RESUMO
CTX-M-131 is a natural Asp240Gly variant from the CTX-M-2 group detected in a Providencia rettgeri clinical strain from Brazil. Molecular analysis showed that blaCTX-M-131 was inserted in a complex class 1 integron harbored by a 112-kb plasmid, which has not been previously described as a platform for CTX-M-encoding genes with the Asp240Gly mutation. Steady-state kinetic parameters showed that the enzyme has a typical cefotaximase catalytic profile and an enhanced activity against ceftazidime.
Assuntos
Providencia/genética , beta-Lactamases/genética , Antibacterianos/farmacologia , Brasil , Ceftazidima/farmacologia , Integrinas/genética , Cinética , Plasmídeos/genética , beta-Lactamases/metabolismoRESUMO
Giardia duodenalis is a protozoan that parasitizes humans and other mammals and causes giardiasis. Although its isolates have been divided into seven assemblages, named A to G, only A and B have been detected in human faeces. Assemblage A isolates are commonly divided into two genotypes, Al and All. Even though information about the presence of this protozoan in water and sewage is available in Brazil, it is important to verify the distribution of different assemblages that might be present, which can only be done by genotyping techniques. A total of 24 raw and treated sewage, surface and spring water samples were collected, concentrated and purified. DNA was extracted, and a nested PCR was used to amplify an 890 bp fragment of the gdh gene of G. duodenalis, which codes for glutamate dehydrogenase. Positive samples were cloned and sequenced. Ten out of 24 (41.6%) samples were confirmed to be positive for G. duodenalis by sequencing. Phylogenetic analysis grouped most sequences with G. duodenalis genotype All from GenBank. Only two raw sewage samples presented sequences assigned to assemblage B. In one of these samples genotype All was also detected. As these assemblages/genotypes are commonly associated to human giardiasis, the contact with these matrices represents risk for public health.
Assuntos
Giardia/classificação , Giardia/genética , Giardíase/parasitologia , Esgotos/microbiologia , Microbiologia da Água , Animais , Brasil , DNA de Protozoário/genética , Genótipo , Giardia/isolamento & purificação , Giardíase/epidemiologia , Glutamato Desidrogenase/genética , Humanos , Dados de Sequência Molecular , Reação em Cadeia da PolimeraseRESUMO
Since Staphylococcus aureus can cause several types of diseases, the development of antibiotic resistance poses an even greater threat to public health. S. aureus is known to possess the adaptive capability to promptly respond to antibiotics, making it resistant and increasingly difficult to treat; methicillin-resistant strains of S. aureus are a major concern with regard to this species. Previous studies reported the identification of methicillin-resistant S. aureus in food, demonstrating that this can represent a source of S. aureus which may carry the mecA gene. Fifty-seven S. aureus isolates, previously obtained from different types of food, were screened by polymerase chain reaction with specific primers for the mecA gene, which mediates methicillin resistance. Five (9%) isolates showed the presence of mecA gene, demonstrating that food may contain microorganisms possessing resistance genes. This study emphasizes the need to include food as a possible source of S. aureus carrying mecA gene and the need to monitor these products. Moreover, this is the first report of the presence of mecA genes in S. aureus isolated from ready-to-eat food in Brazil and Latin America.
Assuntos
Proteínas de Bactérias/genética , Fast Foods/microbiologia , Microbiologia de Alimentos , Resistência a Meticilina , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , Brasil , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Farmacorresistência Bacteriana , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Tipagem Molecular , Proteínas de Ligação às Penicilinas , Reação em Cadeia da Polimerase , Staphylococcus aureus/classificação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificaçãoRESUMO
A high incidence of waterborne diseases is observed worldwide and in order to address contamination problems prior to an outbreak, quantitative microbial risk assessment is a useful tool for estimating the risk of infection. The objective of this paper was to assess the probability of Giardia infection from consuming water from shallow wells in a peri-urban area. Giardia has been described as an important waterborne pathogen and reported in several water sources, including ground waters. Sixteen water samples were collected and examined according to the US EPA (1623, 2005). A Monte Carlo method was used to address the potential risk as described by the exponential dose response model. Giardia cysts occurred in 62.5% of the samples (<0.1-36.1 cysts/l). A median risk of 10⻹ for the population was estimated and the adult ingestion was the highest risk driver. This study illustrates the vulnerability of shallow well water supply systems in peri-urban areas.
