Please use this identifier to cite or link to this item:
http://hdl.handle.net/10773/11119
Title: | PCR-DGGE-based methodologies to assess diversity and dynamics of Aeromonas communities |
Author: | Calhau, V. Tacão, M. Morgado, F. Correia, A. Henriques, I. |
Keywords: | Aeromonas Aeromonas - genetics Base Sequence Cluster Analysis DNA Primers DNA, Bacterial DNA, Bacterial - genetics Electrophoresis, Gel, Two-Dimensional Electrophoresis, Gel, Two-Dimensional - methods Genes, Bacterial Genetic Variation Humans Phylogeny Polymerase Chain Reaction Polymerase Chain Reaction: methods Portugal Seasons Water Microbiology Genus-specific primers gyrB PCR–DGGE rpoD sodB |
Issue Date: | Feb-2010 |
Publisher: | Wiley-Blackwell |
Abstract: | Aims: Aeromonas is ubiquitous in aquatic environments and may cause infectious diseases in fish and humans. However, reliable and specific methods to evaluate the diversity and dynamics of Aeromonas populations are currently unavailable. This study aimed to develop PCR–DGGE methodologies for culture-independent analysis of Aeromonas populations in water systems. Methods and Results: Three primer sets were designed to amplify selected sections of genes gyrB, rpoD and sodB from Aeromonas . Their specificity was confirmed by in silico analysis and by PCR on DNA from pure cultures. Estuarine water samples were analyzed by PCR–DGGE using those primers. DGGE patterns clearly clustered according to seasonal factors, and Aeromonas communities were surprisingly stable along a salinity gradient. Sequences of cloned amplicons affiliated to sequences belonging to seven Aeromonas species previously isolated from the same environment. Conclusions: The three systems used showed to be useful to describe the diversity of Aeromonas communities. However, the combined use of more than one primer set is advisable. Significance and Impact of the Study: The methods presented here can be applied to understand the natural pool of Aeromonas and also to monitor and control these bacteria in aquatic reservoirs. |
Peer review: | yes |
URI: | http://hdl.handle.net/10773/11119 |
DOI: | 10.1111/j.1365-2672.2009.04452.x |
ISSN: | 1364-5072 |
Appears in Collections: | CESAM - Artigos DBio - Artigos PT Mar - Artigos Ria de Aveiro - Artigos |
Files in This Item:
File | Description | Size | Format | |
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113_PCR–DGGE-based methodologies.pdf | 408.41 kB | Adobe PDF |
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