A homolog of the O157 urease-encoding O island 48 is present in porcine O149:H10 enterotoxigenic Escherichia coli

The relationship of the urease operon in the highly virulent O149 porcine enterotoxigenic Escherichia coli (ETEC) strain Ro8 to a genomic island (GI) homologous to O island (OI) 48 of O157 enterohemorrhagic E. coli (EHEC) strain EDL933 was investigated. Eighty-four of 84 O149:H10 strains were urease positive whereas 44 of 44 O149:H43 porcine ETEC strains were urease-negative. Seventeen of 17 O149:H10 strains that were tested possessed the OI-48 homolog whereas 24 of 24 O149:H43 strains lacked this OI. Transposon insertions in lipB or guaA genes in strain Ro8 eliminated urease activity while insertions in the caiF gene increased urease activity. When the O149 ure operon was cloned on a high copy number plasmid, urease expression was increased approximately 11-fold in Ro8 and 83-fold in O157 strain EDL933 compared with that in the wild type Ro8. The O149 urease activity was expressed despite the presence of the same premature stop codon in ureD that is present in ure+ O157:H7 strains that are urease-negative. The ure operon in Ro8 consists of 4 893 nucleotides with 99% identity with the ure operons in EHEC O157:H7 strains EDL933 and Sakai, and is part of a GI similar to GI-48 of strain EDL933. This OI, designated OI-48149 , is inserted in the serX tRNA gene in strain Ro8 and contains genes for urease, tellurite resistance, iha and an AIDA-I-like adhesin. The presence of a homolog of the O157:H7 OI-48 in highly virulent O149 porcine ETEC suggests that this OI may contribute to establishment of the bacteria in the intestine.     

Ulcerative enterocolitis in two goats associated with enterotoxin- and beta2 toxin-positive Clostridium perfringens type D

Enterotoxemia caused by Clostridium perfringens type D in sheep is believed to result from the action of epsilon toxin (ETX). However, the sole role of ETX in the intestinal changes of the acute and chronic forms of enterotoxemia in goats remains controversial, and the synergistic action of other C. perfringens toxins has been suggested previously. The current study examined 2 goats that were found dead without premonitory clinical signs. Gross lesions at necropsy consisted of multifocal fibrinonecrotic enterocolitis, edematous lungs, and excess pleural fluid. Histologically, there were multifocal fibrinonecrotic and ulcerative ileitis and colitis, edema of the colonic serosa, and proteinaceous interstitial edema of the lungs. Clostridium perfringens type D carrying the genes for enterotoxin (CPE) and beta2 toxin (CPB2) was cultured from intestinal content and feces of 1 of 2 goats, while C. perfringens type D CPB2-positive was isolated from the other animal. When multiple colonies of the primary isolations from both animals were tested by Western blot, most of the isolates expressed CPB2, and only a few isolates from the first case expressed CPE. Alpha toxin and ETX were detected in ileal and colonic contents and feces of both animals by antigen capture enzyme-linked immunosorbent assay. CPB2, but not CPE, was identified in the small and large intestines of both goats by immunohistochemistry. These findings indicate that CPB2 may have contributed to the necrotic changes observed in the intestine, possibly assisting ETX transit across the intestinal mucosa.     

Extensive cage culture of pejerrey (Odontesthes bonariensis) in a shallow pampean lake in Argentina

Pejerrey is an important zooplanktivorous native fish of the Argentinean inland waters. It has been traditionally propagated for stocking purposes by relatively costly semi‐intensive and intensive methods. In this study, we evaluated the implementation of an extensive culture method by using floating cages in a shallow pampean lake. Four cages were installed in the Lacombe Lake and stocked with juveniles (16.24 ± 1.69 mm length) at 50 fish m^?3 density for growing until the size of 150 mm, which is considered as a suitable size for stocking. Throughout the experiment, the temperature ranged between 10 and 26 °C and the zooplankton biomass ranged between 12 and 3269 μg dw L^?1. The growth patterns in the length were similar in the four cages and directly related to the lake thermal conditions and zooplankton availability. The average final length after 315 days was 154.4 ± 8.8 mm. The survival rates ranged between 53.5% and 64.7% during the first 110 days and 11.1–25.7% at termination. Growth rate for the first 2 months was the highest documented for pejerrey culture. This simple technique offers the possibility to produce juvenile pejerrey at a low cost and provides the alternative of reinforcing the natural populations with fish already adapted to the natural environmental conditions.     

The Caldenal ecosystem: Effects of a prescribed burning on soil chemical properties

Before and after a prescribed burning, the upper 2.5 cm of soil in patches of forage, nonforage, and shrub vegetation were sampled. Then, bulk density, gravimetric moisture, pH in water, available phosphorus, exchangeable cations, cationic exchange capacity, total nitrogen, total, particulate, and mineral organic carbon contents were determined. Maximum temperature ranges reached over the ground during burning were measured and the highest values were registered in shrubs. Results showed that total, particulate, and mineral organic carbon and total nitrogen contents and percent base saturation increased in all soils after burning, without detecting differential effects between vegetation patches. After burning, available phosphorus content increased both for forage and shrub patches. A Na⁺ content decrease was observed for all soils in the different patches while Mg²⁺ content increased. No changes in the other analyzed variables were observed. Prescribed burning, as studied in the present work, might improve the Caldenal soils’ chemical fertility, mainly due to an increase in labile organic matter and some nutrients essential for plant growth.     