Assuntos
Banhos , Ingestão de Líquidos , Giardia/isolamento & purificação , Giardíase/microbiologia , Microbiologia da Água , Abastecimento de Água/análise , Adulto , Brasil/epidemiologia , Giardia/imunologia , Giardíase/epidemiologia , Giardíase/imunologia , Humanos , Medição de Risco/métodos , População Rural , População Suburbana , Fatores de TempoRESUMO
It is known that Aeromonas spp. possess different chromosomal ß-lactamase genes. Presence and phenotypic expression of bla TEM, bla SHV, and bla CTX-M ESBL-encoding genes were investigated in environmental water isolates of Aeromonas hydrophila and Aeromonas jandaei. Presence of bla SHV and bla CTX-M genes was not observed, and bla TEM gene was verified in 91% of the isolates. Sequencing of 10 fragments showed the occurrence of bla TEM-116.
RESUMO
Cryptosporidium and Giardia are associated with cases of water and foodborne outbreaks in the world. This study included 50 samples of surface raw water collected from two watersheds in the state of São Paulo, Brazil. The isolation of (oo)cysts was performed in accordance with the U.S. Environmental Protection Agency's methods 1623 and genotypic characterization and quantification were carried out by Nested PCR and qPCR assays based on 18S rRNA and gdh genes, respectively. U.S. EPA 1623 method showed the presence of (oo)cysts in 40% ([Formula: see text] = 0.10 oocysts/L) and 100% ([Formula: see text] = 7.6 cysts/L) of samples from São Lourenço River, respectively, and 24% ([Formula: see text] = 0.8 oocysts/L) and 60% ([Formula: see text] = 1.64 cysts/L) of Guarapiranga Reservoir, respectively. The qPCR assay detected C. hominis/parvum in 52% (0.06 to 1.85 oocysts/L) of São Lourenço River and 64% (0.09 to 1.4 oocysts/L) of Guarapiranga Reservoir samples. Presence/absence test for Giardia intestinalis was positive in 92% of São Lourenço River and 8% of Guarapiranga Reservoir samples. The assemblage A was detected in 16% (0.58 to 2.67 cysts/L) in São Lourenço River and no positive samples were obtained for assemblage B in both water bodies. The characterization of anthroponotic species C. parvum/hominis, G. intestinalis, and assemblage A was valuable in the investigation of possible sources of contamination in the watersheds studied confirming the need of expanding environmental monitoring measures for protection of these water sources in our country.
Assuntos
Cryptosporidium/isolamento & purificação , Monitoramento Ambiental/métodos , Giardia/isolamento & purificação , Rios/parasitologia , Animais , Brasil , Cidades , Cryptosporidium/genética , Genótipo , Giardia/genética , Oocistos/isolamento & purificação , RNA Ribossômico 18S/genéticaRESUMO
The objectives of the study were to detect and genotype Cryptosporidium spp. and Giardia intestinalis in wastewater samples obtained from five cities with high transit of people in the State of São Paulo, Brazil, and at the entrance of a Wastewater Treatment Plant (WWTP) in Lima, Peru. Samples were collected and concentrated by centrifugation. The genomic DNA was extracted for molecular characterization by nested PCR for Cryptosporidium and double nested PCR for Giardia, followed by sequencing and phylogenetic analysis. G. intestinalis was found in 63.6 % of the samples, and the human assemblages A and B were identified. Cryptosporidium sp. was found in 36.4 % of the samples, and the species were corresponding to Cryptosporidium hominis, Cryptosporidium cuniculus, and Cryptosporidium muris. Results revealed the presence of human pathogenic Cryptosporidium species and G. intestinalis human pathogenic assemblages. Molecular tools highlight the importance to map the genetic diversity of these parasites, as well as to detect their epidemiological circulation pathway in the environment.
