RESUMO
Vibrio mimicus is a gram-negative bacterium responsible for diseases in humans. Three strains of V. mimicus identified as V. mimicus 87, V. mimicus 92 and V. mimicus 93 were isolated from a shrimp processing facility in Guaymas, Sonora, Mexico. The strains were analyzed using several molecular techniques and according to the cluster analysis they were different, their similarities ranged between 51.3% and 71.6%. ERIC-PCR and RAPD (vmh390R) were the most discriminatory molecular techniques for the differentiation of these strains. The complete genomes of two strains (V. mimicus 87, renamed as CAIM 1882, and V. mimicus 92, renamed as CAIM 1883) were sequenced. The sizes of the genomes were 3.9 Mb in both strains, with 2.8 Mb in ChI and 1.1 Mb in ChII. A 12.7% difference was found in the proteome content (BLAST matrix). Several virulence genes were detected (e.g. capsular polysaccharide, an accessory colonization factor and genes involved in quorum-sensing) which were classified in 16 categories. Variations in the gene content between these genomes were observed, mainly in proteins and virulence genes (e.g., hemagglutinin, mobile elements and membrane proteins). According to these results, both strains were different, even when they came from the same source, giving an insight of the diversity of V. mimicus. The identification of various virulence genes, including a not previously reported V. mimicus gene (acfD) in ChI in all sequenced strains, supports the pathogenic potential of this species. Further analysis will help to fully understand their potential virulence, environmental impact and evolution.
Assuntos
Manipulação de Alimentos , Microbiologia de Alimentos , Penaeidae/microbiologia , Vibrio mimicus/isolamento & purificação , Microbiologia da Água , Animais , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Impressões Digitais de DNA/métodos , DNA Bacteriano/análise , DNA Bacteriano/genética , Contaminação de Alimentos , Congelamento , Genes Bacterianos , Proteínas Hemolisinas/genética , México , Técnica de Amplificação ao Acaso de DNA Polimórfico , Ribotipagem , Alinhamento de Sequência , Análise de Sequência de DNA , Especificidade da Espécie , Vibrio mimicus/classificação , Vibrio mimicus/genética , Vibrio mimicus/patogenicidade , Virulência/genéticaRESUMO
This report describes Vibrio seventh pandemic island II (VSP-II) and three novel variants revealed by comparative genomics of 23 Vibrio cholerae strains and their presence among a large and diverse collection of V. cholerae isolates. Three VSP-II variants were reported previously and our results demonstrate the presence of three novel VSP-II in clinical and environmental V. cholerae marked by major deletions and genetic rearrangements. A new VSP-II cluster was found in the seventh pandemic V. cholerae O1 El Tor strain CIRS101, which is dominant (95%) among the recent (2004-2007) seven pandemic V. cholerae O1 El Tor isolates from two endemic sites, but was not found in older strains from the same region. Two other variants were found in V. cholerae TMA21 and RC385, two environmental strains from coastal Brazil and the Chesapeake Bay, respectively, the latter being prevalent among environmental V. cholerae non-O1/non-O139 and Vibrio mimicus. The results of this study indicate that the VSP-II island has undergone significant rearrangement through a complex evolutionary pathway in V. cholerae. Interestingly, one of the new VSP-II revealed the presence of 'old' and 'new' V. cholerae O1 El Tor pandemic clones circulating in some of the areas where cholera is endemic.
Assuntos
Ilhas Genômicas , Genômica , Vibrio cholerae O1/genética , Vibrio cholerae não O1/genética , Vibrio mimicus/genética , Brasil , Cólera/microbiologia , DNA Bacteriano/genética , Microbiologia Ambiental , Ordem dos Genes , Humanos , Reação em Cadeia da Polimerase , Estados Unidos , Vibrio cholerae O1/isolamento & purificação , Vibrio cholerae O1/patogenicidade , Vibrio cholerae não O1/isolamento & purificação , Vibrio cholerae não O1/patogenicidade , Vibrio mimicus/isolamento & purificação , Vibrio mimicus/patogenicidadeRESUMO
Vibrio cholerae and Vibrio mimicus have similar phenotypes and genomes making rapid differentiation of these two species difficult. The first standard multilocus sequence analysis (MLSA) scheme for the identification of these species is described. A collection of 45 representative isolates from different geographical regions and hosts was examined using segments of the housekeeping genes pyrH, recA and rpoA. Overall, the closest phylogenetic neighbours of these species were Vibrio furnissii and Vibrio fluvialis. V. cholerae and V. mimicus formed separate species clusters on the basis of each gene, suggesting that these genes are useful as identification markers. These species clusters arose by the accumulation of point mutations. The pyrH gene showed the highest resolution for differentiating V. cholerae and V. mimicus. The maximum interspecies pyrH gene sequence similarity was 91 %. Clearly, V. mimicus strains were more heterogeneous than V. cholerae strains at the three loci. It is suggested that vibrio species may be defined on the basis of MLSA data. A vibrio species was defined as a group of strains forming a monophyletic group on the basis of these loci and with an intraspecific sequence similarity of at least 95 %. V. cholerae and V. mimicus isolates can be readily identified through the open database resource 'The Taxonomy of Vibrios' (http://www.taxvibrio.lncc.br/).
Assuntos
RNA Polimerases Dirigidas por DNA/genética , Núcleosídeo-Fosfato Quinase/genética , Recombinases Rec A/genética , Análise de Sequência de DNA/métodos , Vibrio cholerae/classificação , Vibrio mimicus/classificação , Técnicas de Tipagem Bacteriana , DNA Bacteriano/análise , DNA Bacteriano/genética , Humanos , Dados de Sequência Molecular , Filogenia , Vibrio cholerae/genética , Vibrio mimicus/genéticaRESUMO
De forma similar ao V. cholerae, outros vibrios potencialmente patogênicos podem causar doença no homem em uma variedade de formas de quadros auto-limitantes até diarréia severa a cólera, septicemia, celulite e infecções de pele. Com o advento da biologia molecular, novas técnicas vêm sendo empregadas para o estudo do gênero Vibrio com o intuito de determinar presença de fatores de virulência, traçar rotas de transmissão, ou ainda como ferramenta no estudo epidemiológico destes organismos. O presente estudo visou pesquisar a presença de genes codificadores de virulência em cepas de V. cholerae provenientes de vários países, e ainda estudar do polimorfismo genético, com o emprego de iniciadores para seqüências ERIC e BOX e eletroforese em campo pulsado (PFGE), para determinar a relação entre cepas de V. cholerae, V. mimicus, V. parahemolyticus, V. fluviais e V. alginolyticus isolados de amostras de ostras e mexilhões no Brasil, e V. metschnikovii isolados de amostras de peixe isolados de outros países bem como com o Padrão ATCC de cada espécie. Foi realizada também, a confirmação da posição taxonômica de cepas de V. metschnikovii isolados de amostras de peixes, e a comparação dos padrões de bandas com isolados de outros países e cepa padrão da espécie. A posição taxonômica de cepas de V. metschnikovii foi confirmada através da Hibridização DNA/DNA, e o estudo do polimorfismo genético através de ERIC PCR, BOX PCR e eletroforese em campo pulsado demonstrou que essas são poderosas ferramentas para estudos epidemiológicos do gênero Vibrio.