Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Free and total GMP (glycomacropeptide) contents of milk during bovine lactation

Amostras quinzenais, desde o parto até o final do período de lactação, obtidas de 34 vacas de três diferentes raças e propriedades, foram analisadas quanto à presença de GMP livre. Um pool das amostras quinzenais de cada rebanho foi analisada tanto para o conteúdo de GMP livre quanto para o GMP total (liberado da k-caseína pela ação da renina), correlacionando-os com as condições sanitárias do animal e do úbere, à fase da lactação e à produção de leite. A maioria dos problemas sanitários concentrou-se próximo ao parto, com poucas e espaçadas ocorrências de mastites clínicas. Os resultados do teste de CMT mostraram reações compatíveis às fases da lactação. Para o GMP livre as maiores variações ocorreram em função do período de lactação e em conseqüência de mastites clínicas e subclínicas. Valores elevados foram observados no início da lactação (5,87mg de ácido siálico/L de leite), normalizando para valores próximos de 3,30mg/L já ao final do segundo mês e voltando a elevar-se no terço final da lactação. em média, as mesmas tendências foram observadas para o teor de GMP total liberado pela ação de coalho comercial, iniciando com valores ligeiramente elevados (35,59mg/L), tornando-se normal e assim se mantendo até o sexto mês com valores próximos a 27,15mg/L, e novamente elevando-se gradualmente até o final da lactação, com 58,35mg de ácido siálico/L de leite. Esses resultados mostram-se úteis para a correta interpretação de métodos aplicados à seleção do leite, seja em relação ao status proteolítico da matéria-prima ou mesmo para coibição de fraudes por adição de soro ao leite.

Avanços metodológicos na seleção do sexo de espermatozóides bovinos para utilização no melhoramento genético e na produção animal

O mercado demanda, desde final da década de 20, uma tecnologia para a sexagem de espermatozóides que possa ser inserida na indústria de produção de sêmen congelado e que tenha as seguintes características: a) não altere a viabilidade espermática; b) seja compatível com a congelação do espermatozóide sexado; c) permita a sexagem de espermatozóides previamente congelados e descongelados; d) permita a produção de várias doses de sêmen sexado congelado por dia, com custo compatível ao mercado. A importância dessa tecnologia para maximizar a produção animal a um custo baixo tem sido um desafio da pesquisa a vários anos. A possibilidade de produzir, em escala comercial, doses de sêmen enriquecidas com espermatozóides X ou Y aumentará os benefícios do uso da inseminação artificial no seu papel de maximizar o progresso genético entre gerações de acordo com os requerimentos de cada programa de melhoramento animal. Diferentes rotas tecnológicas são percorridas na tentativa de selecionar-se o sexo em mamíferos, tanto nas espécies de interesse zootécnico quanto em espécies ameaçadas de extinção, animais de companhia. Neste sentido, existem duas alternativas: a separação de espermatozóides portadores do cromossomo X, daqueles portadores do cromossomo Y; ou a sexagem de embriões pré-implantados. A viabilidade da sexagem de espermatozóides em bovinos é esperada por muitos anos e os desenvolvimentos recentes tornaram essa tecnologia de aplicação commercial. Entretanto, muitas limitações ainda existem, principalmente, referente à taxa de gestação em condições de campo. Isso restringe a utilização dessa tecnologia no melhoramento genético e produção animal. Nessa palestra abordaremos os potenciais sistemas de criação e produção que poderão beneficiar-se com a sexagem de espermatozóides, quando essas limitações forem solucionadas.

Characterization of bovine transcripts preferentially expressed in testis and with a putative role in spermatogenesis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

Restriction fragment length polymorphism (RFLP) in Exon 2 of the BoLA-DRB3 gene in South American cattle

The Bola-DRB3 gene participates in the development of the immune response and is highly polymorphic. For these reasons, it has been a candidate gene in studies of the genetic basis of disease resistance and in population genetic analysis. South American native cattle breeds have been widely replaced by improved exotic breeds leading to a loss of genetic resources. In particular South American native breeds have high levels of fertility and disease resistance. This work describes genetic variability in the BoLA-DRB3 gene in native (Caracu, Pantaneiro, Argentinean Creole) and exotic (Holstein, Jersey, Nelore, Gir) cattle breeds in Brazil and Argentina. PCR-RFLP alleles were identified by combining the restriction patterns for the BoLA-DRB3.2 locus obtained with RsaI, BstY, and HaeIII restriction enzymes. Allelic frequencies and deviations from the Hardy-Weinberg equilibrium were also calculated. Analysis of the 24 BoLA-DRB3 PCR-RFLP alleles identified showed differences in the allele distributions among breeds.

