Current Microbiology

, Volume 75, Issue 7, pp 827–834 | Cite as

Rapid Detection of Escherichia coli in Water Using Sample Concentration and Optimized Enzymatic Hydrolysis of Chromogenic Substrates

  • Jianyong WuEmail author
  • Jill R. Stewart
  • Mark D. Sobsey
  • Chris Cormency
  • Michael B. Fisher
  • Jamie K. Bartram


Methods for rapid detection of fecal indicator bacteria in water are important to ensure that water is safe for drinking, bathing, recreation, fishing and shellfish harvesting. In this study, we tested experimental conditions for bacterial hydrolysis of two promising enzymatic substrates, 5-Bromo-4-chloro-3-indolyl β-d-glucuronide (X-Gluc) and Resorufin β-d-glucuronide (REG), and optimized parameters such as temperature and pH to determine conditions for rapid reactions. We then innovated a membrane filter-based approach to facilitate more rapid enzyme-based detection of Escherichia coli in water based on the combination of an initial concentration step and optimized test conditions. For this approach, a water sample (10‒100 mL) is filtered through a 0.45-µm pore size filter with a diameter of 4 or 13 mm. After filtration, a newly designed rapid detection broth is added containing the enzymatic inducer Methyl-beta-d-Glucuronide sodium (MetGlu) and the substrate REG or X-Gluc. After a few (1‒7) hours of incubation at 35 °C, the filter shows pink color (for REG-containing broth) or green color (for X-Gluc containing broth) if E. coli is present. The study provides insights and approaches towards developing a simple, fast, and low-cost method to detect fecal indicator bacteria in water.



We would like to thank UNICEF (No. 43171142) and the North Carolina Sea Grant (No. 2014-1751-02) for financial support.

Compliance with Ethical Standards

Conflict of interest

The authors have no any conflict of interest with these awards. Jamie Bartram has served on UNICEF advisory committees unconnected to the subject of the award.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Jianyong Wu
    • 1
    Email author
  • Jill R. Stewart
    • 1
  • Mark D. Sobsey
    • 1
  • Chris Cormency
    • 2
  • Michael B. Fisher
    • 1
  • Jamie K. Bartram
    • 1
  1. 1.Department of Environmental Sciences and Engineering, Gillings School of Global Public Health, The Water InstituteUniversity of North Carolina at Chapel HillChapel HillUSA
  2. 2.Water, Sanitation and Education CenterUNICEF Supply DivisionCopenhagenDenmark

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