Mitochondrial genotype segregation and effects during mammalian development: Applications to biotechnology

Mitochondria are endosymbiotic organelles responsible for energy production in practically every eukaryotic cell. Their uniparental fashion of inheritance, maternally inherited in mammals, and the homogeneity of mitochondrial DNA (mtDNA) within individuals and matrilineages, are biological phenomena that remain unexplained. This paper reviews some of the recent findings on mitochondrial influences on the manner in which embryos develop and how their genotypes are inherited in mammals, with particular emphasis on the genetic bottleneck effect. Animal models carrying a mix of mtDNAs (heteroplasmic) have been produced by karyoplast and cytoplast transplantation to analyze the segregation patterns at different stages during embryogenesis, in fetuses and offspring. Comparisons performed between murine and bovine reveal interesting changes in segregation and replication of transplanted mtDNAs. We have recently obtained Bos indicus and Bos taurus fetuses and calves from embryos reconstructed using enucleated polymorphic oocytes of Bos taurus origin. These and other findings on mitochondrial biology will have important implications in determining the cytoplasmic genotype of clones and in the preservation of endangered breeds and species. (C) 1999 by Elsevier B.V.

Simulação e análise econômica da produção in vivo e in vitro de embriões em bovinos

The objective of this work was to evaluate the effect of the variables number of recipients, synchronization protocol, reproductive efficiency indicators and pregnancy cost, in the economic effectiveness of in vivo and in vitro bovine embryo production. A simulation application was elaborated to allow the user to insert the input variable parameters. A basic scenario, from the efficiency traditional rates of in vivo (ET) and in vitro production (IVP) techniques of bovine embryos, was introduced in the software as a criterion to compare the results. This software was able to reproduce both ET and IVP scenarios. The embryo production was simulated through stochastic simulation. The optimal number of recipients using sensitivity analysis was determined. The net present value and cost per pregnancy were used as a decision parameter. The synchronization for fixed-time embryo transfer decreased the recipient idleness and, consequently, the final cost of pregnancy, in comparison to the traditional methodology. Foetal sexing must be associated to IVP of bovine embryos. In addition, the optimal recipient number per donor is variable and depends on data inserted in the system.

Polimorfismo da transferrina e albumina e as associações com precocidade sexual em bovinos da raça Nelore doadores de sêmen

O presente estudo utilizou 16 animais Bos taurus indicus da raça Nelore doadores de sêmen. Estes animais foram divididos em grupos de acordo com a idade em que o sêmen congelou pela primeira vez. O grupo I, considerado precoce, apresentou animais com sêmen passível de congelação com idade inferior a 20 meses. O grupo II, composto por animais que tiveram o sêmen congelado com idade entre 21 e 26 meses. E o grupo III, tido como tardio, composto por animais com sêmen congelável com idade superior a 27 meses. Para análise dos padrões eletroforéticos da transferrina e albumina, amostras de sangue foram colhidas em tubos heparinizados e submetidos a centrifugação de 2.500 G por 15 minutos para separação do plasma sangüíneo. As amostras de plasma sangüíneo foram processadas para que a corrida eletroforética em gel de poliacrilamida pudesse ser realizada. Para a coloração do gel, usou-se Coomasie Brilliant Blue. Após análise dos padrões eletroforéticos da transferrina e albumina, observou-se que não houve relação detectável entre os fenótipos da albumina e precocidade sexual de touros doadores. Entretanto, em relação à transferrina, foi possível sugerir uma associação entre o alelo TfD com touros portadores de sêmen congelável precocemente ou medianamente em termos de idade à congelação.

Sexagem de espermatozoides bovinos por centrifugação em gradiente descontínuo de densidade de Percoll

