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

Rivera, Irma Nelly Gutierrez; Lipp, Erin K.; Gil, Ana I.; Choopun, Nipa; Huq, Anwar; Colwell, Rita R.

doi:10.1046/j.1462-2920.2003.00443.x

Cited by 89 publications

(96 citation statements)

References 58 publications

(66 reference statements)

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“…DNA was extracted using a modification of the method of Rivera et al (47). The differences were as follows: the cell pellet was resuspended in 360 l SET buffer (20% [wt/vol] sucrose, 50 mM Tris-HCl [pH 8.0], 50 mM EDTA), and the volumes of all solutions added thereafter were one-fifth the original volumes described (47). The quality and quantity of the DNA extracted from the surface-colonizing microbes were determined by electrophoresis on 0.8% (wt/vol) agarose gels in 0.5ϫ Tris-borate-EDTA.…”

Section: Methodsmentioning

confidence: 99%

Cross-Ocean Distribution of Rhodobacterales Bacteria as Primary Surface Colonizers in Temperate Coastal Marine Waters

Dang

Chen

et al. 2008

Appl Environ Microbiol

349

279

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Bacterial surface colonization is a universal adaptation strategy in aquatic environments. However, neither the identities of early colonizers nor the temporal changes in surface assemblages are well understood. To determine the identities of the most common bacterial primary colonizers and to assess the succession process, if any, of the bacterial assemblages during early stages of surface colonization in coastal water of the West Pacific Ocean, nonnutritive inert materials (glass, Plexiglas, and polyvinyl chloride) were employed as test surfaces and incubated in seawater off the Qingdao coast in the spring of 2005 for 24 and 72 h. Phylogenetic analysis of the 16S rRNA gene sequences amplified from the recovered surface-colonizing microbiota indicated that diverse bacteria colonized the submerged surfaces. Multivariate statistical cluster analyses indicated that the succession of early surface-colonizing bacterial assemblages followed sequential steps on all types of test surfaces. The Rhodobacterales, especially the marine Roseobacter clade members, formed the most common and dominant primary surface-colonizing bacterial group. Our current data, along with previous studies of the Atlantic coast, indicate that the Rhodobacterales bacteria are the dominant and ubiquitous primary surface colonizers in temperate coastal waters of the world and that microbial surface colonization follows a succession sequence. A conceptual model is proposed based on these findings, which may have important implications for understanding the structure, dynamics, and function of marine biofilms and for developing strategies to harness or control surface-associated microbial communities.

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Section: Methodsmentioning

confidence: 99%

Cross-Ocean Distribution of Rhodobacterales Bacteria as Primary Surface Colonizers in Temperate Coastal Marine Waters

Dang

Chen

et al. 2008

Appl Environ Microbiol

349

279

View full text Add to dashboard Cite

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“…In vitro and in vivo studies showed that VBNC cells continue to be metabolically active, retain pathogenic potential but do not grow and do not form a colony on traditional culture media (57). VBNC bacteria can be detected in biological samples by culture-independent methods only (e.g., immunofluorescence, PCR) (58,59,60). Under favorable conditions (such as a rise in temperature), reversion from the VBNC to the culturable state can take place (56,61).…”

Section: Effect Of Increasing Sst On Vibrio Growth and Biology: Fieldmentioning

confidence: 99%

Effects of Global Warming onVibrioEcology

et al. 2015

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Vibrio-related infections are increasing worldwide both in humans and aquatic animals. Rise in global sea surface temperature (SST), which is approximately 1°C higher now than 140 years ago and is one of the primary physical impacts of global warming, has been linked to such increases. In this chapter, major known effects of increasing SST on the biology and ecology of vibrios are described. They include the effects on bacterial growth rate, both in the field and in laboratory, culturability, expression of pathogenicity traits, and interactions with aquatic organisms and abiotic surfaces. Special emphasis is given to the effect of ocean warming on Vibrio interactions with zooplankters, which represent one of the most important aquatic reservoirs for these bacteria. The reported findings highlight the biocomplexity of the interactions between vibrios and their natural environment in a climate change scenario, posing the need for interdisciplinary studies to properly understand the connection between ocean warming and persistence and spread of vibrios in sea waters and the epidemiology of the diseases they cause.

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“…Samples were thawed at room temperature, and total nucleic acids were extracted and purified using methods described previously by Rivera et al [36]. Crude DNA was purified with the WIZARD DNA Clean Up System (Promega).…”

Section: Extraction Of Genomic Dna and Pcr Amplification Of Bacterialmentioning

confidence: 99%

Diversity of Bacterial Communities in Container Habitats of Mosquitoes

Ponnusamy

Stav

et al. 2008

Microb Ecol

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We investigated the bacterial diversity of microbial communities in water-filled, human-made and natural container habitats of the mosquitoes Aedes aegypti and Aedes albopictus in suburban landscapes of New Orleans, Louisiana in 2003. We collected water samples from three classes of containers, including tires (n=12), cemetery urns (n=23), and miscellaneous containers that included two tree holes (n=19). Total genomic DNA was extracted from water samples, and 16S ribosomal DNA fragments (operational taxonomic units, OTUs) were amplified by PCR and separated by denaturing gradient gel electrophoresis (DGGE). The bacterial communities in containers represented diverse DGGE-DNA banding patterns that were not related to the class of container or to the local spatial distribution of containers. Mean richness and evenness of OTUs were highest in water samples from tires. Bacterial phylotypes were identified by comparative sequence analysis of 90 16S rDNA DGGE band amplicons. The majority of sequences were placed in five major taxa: Alpha-, Beta-and Gammaproteobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, and an unclassified group; Proteobacteria and Bacteroidetes were the predominant heterotrophic bacteria in containers. The bacterial communities in human-made containers consisted mainly of undescribed species, and a phylogenetic analysis based on 16S rRNA sequences suggested

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

Cited by 89 publications

References 58 publications

Cross-Ocean Distribution of Rhodobacterales Bacteria as Primary Surface Colonizers in Temperate Coastal Marine Waters

Cross-Ocean Distribution of Rhodobacterales Bacteria as Primary Surface Colonizers in Temperate Coastal Marine Waters

Effects of Global Warming onVibrioEcology

Diversity of Bacterial Communities in Container Habitats of Mosquitoes

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