Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1988 Jun;54(6):1595–1601. doi: 10.1128/aem.54.6.1595-1601.1988

National field evaluation of a defined substrate method for the simultaneous enumeration of total coliforms and Escherichia coli from drinking water: comparison with the standard multiple tube fermentation method.

S C Edberg 1, M J Allen 1, D B Smith 1
PMCID: PMC202702  PMID: 3046490

Abstract

A defined substrate method was developed to simultaneously enumerate total coliforms and Escherichia coli from drinking waters without the need for confirmatory or completed tests. It is a new method based on technology that uses a hydrolyzable substrate as a specific indicator-nutrient for the target microbes. No equipment other than a 35 degrees C incubator and long-wavelength (366-nm) light is necessary. To perform the test, one only has to add water to the powdered ingredients in a tube or flask. If total coliforms are present in the water sample, the solution will change from its normal colorless state (no target microbes present) to yellow. The specific presence of E. coli will cause the same tube to fluoresce under a longwave (366-nm) UV lamp. The test, called Autoanalysis Colilert (AC), was compared with Standard Methods for the Examination of Water and Wastewater 10-tube multiple tube fermentation (MTF) in a national evaluation. Five utilities, representing six U.S. Environmental Protection Agency regions, participated. All water samples came from distribution systems. Split samples from a wide variety of water sources were analyzed for the MPN-versus-MPN comparison. A total of 1,086 tubes were positive by MTF, and 1,279 were positive by AC. There was no statistical difference between MTF and AC. Species identifications from positive tubes confirmed the sensitivity of the AC. A national evaluation of the AC test showed that it: (i) was as sensitive as Standard Methods MTF, (ii) specifically enumerated 1 total coliform per 100 ml, in a maximum of 24 h, (iii) simultaneously enumerated 1 E. coli per 100 ml in the same analysis, (iv) was not subject to false-positive or false-negative results by heterotrophic bacteria, (v) did not require confirmatory tests, (vi) grew injured coliforms, (vii) was easy to inoculate, and (viii) was very easy to interpret.

Full text

PDF
1595

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bissonnette G. K., Jezeski J. J., McFeters G. A., Stuart D. G. Evaluation of recovery methods to detect coliforms in water. Appl Environ Microbiol. 1977 Mar;33(3):590–595. doi: 10.1128/aem.33.3.590-595.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Edberg S. C., Atkinson B., Chambers C., Moore M. H., Palumbo L., Zorzon C. F., Singer J. M. Clinical evaluation of the MICRO-ID, API 20E, and conventional media systems for identification of Enterobacteriacea. J Clin Microbiol. 1979 Aug;10(2):161–167. doi: 10.1128/jcm.10.2.161-167.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Edberg S. C., Piscitelli V., Cartter M. Phenotypic characteristics of coliform and noncoliform bacteria from a public water supply compared with regional and national clinical species. Appl Environ Microbiol. 1986 Sep;52(3):474–478. doi: 10.1128/aem.52.3.474-478.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Evans T. M., LeChevallier M. W., Waarvick C. E., Seidler R. J. Coliform species recovered from untreated surface water and drinking water by the membrane filter, standard, and modified most-probable-number techniques. Appl Environ Microbiol. 1981 Mar;41(3):657–663. doi: 10.1128/aem.41.3.657-663.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Evans T. M., Waarvick C. E., Seidler R. J., LeChevallier M. W. Failure of the most-probable-number technique to detect coliforms in drinking water and raw water supplies. Appl Environ Microbiol. 1981 Jan;41(1):130–138. doi: 10.1128/aem.41.1.130-138.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jacobs N. J., Zeigler W. L., Reed F. C., Stukel T. A., Rice E. W. Comparison of membrane filter, multiple-fermentation-tube, and presence-absence techniques for detecting total coliforms in small community water systems. Appl Environ Microbiol. 1986 May;51(5):1007–1012. doi: 10.1128/aem.51.5.1007-1012.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. LeChevallier M. W., Cameron S. C., McFeters G. A. New medium for improved recovery of coliform bacteria from drinking water. Appl Environ Microbiol. 1983 Feb;45(2):484–492. doi: 10.1128/aem.45.2.484-492.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. McFeters G. A., Cameron S. C., LeChevallier M. W. Influence of diluents, media, and membrane filters on detection fo injured waterborne coliform bacteria. Appl Environ Microbiol. 1982 Jan;43(1):97–103. doi: 10.1128/aem.43.1.97-103.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. McFeters G. A., Stuart D. G. Survival of coliform bacteria in natural waters: field and laboratory studies with membrane-filter chambers. Appl Microbiol. 1972 Nov;24(5):805–811. doi: 10.1128/am.24.5.805-811.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Morgan G. B., Gubbins P., Morgan V. A critical appraisal of the membrane filter technic. Health Lab Sci. 1965 Oct;2(4):227–237. [PubMed] [Google Scholar]
  11. Petzel J. P., Hartman P. A. A note on starch hydrolysis and beta-glucuronidase activity among flavobacteria. J Appl Bacteriol. 1986 Nov;61(5):421–426. doi: 10.1111/j.1365-2672.1986.tb04306.x. [DOI] [PubMed] [Google Scholar]
  12. SHIPE E. L., Jr, CAMERON G. M. A comparison of the membrane filter with the most probable number method for coliform determinations from several waters. Appl Microbiol. 1954 Mar;2(2):85–88. doi: 10.1128/am.2.2.85-88.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES