P > 0.01). The GT genotype affects negatively the number and proportion of viable cumulus-oocyte complexes and the proportion of pregnancies at Day 30 after transfer. This finding is an indication of a genetic effect of these characteristics.

105 RETAINED FETAL MEMBRANES: INCIDENCE AND EFFECT ON MILK PRODUCTION AND REPRODUCTIVE PERFORMANCE IN DAIRY COWS

The objectives were to describe the incidence of retained fetal membrane (RFM) and its effect on production and reproduction of dairy cows in tropical environments. Holstein and crossbred Holstein × Gyr cows and heifers calving between May 2012 and May 2013 from 4 dairy farms were enrolled in this study. Retained fetal membranes were defined as the non-expelling of placenta within 12 to 24 h after calving. All cows with RFM were treated with 10 mg kg–1 of oxitetracycline IV and drenching containing 100 g of CaCl2, 100 g of KCl, 200 g of Mg(SO4), and 350 mL of propylene glycol in 25 L of water once per day for 3 days. The incidence of RFM was 14.7% (92/624) in multiparous cows and 11.5% (67/583) in primiparous cows (P = 0.003). The incidence of retained placenta was 14.7% (92/624) during the summer and 14.8% (108/731) during the winter (P = 0.99). Conception rates after first postpartum AI were 32.5% (276/850) and 25.8% (34/132) for healthy cows and cows with RFM, respectively (P = 0.12). More cows that did not have RFM were pregnant at 150 DIM than cows with retained placenta (42.3 v. 29.3%; P = 0.003). Based on predicted 305-day milk yield, cows with retained placenta produced less milk than healthy cows (5924.0 ± 158.0 v. 6315.3 ± 61.0 kg, respectively; P = 0.009). The average peak production was 30.8 ± 0.3 and 29.1 ± 0.7 kg for healthy and RFM cows, respectively (P = 0.02). During the first 60 days in milk, the total milk production for RFM and healthy cows was, respectively, 1305.7 ± 37.3 and 1441.2 ± 15.8 L (P = 0.0008). In conclusion, although season did not influence the incidence of RFM in dairy cows, parity had an effect. Moreover, RFM was associated with a decrease in reproductive efficiency and milk production in dairy cows managed in the tropics. Financial support was received from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) of Brazil.

Effect of somatic cell count and mastitis pathogens on milk composition in Gyr cows

BackgroundGyr cows are well adapted to tropical conditions, resistant to some tropical diseases and have satisfactory milk production. However, Gyr dairy herds have a high prevalence of subclinical mastitis, which negatively affects their milk yield and composition. The objectives of this study were (i) to evaluate the effects of seasonality, mammary quarter location (rear x front), mastitis-causing pathogen species, and somatic cell count (SCC) on milk composition in Gyr cows with mammary quarters as the experimental units and (ii) to evaluate the effects of seasonality and somatic cell count (SCC) on milk composition in Gyr cows with cows as the experimental units. A total of 221 lactating Gyr cows from three commercial dairy farms were selected for this study. Individual foremilk quarter samples and composite milk samples were collected once a month over one year from all lactating cows for analysis of SCC, milk composition, and bacteriological culture.ResultsSubclinical mastitis reduced lactose, nonfat solids and total solids content, but no difference was found in the protein and fat content between infected and uninfected quarters. Seasonality influenced milk composition both in mammary quarters and composite milk samples. Nevertheless, there was no effect of mammary quarter position on milk composition. Mastitis-causing pathogens affected protein, lactose, nonfat solids, and total solids content, but not milk fat content. Somatic cell count levels affected milk composition in both mammary quarters and composite samples of milk.ConclusionsIntramammary infections in Gyr cows alter milk composition; however, the degree of change depends on the mastitis-causing pathogen. Somatic cell count is negatively associated with reduced lactose and nonfat solids content in milk. Seasonality significantly affects milk composition, in which the concentration of lactose, fat, protein, nonfat solids and total solids differs between dry and wet seasons in Gyr cows.

