Simple enumeration of Escherichia coli concentrations in river water samples by measuring β-d-glucuronidase activities in a microplate reader

Abstract

Monitoring of Escherichia coli concentrations in river water (RW) is essential to identify fecal pollution of the river. The objective of this study was to assess the suitability of a novel, simple and high throughput method developed in our laboratory to enumerate E. coli concentrations in RW samples. The method is based on the use of the synthetic substrate specific for the β-d-glucuronidase (GUS) produced by E. coli. GUS activities and E. coli concentrations were monitored at eight selected sites in rivers running through Sapporo, Japan. Because the fluorescence intensities of the synthetic substrate in the RW samples increased linearly over a 4-h incubation period, we could estimate the GUS activities of the RW samples. The GUS activities were highly correlated with E. coli concentrations at >100 most probable numbers 100 mL−1 with a correlation coefficient of 0.87. The GUS activities of the RW samples collected from all sampling sites fitted well to a single correlation equation, which indicates that it was applicable to the estimation of E. coli concentrations regardless of the sampling sites. This method is simple, rapid, reliable, inexpensive, and high throughput, and is therefore useful for monitoring E. coli in RW. HIGHLIGHTS A novel method was applied for enumeration of Escherichia coli in urban rivers.; Response time was <3 h and detection limit was 430 most probable numbers 100 mL−1.; Specific enzyme activities of E. coli were similar in wastewater and river waters.; Correlation between enzyme activity and E. coli concentration was site independent.; E. coli concentrations were monitored at eight different sites over 9 months.;

Keywords

• enzyme
• fecal pollution
• fluctuation
• fluorescence
• sapporo
• wastewater