Estimates of genetic correlations between days to calving and reproductive and weight traits in Nelore cattle

Data comprising 53,181 calving records were analyzed to estimate the genetic correlation between days to calving (DC), and days to first calving (DFC), and the following traits: scrotal circumference (SC), age at first calving (AFC), and weight adjusted for 550 d of age (W550) in a Nelore herd. (Co)variance components were estimated using the REML method fitting bivariate animal models. The fixed effects considered for DC were contemporary group, month of last calving, and age at breeding season (linear and quadratic effects). Contemporary groups were composed by herd, year, season, and management group at birth; herd and management group at weaning; herd, season, and management group at mating; and sex of calf and mating type (multiple sires, single sire, or AI). In DFC analysis, the same fixed effects were considered excluding the month of last calving. For DC, a repeatability animal model was applied. Noncalvers were not considered in analyses because an attempt to include them, attributing a penalty, did not improve the identification of genetic differences between animals. Heritability estimates ranged from 0.04 to 0.06 for DC, from 0.06 to 0.13 for DFC, from 0.42 to 0.44 for SC, from 0.06 to 0.08 for AFC, and was 0.30 for W550. The genetic correlation estimated between DC and SC was low and negative (-0.10), between DC and AFC was high and positive (0.76), and between DC and W550 was almost null (0.07). Similar results were found for genetic correlation estimates between DFC and SC (-0.14), AFC (0.94), and W550 (-0.02). The genetic correlation estimates indicate that the use of DC in the selection of beef cattle may promote favorable correlated responses to age at first mating and, consequently, higher gains in sexual precocity can be expected.

Effects of age at slaughter and sire genotype on fatness, muscularity, and the quality of meat from ram lambs born to Romney ewes

Ram lambs (n = 356) born to Romney ewes and sired by a range of straightbred or crossbred sires were evaluated for carcass composition and, for a subsample (n = 236), some meat quality characteristics over a period of 4 years. Within each year, 2 to 4 sire genotypes, including Romney controls, were evaluated at 2 slaughter ages (5-9 months and 10-11 months). Weight-adjusted carcass length was greater for lambs with some East Friesian or Finn genes, but no differences were found in dressing-out percent. At a constant carcass weight, carcass fatness (fat depths and leg dissectible fat) was lower in lambs with East Friesian, Texel, or Finn genes. Meat yield at a constant weight tended to be greatest in sire groups that were least fat, and M:B ratio in the femur region was highest in lambs with Texel genes and, to a lesser extent, those with Poll Dorset genes. Leg muscularity in the femur region generally paralleled differences in M:B, except in a comparison of East Friesian-cross and Romney lambs where the former had lower muscularity but slightly better M:B. Lambs slaughtered at the older age for each year had longer and leaner carcasses at the same weight, possibly because they had been taken part way through a winter or because of slower growth rates leading up to slaughter. Meat quality characteristics differed little between the genotype groups, but for 2 of 3 years, meat from the semimembranosus muscle of lambs in the older slaughter lot was significantly tougher.

Suckling behaviour of Nelore, Gir and Caracu calves and their crosses

It is reasonable to assume that the knowledge of suckling behaviour contributes to optimal management and selection of beef cattle. However, there is little information about suckling behaviour of some beef cattle breeds. The aim of this study was to describe the suckling behaviour of two zebu (Bos indicus) and one criollo (Bos taurus) breeds, analysing the potential effects of breed and some environmental factors on suckling frequency and duration. Forty cows, 17 Nelore, 14 Gir (both zebu) and 9 Caracu (criollo) were bred in a diallelic crossing design. The cows and resulting calves were kept on pasture from birth to weaning. Their behaviour was recorded weekly during daylight. Three behavioural traits were considered: number of suckling meals (NSM), duration of each suckling meal (DSM) and total suckling duration (TSD). Allosuckling was not observed. The calves suckled at any time during the daylight and the overall means were: NSM = 2.57 +/- 0.05 meals/12 h (from back transformed data), DSM = 9.25 +/- 0.11 min/suckling meal and TSD = 23.76 +/- 0.47 min/12 h. There was an effect of dam's breed on NSM and DSM; the calf's genetic group within breed of cow influenced NSM and TSD when the dams were from the Nelore breed. The age of calf had significant effects on all traits. Males averaged higher NSM and TSD (2.60 +/- 0.03 meals and 25.05 +/- 1.37 min/12 h, respectively) than females (2.12 +/- 0.04 meals and 21.51 +/- 1.55 min/12 h, respectively). The differences in suckling behaviour seem to be produced by a complex combination of genetic and environmental factors, which result in a particular behavioural relationship within mother-offspring pairs. (c) 2006 Elsevier B.V. All rights reserved.

Ruminal methane emission by dairy cattle in Southeast Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estimation of genetic parameters for body weights, scrotal circumference, and testicular volume measured at different ages in Nellore cattle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Predicting the chemical composition of the body and the carcass of 3/4Boer x 1/4Saanen kids using body components

The determination of the chemical composition of body and carcass is important in nutritional and growth regulation studies. The purpose of this study was to develop equations to predict the chemical composition of body and carcass using chemical composition of body components. Twenty 3/4Boer x 1/4Saanen crossbred male kids, weighing from 20 to 35 kg, were used in this study. The empty body chemical composition was measured by grinding all body components and sampling for chemical analyses. The body components used to estimate body and carcass composition were: neck, fore leg, ribs, loin, hind leg, 9-11 th rib section, non-carcass components (head plus feet, organs plus blood, and hide), visceral fat, and kidney fat. The chemical composition of organs plus blood and 9-11 th rib section had the highest precision to estimate percentage of fat, protein, and water in the body (r(2) of 0.94, 0.82, and 0.90, respectively). For carcass composition, the chemical composition of ribs was the best component to predict all carcass chemical components; however, the equations to estimate the percentages of protein and ash showed a low precision (r(2) = 0.48, 0.44, respectively). The 9-11 th rib section was accurate and precise to estimate carcass fat percentage. We concluded the chemical composition of the body of 3/4Boer x 1/4Saanen crossbred male kids was highly correlated with the composition of body parts, specifically organs plus blood and 9-11 th rib section. Further studies should focus on evaluating these body parts for different breeds and genders under different production scenarios. (C) 2007 Elsevier B.V. All rights reserved.

Habitat use and home range of brown-nosed coati, Nasua nasua (Carnivora: Procyonidae) in the Brazilian Cerrado biome

The brown-nosed coati (Nasua nasua) is a carnivorous species found in all the Brazilian biomes, some of which are endangered areas. The aim of this work was to determine the habitat use and selection, home range and core area of N. nasua in the Cerrado biome, central region of Tocantins, Brazil. The study was carried out in an area of approximately 20 000ha from May 2000 to July 2002. A total of seven box traps were placed in the area for 13 months, three of 11 captured animals were followed and monitored by radio-tracking during 13 months. The monitoring was conducted once a day, three times a week using a car and walking through the study area (radio-tracking and visual contact). The results demonstrate that these three males used more frequently the gallery forest formation, followed by cerrado and wetlands. The use of gallery forest by these animals indicated an habitat selection (Proportion test, z=12.98, p< 0.01). Besides, adult males used the gallery forest more frequently (Fisher's exact test, p<0.01) and wetlands less frequently (Fisher's exact test, p<0.01) than juvenile males, without significant differences between animal ages for cerrado percentage of habitat use. Besides, results also showed a gallery forest selection by adult (Proportion test z= 13.62, p<0.01) and juvenile (Proportion test z=2.68, p<0.01) males, and a wetland selection by the juvenile male (Proportion test z=3.90, p<0.01). The home ranges varied from 2.20 to 7.55km2 for the Minimum Convex Polygon 100% (MCP 100%) and from 4.38 to 13.32km2 for the Harmonic Mean 95% (HM 95%). The smallest home range overlap occurred between the adult males (Nm1 and Nm3), and the greatest between the juvenile Njm2 and the adult Nm1. The average of the core area (HM 75%) for the three monitored animals represented 21.29% of the home range calculated with HM 95%. No overlap between core areas was observed for adult males, but, it was an overlap between the core area of the juvenile male and its band with that of the two adult males. The present study provides new data on core area size and frequency habitat use by adult and juvenile males of N. nasua in the Brazilian Cerrado, that may support conservation efforts. Rev. Biol. Trop. 58 (3): 1069-1077. Epub 2010 September 01.

Associations of FASN gene polymorphisms with economical traits in Nellore cattle (Bos primigenius indicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A technique for the capture of free-ranging marsh deer (blastocerus dichotomus)

The marsh deer (Blastocerus dichotomus) is an endangered species froth the marshlands of central South America. Its population has declined in several regions due to the loss of available habitat caused by human activities, especially the construction of hydroelectric darns. The capture of individual deer is critical for research programs and population management. This report describes a novel live-capture technique, which uses a helicopter to drive the animals into a terrain that restricts their movement such as thick vegetation or deep water (60-120 cm in depth). Following confinement, animals are manually restrained. The short pursuit time (median = 2 thin), low mortality rate (0.82-3.28%), and the absence of injury to both the capture team and animals suggest that this method is appropriate for the safe capture of this species. Body temperature correlated with the pursuit time (R-2 = 0.15) but was not significantly altered with pursuit times <3 min.