Assuntos
Cryptosporidium/isolamento & purificação , Giardia lamblia/isolamento & purificação , Águas Residuárias/parasitologia , Brasil , Cidades , Cryptosporidium/genética , Genótipo , Giardia lamblia/genética , Peru , Filogenia , Reação em Cadeia da Polimerase , RNA de Protozoário/genética , RNA Ribossômico 18S/genética , Análise de Sequência de DNARESUMO
INTRODUCTION: CTX-M enzymes are the most prevalent extended-spectrum beta-lactamases (ESBLs) in Brazil and around the world. The spread of CTX-M lies in their ability to be mobilized by insertion sequences and integrons. This study aimed to identify the mobile genetic structures associated with bla(CTX-M) genes from clinical Enterobacteriaceae strains. METHODOLOGY: Twenty-eight clinical non-clonal Enterobacteriaceae were screened by PCR for the presence of bla(CTX-M) genes and class 1 integrase (int1), and for the association of bla(CTX-M) with class 1 integrons. Plasmid incompatibility groups were assessed by PBRT. Wild-type plasmids were transformed into electrocompetent E. coli, and the S1-PFGE technique was used to verify the presence of high-molecular-weight plasmids in both wild-type strains and E. coli transformants. RESULTS: Sequencing showed that strains carried bla(CTX-M-2) (n = 25) and bla(CTX-M-59) (n = 3) genes inserted into the 3'-end of complex class 1 integrons. Thirteen strains also carried bla(TEM) and bla(SHV) genes. CTX-M-2/59-containing complex class 1 integrons were also present in E. coli transformants. The most frequent Inc groups were IncA/C (n = 10) and IncF (n = 8). Heavy plasmids were observed in both wild-type strains and E. coli transformants. CONCLUSIONS: The presence of the same bla(CTX-M-2-group)-containing genetic structure in seven Enterobacteriaceae species isolated at seven hospital wards shows the great mobility potential of complex class 1 integrons. Also, this is the first report of TEM-15, SHV-45, and SHV-55 in Latin America. The genetic environment of bla(CTX-M-2) accounts for their maintenance and spread among Gram-negative bacteria.
Assuntos
Proteínas de Bactérias/genética , Farmacorresistência Bacteriana , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/isolamento & purificação , beta-Lactamases/genética , Proteínas de Bactérias/classificação , Brasil/epidemiologia , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Primers do DNA , DNA Bacteriano/análise , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/microbiologia , Humanos , Reação em Cadeia da Polimerase , beta-Lactamases/classificaçãoRESUMO
Vibrio fluvialis is a halophilic bacterium found in many environments and is mainly associated with sporadic cases and outbreaks of gastroenteritis in humans. Here, we describe the genome sequences of environmental strains of V. fluvialis 560 (Vf560) and V. fluvialis 539 (Vf539) possessing a variant of the integrative and conjugative element (ICE) SXT for the first time in Brazil and South America.
RESUMO
In the last decade, atypical Listeria monocytogenes and L. innocua strains have been detected in food and the environment. Because of mutations in the major virulence genes, these strains have different virulence intensities in eukaryotic cells. In this study, we performed phenotypic and genotypic characterization of atypical L. monocytogenes and L. innocua isolates obtained from swine slaughterhouses and meat markets. Forty strains were studied, including isolates of L. monocytogenes and L. innocua with low-hemolytic activity. The isolates were characterized using conventional phenotypic Listeria identification tests and by the detection and analysis of L. monocytogenes-specific genes. Analysis of 16S rRNA was used for the molecular identification of the Listeria species. The L. monocytogenes isolates were positive for all of the virulence genes studied. The atypical L. innocua strains were positive for hly, plcA, and inlC. Mutations in the InlC, InlB, InlA, PI-PLC, PC-PLC, and PrfA proteins were detected in the atypical isolates. Further in vitro and transcriptomic studies are being developed to confirm the role of these mutations in Listeria virulence.
