The present study was designed to develop a novel, rapid, direct DNA-based protocol to enumerate aeromonads in recreational waters. An Aeromonas genus-specific real-time quantitative polymerase chain reaction (Q-PCR) protocol was developed and optimized using newly designed genus-specific oligonucleotide primers derived from the gyrase B subunit (GyrB) gene. A standard curve was developed based on the PCR protocol with a minimum quantification limit of 10 cell equivalents ml−1 achieved using an autoclaved water sample from recreational water spiked with known quantities of an Aeromonas ATCC strain. The Q-PCR protocol was validated and applied to detect and quantify the total number of aeromonads in water samples collected from two fresh water beaches on Lake Ontario. The Q-PCR protocol revealed significantly higher numbers of aeromonads in all water samples than a culture-based assay at both beaches. Foreshore sand was found to serve as a reservoir of high concentrations of Aeromonas similar to this phenomenon noted for enteric bacteria like Eschershia coli. The new real-time Q-PCR protocol facilitated the rapid quantification of total numbers of Aeromonas cells present in recreational water samples in <3 hours without culturing.
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Research Article|
February 01 2009
DNA-based real-time detection and quantification of aeromonads from fresh water beaches on Lake Ontario
Izhar U. H. Khan;
Izhar U. H. Khan
1National Water Research Institute, Water Science and Technology Directorate, Environment Canada, 867 Lakeshore Rd, Burlington, ON L7R 4A6, Canada
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Alyssa Loughborough;
Alyssa Loughborough
1National Water Research Institute, Water Science and Technology Directorate, Environment Canada, 867 Lakeshore Rd, Burlington, ON L7R 4A6, Canada
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Thomas A. Edge
1National Water Research Institute, Water Science and Technology Directorate, Environment Canada, 867 Lakeshore Rd, Burlington, ON L7R 4A6, Canada
Tel.: +905-319-6932 Fax: +905-336-6430; E-mail: [email protected]
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J Water Health (2009) 7 (2): 312–323.
Article history
Received:
April 17 2008
Accepted:
July 30 2008
Citation
Izhar U. H. Khan, Alyssa Loughborough, Thomas A. Edge; DNA-based real-time detection and quantification of aeromonads from fresh water beaches on Lake Ontario. J Water Health 1 June 2009; 7 (2): 312–323. doi: https://doi.org/10.2166/wh.2009.041
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