A case of yellow fever in a brown howler (Alouatta fusca) in Southern Brazil.

Many brown howlers (Alouatta fusca) have died in a 3-month period in a subtropical forest in Southern Brazil. One was examined after a systemic illness. According to clinical signs, and necropsy and histopathology findings, yellow fever virus (YFV) infection was suspected. Tissue sections from liver, kidney, and lymphoid organs were screened by immunohistochemistry for YFV antigens. Cells within those tissues stained positively with a polyclonal antibody against YFV antigens (1:1,600 dilution), and yellow fever was diagnosed for the first time in the brown howler in the area.     

Matricaria chamomilla CH12 decreases handling stress in Nelore calves

Matricaria chamomilla CH₁₂ is a phytotherapeutic or homeopathic product, which has been used to reduce stress. Here, we examined its effect on preventing handling stress in bovines. Sixty Nelore calves were randomly distributed into two equal groups. One group was administered Matricaria chamomilla CH₁₂ in diet and the other the ''control'' was not. Animals in both groups were maintained unstressed for 30 days to adjust to the feeding system and pasture, and were then stressed by constraint on the 31th, 38th, 45th and 60th experimental days. Blood samples were taken on these days after animals had been immobilization in a trunk contention for 5 min. Stress was followed by analyzing serum cortisol levels. These peaked on the 45th day and then decreased, but not to baseline, on the 60th day. On the 45th day cortisol levels were significantly lower in animals fed Matricaria chamomilla CH₁₂, suggesting that this product reduces stress. These effects may be a consequence of its inhibiting cortisol production and its calming and anxiolytic effects.     

Effects of human recombinant granulocyte colony stimulating factor (HR-GCSF) on the hemogram of lactating dairy cattle

Mastitis commonly occurs in the dairy cow and results in an influx of granulocytes into the mammary gland. Presently, colony stimulating factors have been isolated. One factor, granulocyte colony stimulating factor (GCSF), has been identified and reproduced using recombinant DNA technologies. In an experiment, lactating bovine were given a bacterially-synthesized human recombinant granulocyctic colony stimulating factor (Hr-GCSF) at a specified dose rate to monitor and characterize their hematological responses. Doses of HR-GCSF were administered subcutaneously and blood samples collected from the tail vein into vacutainer tubes. Results of the study indicated a toleration by the bovine for the HR-GCSF for the tested period, and that the HR-GCSF can stimulate a sustained elevation of circulating neutrophils.     

Digestive peptidases and proteinases in the midgut gland of the pink shrimp Farfantepenaeus paulensis (Crustacea, Decapoda, Penaeidae)

Proteases from the midgut gland of the Farfantepenaeus paulensis juveniles were assessed. Enzyme activity was determined using protease substrates and inhibitors. The effect of pH, temperature and calcium on proteolytic activity was assayed. Caseinolytic activity was analysed in substrate-sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Trypsin, chymotrypsin and leucine aminopeptidase activity was detected. Proteolytic activity was strongly inhibited by the specific trypsin inhibitors. Tosyl-phenylalanine chloromethyl ketone inhibited 59.3% of chymotrypsin activity. The greatest trypsin-like activity occurred at pH 8.0 and 45 °C. Chymotrypsin-like activity reached maximal values at alkaline pH (7.2–9.0) and 55 °C. CaCl₂ did not increase trypsin-like activity, but rather inhibited it at concentrations of 30 (20%), 50 (30%) and 100 mM (50%). The substrate-SDS-PAGE zymogram revealed eight proteinase bands. Two possibly thermal-resistant (85 °C, 30 min) chymotrypsin isoforms were found, which were inhibited by phenyl-methyl-sulphonyl-fluoride. Aminopeptidase activity of enzyme extracts (Arg, Leu, Lys, Phe and Val) and the recommended concentrations of these essential amino acids in penaeid shrimp diets were positively correlated ( P <0.05). Beause protein digestion involves the combined action of different enzymes, adequate knowledge of shrimp digestion and enzyme characteristics is required for the assessment of the digestive potential of different feed sources and development of in vitro digestibility protocols.     

A time series analysis of brown eye spot progress in conventional and organic coffee production systems

Brown eye spot (BES), caused by Cercospora coffeicola, is an important coffee disease in Brazil. Losses related to this disease have increased over the last few years. Because the factors associated with the disease dynamics are not fully understood, it is important to gather information about this in different cropping systems. BES epidemics were compared from three production systems: organic (OS), organic under shade (OSS) and conventional under full sun (CS). This study was conducted in Ervália‐MG, Brazil, from November 2004 to October 2008. Disease progress was modelled with all 4 years of data in a time series analysis. Disease intensity (severity (SEV) and incidence (INC)), leaf setting (LS) and leaf fall (LF) were assessed on a monthly basis. The highest values for SEV, INC, LF, LS, as well as a larger area under the disease progress curve and maximum disease occurred in CS and in the upper branches, whereas the lowest values for all variables occurred in OSS and in the lower branches. The highest values occurred for SEV and INC from May to July, for LF from July to September, and for LS from October to January. The disease progress was successfully modelled via a time series analysis. The seasonal behaviour of disease progress for all years and production systems was modelled with a nonlinear sinusoidal model with autoregressive moving average (ARMA) errors. Estimated parameters were generated which could be useful for comparative epidemiology, and it was shown that shade could be used in the field for BES management.     

