Detection of microcystin-producing cyanobacteria in Missisquoi Bay, Quebec, Canada, using Quantitative PCR

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DOIResolve DOI: http://doi.org/10.1128/AEM.00183-10
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TypeArticle
Journal titleApplied and environmental microbiology
Volume76
Issue15
Pages51055112; # of pages: 8
AbstractToxic cyanobacterial blooms, as well as their increasing global occurrence, pose a serious threat to public health, domestic animals, and livestock. In Missisquoi Bay, Lake Champlain, public health advisories have been issued from 2001 to 2009, and local microcystin concentrations found in the lake water regularly exceeded the Canadian drinking water guideline of 1.5 µg liter-1. A quantitative PCR (Q-PCR) approach was developed for the detection of blooms formed by microcystin-producing cyanobacteria. Primers were designed for the ?-ketoacyl synthase (mcyDKS) and the first dehydratase domain (mcyDDH) of the mcyD gene, involved in microcystin synthesis. The Q-PCR method was used to track the toxigenic cyanobacteria in Missisquoi Bay during the summers of 2006 and 2007. Two toxic bloom events were detected in 2006: more than 6.5 x 104 copies of the mcyDKS gene ml-1 were detected in August, and an average of 4.0 x 104 copies ml-1 were detected in September, when microcystin concentrations were more than 4 µg liter-1 and approximately 2 µg liter-1, respectively. Gene copy numbers and total microcystin concentrations (determined by enzyme-linked immunosorbent assay [ELISA]) were highly correlated in the littoral (r = 0.93, P < 0.001) and the pelagic station (r = 0.87, P < 0.001) in 2006. In contrast to the situation in 2006, a cyanobacterial bloom occurred only in late summer-early fall of 2007, reaching only 3 x 102 mcyDKS copies ml-1, while the microcystin concentration was barely detectable. The Q-PCR method allowed the detection of microcystin-producing cyanobacteria when toxins and toxigenic cyanobacterial abundance were still below the limit of detection by high-pressure liquid chromatography (HPLC) and microscopy. Toxin gene copy numbers grew exponentially at a steady rate over a period of 7 weeks. Onshore winds selected for cells with a higher cell quota of microcystin. This technique could be an effective approach.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Biotechnology Research Institute
Peer reviewedYes
NRC number53334
NPARC number16005528
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Record identifierbd16fcb8-7d4f-4340-a5cc-ed1dc8c610cc
Record created2010-11-05
Record modified2016-05-09
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