Molecular cloning and characterization of sea bass (Dicentrarchus labrax L.) CD8alpha

In this work, the gene and cDNA of the sea bass CD8alpha have been isolated and characterized. The coding sequence has an ORF of 666 bp. It retains the Ig motif that interacts with MHC and the two cysteines responsible for an intra-chain disulfide bridge. The hinge region contains the two essential cysteines involved in dimerization. The transmembrane region is well conserved in all analysed sequences. Similar to other teleosts, the cytoplasmic region lacks the consensus p56(lck) motif common in higher vertebrates. Analysis of the expression pattern using RT-PCR shows the highest expression in the thymus. Like in the human gene, the sea bass CD8alpha genomic structure is organized into six exons, which roughly correspond to separate functional domains of the protein. Southern blotting shows that CD8alpha exists as a single copy gene. Together, these results support the concept that the basic structure of CD8alpha has been maintained through evolution.     

Evaluation of antiviral activity of phenolic compounds and derivatives against rabies virus

Human rabies is a viral disease with a great impact on public health, mainly on account of its fatal course in the majority of cases. Despite the well-established prophylaxis by immunization, rabies is believed to be responsible for 40,000-70,000 human deaths per year, mostly in endemic areas. Palliative support and experimental protocols to avoid death have been employed with no expressive results, with the exception of a recent human case of recovery from rabies. No antiviral drugs are currently available to fight against this infection. In combination with the prophylaxis, an antiviral drug would be useful for human rabies treatment, providing enhanced protection against the encephalitis caused by the virus. Phenolic compounds are derived from the secondary plant metabolism, although they can also be obtained by synthetic processes. Many studies have shown a great range of pharmacological effects for these substances, including vasodilatation, antiallergenic, antiinflammatory and antiviral properties, among others. In this study, the potential in-vitro anti-rabies activity of 24 synthetic phenolic compounds was evaluated using McCoy cells and PV rabies strain. The cytotoxicity (CC50) was assayed by the MTT method and the antiviral activity (IC50) was estimated by the inhibition of viral cytopathic effects. Isoprinosine and ketamine were used as positive controls. The tested compounds showed selectivity indices (SI=CC50/IC50) ranging from 1.0 to 3.9. Six phenolic compounds failed to inhibit the cytopathic effect to any degree, and four showed SI > or = 3.0. According to these results, some probable structure-activity relationships are suggested. It was observed that the presence of free hydroxyl and ether groups influenced the anti-rabies activity. However, additional studies are required to establish these relationships.     

Investigating phenotypic variability in Colletotrichum lindemuthianum populations

Colletotrichum lindemuthianum, causal agent of anthracnose in the common bean, has wide genetic variability. Differential bean cultivars and morphological and physiological characteristics were used to analyze 74 isolates of C. lindemuthianum collected in two counties in the state of Minas Gerais, Brazil. Six different races were found, with a predominance of race 65 at both locations. Isolates were classified according to their sensitivities to the fungicide thiophanate-methyl, normally used in the control of common bean anthracnose. In all, ≈10% of isolates were resistant to the fungicide in vitro. Characteristics such as indexes of mycelia growth rate, colony diameter, sporulation capacity, and percentage of germination demonstrated the high genetic variability of C. lindemuthianum. We also observed variation in conidial cytology. The conidia of most isolates showed septa formation after germination, in contrast to septa absence, previously reported in the literature. Sexual and asexual reproduction were evaluated for mechanisms that may contribute in the generation of variability in C. lindemuthianum. Conidial anastomosis tubes were commonly found, indicating that asexual reproduction can help increase variability in this species. Information from this study confirmed high variability in C. lindemuthianum and will guide future studies in basic knowledge and applied technologies.     

Modeling the Potential Geographic Distribution of Black Pepper（Piper nigrum）in Asia Using GIS Tools

Known as the "king of spices",black pepper (Piper nigrum),a perennial crop of the tropics,is economically the most important and the most widely used spice crop in the world.To understand its suitable ...     

The thermostable β-1,3-1,4-glucanase from Clostridium thermocellum improves the nutritive value of highly viscous barley-based diets for broilers

1. Microbial β-1,3-1,4-glucanases improve the nutritive value of barley-based diets for poultry by effectively decreasing the degree of polymerisation of the anti-nutritive soluble β-glucans. Glycoside hydrolases (GHs) acting on recalcitrant polysaccharides display a modular architecture comprising a catalytic domain linked to one or more non-catalytic Carbohydrate-Binding Modules (CBMs).

