Evidence of evolving extraintestinal enteroaggregative Escherichia coli ST38 clone

BACKGROUND: Several clones of extended-spectrum β-lactamase (ESBL)–producing extraintestinal pathogenic Escherichia coli (ExPEC) have globally expanded their distribution. ExPEC infections often originate from the patient’s own intestinal flora, although the degree of overlap between diarrheagenic E. coli and ExPEC pathotypes is unclear. Relatively little is known about antimicrobial drug resistance in the most common diarrheagenic E. coli groups, including enteroaggregative E. coli (EAEC), and bacterial gastroenteritis is generally managed without use of antimicrobial drugs. APPROACHES: We conducted this study to establish the presence and characteristics of ESBL-producing EAEC in a well-defined collection of ESBL-producing isolates. The isolates were from human and animal sources in Germany, the Netherlands, and the United Kingdom. DNA from 359 ESBL isolates was screened for the presence of the EAEC transport regulator gene (aggR), located on the EAEC plasmid, using a real-time PCR assay and the phylogroup was determined for each positive isolate. A microarray was used to detect ESBL genes, such as blaCTX-M, at the group level, as previously described. The antimicrobial drug susceptibilities of EAEC isolates were determined and virulence factors associated with intestinal and extraintestinal infection and with EAEC were investigated . RESULTS AND CONCLUSIONS: We assigned a virulence score (total number of virulence factor genes detected; maximum possible score 22) and a resistance score (total number of drug classes; maximum score 11) to each isolate. We isolated 11 EAEC from humans. Eight of the EAEC were isolated from urine specimens, and 1 was isolated from a blood culture; 63% belonged to phylogroup D (Table). EAEC ST38, the most common (55%) ST, was significantly associated with extraintestinal sites in the subset of 140 human isolates (Fisher exact test, p<0.0001)

OCCURRENCE OF PATHOGENIC MICROORGANISMS IN FISH SOLD IN SAO PAULO, BRAZIL

This study investigated the presence of potentially human pathogenic strains of Vibrio spp., Aeromonas spp., Escherichia coli, Salmonella spp. and Staphylococcus aureus in fish commercialized in street markets of Sao Paulo city, Brazil. Twenty fish of different species were analyzed for foodborne pathogens using conventional methods. High levels of fecal contamination were detected in 25% of samples. S. aureus was isolated from 10% of samples. All were negative for Salmonella. Vibrio species, including Vibrio cholerae non-O1/non-O139, were observed in 85% of samples although Vibrio parahaemolyticus was not found in this study. Aeromonas spp., including A. hydrophila, was isolated from 50% of fish samples. The occurrence of these pathogens suggests that the fish commercialized in Sao Paulo may represent a health risk to the consumers.

Genotyping, physiological features and proteolytic activities of a potentially pathogenic Acanthamoeba sp isolated from tap water in Brazil

Acanthamoeba spp., known to cause keratitis and granulomatous encephalitis in humans, are frequently isolated from a variety of water sources. Here we report for the first time the characterization of an Acanthamoeba sp. (ACC01) isolated from tap water in Brazil. This organism is currently being maintained in an axenic growth medium. Phylogenetic analysis based on SSU rRNA gene sequences positioned the new isolate in genotype T4, closest to the keratitis-causing isolate, A. polyphaga ATCC 30461 (similar to 99% similarity). Acanthamoeba ACC01 and A. polyphaga 30461 both grew at 37 degrees C and were osmotically resistant, multiplying in hyperosmolar medium. Both isolates secreted comparable amounts of proteolytic enzymes, including serine peptidases that were optimally active at a near neutral/alkaline pH and resolved identically in gelatin gels. Incubation of gels at pH 4.0 with 2 mM DTT also indicated the secretion of similar cysteine peptidases. Altogether, the results point to the pathogenic potential of Acanthamoeba ACC01. (C) 2009 Elsevier Inc. All rights reserved.

Melanin as a virulence factor of Paracoccidioides brasiliensis and other dimorphic pathogenic fungi: a minireview

Melanin pigments are substances produced by a broad variety of pathogenic microorganisms, including bacteria, fungi, and helminths. Microbes predominantly produce melanin pigment via tyrosinases, laccases, catecholases, and the polyketide synthase pathway. In fungi, melanin is deposited in the cell wall and cytoplasm, and melanin particles (""ghosts"") can be isolated from these fungi that have the same size and shape of the original cells. Melanin has been reported in several human pathogenic dimorphic fungi including Paracoccidioides brasiliensis, Sporothrix schenckii, Histoplasma capsulatum, Blastomyces dermatitidis, and Coccidioides posadasii. Melanization appears to contribute to virulence by reducing the susceptibility of melanized fungi to host defense mechanisms and antifungal drugs.