Milk yield persistency in Brazilian Gyr cattle based on a random regression model

With the objective of evaluating measures of milk yield persistency, 27,000 test-day milk yield records from 3362 first lactations of Brazilian Gyr cows that calved between 1990 and 2007 were analyzed with a random regression model. Random, additive genetic and permanent environmental effects were modeled using Legendre polynomials of order 4 and 5, respectively. Residual variance was modeled using five classes. The average lactation curve was modeled using a fourth-order Legendre polynomial. Heritability estimates for measures of persistency ranged from 0.10 to 0.25. Genetic correlations between measures of persistency and 305-day milk yield (Y305) ranged from -0.52 to 0.03. At high selection intensities for persistency measures and Y305, few animals were selected in common. As the selection intensity for the two traits decreased, a higher percentage of animals were selected in common. The average predicted breeding values for Y305 according to year of birth of the cows had a substantial annual genetic gain. In contrast, no improvement in the average persistency breeding value was observed. We conclude that selection for total milk yield during lactation does not identify bulls or cows that are genetically superior in terms of milk yield persistency. A measure of persistency represented by the sum of deviations of estimated breeding value for days 31 to 280 in relation to estimated breeding value for day 30 should be preferred in genetic evaluations of this trait in the Gyr breed, since this measure showed a medium heritability and a genetic correlation with 305-day milk yield close to zero. In addition, this measure is more adequate at the time of peak lactation, which occurs between days 25 and 30 after calving in this breed.

17 CELL CYCLE SYNCHRONIZATION OF BOVINE FIBROBLASTS BY AZADIRACHTA INDICA EXTRACTS

One requirement for somatic cell nuclear transfer (NT) is the coordination between donor cell cycle and recipient cytoplasm. As an alternative to commercially available substances to synchronize the cell cycle in G0/G1, we tested 2 extracts, aqueous and hexane obtained from the plant Azadirachta indica A. Juss (popularly called Neem). Extracts from this plant have shown antiviral, antibacterial and anticancer activities, widely described in the literature (Kumar et al. 2009 Invest. New Drugs 27, 246–252). The hexane extract was prepared in the Soxhlet apparatus until total collapse and then submitted to rotary evaporation. The aqueous extract was prepared by dynamic maceration and was subsequently lyophilized. Bovine fibroblasts collected from Gyr cows were cultured in DMEM (Sigma) supplemented with 10% fetal cow serum (FCS) and incubated at 37°C, 5% CO2 and 95% humidity. After obtaining 70% of cell confluence, the extract was added to cells at the following concentrations: 0 μg mL–1 (negative control), 50 μg mL–1, 100 μg mL–1, 200 μg mL–1 and 300 μg mL–1, for 12 and 24 h. Simultaneously, a group with serum starvation (positive control; cells cultured in DMEM plus 0.5% FBS for 3 days) was prepared. Three repetitions were performed in triplicate for each concentration and control groups. Cell cycle readings were performed by flow cytometry (Facs Callibur, Becton Dickinson, San Jose, CA, USA) and DNA histograms were analysed by WinMDI software to determine the percentage of cells in G0/G1 phase, S and G2 cell cycle, so that 10 000 cells were analysed in each reading on a flow cytometer. Data were analysed by analysis of variance and means were compared by Student-Newman-Keuls test. The percentages of cells at G0/G1 phase for aqueous extracts were lower (P < 0.05), regardless of the concentration and exposure time, than the 0 μg mL–1 (83.73 ± 1.14%) and serum starvation (86.64 ± 1.44%). In contrast, the percentages of cells synchronized at G0/G1 with 50 μg mL–1 for 12 h (84.23 ± 0.56%), 50 μg mL–1 for 24 h (85.66 ± 0.57%), 100 μg mL–1 for 12 h (87.85 ± 0.51%) and 200 μg mL–1 for 12 h (85.87 ± 0.45%) using hexane extracts were higher (P < 0.05) than with 0 μg mL–1 (81.44 ± 0.29%), but lower (P < 0.05) than the serum starvation (91.33 ± 0.31%). In conclusion, hexane extracts from the plant Azadirachta indica A. Juss can synchronize mammalian cell cycle at G0/G1 despite the low proportion when compared with serum starvation. Studies to evaluate efficiency of cell cycle resuming and viability after somatic cell nuclear transfer are ongoing. Financial support: FAPEMIG, CNPq and Project 01.07.01.002.

169 THE RELATIONSHIP BETWEEN OOCYTE RECOVERY AND EMBRYO PRODUCTION IN BOS INDICUS

The ovarian follicular population has been used as a parameter to evaluate fertility and also the potential of donors undergoing assisted reproductive procedures in both human medicine and animal practice. There is a high correlation between follicular population and oocyte recovery by ovum pickup (OPU), but the relationship between oocyte recovery, embryo production and pregnancy rates may not be fully understood. The aim of the present study was to evaluate whether the conversion rate of oocytes to embryos and further pregnancies could be positively related to the number of cumulus–oocyte complexes (COC) recovered after OPU in cattle. For this purpose, records of 626 OPU sections from 251 nonlactating Gyr cows (dairy Zebu breed) were analysed. The animals had a good body condition score, were kept in a good feeding pasture (Brachiaria spp.) and were supplemented with corn silage and a mixture of corn, soybeans and vitamin and minerals, according to their nutritional requirements. For each ovarian aspiration, the ovarian follicular wave was previously synchronized with an auricular implant (Norgestomet-Crestar®), IM injections of 2 mg of oestradiol benzoate (Gonadiol®) and 0.25 mg of D-cloprostenol (Sincrocio®). The OPU procedures were performed using an ultrasound device (Aquila Vet, Esaote, São Paulo, Brazil) equipped with a vaginal sector 7.5-MHz probe, disposable 20 G needles and a vacuum pressure of 80 mmHg. The cows were ranked in quartiles regarding the total number of COC recovered. To reduce bias related to the eventual fluctuation of OPU results, for the present analysis the authors used only the recorded OPU session of each cow with the highest number of COC recovered. Viable COC were fertilized with sex-sorted (X) semen of Gyr bulls previously tested for in vitro embryo production. Conversion rates (%) of the total and viable oocytes to embryos, viable oocytes to pregnancy and embryo to pregnancy were evaluated for each quartile. Differences between the first and fourth quartiles were accessed by Fisher's exact test. In the 251 OPU, 4246 total and 3173 viable COC were recovered, resulting in the production of 1001 embryos (31.5%) and 453 pregnancies (45.3%). The cows ranked in the first, second, third and fourth quartiles produced >30 (41.6 ± 10.6), 21 to 30 (25.2 ± 3.0), 12 to 20 (15.9 ± 2.6) and <12 (6.7 ± 3.1) total oocytes. The average viable oocyte (29.1 ± 11.0, 18.1 ± 5.3, 11.1 ± 3.7 and 4.5 ± 2.7, respectively) and embryo production (8.6 ± 5.7, 5.2 ± 3.6, 3.8 ± 2.8 and 1.8 ± 1.8, respectively) were different (P < 0.0001) among all quartiles. Pregnancy rates, however, did not differ (46.0, 44.9, 43.9 and 45.6%, respectively; P > 0.05). Interestingly, the conversion rates (viable oocytes to embryos and viable oocytes to pregnancies) were higher (P < 0.0001 and 0.002) in cows from the last quartile (51.1 and 31.9%) compared with those from the first quartile (23.7 and 14.7%). In conclusion, the number of COC recovered by OPU (and consequently the ovarian follicular count) can further predict the total number of embryos and pregnancies produced, but it is not directly related to the oocyte development potential. Biotran and Fapemig Project CVZ APQ 01654/09 and BPD 0007/10.

Evaluation of somatic cell count thresholds to detect subclinical mastitis in Gyr cows

The objectives of this study were (1) to determine the sensitivity (Se) and specificity (Sp) of somatic cell count (SCC) thresholds to identify subclinical mastitis in Gyr cows caused by major and minor pathogens; (2) to study the effects of month of sampling, rear or front mammary quarters, herd, intramammary infection (IMI), and bacterial species on SCC at quarter level; and (3) to describe the prevalence of IMI in Gyr cows in commercial dairy herds. In total, 221 lactating Gyr cows from 3 commercial dairy farms were selected. Milk samples were collected from individual quarters once a month for 1 yr from all lactating cows for SCC and bacteriological analysis. Mammary quarters were considered the experimental units and the SCC results were log10-transformed. Four SCC thresholds (100, 200, 300 and 400 × 103 cells/mL) were used to determine Se and Sp to identify infected mammary quarters. The overall prevalence of IMI in quarter milk samples of Gyr cows was 49.8%, and the prevalence of minor pathogens was higher (31.9%) than that of major pathogens (17.8%). Quarter samples with microbial isolation presented higher SCC compared with negative samples. Sensitivity and Sp of selected SCC thresholds varied according to the group of pathogen (major and minor) involved in the IMI definition. Sensitivity increased and Sp decreased when mammary quarters with only major pathogens isolation were considered positive. The use of a single SCC analysis to classify quarters as uninfected or infected in Gyr cows may not be a useful test for this breed because Se and Sp of SCC at the studied thresholds were low. The occurrence of IMI and the bacterial species are the main factors responsible for SCC variation in mammary quarters of Gyr cows. Milk samples with major pathogens isolation elicited higher SCC than those with minor pathogens.

