An alternative genotyping method using dye-labeled universal primer to reduce unspecific amplifications

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

Intestinal colonization of a human subject by vancomycin-resistant Enterococcus faecium

Objective: To study the ability of two strains of vancomycin-resistant Enterococcus faecium to colonize the human intestine. Methods: A single human subject ingested separately two strains of vancomycin-resistant E. faecium isolated from a pig and a chicken. The feces were cultured on selective medium. Prior to ingestion no vancomycin-resistant cocci were present in the feces. Ingestion of 10 4-10 5 CFU resulted in either no colonization or isolation only after enrichment. Ingestion of 10 7 CFU of one strain resulted in colonization for a period of nearly 3 weeks, with fecal counts at times in excess of 10 6 CFU/g. Ingestion of similar numbers of the other strain and reingestion of the first strain resulted in excretion in the feces for much shorter periods. When the fecal count of the ingested strains was greater than 10 4-10 5 CFU/g, the strains were isolated from swabs taken from perianal skin. Conclusions: Vancomycin-resistant E. faecium strains from pigs and poultry a re able to colonize the human gut and the perianal skin.

Pheno and genotyping of Staphylococcus aureus, isolated from bovine milk samples from São Paulo State, Brazil

In the present study, 87 Staphylococcus aureus isolates obtained from milk samples of 87 cows with mastitis in 6 different municipal districts of 2 regions of São Paulo State, Brazil, were compared pheno and genotypically. Pulsed-field gel electrophoresis (PFGE) analysis of the strains was performed, and PCR was carried out to detect genes for a number of staphylococcal cell surface proteins, exoproteins, and 3 classes of agr genes. Nine distinct S. aureus lineages (LA-LI) were identified by PFGE. The lineages LA and LE, which accounted together for 63 strains (72.2%), were prevalent and had been collected from all of the 6 municipal districts, indicating a broad geographic distribution of these lineages; LB, LC, LD, LF, LG, LH, and LI, however, were isolated sporadically and accounted for 24 strains (27.8%). Some characteristics, like penicillin resistance and the presence of cap8 and agr class II genes, were associated with the prevalent lineages (LA and LE), and penicillin susceptibility and the presence of cna and cap5 genes were associated with sporadic lineages. According to the present results, some S. aureus lineages possess a combination of genes that confer the propensity to cause and disseminate infection, and only a limited number of clones are responsible for the cases of bovine mastitis on the various farms. © 2004 NRC Canada.

Toxoplasma gondii genotyping in a dog co-infected with distemper virus and ehrlichiosis rickettsia

This paper reports a toxoplasmosis, erhlichiosis and distemper co-infection in a dog with an exuberant neuropathological clinical picture. Primary involvement was discussed based on information collected in the analysis of the clinical case, such as neurological impairment, epidemiological data, poor immunoprophylactic scheme of the dog affected and the role of these diseases on immunosuppression. Canine distemper and ehrlichiosis were diagnosed based on epidemiologic data, clinical signs, hematological and cytological evaluation. Toxoplasma gondii was isolated and genetically characterized as Type I using restriction analysis (RFLP) with SAG-2 genes. Immunosuppression features of both dogs and human beings are discussed, as well as implications on animal and public health. This is the first report on toxoplasmosis, ehrlichiosis and distemper co-infection in a dog in Brazil, associated with genotyping determination of the T. gondii strain involved.

Densidade de Enterococcus sp em águas recreacionais e areias de praias do município de São Vicente-SP, Brasil e sua relação com parâmetros abióticos

Coastal cities attract a large number of tourists for their beautiful landscape and recreational activities, increasing the municipalities' income source. Thus, a need is clear for the establishment of beach quality monitoring programs to ensure bathers health. Although there is an effective monitoring program for recreational waters in Brazil there are no programs to certify the quality of beach sands. In this sense, the aim of this work was to analyze the density of bacteria from the genus Enterococcus in both sand and water from two beaches from São Vicente, São Paulo (Brazil) and correlate these densities to abiotic parameters such as: temperature, salinity, particle size, organic matter and tides). Water and sand samples were collected during February 2006 on the beaches of Gonzaguinha e Ilha Porchat and bacterial densities were determined by membrane filter technique. Temperature and salinity were measured in situ with a thermometer and a refractometer while particle size and organic matter were determined according to methods described by Suguio and Dean. There were significant differences between densities found in water and sand (p=0.004), being approximately 20 times higher in Gonzaguinha's beach sands. Similar results were found for Ilha Porchat beach samples, being the densities found in sand 3 times greater than those found in water. Both beaches showed a negative correlation between bacterial densities and salinity and temperature, suggesting a deleterious effect of these parameters on the bacterial community. On the other hand, no correlation was found between organic matter and particle size and bacterial densities. This work demonstrates that there is a great necessity for monitoring beach sand since the evaluation of beach quality is an important part of integrated coastal management programs.