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

Effect of breed and corpus luteum on pregnancy rate of bovine embryo recipients

The objective of this study was to evaluate pregnancy rates of recipients of different breed groups (Nellore and crossbreed), as well as the effects of size and type of the corpus luteum (CL) on plasmatic concentrations of progesterone and pregnancy rates of embryo recipients. A total of 152 heifers were synchronized with progesterone implants and on the day of embryo transfer, previously obtained by superovulation and frozen in ethylene glycol, the diameter and type of the corpus luteum (cavitary and compact) was measured and blood was collected for progesterone measurement. The pregnancy rate was 44.1%, with a diameter of corpus luteum higher in recipients that became pregnant (2.03±0.41) compared with non-pregnant ones (1.86±0.34 cm). Plasmatic concentrations of progesterone did not differ between pregnant (1.50±1.05) and non-pregnant (1.31±0.91 ng/mL) animals. The type of corpus luteum did not influence the pregnancy rates. Only Angus and crossbred Marchigiana differ among themselves in pregnancy rates (33.3 and 59.2%, respectively). The pregnancy probability was affected only by CL diameter, but not by P4 plasmatic concentration. Selection of the corpus luteum size at the time of embryo transfer is an important factor to increase pregnancy rates in recipients, and compact and cavitary corpora lutea do not influence the pregnancy rates of bovine embryo recipients. Nellore recipients have pregnancy rates that are satisfactory and comparable to crossbred (Bos taurus × Bos indicus) recipients.

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.

Heteroses materna e individual para ganho de peso pré-desmama em bovinos Nelore × Hereford e Nelore × Angus

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

Gastrointestinal nematode infection in beef cattle of different genetic groups in Brazil

Resistance to natural infection by gastrointestinal nematodes was compared in 67 female calves of the following genetic groups: Nelore (NX); 1/2 Senepol + 1/2 Nelore (SN); and 1/2 Aberdeen Angus + 1/2 Nelore (AN). The NX (n = 26), SN (n = 23) and AN (n = 18) animals were monitored for 14 months, during which they remained without treatment, allowed to graze in a tropical environment. Eggs per gram of feces (EPG), coprocultures and packed cell volume (PCV) were carried out monthly. No significant effects of the interaction between the genetic groups and month/year of collection and the genetic group on the EPG were found, but there was a significant influence of the month of collection (P < 0.01). The monthly PCV measurements did not differ for the animals of the three genetic groups and there was no association found between the EPG and PCV. The animals of the SN and NX groups showed similar numbers of EPG with results zero, while for the AN group these numbers were significantly lower (P < 0.05). Although the NX group had a large number of EPG with results zero, it also contained many animals with high counts, meaning this group had higher averages during the entire study period. The following nematode genera were found in the coprocultures: Haemonchus, Cooperia, Oesophagostomum and Trichostrongylus, the latter in smallest proportion. There was no significant difference between the genetic groups for averages of all parasites identified, except Cooperia, which were present in higher numbers in the animals of the NX group (P < 0.05). The results obtained in this experiment suggest that the use of Bos taurus x Bos indicus crossbreeds can be a good strategy to reduce the use of chemical control in Brazil. (C) 2009 Elsevier B.V. All rights reserved.

Características da carcaça e da carne de novilhos mantidos em pastagem de capim-marandu submetidos a diferentes estratégias de suplementação

Este trabalho foi realizado com o objetivo de avaliar as características quantitativas e qualitativas da carcaça e da carne de 24 novilhos submetidos a três estratégias de suplementação em pastagem: SD - suplementação diária; da - suplementação em dias alternados; e FS - suplementação oferecida de segunda à sexta-feira e suspensa aos sábados e domingos. Foram utilizados 24 bovinos mestiços (Bos indicus x Bos taurus) com peso inicial de 230 kg mantidos em pastagem de Brachiaria brizantha cv. Marandu no período das águas de 2003 e nos períodos de seca e das águas de 2004, quando atingiram o peso de abate. O delineamento experimental utilizado foi o inteiramente casualizado, com três tratamentos e oito repetições. As características quantitativas e qualitativas da carcaça e da carne não foram influenciadas pelas diferentes estratégias de suplementação, mesmo quando o suplemento foi fornecido apenas em dias alternados ou quando não foi fornecido nos finais de semana. Na média, os animais apresentaram peso de abate de 468,21 kg de PV, rendimento de carcaça quente de 50,26%, área de olho-de-lombo de 59,67 cm² e espessura de gordura de 3,3 mm. A carne foi classificada como macia, com suculência e palatabilidade levemente acima da média.