Concentrate and crude protein levels in diets for dairy Gyr lineage cows grazing elephant-grass during the rainy season

The objective of this work was to evaluate the effects of three levels of concentrate (2.0, 4.0 and 6.0 kg/cow/day) and two levels of crude protein (CP) (14 and 16% total dietary dry matter), in comparison to mineral mixture (control) on the intake, apparent digestibility, milk composition and yield and on feed efficiency and use of concentrates of cows grazing elephant grass (Pennisetum purpureum, Schum) in the rainy season. Twenty-one milking Gyr cows with average body weight of 426 kg and yield of 13.0 kg of milk/cow/day at 55 days of lactation were distributed in randomized blocks design, with seven diets (treatments) in a 3 × 2 + 1 factorial arrangement and three replications, in a period of 84 days. Forage dry matter intake was not influenced by the diets, but total dietary dry matter intake increased by 45% with the inclusion of concentrate in the diet. However, milk yield increased by only 17% (1.76 kg more milk per day) with the use of concentrate. For dairy Gyr cows grazing elephant-grass during the rainy season, 2 kg of concentrate/cow/day and 14% of CP in the total diet provided the best productive response without harming body weight.

143 TREATMENT WITH VITAMINS A AND E IMPROVE OOCYTE QUALITY AND IN VITRO EMBRYO DEVELOPMENT IN BOS INDICUS COWS

We assessed the impact of a single intra-muscular injection of vitamins A and E on oocyte quality and yield and early embryo development in Bos indicus cows. Twenty Bos indicus cows, of Gyr, Brahman, and Nellore breeds, weighing between 450 and 600 kg were subjected to ovum pick-up (OPU). Oocytes were collected in Dulbecco’s PBS (DBPS) containing heparin and antibiotics, counted, and morphologically classified. Viable oocytes were taken to the laboratory, in vitro matured for 24 h and in vitro fertilized using 25 million sperm mL–1. After 168 h of incubation (39°C, 5% CO2), viable embryos were counted and classified. Then, after 10 days, the cows received an intra-muscular injection of 1 000 000 UI of vitamin A and 1 g of vitamin E, and, after 12 days, were again subjected to the same procedure described above. Differences in oocyte yield and embryo development were analysed using paired t-test. The 40 OPUs from 20 cows yielded a total of 520 oocytes. Nellore and Brahman cows produced more embryos/cow (P < 0.01) compared with Gyr. After vitamin treatment, the cows produced more oocytes (n = 303; P < 0.01) compared with the previous OPU (n = 217), resulting in 95 more viable oocytes (31%). Brahman, Gyr, and Nellore cows yielded 11.2 ± 1.8, 8.5 ± 1.5, and 12.0 ± 2.6 oocytes before vitamin treatment, respectively. From those oocytes, 224 embryos were obtained, 89 before vitamin injection and 135 after treatment (P < 0.02), with 36 more embryos (40%) produced. Irrespective of breed, cows responded equally to vitamin injection. A single parenteral injection of vitamins A and E had a significant positive effect on oocyte yield after OPU and in vitro embryo development on Bos indicus cows. We suggest that this effect is probably due to the positive influence of retinoids on oocyte and embryo development.

Fertility in Holstein × Gyr cows in a subtropical environment after insemination with Gyr sex-sorted semen

Breeding records, including 649 inseminations during fall and winter at a dairy farm in a subtropical area of Western Mexico (24 masculine N; 24 degrees C, mean annual temperature 24 degrees C) were analyzed to document effects of sex-sorted semen from commercial Gyr bullls, estrus synchronization protocol, inseminator, sire and environmental conditions on fertility of crossbred cows (Holstein x Gyr). Percentage of services resulting in pregnancies decreased sharply when sex-sorted semen was used (22.7 vs. 37.7%; P < 0.01). Although statistically not significant (P = 0.31), cows whose first insemination was in November experienced a numerically greater reduction (21 percentage points) in pregnancy rate compared to cows whose first insemination occurred in December. Substantial increases in services per pregnancy (4.71 +/- 1.35 vs. 2.13-2.43; P < 0.01) were associated with the warmer month of the study period, November, compared to other winter months. Pregnancy rates of cows regardless of semen category (33%) were not affected by sire, temperature-humidity index and estrus synchronization protocol. Cows inseminated by one inseminator had higher pregnancy rates (P < 0.01) than cows inseminated by other two technicians. The sorted sperm produced 91% (142/156) female offspring. It was concluded that, under the field conditions of the present study, pregnancy rate with sexed semen was 15 percentage points lower than pregnancy rates using conventional semen, with 91% of female calves derived from sexed sperm.