Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNAs from Vibrio cholerae O1, O139, and non-O1 strains

Rivera, I. G.; Chowdhury, M. A. R.; Huq, Anwar; Jacobs, Dan N.; Martins, Maria Therezinha; Colwell, Rita R.

doi:10.1128/aem.61.8.2898-2904.1995

Cited by 148 publications

(87 citation statements)

References 49 publications

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“…Shewanella putrefaciens was observed in the intestine of common carp (Al-Harbi & Uddin, 2008). Reddacliff et al (1993) isolated V. cholerae from septicaemic goldfish, but the species is a common inhabitant of aquatic environments, with many nontoxigenic strains (Rivera et al, 1995). Vibrio spp.…”

Section: Discussionmentioning

confidence: 99%

Influence of the diet on the microbial diversity of faecal and gastrointestinal contents in gilthead sea bream (Sparus aurata) and intestinal contents in goldfish (Carassius auratus)

Silva

Nicoli

Zambonino-Infante

et al. 2011

FEMS Microbiology Ecology

105

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Fish intestinal microbiota changes with the diet and this effect is of particular interest considering the increasing substitution of fish meal by plant protein sources. The objective of this work was to study the effects of partial substitution of fish meal with lupin and rapeseed meals on gut microbiota of the gilthead sea bream (Sparus aurata) and in goldfish (Carassius auratus). Faecal, gastrointestinal and intestinal contents were characterized using culture-based and molecular methods. Vibrionaceae was high in faeces and in the intestine of sea bream, while a more diverse microbiota was retrieved from the stomach, where Bacillales and Flavobacteriaceae appeared to be influenced by the diet. PCR-denaturing gradient gel electrophoresis profiles revealed a high diversity of the microbiota transiting in the sea bream digestive tract, with a shift between gastric and intestinal communities, especially in the group fed with lupin meal. The goldfish was different, with a predominance of Aeromonas spp., Shewanella putrefaciens and Staphylococcus spp. among the aerotolerant-cultivable bacteria. The culture-independent methods revealed the presence of anaerobes like Cetobacterium somerae, and that of Vibrio spp., likely in a viable, but noncultivable state. There was a trend towards decreasing diversity in goldfish microbiota with the partial substitution by lupin, which seemed to inhibit some taxa.

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Section: Discussionmentioning

confidence: 99%

Influence of the diet on the microbial diversity of faecal and gastrointestinal contents in gilthead sea bream (Sparus aurata) and intestinal contents in goldfish (Carassius auratus)

Silva

Nicoli

Zambonino-Infante

et al. 2011

FEMS Microbiology Ecology

105

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“…One aspect of preventing cholera outbreaks is determining the microbiological risk of ballast water from ships coming from endemic areas that could transport and spread pathogenic V. cholerae. Although detection of bacteria using DNA extracted from sea water depends on their concentration in the sea water sample, the limit of detection observed in this and previous studies (Rivera et al, 1995a; was such that we could establish the presence of V. cholerae O1 and O139 in 100 ng of total DNA from aquatic samples. The same approach applied to plankton samples yielded only 57.6% success.…”

Section: Discussionmentioning

confidence: 99%

“…DNA extraction was performed according to the method described previously (Rivera et al, 1995b). Approximately 100 ng ml -1 DNA template was used per PCR (Rivera et al, 1995a;. DNA and PCR amplicons were analysed by gel electrophoresis on a 1.4% (w/v) agarose gel as described before.…”

Section: Evaluation Of the Multiplex O1/o139 Pcrmentioning

confidence: 99%

Method of DNA extraction and application of multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae O1 and O139 from aquatic ecosystems

Rivera

Lipp

Gil

et al. 2003

Environmental Microbiology

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Vibrio cholerae is a free-living bacterium found in water and in association with plankton. V. cholerae non-O1/non-O139 strains are frequently isolated from aquatic ecosystems worldwide. Less frequently isolated are V. cholerae O1 and V. cholerae O139, the aetiological agents of cholera. These strains have two main virulence-associated factors, cholera toxin (CT) and toxin co-regulated pilus (TCP). By extracting total DNA from aquatic samples, the presence of pathogenic strains can be determined quickly and used to improve a microbiological risk assessment for cholera in coastal areas. Some methods suggested for DNA extraction from water samples are not applicable to all water types. We describe here a method for DNA extraction from coastal water and a multiplex polymerase chain reaction (PCR) for O1 and O139 serogroups. DNA extraction was successfully accomplished from 117 sea water samples collected from coastal areas of Perú, Brazil and the USA. DNA concentration in all samples varied from 20 ng to 480 micro g micro l-1. The sensitivity of the DNA extraction method was 100 V. cholerae cells in 250 ml of water. The specificity of multiplex O1/O139 PCR was investigated by analysing 120 strains of V. cholerae, Vibrio and other Bacteria species. All V. cholerae O1 and O139 tested were positive. For cholera surveillance of aquatic environments and ballast water, total DNA extraction, followed by V. cholerae PCR, and O1/O139 serogroup and tcpA/ctxA genes by multiplex PCR offers an efficient system, permitting risk analysis for cholera in coastal areas.

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“…Both REP and ERIC sequences are based on short extragenic repetitive sequences that are conserved in the Enterobacteriaceae [31,35]. These sequences vary in location throughout the chromosome depending on the species examined [3,38,39,42] and have been used to ¢ngerprint a number of bacterial species [31,36,37]. PGRS-PCR targets GC-rich repetitive sequences and was developed to type M. tuberculosis complex bacteria [30].…”

Section: Discussionmentioning

confidence: 99%

Genotyping of clinical Serratia marcescens isolates: a comparison of PCR-based methods

Patton¹

2001

FEMS Microbiology Letters

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The polymerase chain reaction (PCR)-based procedures of randomly amplified polymorphic DNA (RAPD) and repetitive element (RE)-based PCR were used to amplify total DNA prepared from each of 62 clinical Serratia marcescens isolates. Three different random primers, designated 1060, 1254 and 1283, were used individually in RAPD-PCR. Primers representing enterobacterial repetitive intergenic consensus (ERIC) sequences, extragenic palindromic (REP) elements, and polymorphic GC-rich repetitive sequences (PGRS) constituted the repetitive element-PCR. We were able to generate 40, 40 and 58 genotypic groupings using the 1060, 1254 and 1283 RAPD primers, respectively. Using the ERIC, REP and PGRS primers, 19, 54 and 60 unique genotypic profiles were yielded, respectively. The PGRS primers, which were developed to amplify GC-rich repetitive sequences in the genome of Mycobacteria, were the most discriminatory. These data indicate that both of these PCR-based approaches are a valid means of discriminating strain differences among isolates of S. marcescens and the amount of differentiation depends on the primer used. These techniques should prove useful for routine surveillance or in examining outbreaks of S. marcescens in clinical settings. ß

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Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNAs from Vibrio cholerae O1, O139, and non-O1 strains

Cited by 148 publications

References 49 publications

Influence of the diet on the microbial diversity of faecal and gastrointestinal contents in gilthead sea bream (Sparus aurata) and intestinal contents in goldfish (Carassius auratus)

Influence of the diet on the microbial diversity of faecal and gastrointestinal contents in gilthead sea bream (Sparus aurata) and intestinal contents in goldfish (Carassius auratus)

Method of DNA extraction and application of multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae O1 and O139 from aquatic ecosystems

Genotyping of clinical Serratia marcescens isolates: a comparison of PCR-based methods

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