Assuntos
Matadouros , Microbiologia de Alimentos , Genótipo , Listeria monocytogenes , Carne/microbiologia , Mutação , Fatores de Virulência/genética , Animais , Humanos , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , SuínosRESUMO
Vibrio cholerae O1 is the causative agent of cholera and is ubiquitous in the aquatic environment, while V. cholerae strains non-O1 and non-O139 are recognized as causative agents of sporadic and localized outbreaks of diarrhea. Here, we report the complete sequence of a non-O1 and non-O139 V. cholerae strain (VCC19), which was isolated from the environment in Brazil. The sequence includes the integrative conjugative element (ICE). This paper is the first report of the presence of such an element in a V. cholerae strain isolated in Brazil.
RESUMO
Vibrio parahaemolyticus is a marine bacterium, responsible for gastroenteritis in humans. Most of the clinical isolates produce thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) encoded by tdh and trh genes respectively. In this study, twenty-three V. parahaemolyticus, previously isolated from oysters and mussels were analyzed by PCR using specific primers for the 16S rRNA and virulence genes (tdh, trh and tlh) and for resistance to different classes of antibiotics and PFGE. Nineteen isolates were confirmed by PCR as V. parahaemolyticus. The tlh gene was present in 100% of isolates, the tdh gene was identified in two (10.5%) isolates, whereas the gene trh was not detected. Each isolate was resistant to at least one of the nine antimicrobials tested. Additionally, all isolates possessed the blaTEM-116 gene. The presence of this gene in V. parahaemolyticus indicates the possibility of spreading this gene in the environment. Atypical strains of V. parahaemolyticus were also detected in this study.
Assuntos
Ostreidae/microbiologia , Frutos do Mar/microbiologia , Vibrio parahaemolyticus/isolamento & purificação , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Brasil , Proteínas Hemolisinas/genética , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Vibrio parahaemolyticus/efeitos dos fármacos , Vibrio parahaemolyticus/patogenicidade , Fatores de Virulência/genéticaRESUMO
The study aimed to analyze the themes related to the area of food safety surveillance that were approached in scientific research studies from postgraduate programs, with potential in-service application. A total of 337 theses and dissertations submitted to Universidade de São Paulo between 1993 and 2007 was analyzed. The results showed that research developed in universities can be applied to health surveillance, mainly regarding orientation to workers in this area in terms of updated practices.
Assuntos
Dissertações Acadêmicas como Assunto , Bibliometria , Inocuidade dos Alimentos , Pesquisa/estatística & dados numéricos , Universidades/estatística & dados numéricos , Brasil , Humanos , Vigilância da PopulaçãoRESUMO
O objetivo deste estudo foi investigar procedimentos de biossegurança adotados por profissionais de manicure, pedicure, tatuagem, piercing e maquiagem definitiva em Jacareí-SP. Utilizou-se abordagem descritiva e observacional de corte transversal. A pesquisa de campo foi realizada entre maio e junho de 2011. Foram feitas quarenta entrevistas com profissionais de estabelecimentos localizados no centro e nos dez bairros mais populosos. Utilizou-se questionário para avaliar conhecimentos e atitudes dos profissionais e um formulário para a observação de seus procedimentos e estrutura física dos estabelecimentos. Verificou-se falta de conhecimento sobre biossegurança pelos profissionais e deficiência na regulamentação desses serviços. Embora 55 por cento dos profissionais tenham realizado treinamento, seus procedimentos e a infraestrutura dos estabelecimentos foram favoráveis à transmissão de doenças. Sobre os processos de limpeza, desinfecção e esterilização de instrumentais, nenhum dos entrevistados sabia o tempo e a temperatura ideal para a esterilização, 57,5 por cento tinham equipamento inadequado para sua realização e 80 por cento não tinham termostato ou termômetro no equipamento para a conferência da temperatura. Apenas 57,5 por cento acreditavam que poderiam transmitir doenças durante sua prática profissional. Quarenta e cinco por cento dos entrevistados relataram ter tido contato com sangue sem luvas. Outro problema observado foi a reutilização de materiais descartáveis. Somente 10 por cento possuíam área específica para a esterilização de instrumentais. Evidenciou-se a necessidade de formação de qualidade sobre boas práticas de biossegurança a esses profissionais, além de normas e diretrizes pormenorizadas para a prevenção de infecções nesses serviços, bem como a melhoria da vigilância nesses estabelecimentos.
