Assessment of DGAT1 and LEP gene polymorphisms in three Nelore (Bos indicus) lines selected for growth and their relationship with growth and carcass traits

The aim of this study was to analyze LEP and DGAT1 gene polymorphisms in 3 Nelore lines selected for growth and to evaluate their effects on growth and carcass traits. Traits analyzed were birth, weaning, and yearling weight, rump height, LM area, backfat thickness, and rump fat thickness obtained by ultrasound. Two SNP in the LEP gene [LEP 1620(A/G) and LEP 305(T/C)] and the K232A mutation in the DGAT1 gene were analyzed. The sample consisted of 357 Nelore heifers from 2 lines selected for yearling weight and a control line, established in 1980, at the Estacao Experimental de Zootecnia de Sertaozinho (Sertaozinho, Brazil). Three genotypes were obtained for each marker. Differences in allele frequencies among the 3 lines were only observed for the DGAT1 K232A polymorphism, with the frequency of the A allele being greater in the control line than in the selected lines. The DGAT1 K232A mutation was associated only with rump height, whereas LEP 1620(A/G) was associated with weaning weight and LEP 305(T/C) with birth weight and backfat thickness. However, more studies, with larger data sets, are necessary before these makers can be used for marker-assisted selection.

Human-animal interaction, stress, and embryo production in Bos indicus embryo donors under tropical conditions

This study investigated the effect of human-animal interaction (HAI) and the stress response on the quality of embryo production in superovulated Nelore (Bos indicus) cattle, under tropical conditions. Thirty-two females underwent a superovulation protocol for 5 days. Cortisol concentrations were determined in blood plasma collected on days 0, 4, and 5. Artificial insemination was performed on days 4 and 5, and nonsurgical embryo flushing on day 11. Embryo production and viability were determined. Human stimulation, animal behaviors, accidents, and handling time were recorded to assess HAI. Cattle age was negatively correlated with accidents, frequency of aversive behaviors, and negative stimuli by stockperson during transit through corral compartments to receive superovulation treatments. The factor analysis revealed two distinct groups. The first group was called stressed and had higher cortisol concentration than the nonstressed group, 16.0 +/- 2.1 and 12.5 +/- 1.0 ng/mL, respectively. Comparisons between these groups showed that the frequency of voice emissions by the stockperson and the number of accidents were higher in the stressed group, and also, the mean handling time was longer in the stressed group than for the nonstressed. As a result, viability rate of the embryos was 19% lower in the stressed group (P < 0.05). This indicates that intensive negative HAI is likely related to stress, which affects embryo production in a superovulation program.

Hormone release and behavior during suckling and milking in Gir, Gir x Holstein, and Holstein cows

There are several different milking management systems in Latin America, because Gir cattle are reputed to be easily stressed and not well adapted to machine-milking. This paper, therefore, provides an overview of hormone release and behavior during suckling and milking in Gir cows and their crossbred offspring. Several experiments were performed to study oxytocin release during exclusive suckling or exclusive hand- and machine-milking, oxytocin, and prolactin release during a mixed suckling-milking system and oxytocin release after weaning. Cortisol concentrations and behavior were also examined. Concentration of oxytocin, released during suckling, and both types of milking were high, but the maximum concentration measured during suckling was significantly greater than that observed during exclusive milking. In the mixed suckling-milking system, the greatest oxytocin and prolactin releases were measured during suckling. Cortisol concentrations measured before, during, and after milking demonstrated that Gir x Holstein and Holstein cows were not stressed. On the other hand, although Gir had greater concentrations of cortisol, the percentage of residual milk for Gir cows was less than for dairy cows exposed to different stressful situations. In general, Gir cows and their crossbred offspring adapted to machine-milking, although these breeds can react negatively to milkers. Gir, Gir x Holstein, and Holstein cows all had similar cortisol levels during and after milking.