Mapping of quantitative trait loci controlling tick [Riphicephalus (Boophilus) microplus] resistance on bovine chromosomes 5, 7 and 14

Differences in domestication and selection processes have contributed to considerable phenotypic and genotypic differences between Bos taurus and Bos indicus cattle breeds. of particular interest in tropical and subtropical production environments are those genetic differences between subspecies that underlie the phenotypic extremes in tolerance and susceptibility to parasite infection. In general, B. taurus cattle are more susceptible to ectoparasites than B. indicus cattle in tropical environments, and much of this difference is under genetic control. To identify genomic regions involved in tick resistance, we developed a B. taurus x B. indicus F-2 experimental population to map quantitative trait loci (QTL) for resistance to the Riphicephalus (Boophilus) microplus tick. About 300 individuals were measured for parasite load in two seasons (rainy and dry) and genotyped for 23 microsatellite markers covering chromosomes 5, 7 and 14. We mapped a suggestive chromosome-wide QTL for tick load in the rainy season (P < 0.05) on chromosome 5. For the dry season, suggestive (P < 0.10) chromosome-wide QTL were mapped on chromosomes 7 and 14. The additive effect of the QTL on chromosome 14 corresponds to 3.18% of the total observed phenotypic variance. Our QTL-mapping study has identified different genomic regions controlling tick resistance; these QTL were dependent upon the season in which the ticks were counted, suggesting that the QTL in question may depend on environmental factors.

Identification, characterization and expression analysis of hepcidin gene in sheep

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR

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

Babesia spp. infection in Boophilus microplus engorged females and eggs in São Paulo State, Brazil

Babesia spp. infections were investigated in Bos taurus x Bos indicus dairy cows and calves and in Boophilus microplus engorged female ticks and eggs. Blood samples and engorged female ticks were collected from 25 cows and 27 calves. Babesia spp. was detected in ticks by microscopic examination of hemolymph of engorged female and by squashes of egg samples. Cattle infection was investigated in blood thin smears and by DNA amplification methods (PCR and nested PCR), using specific primers for Babesia bovis and Babesia bigemina. Merozoites of B. bovis (3 animals) and B. bigemina (12 animals) were detected exclusively in blood smears of calves. DNA amplification methods revealed that the frequency of B. bigemina infection in calves (92.6%) and in cows (84%) and of B. bovis in calves (85.2%) and in cows (100%) did not differ significantly (P > 0.05). Babesia spp. infection was more frequent in female ticks and eggs collected from calves (P < 0.01) than from cows, especially in those which had patent parasitemia. Hatching rates of B. microplus larvae were assessed according to the origin of engorged females, parasiternia of the vertebrate host, frequency and intensity of infection in engorged female tick, and frequency of egg infection. Hatching rate was lower in samples collected from calves (P < 0.01) than from cows, and in those in which Babesia spp. was detected in egg samples (P < 0.01). Published by Elsevier B.V.

Características produtivas de novilhas submetidas à restrição alimentar e dieta suplementada com vitaminas D e E

O objetivo deste trabalho foi avaliar os efeitos da restrição alimentar e de dieta de suplementação com vitaminas D e E sobre o desempenho e características de carcaça de novilhas Canchim. Vinte e quatro fêmeas, com peso vivo inicial médio de 251,95±18,49 kg, foram distribuídas em quatro tratamentos: alimentação em quantidade restrita e não restrita, com e sem suplementação de vitaminas D e E. Utilizou-se o delineamento inteiramente casualizado, em arranjo fatorial 2x2, com seis repetições. Os animais com alimentação restrita receberam, por 48 dias, 70% da dieta fornecida ao grupo de alimentação não restrita. Após o período de restrição (peso médio de 300,10 kg), eles voltaram a receber ração à vontade por mais 77 dias, até atingirem peso de abate de 380 kg. As avaliações foram feitas ao final dos períodos de restrição e de realimentação. Não foi observado efeito da restrição alimentar sobre o desempenho animal. Novilhas que não receberam suplementação vitamínica apresentaram maiores índices de fragmentação miofibrilar, luminosidade e mastigabilidade, e menor força de cisalhamento. A restrição alimentar não afeta o desempenho nem as características de carcaça das novilhas Canchim, e a suplementação com vitaminas D e E não melhora a qualidade da carne.

Characterization and transcriptional analysis of the promoter region of the Duffy blood group, chemokine receptor (DARC) gene in cattle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)