The objective of this study was to investigate biosafety procedures adopted by manicure, pedicure, tattoo, piercing and permanent makeup professionals in Jacareí-SP. We used a descriptive, observational and cross-sectional approach. The field research was conducted between May and June 2011. Forty professionals were surveyed in the downtown area and in the ten most populated districts of the municipality. We used a questionnaire to assess knowledge and attitudes of professionals as well as a formulary for the observation of professional procedures and physical structure of establishments. It has been found that professionals lack knowledge on biosafety procedures, and that the regulation of these services was deficient. Although 55% of professionals have attended to training courses, their procedures and establishments' infrastructure were favorable to disease transmission. Regarding the processes for cleaning, disinfecting and sterilizing instruments, none of interviewed knew the ideal time and temperature for sterilization, 57.5% had inadequate equipment for it and 80% had no thermostat or thermometer in the equipment for checking the temperature. Only 57.5% believed they could transmit infectious diseases during their professional practice. Forty-five percent of respondents reported having had contact with blood without wearing gloves. Another problem observed was the reuse of disposable material. Only 10% had a specific area for sterilizing instruments. The results demonstrated the need for providing to these professionals quality training on good biosafety practices and standards, detailed guidelines for the prevention of infections in these services as well as improvement on these establishments' surveillance.
Assuntos
Humanos , Masculino , Feminino , Centros de Embelezamento e Estética , Conhecimentos, Atitudes e Prática em Saúde , Infecções/transmissão , Tatuagem , Transmissão de Doença Infecciosa , Vigilância Sanitária/métodos , Demografia , Entrevistas como AssuntoRESUMO
A biotecnologia avançou consideravelmente nos séculos XX e XXI, de tal forma que, atualmente, 29 países são responsáveis pela produção de 160 milhões de hectares de organismos geneticamente modificados (OGMs). Esse desenvolvimento científico, tecnológico e produtivo requer medidas de monitoramento e controle para impedir grandes e futuros danos da introdução desses produtos no meio ambiente e no mercado. O presente trabalho teve como objetivo apresentar o cenário brasileiro dos alimentos geneticamente modificados e a evolução do corpo legislativo nacional relativo a eles, analisando-o com base nas políticas internacionais de produção e comércio de transgênicos. Para tanto, foi realizado um levantamento no que tange à legislação e literatura, reportadas até o momento, sobre a evolução do cenário dos OGMs de origem vegetal no Brasil e no mundo de 2007 a 2011, relacionando os dados obtidos com os modelos normativos europeu e estadunidense. As ações de vigilância sanitária no campo dos organismos geneticamente modificados são fundamentalmente de controle, monitoramento e fiscalização das etapas de desenvolvimento, plantio e comércio dos produtos transgênicos. A legislação brasileira segue os padrões internacionais de boas condutas relacionadas aos organismos geneticamente modificados, com normativas protetoras e com agências fiscalizadoras das atividades com esses produtos. No entanto, encontra-se em meio à dualidade das divergentes condutas europeia e americana, nas quais prevalece, atualmente, um posicionamento liberal.