Photodynamic and Antibiotic Therapy Impair the Pathogenesis of Enterococcus faecium in a Whole Animal Insect Model

Enterococcus faecium has emerged as one of the most important pathogens in healthcare-associated infections worldwide due to its intrinsic and acquired resistance to many antibiotics, including vancomycin. Antimicrobial photodynamic therapy (aPDT) is an alternative therapeutic platform that is currently under investigation for the control and treatment of infections. PDT is based on the use of photoactive dye molecules, widely known as photosensitizer (PS). PS, upon irradiation with visible light, produces reactive oxygen species that can destroy lipids and proteins causing cell death. We employed Galleria mellonella (the greater wax moth) caterpillar fatally infected with E. faecium to develop an invertebrate host model system that can be used to study the antimicrobial PDT (alone or combined with antibiotics). In the establishment of infection by E. faecium in G. mellonella, we found that the G. mellonella death rate was dependent on the number of bacterial cells injected into the insect hemocoel and all E. faecium strains tested were capable of infecting and killing G. mellonella. Antibiotic treatment with ampicillin, gentamicin or the combination of ampicillin and gentamicin prolonged caterpillar survival infected by E. faecium (P = 0.0003, P = 0.0001 and P = 0.0001, respectively). In the study of antimicrobial PDT, we verified that methylene blue (MB) injected into the insect followed by whole body illumination prolonged the caterpillar survival (P = 0.0192). Interestingly, combination therapy of larvae infected with vancomycin-resistant E. faecium, with antimicrobial PDT followed by vancomycin, significantly prolonged the survival of the caterpillars when compared to either antimicrobial PDT (P = 0.0095) or vancomycin treatment alone (P = 0.0025), suggesting that the aPDT made the vancomycin resistant E. faecium strain more susceptible to vancomycin action. In summary, G. mellonella provides an invertebrate model host to study the antimicrobial PDT and to explore combinatorial aPDT-based treatments.

O emprego da metodologia de superfície de resposta no desenvolvimento de um novo produto simbiótico, fermentado com Enterococcus faecium CRL 183 e Lactobacillus helveticus ssp jugurti 416, à base de extratos aquosos de soja e de Yacon (Smallanthus sonchifolius)

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

Avaliação do simbiótico fermentado com Enterococcus faecium CRL 183 e Lactobacillus helveticus ssp jugurti 416, à base de extratos aquosos de soja e de yacon (Smallanthus sonchifolius) no controle do desenvolvimento do Diabetes Mellitus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Densidade e resistência a antimicrobianos de Enterococcus sp e Escherichia coli isoladas de águas, areias e algas do gênero Sargassum de praias recreacionais do Litoral Norte do Estado de São Paulo

Pós-graduação em Ciências Biológicas (Microbiologia Aplicada) - IBRC

Avaliação in vitro de caracteristicas probióticas do enterococcus faecium CRL 183 e do Lactobacillus helveticus ssp jugurti 416

Pós-graduação em Alimentos e Nutrição - FCFAR

Influência do consumo de Iogurte de soja fermentado com Enterococcus faecium na microbiota intestinal de animais e humanos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Alterações comportamentais, psicológicas, de higiene bucal e presença de microrganismos superinfectantes e viruses da família herpesviridae em pacientes mantidas em programa de desintoxicação para dependentes químicos

Pós-graduação em Odontologia - FOA

Ocorrência de microrganismos oportunistas e superinfectantes em crianças com 6, 12 e 18 meses de idade e suas mães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Prevalence and genotyping of hepatitis C virus in blood donors in the state of Pará, Northern Brazil

Given the scarcity of epidemiological information on hepatitis C virus (HCV) infection in Northern Brazil, we determined the prevalence and genotypic frequency in blood donors in the state of Pará (PA). Blood samples from all of the blood donors at the Fundação HEMOPA (blood bank of PA) from 2004-2006 were screened for the presence of antibodies to anti-HCV and samples seroreactive to anti-HCV were further tested for HCV RNA using real-time PCR. In total, 116 HCV-RNA samples were genotyped, based on maximum likelihood phylogenetic analyses, using BioEdit, Modelgenerator, PHYML and FigTree software. The population consisted of 242,726 volunteers who donated blood from 2004-2006; the most common subgroup was males between the ages of 18-29 years old (37.30%). Within the whole group, 1,112 blood donors (0.46%) had indeterminate or positive serology; among these, 28.78% were males whose ages ranged from 18-29 years. A diagnosis of chronic HCV infection was confirmed for 304 donors (60.20% males; 66.45% were 30-49 years old), resulting in a prevalence of HCV RNA in 0.13% of the samples (304 of 242,726). HCV genotyping revealed a high frequency of genotype 1 (108/116) followed by genotype 3 (8/116). This study found HCV infection to be relatively infrequent in PA; genotype 1 was most commonly isolated. This information can help guide prevention and control policies aimed at efficient diagnosis and control measures.

Efeito de uma bebida probiótica (Enterococcus faecium CRL 183 e Bifidobacterium longum ATCC 15707) à base de extrato aquoso de soja no desenvolvimento de colite quimicamente induzida em ratos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)