Comparison of Enterococcus measurements in freshwater at two recreational beaches by quantitative polymerase chain reaction and membrane filter culture analysis.
Cell densities of the fecal pollution indicator genus, Enterococcus, were determined by a rapid (3 h or less) quantitative polymerase chain reaction (QPCR) analysis method in 100 ml water samples collected from recreational beaches on Lake Michigan and Lake Erie during the summer of 2003. Measurements by this method were compared with counts of Enterococcus colony-forming units (CFU) determined by Method 1600 membrane filter (MF) analysis using mEI agar. The QPCR method had an estimated 95% confidence, minimum detection limit of 27 Enterococcus cells per sample in analyses of undiluted DNA extracts and quantitative analyses of multiple lake water samples, spiked with known numbers of these organisms, gave geometric mean results that were highly consistent with the spike levels. At both beaches, the geometric means of ambient Enterococcus concentrations in water samples, determined from multiple collection points during each sampling visit, showed approximately lognormal distributions over the study period using both QPCR and MF analyses. These geometric means ranged from 10 to 8548 cells by QPCR analysis and 1-2499 CFU by MF culture analysis in Lake Michigan (N=56) and from 8 to 8695 cells by QPCR and 3-1941 CFU by MF culture in Lake Erie (N=47). Regression analysis of these results showed a significant positive correlation between the two methods with an overall correlation coefficient (r) of 0.68.
Haugland
Richard A
RA
US Environmental Protection Agency, National Exposure Research Laboratory, 26 W. Martin Luther King Dr., Cincinnati, OH 45268-1314, USA. haugland.rich@epa.gov
Siefring
Shawn C
SC
Wymer
Larry J
LJ
Brenner
Kristen P
KP
Dufour
Alfred P
AP
D000362:Agar / Q000737:chemistry; D003470:Culture Media; D004247:DNA / Q000737:chemistry; D016983:Enterococcus / Q000302:isolation & purification*; D005374:Filtration; D016133:Polymerase Chain Reaction / Q000379:methods; D012107:Research Design; D012623:Seawater / Q000382:microbiology*; D013997:Time Factors; D014871:Water Microbiology
D003470:Culture Media; D000362:Agar; D004247:DNA