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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
Polymerase Chain Reaction
Polymerase Chain Reaction: methods
Water Microbiology
Genus-specific primers
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
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

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