Structure and Calcium-Binding Activity of LipL32, the Major Surface Antigen of Pathogenic Leptospira sp.

Leptospixosis, a spirochaetal zoonotic disease caused by Leptospira, has been recognized as an important emerging infectious disease. LipL32 is the major exposed outer membrane protein found exclusively in pathogenic leptospires, where it accounts for up to 75% of the total outer membrane proteins. It is highly immunogenic, and recent studies have implicated LipL32 as an extracellular matrix binding protein, interacting with collagens, fibronectin, and laminin. In order to better understand the biological role and the structural requirements for the function of this important lipoprotein, we have determined the 2.25-angstrom-resolution structure of recombinant LipL32 protein corresponding to residues 21-272 of the wild-type protein (LipL32(21-272)). The LipL32(21-272) monomer is made of a jelly-roll fold core from which several peripheral secondary structures protrude. LipL32(21-272) is structurally similar to several other jelly-roll proteins, some of which bind calcium ions and extracellular matrix proteins. Indeed, spectroscopic data (circular dichroism, intrinsic tryptophan fluorescence, and extrinsic 1-amino-2-naphthol-4-sulfonic acid fluorescence) confirmed the calcium-binding properties of LipL32(21-272). Ca(2+) binding resulted in a significant increase in the thermal stability of the protein, and binding was specific for Ca(2+) as no structural or stability perturbations were observed for Mg(2+), Zn(2+), or Cu(2+). Careful examination of the crystal lographic structure suggests the locations of putative regions that could mediate Ca(2+) binding as well as binding to other interacting host proteins, such as collagens, fibronectin, and lamixidn. (C) 2009 Elsevier Ltd. All rights reserved.

Biosurfactants as Agents to Reduce Adhesion of Pathogenic Bacteria to Polystyrene Surfaces: Effect of Temperature and Hydrophobicity

Polystyrene surfaces were conditioned with surfactin and rhamnolipid biosurfactants and then assessed regarding the attachment of Staphylococcus aureus, Listeria monocytogenes, and Micrococcus lute us. The effect of different temperatures (35, 25, and 4 degrees C) on the anti-adhesive activity was also studied. Microbial adhesion to solvents and contact angle measurements were performed to characterize bacteria and material surfaces. The results showed that surfactin was able to inhibit bacterial adhesion in all the conditions analyzed, giving a 63-66% adhesion reduction in the bacterial strains at 4 degrees C. Rhamnolipid promoted a slight decrease in the attachment of S. aureus. The anti-adhesive activity of surfactin increased with the decrease in temperature, showing that this is an important parameter to be considered in surface conditioning tests. Surfactin showed good potential as an anti-adhesive compound that can be explored to protect surfaces from microbial contamination.

Surfactin reduces the adhesion of food-borne pathogenic bacteria to solid surfaces

Aims: To investigate the effect of the biosurfactants surfactin and rhamnolipids on the adhesion of the food pathogens Listeria monocytogenes, Enterobacter sakazakii and Salmonella Enteritidis to stainless steel and polypropylene surfaces. Methods and Results: Quantification of bacterial adhesion was performed using the crystal violet staining technique. Preconditioning of surfaces with surfactin caused a reduction on the number of adhered cells of Ent. sakazakii and L. monocytogenes on stainless steel. The most significant result was obtained with L. monocytogenes where number of adhered cells was reduced by 10(2) CFU cm(-2). On polypropylene, surfactin showed a significant decrease on the adhesion of all strains. The adsorption of surfactin on polystyrene also reduces the adhesion of L. monocytogenes and Salm. Enteritidis growing cells. For short contact periods using nongrowing cells or longer contact periods with growing cells, surfactin was able to delay bacterial adhesion. Conclusions: The prior adsorption of surfactin to solid surfaces contributes on reducing colonization of the pathogenic bacteria. Significance and Impact of the Study: This is the first work investigating the effect of surfactin on the adhesion of the food pathogens L. monocytogenes, Ent. sakazakii and Salm. Enteritidis to polypropylene and stainless steel surfaces.