Testicular traits as selection criteria for young Nellore bulls

The objective of this study was to evaluate the possible use of biometric testicular traits as selection criteria for young Nellore bulls using Bayesian inference to estimate heritability coefficients and genetic correlations. Multitrait analysis was performed including 17,211 records of scrotal circumference obtained during andrological assessment (SCAND) and 15,313 records of testicular volume and shape. In addition, 50,809 records of scrotal circumference at 18 mo (SC18), used as an anchor trait, were analyzed. The (co) variance components and breeding values were estimated by Gibbs sampling using the Gibbs2F90 program under an animal model that included contemporary groups as fixed effects, age of the animal as a linear covariate, and direct additive genetic effects as random effects. Heritabilities of 0.42, 0.43, 0.31, 0.20, 0.04, 0.16, 0.15, and 0.10 were obtained for SC18, SCAND, testicular volume, testicular shape, minor defects, major defects, total defects, and satisfactory andrological evaluation, respectively. The genetic correlations between SC18 and the other traits were 0.84 (SCAND), 0.75 (testicular shape), 0.44 (testicular volume), -0.23 (minor defects), -0.16 (major defects), -0.24 (total defects), and 0.56 (satisfactory andrological evaluation). Genetic correlations of 0.94 and 0.52 were obtained between SCAND and testicular volume and shape, respectively, and of 0.52 between testicular volume and testicular shape. In addition to favorable genetic parameter estimates, SC18 was found to be the most advantageous testicular trait due to its easy measurement before andrological assessment of the animals, even though the utilization of biometric testicular traits as selection criteria was also found to be possible. In conclusion, SC18 and biometric testicular traits can be adopted as a selection criterion to improve the fertility of young Nellore bulls.

Delivery of cloned offspring: experience in Zebu cattle (Bos indicus)

The production of a healthy cloned calf is dependent on a multitude of successful steps, including reprogramming mediated by the oocyte, the development of a functional placenta, adequate maternal-fetal interaction, the establishment of a physiological metabolic setting and the formation of a complete set of well-differentiated cells that will eventually result in well-characterised and fully competent tissues and organs. Although the efficiency of nuclear transfer has improved significantly since the first report of a somatic cell nuclear transfer-derived animal, there are many descriptions of anomalies concerning cloned calves leading to high perinatal morbidity and mortality. The present article discusses some our experience regarding perinatal and neonatal procedures for cloned Zebu cattle (B. indicus) that has led to improved survival rates in Nellore cloned calves following the application of such `labour-intensive technology`.

Fluctuating abundance of humpback whales (Megaptera novaeangliae) in a calving ground off coastal Brazil

The humpback whale (Megaptera novaeangliae) population that uses Abrolhos Bank, off the east coast of Brazil as a breeding ground is increasing. To describe temporal changes in the relative abundance of humpback whales around Abrolhos, seven years (1998-2004) of whale count data were collected during July through to November. During one-hour-scans, observers determined group size within 9.3 km (5 n.m.) of a land-based observing station. A total Of 930 scans, comprising 7996 sightings of adults and 2044 calves were analysed using generalized linear models that included variables for time of day, day of the season, years and two-way interactions as possible predictors. The pattern observed was the gradual build-up and decline in whale counts within seasons. Patterns and peaks of adult and calf counts varied among years. Although fluctuation was observed, there was generally an increasing trend in adult counts among years. Calf counts increased only in 2004. These fluctuations may have been caused by some environmental conditions in humpback whales` summering grounds and also by changes in spatial-temporal concentrations in Abrolhos Bank. The general pattern observed within the study area mirrored what was observed in the whole Abrolhos Bank. Knowledge of the consistency with which humpback whales use this important nursing area should prove beneficial for designing future monitoring programmes especially related to whale watching activities around Abrolhos Archipelago.

Efficiency of wear and decalcification technique for estimating the age of estuarine dolphin Sotalia guianensis

Most techniques used for estimating the age of Sotalia guianensis (van B,n,den, 1864) (Cetacea; Delphinidae) are very expensive, and require sophisticated equipment for preparing histological sections of teeth. The objective of this study was to test a more affordable and much simpler method, involving of the manual wear of teeth followed by decalcification and observation under a stereomicroscope. This technique has been employed successfully with larger species of Odontoceti. Twenty-six specimens were selected, and one tooth of each specimen was worn and demineralized for growth layers reading. Growth layers were evidenced in all specimens; however, in 4 of the 26 teeth, not all the layers could be clearly observed. In these teeth, there was a significant decrease of growth layer group thickness, thus hindering the layers count. The juxtaposition of layers hindered the reading of larger numbers of layers by the wear and decalcification technique. Analysis of more than 17 layers in a single tooth proved inconclusive. The method applied here proved to be efficient in estimating the age of Sotalia guianensis individuals younger than 18 years. This method could simplify the study of the age structure of the overall population, and allows the use of the more expensive methodologies to be confined to more specific studies of older specimens. It also enables the classification of the calf, young and adult classes, which is important for general population studies.