2. GHs and CBMs have been classified in different families based on primary structure similarity (see CAZy webpage at http://www.cazy.org). The role of CBMs is to anchor the appended GHs into their target substrates, therefore eliciting the efficient hydrolysis of structural carbohydrates.

3. Here we describe the biochemical properties of the family 16 GH from Clostridium thermocellum, termed CtGlc16A. CtGlc16A is a thermostable enzyme that specifically acts on β-1,3-1,4-glucans with a remarkable catalytic activity (38000?U/mg protein).

4. CtGlc16A, individually or fused to the family 11 β-glucan-binding domain of cellulase CtLic26A-Cel5E of C. thermocellum, was used to supplement a highly viscous barley-based diet for broilers.

5. The data showed that birds fed on diets supplemented with the recombinant enzymes displayed an improved performance when compared with birds given diets not supplemented with exogenous enzymes. However, inclusion of the non-catalytic CBMs had no influence on the capacity of CtGlc16A to reduce the anti-nutritive effects of soluble β-1,3-1,4-glucans.

6. The data suggest that at elevated dosage rates, CBMs might be unable to potentiate the catalytic activity of appended catalytic domains; this effect may only be revealed when feed enzymes are incorporated at low levels.     

Prediction of physical and chemical body compositions of purebred and crossbred Nellore cattle using the composition of a rib section

The goal of this research was to develop empirical equations to predict chemical and physical compositions of the carcass and the body using the composition of the 9th- to 11th-rib section (rib(9-11)) and other measurements. A database (n = 246) from 6 studies was developed and comprised 37 bulls (BU), 115 steers (STR), and 94 heifers (HF), of which 132 were Nellore (NEL), 76 were NEL × Angus crossbreds (NA), and 38 were NEL × Simmental crossbreds (NS). The right half carcass and the rib(9-11) from the left half carcass were analyzed for ether extract (EE), CP, and water. The remaining components were chemically analyzed to determine the composition of the body. A stepwise procedure was used to determine the variable inclusion in the regression models. The variables included were EE in the rib(9-11) (EER; %), CP in the rib(9-11) (CPR; %), water in the rib(9-11) (WR; %), visceral fat (VF; %; KPH and mesenteric fats), organs plus viscera (OV; %), carcass dressing percentage (CD; %), cold carcass weight (kg), and empty BW (EBW; kg). No sex or breed effects were found on EE and CP compositions of the carcass (C(EE) and C(CP), respectively; %); the equations were as follows: C(EE) = 4.31 + 0.31 × EER + 1.37 × VF [n = 241; R(2) = 0.83; mean square error (MSE) = 4.53] and C(CP) = 17.92 + 0.60 × CPR - 0.17 × CD (n = 238; R(2) = 0.50; MSE = 1.58). Breed affected water content in the carcass (C(W), %); the equations were as follows: C(W) = 48.74 + 0.28 × WR - 0.017 × EBW for NEL; C(W) = 46.69 + 0.32 × WR - 0.017 × EBW for NA; and C(W) = 38.06 + 0.48 × WR - 0.017 × EBW for NS (n = 243; R(2) = 0.67; MSE = 5.17). A sex effect was found on body chemical EE composition (BW(EE)); the equations were as follows: BW(EE) = 2.75 + 0.33 × EER + 1.80 × VF for BU; BW(EE) = 1.84 + 0.33 × EER + 1.91 × VF for STR; and BW(EE) = 4.77 + 0.33 × EER + 1.28 × VF for HF (n = 243; R(2) = 0.89; MSE = 3.88). No sex or breed effects were found on CP composition in the body (BW(CP)); the equation was as follows: BW(CP) = 14.38 + 0.24 × CPR (n = 240; R(2) = 0.59; MSE = 1.06). A sex effect was found for body water content (BW(W)); the equations were as follows: BW(W) = 38.31 + 0.33 × WR - 1.09 × VF + 0.50 × OV for BU; BW(W) = 45.67 + 0.25 × WR - 1.89 × VF + 0.50 × OV for STR; and BW(W) = 31.61 + 0.47 × WR - 1.06 × VF + 0.50 × OV for HF (n = 241; R(2) = 0.81; MSE = 3.84). The physical carcass composition indicated a breed effect on all components and a sex effect for fat in the carcass. We conclude that body and carcass compositions can be estimated with rib(9-11) for purebred and crossbred NEL animals, but specific equations have to be developed for different groups of animals.     