The Isolation and Characterization of Multiply Antibiotic Resistant Strains of Fish Pathogenic Flavobacterium Species

Since the development of the first antibiotics in the 1940’s, there has been widespread overuse in both clinical and agricultural applications. Antibiotic resistance has become a significant problem as a result of subsequent dissemination of antibiotics into the environment, and multiply-resistant strains of bacteria are now a major pathogenic threat. In this study eight separate strains of Flavobacterium responsible for recent disease outbreaks in fish hatcheries throughout Maine were collected and analyzed. All eight strains were found to be resistant to high levels of a number of different antibiotics, including those used for aquaculture as well as human chemotherapeutic applications. Flavobacterium isolates were also shown phenotypically to transfer antibiotic resistance determinants using a conjugation mating system in which Flavobacterium was the donor and Escherichia coli DH5- alpha was the recipient. This experiment suggests that it may be possible for Flavobacterium strains to transfer their multiple antibiotic resistance determinants to human pathogenic bacterial strains. Importantly, none of the hatcheries from which the Flavobacterium isolates were obtained had ever used antibiotics to treat their fish stock. It is possible that there is another selective agent responsible for the development of antibiotic resistance in the absence of antibiotic pressure. Mercury is one possible candidate, as all of the strains tested were resistant to mercuric chloride and it is known that genes encoding antibiotic resistance can be carried on the same mobile genetic elements that encode for mercury resistance. Preliminary data also suggest that the majority of the Flavobacterium isolates contain genes for mercuric ion reduction, which would confirm the mercury resistance genotype.

A guerra dos clones: a replicação de modelos de negócio na legislação brasileira e seu impacto no desenvolvimento do setor de internet e tecnologia

O presente artigo busca trazer uma perspectiva jurídica ao debate sobre o impacto da replicação de modelos de negócio na Internet. O objetivo é oferecer uma análise sobre os copycats – empresas que reproduzem modelos de negócio inovadores desenvolvidos por empresas terceiras – em conjunto com os institutos da liberdade de concorrência e propriedade intelectual, examinando a contraposição existente entre esses no arcabouço jurídico nacional e buscando responder, como pergunta de trabalho, se esses mecanismos podem ser interpretados, à luz do ordenamento brasileiro, como mecanismos de incentivo ou restrição ao desenvolvimento de copycats no país.

Leaf anatomy of rubber-tree clones

A seringueira é uma planta de fácil reconhecimento por ser lenhosa, de porte mediano a grande, que apresenta um padrão característico de desfolha e reenfolhamento e, sobretudo, pela produção de látex. O objetivo do trabalho foi efetuar um estudo anatômico e morfológico foliar, comparando os clones RRIM 600 e GT 1 de seringueira &91;Hevea brasiliensis (Wild. ex Adr. de Juss.) Muell.- Arg&93;, desenvolvidos sob as mesmas condições edáficas e climáticas, para obtenção de informações que possam fornecer subsídios para correlações com dados fisiológicos e também diferenciar os clones em relação ao conteúdo de fibras, espessamento de tecidos do parênquima paliçádico e do parênquima lacunoso, caracterização anatômica do pecíolo, número e tamanho de estômatos e fornecer dados referentes a morfologia foliolar. Foram realizadas secções transversais na região do mesófilo, nervura central e pecíolo, seguindo-se os métodos usuais de preparação de lâminas permanentes. Foram realizadas análises biométricas de extensões de tecidos dos parênquimas paliçádico e lacunoso e contagem do número de células do parênquima lacunoso. Paralelamente foram realizadas análises biométricas para aferições de estômatos. Não houve diferenças para a altura das células epidérmicas, altura e número de camadas do parênquima lacunoso e para o comprimento e para a maior largura do limbo foliolar. Porém houve variação para a espessura das células do parênquima paliçádico, sendo que GT 1 apresentou maior espessura em relação a RRIM 600. GT1 apresentou maior número de estômatos em relação a RRIM 600, porém com menor tamanho. GT1 apresentou maior diâmetro da nervura central da folha e do pecíolo e maior quantidade de fibras de esclerênquima que RRIM 600.

Serological investigation and PCR in detection of pathogenic leptospires in snakes

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

Respostas fisiológicas de diferentes clones de eucalipto sob diferentes regimes de irrigação

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

Consequências da deriva de clomazone e sulfentrazone em clones de E. grandis x E. urophylla

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Isolation of extraintestinal pathogenic Escherichia coli from diarrheic dogs and their antimicrobial resistance profile

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