First molecular estimate of sex-ratio of southern right whale calves, Eubalaena australis, for Brazilian waters

The southern right whale (Eubalaena australis) was one of the most intensively hunted whales between the 17th and 20th centuries in the southern hemisphere. Recent estimates indicate that today there are around 7000 whales, representing 5 to 10% Of its original population. On the other hand, recent studies estimated that the population that migrates to the Brazilian coast grew by 14% from 1987 to 2003. However, there is no information about sex-ratio for adults or for calves in this region, which is an important parameter for understanding the biology of the species. We present here the first estimate Of calves` sex-ratio of southern right whales found along the southern Brazilian coast, one of the most important wintering grounds for the species. Sex was molecularly indentified for 21 biopsies collected from calves between 1998 and 2002, along the coast of Rio Grande do Sul and Santa Catarina States, in southern Brazil. The sex-ratio was two females for one male, however, it was not statistically different (chi(2) test, alpha = 0.05; df = 1) from the expected ratio of 1:1. This result is in accordance with the sex-ratio estimated for the species of all ages using external morphology (and behaviour in formation), (is well as for most species of baleen whales.

Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis

Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of X (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data. suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens. (c) 2009 Elsevier Ltd. All rights reserved.

Exogenous insulin stimulates glycogen accumulation in Rhipicephalus (Boophilus) microplus embryo cell line BME26 via PI3K/AKT pathway

Ticks are obligatory blood-feeding arthropods and important vectors of both human and animal disease agents. Besides its metabolic role, insulin signaling pathway (ISP) is widely described as crucial for vertebrate and invertebrate embryogenesis, development and cell survival. In such cascade, Phosphatidylinositol 3-OH Kinase (PI3K) is hierarchically located upstream Protein Kinase B (PKB). To study the insulin-triggered pathway and its possible roles during embryogenesis we used a culture of embryonic Rhipicephalus microplus cells (BME26). Exogenous insulin elevated cell glycogen content in the absence of fetal calf serum (FCS) when compared to cells without treatment. Moreover, in the presence of PI3K inhibitors (Wortmannin or LY294002) these effects were blocked. We observed an increase in the relative expression level of PI3K`s regulatory subunit (p85), as determined by qRT-PCR. In the presence of PI3K inhibitors these effects on transcription were also reversed. Additionally, treatment with Wortmannin increased the expression level of the insulin-regulated downstream target glycogen synthase kinase 3 beta (GSK3 beta). The p85 subunit showed elevated transcription levels in ovaries from fully engorged females, but was differentially expressed during tick embryogenesis. These results strongly suggest the presence of an insulin responsive machinery in BME26 cells, and its correlation with carbohydrate/glycogen metabolism also during embryogenesis. (C) 2009 Published by Elsevier Inc.

Lu-Hf and U-Pb age determination of Capivarita Anorthosite in the Dom Feliciano Belt, Brazil

The Capivarita Anorthosite, formed in an intraplate environment and later metamorphosed under amphibolites fades conditions, is exposed in the Dom Feliciano Belt as part of the Brasiliano magmatic arc and occurs as a roof-pedant in, or is even intruded by, 0.6 Ga post-collisional granites. In this work, magmatic and metamorphic minerals were dated using the LA-MC-ICP-MS in situ method. U-Pb magmatic and metamorphic zircon dating yielded an age of 1573 +/- 21 Ma and of 606 +/- 6 Ma, respectively, whereas the igneous titanite dating yielded an age of 1530 +/- 33 Ma and the metamorphic ages were 651 +/- 9 Ma and 601 +/- 5 Ma. The Lu-Hf model ages showed two clusters from 1.81 to 2.03 Ga (calf from +2.21 to +6.42) and 2.55-2.62 Ga (epsilon epsilon Hf from -4.59 to -5.64). This intraplate magmatism can be connected to a very important episode of continental accretion in an extensional setting from the fragmentation of the supercontinent during the Early Mesoproterozoic. (C) 2011 Elsevier B.V. All rights reserved.