Adaptation and Validation of Colorimetric Methods in Determining Ammonium and Nitrate on Tropical Soils

Inorganic nitrogen (N) determination in soil extracts is a useful tool in studying soil–N interactions. The main method used worldwide is the steam distillation, but colorimetric methodologies show advantages of being quick, simple, sensitive, and higher yield. However, they are subject to disruption when the extract has interfering organic N, limiting its application in tropical soil. The aim of this study is to evaluate colorimetric methods for the determination of NH₄⁺ (through Berthelot reaction) and NO₃^? + NO₂^? (through Griess-Ilosvay reaction) in saline extracts of tropical soils by addition of activated charcoal during the extraction process. It is recommended to use extraction solution of 2 mol L^?1 potassium chloride (KCl) and reaction time of 30 min. Validation parameters confirmed the best performance of the colorimetric methods, revealing results favorable to the addition of the activated charcoal in the extraction process to increase the accuracy and precision of the colorimetric methods.     

Amaranth Peptides from Simulated Gastrointestinal Digestion: Antioxidant Activity Against Reactive Species

Plant Foods for Human Nutrition - We evaluated the capacity of simulated gastrointestinal digests or alcalase hydrolysates of protein isolates from amaranth to scavenge diverse physiologically...     

Chalinulasterol, a chlorinated steroid disulfate from the Caribbean sponge Chalinula molitba. Evaluation of its role as PXR receptor modulator

Chalinulasterol (1) a new chlorinated sterol disulfate was isolated from the Caribbean sponge Chalinula molitba. Its structure was elucidated using mass spectrometry and NMR experiments. The possible role of chalinulasterol as modulator of the PXR nuclear receptor was investigated but, in spite of the close structural relationship with the PXR agonist solomonsterol A (2), it showed no activity. The structural requirements for the PXR nuclear receptor activity were discussed.     

Proteomic analysis reveals suppression of bark chitinases and proteinase inhibitors in citrus plants affected by the citrus sudden death disease

Citrus sudden death (CSD) is a disease of unknown etiology that greatly affects sweet oranges grafted on Rangpur lime rootstock, the most important rootstock in Brazilian citriculture. We performed a proteomic analysis to generate information related to this plant pathogen interaction. Protein profiles from healthy, CSD-affected and CSD-tolerant stem barks, were generated using two-dimensional gel electrophoresis. The protein spots were well distributed over a pI range of 3.26 to 9.97 and a molecular weight (MW) range from 7.1 to 120 kDa. The patterns of expressed proteins on 2-DE gels made it possible to distinguish healthy barks from CSD-affected barks. Protein spots with MW around 30 kDa and pI values ranging from 4.5 to 5.2 were down-regulated in the CSD-affected root-stock bark. This set of protein spots was identified as chitinases. Another set of proteins, ranging in pI from 6.1 to 9.6 with an MW of about 20 kDa, were also suppressed in CSD-affected rootstock bark; these were identified as miraculin-like proteins, potential trypsin inhibitors. Down-regulation of chitinases and proteinase inhibitors in CSD-affected plants is relevant since chitinases are well-known pathogenesis-related protein, and their activity against plant pathogens is largely accepted.     

Effects of Titanium Dioxide Nanoparticles in Different Metabolic Pathways in the Freshwater Microalga <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis> (Trebouxiophyceae)

The products that employ nanoparticles (NPs) in their composition have increased since the beginning of NP production; hence, their availability in the environment, especially in aquatic ecosystems, tends to increase. In these ecosystems, the phytoplankton is immersed in a complex matrix of nutrients, excreted materials, and other chemical compounds, which can influence the metabolic strategy of microalgae. One of the metabolic ways is mixotrophy, a situation whereby microalgae perform photosynthesis and use dissolved organic carbon at the same time. Most toxicity evaluations do not consider such a metabolic route, but this can represent a preferential metabolism in natural environments. The present study aimed at evaluating the effects of NP-TiO₂ at a log concentration range of ??3.10 to 0.89, on photosynthesis, growth, viability, and biochemical composition of the microalgae Chlorella sorokiniana during photoautotrophic and mixotrophic growth (glucose as the organic carbon source). The results showed lower chlorophyll a and photosynthetic activity in mixotrophy than in photoautotrophy, which can be due to a decreased need for photosynthesis in mixotrophy. Photoautotrophy cultures were sensitive to NPs, reaching 39% of viability at log 0.89, while in mixotrophy, cell viability was not affected by NPs. The biochemical composition and cell density changed as a function of NP concentrations, with increase in the protein/carbohydrate ratio in both treatments. The results showed that C. sorokiniana is more resistant to NPs during mixotrophic growth, but with changes in biochemical composition, whereas the photoautotrophic cultures were more sensitive to the increase in NP concentrations.