Vaccine immune response and interference of colostral antibodies in calves vaccinated against rabies at 2, 4 and 6 months of age born from antirabies revaccinated females

Considering the high prevalence of rabies in cattle, we aimed to evaluate the interference of colostral antibodies transferred to calves after birth and the benefit of administering an antirabies vaccination in two-month-old calves compared to vaccinating at 4 and 6 months of age. Calves born from females revaccinated against rabies during the third trimester of pregnancy were studied. Forty-eight hours after parturition, blood samples from dams and offspring were collected, and antirabies neutralizing antibody titers were analyzed using the Rapid Focus Fluorescent Inhibition Test. We found that all calves had similar titers of antibodies transferred through the colostrum. Furthermore, none of the calves presented a satisfactory serological response after the first vaccination, but all had an appropriate response after revaccination. This study demonstrates that antirabies vaccination should be recommended for calves at two months of age in endemic and epizootic situations.     

Voluntary intake and milk production in F1 Holstein × zebu cows in confinement

The objective of this study was to evaluate the nutrient intake and milk production in Holstein × zebu (F1) cows in feedlot. Eighteen F1 cows were used, divided into three treatments; six were Holstein × Gir (HGI), six were Holstein × Guzerat (HGU), and another six were Holstein × Nelore (HNE), which had recently calved, distributed into simple, random samples, under the same feeding conditions of corn silage and concentrate with 20% crude protein. The three-marker method was used (LIPE, titanium dioxide, and iADF) to estimate the individual intake and digestibility of the nutrients for the cows in group. The mathematical model used to establish the lactation curves was: Y = at(b)e(-ct) by Wood (Nature 216:164-165, 1967). The statistical analyses for the nutrient intake and digestibility, as well as parameters of metabolic efficiency, were performed using multiple linear regression (α = 5%). No effect (P > 0.05) of genetic group was observed for any of the variables studied. The intake and digestibility of the nutrients and the microbial nitrogen presented quadratic curves as a function of the lactation period (P < 0.05). The HGU cows exhibited an accumulated milk production of 4,946.81 kg at 305 days, whereas the HGI cows produced 4,821.78 kg. The HNE cows displayed inferior performance, with a production of 3,674.98 kg. It was concluded that, in confinement, F1 cows from different genetic groups do not exhibit different intake, digestibility, or metabolic efficiency. The HGU and HGI cows have greater cumulative production at 305 days.     

Use of a Real Time PCR for detecting subspecies of Babesia canis

This paper reports the development and use of a Real Time PCR for detection of Babesia canis canis, B. canis rossi, and B. canis vogeli in endemic areas of Brazil. The sequences of the internal transcribed spacer (ITS) of several organisms were aligned and five primers and four probes were designed for amplification of a fragment (around 125 bp) which differentiates subspecies of B. canis. Blood samples collected from dogs living in farms in three distinct rural regions within the State of Minas Gerais (Lavras, Belo Horizonte and Nanuque) were tested. Blood samples had been collected during a dry season (Lavras, n=100; Belo Horizonte, n=50; Nanuque, n=102); the dogs were re-sampled in the subsequent rainy season (Lavras, n=71; Belo Horizonte, n=29; Nanuque, n=66). From each sample, DNA was extracted and Giemsa stained smears were microscopically examined for direct detection of Babesia parasites. B. canis vogeli was the only subspecies found, with an overall prevalence of 9.9% during the dry season and 10.8% during the rainy season. Dogs living in Nanuque and Belo Horizonte showed significantly higher prevalence rates than those living in Lavras (13.7%, 12.0% and 5.0%, respectively). The Real Time PCR developed proved to be appropriate to detect B. canis subspecies in endemic areas.     

Slaframine toxicosis in Brazilian horses causing excessive salivation

Ingestion of food contaminated with slaframine, an alkaloid produced by Rhizoctonia leguminicola, causes a mycotoxicosis, characterised by excessive salivation. Twenty‐eight horses demonstrated this clinical sign after the consumption of alfalfa hay which on inspection showed dark patches on many of the stems. The presence of slaframine (1.5 ppm) in this hay was confirmed by gas chromatography and mass spectroscopy. This is the first equine slaframine toxicosis case reported in Brazil.