Differential colonization patterns of Xylella fastidiosa infecting citrus genotypes

Xylella fastidiosa is a phytopathogenic bacterium that causes disease in many different crops worldwide. In Brazil, X. fastidiosa subsp. pauca causes citrus variegated chlorosis (CVC), which is a disease responsible for economic losses in the citrus agribusiness. Variable host responses to bacterial colonization and disease development have been observed. This work studies the colonization processes of a pathogenic GFP‐labelled X. fastidiosa citrus strain in sweet orange (susceptible) and tangor (resistant) parents and two resulting hybrids that exhibited contrasting responses to CVC. Xylella fastidiosa showed increased populations and movement in the susceptible genotypes, but slower compared to other hosts such as grapevine. Scanning electron microscopy revealed that the predominant pitted stem morphology in citrus makes the bacterial movement difficult. In susceptible genotypes X. fastidiosa can move from the primary to the secondary xylem, whilst it is confined to the primary xylem in resistant plants. Associated with this is an induction of lignification that occurs earlier in the resistant genotypes when in the presence of the pathogen, and represents a genetic mechanism that leads to formation of a physical barrier, impairing bacterial colonization.     

Influence of Habitat and Land Use on the Assemblages of Ephemeroptera,Plecoptera, and Trichoptera in Neotropical Streams

Insects of the orders Ephemeroptera, Plecoptera, and Trichoptera (EPT) are often used to assess the conditions of aquatic environments, but few studies have examined the differences in these communities between riffles and pools. Our objective was to test whether riffles shelter greater richness and abundance of EPT, as well as to assess the sensitivity of these insects for detecting impacts from different land uses in streams in southeastern Brazil. Samples were collected in the dry season of 2012 with a Surber sampler in riffles and pools of nine streams (forest, pasture, and urban areas). Principal component analysis distinguished the streams according to different land uses as a function of percentage of plant cover and water oxygenation level and showed partial distinction between riffles and pools as a function of current speed and percentage of ultrafine sand. Detrended correspondence analysis indicated the distinction in EPT composition between riffles and pools, except in urban streams. The results of this study confirm the expected differences in the EPT fauna structure between riffles and pools, especially in forest and pasture environments. The individual metrics of riffle and pool assemblages showed significantly different responses to land use. Therefore, we suggest individual sampling of riffles and pools, since the metrics of these assemblages’ insects can differ between these habitats and influence the results of assessments in low-order streams.     

Biochemical responses associated with common bean defence against Sclerotinia sclerotiorum

The aim of this study was to investigate changes in defence compounds of common bean cultivars with different levels of resistance to the fungus Sclerotinia sclerotiorum and determine the relation of the compounds to pathogen tolerance. The lines were inoculated with the pathogen and assessed for enzymatic and non-enzymatic parameters related to plant defence: peroxidases (POX), polyphenol oxidases (PPO), catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX), total soluble phenol and lignin contents. Stem tissue samples were collected from two regions of the plant for biochemical analyses. Stem tissue samples were collected from two regions of the plant for biochemical analyses. In the position one, 5 cm of the stem was collected from the region with necrosis caused by the pathogen, and in the position two, 5 cm of the stem was collected from the end of the position one at the times of 12, 24, 48, 72, 96 and 120 h after inoculation (HAI). Greater lignin and total soluble phenol contents and greater induction of POX and SOD activity in inoculated plants in the region near the inoculation (position one) indicate local activation with later signalling for activation of defence mechanisms in other regions of the plant. The genotype with a greater level of resistance was superior to the susceptible one in regard to lignin production and the activities of POX, APX and SOD defence enzymes. These results suggest that a combination of these defence responses in common bean may contribute to greater plant resistance to the pathogen and that these enzymes have potential use in selection of common bean genotypes.     

Studies on the colonization of axenically grown tomato plants by a GFP-tagged strain of Clavibacter michiganensis subsp. michiganensis

In this study, colonization and disease development of axenically-grown tomato plants by Clavibacter michiganensis subsp. michiganensis (Cmm), the causative agent of bacterial wilt and canker, was investigated. For this, a spontaneous rifampicin resistant strain of Cmm was tagged with a marker that expressed a green fluorescent protein (GFP) in a stable way and which possessed a similar virulence to the parental strain. In vitro plants were drop-inoculated at the stem base and the population dynamics was determined by dilution pour-plating in a selective medium. At 3 h after inoculation, Cmm was already present in low densities in roots, stems and leaves. At 16 dpi, Cmm was found throughout the entire plant in high densities of ca. 10¹⁰ cfu g^?1. Symptoms developed in the in vitro plants typical for Cmm, such as canker, wilting and growth reduction. The presence of Cmm in vascular and parenchymatic tissue of in vitro tomato plants was confirmed by epifluorescence stereo- and confocal laser scanning microscopy. This study showed that in vitro tomato plants can be effectively used for detailed studies on interactions between Cmm and its host, in particular if a GFP-tagged strain of the pathogen is used.