Detection of microcystin-producing cyanobacteria in Missisquoi Bay, Quebec, Canada, using quantitative PCR.
Toxic 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 microg 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 beta-ketoacyl synthase (mcyD(KS)) and the first dehydratase domain (mcyD(DH)) 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 10(4) copies of the mcyD(KS) gene ml(-1) were detected in August, and an average of 4.0 x 10(4) copies ml(-1) were detected in September, when microcystin concentrations were more than 4 microg liter(-1) and approximately 2 microg 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 10(2) mcyD(KS) 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 for the routine monitoring of the most at-risk water bodies.
Fortin
Nathalie
N
National Research Council Canada, Biotechnology Research Institute, 6100 Royalmount Ave., Montreal, QC H4P 2R2, Canada.
Aranda-Rodriguez
Rocio
R
Jing
Hongmei
H
Pick
Frances
F
Bird
David
D
Greer
Charles W
CW
D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D015169:Colony Count, Microbial / Q000379:methods; D000458:Cyanobacteria / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006836:Hydro-Lyases / Q000235:genetics; D052998:Microcystins / Q000096:biosynthesis*; D008853:Microscopy; D016133:Polymerase Chain Reaction / Q000379:methods*; D011792:Quebec; D014871:Water Microbiology
D001426:Bacterial Proteins; D017931:DNA Primers; D052998:Microcystins; C078588:microcystin; D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase; D006836:Hydro-Lyases