Biotechnology has advanced much in the XX and XXI century, so that currently 29countries are responsible for the production of 160 million hectares of genetically modified organisms (GMO). This scientific, technological and productive development requires monitoring and control measures to prevent major and future damage from the introduction of these products on the environment and the market. The aim of this paper was to present the Brazilian scenario of genetically modified organisms and the development of the national legislative body relative to them, analyzing it based on the international politics of production and trade of transgenics. It was studied the legislation and literature, reported to date, about the development of genetically modified organismsframework in Brazil and in the world, from 2007 to 2011, comparing these data with the United States and European normative models. The health surveillance activities in the field of genetically modified organisms are fundamentally: control, monitoring and supervision of the stages of development, cultivation and trade of transgenic products. Brazilian legislation follows international standards of good conduct relating to genetically modified organisms, with protective normative and regulatory agencies of the activities with these products. However, Brazilian rules is among the duality of the divergent European and American conducts in which prevails, today, a liberal position
Assuntos
Biotecnologia , Alimentos Geneticamente Modificados , Legislação como Assunto , Vigilância Sanitária , Organismos Geneticamente ModificadosRESUMO
Vibrio parahaemolyticus is a marine bacterium, responsible for gastroenteritis in humans. Most of the clinical isolates produce thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) encoded by tdh and trh genes respectively. In this study, twenty-three V. parahaemolyticus, previously isolated from oysters and mussels were analyzed by PCR using specific primers for the 16S rRNA and virulence genes (tdh, trh and tlh) and for resistance to different classes of antibiotics and PFGE. Nineteen isolates were confirmed by PCR as V. parahaemolyticus. The tlh gene was present in 100 percent of isolates, the tdh gene was identified in two (10.5 percent) isolates, whereas the gene trh was not detected. Each isolate was resistant to at least one of the nine antimicrobials tested. Additionally, all isolates possessed the blaTEM-116 gene. The presence of this gene in V. parahaemolyticus indicates the possibility of spreading this gene in the environment. Atypical strains of V. parahaemolyticus were also detected in this study.
Vibrio parahaemolyticus é uma bactéria marinha, responsável por gastroenterite em humanos. A maioria dos isolados clínicos produzem hemolisina termoestável direta (TDH) e hemolisina TDH-relacionada (TRH) codificadas por genes tdh e trh, respectivamente. Neste estudo, vinte e três V. parahaemolyticus, previamente isolados de ostras e mexilhões foram analisados por PCR utilizando indicadores específicos para o gene 16S rRNA, genes de virulência (tdh, trh e tlh), resistência a diferentes classes de antibióticos, e PFGE. Dezenove isolados foram confirmados por PCR, como V. parahaemolyticus. O gene tlh estava presente em 100 por cento dos isolados, o gene tdh foi identificado em dois (10,5 por cento) dos isolados, enquanto que o gene trh não foi detectado. Cada isolado foi resistente a pelo menos um dos nove antibióticos testados. Além disso, todos os isolados apresentaram resultado positivo para o gene blaTEM-116. A presença deste gene em V. parahaemolyticus indica a possibilidade de propagação desse gene no ambiente. Cepas atípicas de V. parahaemolyticus foram também detectadas neste estudo.
Assuntos
Animais , Ostreidae/microbiologia , Frutos do Mar/microbiologia , Vibrio parahaemolyticus/isolamento & purificação , Antibacterianos/farmacologia , Brasil , Proteínas de Bactérias/genética , Proteínas Hemolisinas/genética , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Vibrio parahaemolyticus/efeitos dos fármacos , Vibrio parahaemolyticus/patogenicidade , Fatores de Virulência/genéticaRESUMO
O objetivo do estudo foi analisar os temas relacionados à área de vigilância sanitária de alimentos abordados em pesquisas científicas de cursos de pós-graduação, com potencial de aplicação no serviço. O total de 337 teses e dissertações apresentadas à Universidade de São Paulo entre os anos de 1993 e 2007 foi analisado. Os resultados mostraram que as pesquisas desenvolvidas nas universidades têm potencial para aplicação em vigilância sanitária, sobretudo no sentido de orientar os profissionais da área em práticas atualizadas.