Biflavonoids from Araucaria angustifolia protect against DNA UV-Induced damage

Ultraviolet radiation is one of the most deleterious forms of radiation to terrestrial organisms and is involved in formation of mutagenic pyrimidine dimers and oxidized nucleotides. The biflavonoid fraction (BFF), extracted from needles of Araucaria angustifolia was capable of protecting calf thymus DNA from damage induced by UV radiation. This occurred through prevention of cyclobutane thymine dimer and 8-oxo-7,8-dihydro-2`-deoxyguanosine formation, this being quantified by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) in a multiple reaction monitoring mode (MRM) and by HPLC-coulometric detection, respectively. (C) 2009 Elsevier Ltd. All rights reserved.

Probing the binding of tetraplatinum(pyridyl)porphyrin complexes to DNA by means of surface plasmon resonance

Tetrapyridylporphyrins containing four chloro(2,2`-bipyridine)platinum(II) complexes attached at the meta (3-H(2)TPtPyP) and para (4-H(2)TPtPyP) positions of the peripheral pyridine ligands were synthesized and their interaction with DNA investigated. The compounds were isolated in the solid state and characterized by means of spectroscopic and analytical techniques. According to molecular simulations, the two isomers exhibit contrasting structural characteristics, consistent with a saddle shape configuration for 3-H(2)TPtPyP and a planar geometry for 4-H(2)TPtPyP. Surface plasmon resonance studies were carried out on the interaction of the complexes with calf thymus DNA, revealing a preferential binding of 3-H(2)TPtPyP, presumably at the DNA major grooves. (C) 2008 Elsevier Inc. All rights reserved.

Mutagenicity and DNA adduct formation of PAH, nitro-PAH, and oxy-PAH fractions of atmospheric particulate matter from Sao Paulo, Brazil

Urban particulate matter (UPM) contributes to lung cancer incidence. Here, we have studied the mutagenic activity and DNA adduct-forming ability of fractionated UPM extractable organic matter (EOM). UPM was collected with a high-volume sampler in June 2004 at two sites, one at street level adjacent to a roadway and the other inside a park within the urban area of the city of Sao Paulo, Brazil. UPM was extracted using dichloromethane, and the resulting EOM was separated by HPLC to obtain PAH, nitro-PAH, and oxy-PAH fractions which were tested for mutagenicity with the Salmonella strains TA98 and YG1041 with and without S9 metabolic activation. The PAH fraction from both sites showed negligible mutagenic activity in both strains. The highest mutagenic activity was found for the nitro-PAH fraction using YG1041 without metabolic activation; however, results were comparable for both sites. The nitro-PAH and oxy-PAH fractions were incubated with calf thymus DNA under reductive conditions appropriate for the activation of nitro aromatic compounds, then DNA adduct patterns and levels were determined with thin-layer chromatography (TLC) (32)p-postlabeling method using two enrichment procedures-nuclease PI digestion and butanol extraction. Reductively activated fractions from both sites produced diagonal radioactive zones (DRZ) of putative aromatic DNA adducts on thin layer plates with both enrichment procedures. No such DRZ were observed in control experiments using fractions from unexposed filters or from incubations without activating system. Total adduct levels produced by the nitro-PAH fractions were similar for both sites ranging from 30 to 45 adducts per 10(8) normal nucleotides. In contrast, the DNA binding of reductively activated oxy-PAH fractions was three times higher and the adduct pattern consisted of multiple discrete spots along the diagonal line on the thin layer plates. However, DNA adduct levels were not significantly different between the sampling sites. Both samples presented the same levels of mutagenic activity. The response in the Salmonella assay was typical of nitroaromatics. Although, more mutagenic activity was related to the nitro-PAH fraction in the Salmonella assay, the oxy-PAH fractions showed the highest DNA adduct levels. More studies are needed to elucidate the nature of the genotoxicants occurring in Sao Paulo atmospheric samples. (C) 2008 Elsevier B.V. All rights reserved.