El objetivo del estudio fue analizar los temas relacionados con el área de vigilancia sanitaria de alimentos abordados en investigaciones científicas de cursos de postgrado, con potencial de aplicación en el servicio. Fue seleccionada la Universidade de São Paulo como base para análisis de 337 tesis y disertaciones en ella presentada entre los años 1993 y 2007. Los resultados mostraron que las investigaciones desarrolladas en las universidades tienen potencial para aplicación en vigilancia sanitaria, sobretodo en el sentido de orientar los profesionales del área en prácticas actualizadas.
Assuntos
Alimentos , Avaliação de Programas e Instrumentos de Pesquisa , Dissertação Acadêmica , Currículo , Vigilância SanitáriaRESUMO
It is known that Aeromonas spp. possess different chromosomal â-lactamase genes. Presence and phenotypic expression of blaTEM, blaSHV, and blaCTX-M ESBL-encoding genes were investigated in environmental water isolates of Aeromonas hydrophila and Aeromonas jandaei. Presence of blaSHV and blaCTX-M genes was not observed, and blaTEM gene was verified in 91 percent of the isolates. Sequencing of 10 fragments showed the occurrence of blaTEM-116.
Assuntos
Humanos , Aeromonas hydrophila/genética , Aeromonas hydrophila/isolamento & purificação , Aeromonas/genética , Aeromonas/isolamento & purificação , Sequência de Bases , Expressão Gênica , Infecções por Bactérias Gram-Negativas , Fenótipo , Meio Ambiente , Técnicas Genéticas , MétodosRESUMO
The protozoan parasite Cryptosporidium has emerged as one of the most important water contaminants, causing outbreaks of waterborne diarrhea worldwide. In order to assess the importance for public health of this pathogens presence in environmental samples, several methods have been developed to isolate and detect Cryptosporidium oocysts. In the present study, a reliable and reproducible method has been standardized for detecting and identifying Cryptosporidium oocysts in water samples in the State of São Paulo, Brazil as the first step for future genotyping studies. Water samples were concentrated by filtration, and then subjected to ultrasound in Tween 80 0.1%, the obtained sediment was transferred into micro tubes containing 1.0 ml of distilled water and stored at -20ºC. DNA was extracted with the addition of 1% PVP in lysis buffer, the organic extraction was performed in Phase Lock GelHeavy®. There was a 214 bp amplification on the expected fragment in five out of the 11 water samples analyzed. The results of this study demonstrated the application usefulness of the standardized test in epidemiological studies and surveillance programs because the technology allowed to increase significantly the amount of amplified product.
O protozoário parasito Cryptosporidium tem emergido como um dos mais importantes contaminantes da água, causando surtos de diarreia de veiculação hídrica em todo mundo. Para avaliar o significado, para a saúde pública,da presença desse agente patogênico em amostras ambientais, vários métodos têm sido desenvolvidos para isolar e detectar oocistos de Cryptosporidium. No presente estudo foi padronizado um método confiável e reprodutível para detectar e identificar oocistos de Cryptosporidium em amostras de água no Estado de São Paulo, Brasil, comoo primeiro passo para futuros estudos de genotipagem. Amostras de água foram concentradas por filtração,submetidas a ultrasom em solução de Tween 80 a 0.1%; o sedimento obtido foi transferido para microtuboscontendo 1,0 ml de água destilada e conservado a -20ºC. O DNA foi extraído com adição de 1% de PVP no tampão de lise; a extração foi realizada em tubo Phase Lock Gel Heavy®. Houve amplificação do fragmento esperado de 214 bp em cinco das 11 amostras de água analisadas. Os resultados deste estudo demonstraram a utilidade deaplicação do teste padronizado em estudos epidemiológicos e em programas de vigilância, em virtude da técnica ter apresentado sensibilidade para incrementar significativamente a quantidade de produto amplificado.
Assuntos
Cryptosporidium , Oocistos , Poluição da Água , Reação em Cadeia da Polimerase , Saúde PúblicaRESUMO
Apresenta as iniciativas feitas para evitar a ocorrência de uma epidemia de cólera no país, como a política de saneamento e as açöes de educaçäo sanitária voltadas para a comunidade. Conclui com as perspectivas relativas à permanência da doença no país (AMSB)