Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1993 Nov;59(11):3534–3544. doi: 10.1128/aem.59.11.3534-3544.1993

New medium for the simultaneous detection of total coliforms and Escherichia coli in water.

K P Brenner 1, C C Rankin 1, Y R Roybal 1, G N Stelma Jr 1, P V Scarpino 1, A P Dufour 1
PMCID: PMC182495  PMID: 8285660

Abstract

A new membrane filter agar medium (MI agar) containing a chromogen, indoxyl-beta-D-glucuronide, and a fluorogen, 4-methylumbelliferyl-beta-D-galactopyranoside, was developed to simultaneously detect and enumerate Escherichia coli and total coliforms (TC) in water samples on the basis of their enzyme activities. TC produced beta-galactosidase, which cleaved 4-methylumbelliferyl-beta-D-galactopyranoside to form 4-methylumbelliferone, a compound that fluoresced under longwave UV light (366 nm), while E. coli produced beta-glucuronidase, which cleaved indoxyl-beta-D-glucuronide to form a blue color. The new medium TC and E. coli recoveries were compared with those of mEndo agar and two E. coli media, mTEC agar and nutrient agar supplemented with 4-methylumbelliferyl-beta-D-glucuronide, using natural water samples and spiked drinking water samples. On average, the new medium recovered 1.8 times as many TC as mEndo agar, with greatly reduced background counts (< or = 7%). These differences were statistically significant (significance level, 0.05). Although the overall analysis revealed no statistically significant difference between the E. coli recoveries on MI agar and mTEC agar, the new medium recovered more E. coli in 16 of 23 samples (69.6%). Both MI agar and mTEC agar recovered significantly more E. coli than nutrient agar supplemented with 4-methylumbelliferyl-beta-D-glucuronide. Specificities for E. coli, TC, and noncoliforms on MI agar were 95.7% (66 of 69 samples), 93.1% (161 of 173 samples), and 93.8% (61 of 65 samples), respectively. The E. coli false-positive and false-negative rates were both 4.3%. This selective and specific medium, which employs familiar membrane filter technology [corrected] to analyze several types of water samples, is less expensive than the liquid chromogen and fluorogen media and may be useful for compliance monitoring of drinking water.

Full text

PDF
3534

Selected References

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

  1. Berg J. D., Fiksdal L. Rapid detection of total and fecal coliforms in water by enzymatic hydrolysis of 4-methylumbelliferone-beta-D-galactoside. Appl Environ Microbiol. 1988 Aug;54(8):2118–2122. doi: 10.1128/aem.54.8.2118-2122.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang G. W., Brill J., Lum R. Proportion of beta-D-glucuronidase-negative Escherichia coli in human fecal samples. Appl Environ Microbiol. 1989 Feb;55(2):335–339. doi: 10.1128/aem.55.2.335-339.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clark D. L., Milner B. B., Stewart M. H., Wolfe R. L., Olson B. H. Comparative study of commercial 4-methylumbelliferyl-beta-D-glucuronide preparations with the Standard Methods membrane filtration fecal coliform test for the detection of Escherichia coli in water samples. Appl Environ Microbiol. 1991 May;57(5):1528–1534. doi: 10.1128/aem.57.5.1528-1534.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Covert T. C., Shadix L. C., Rice E. W., Haines J. R., Freyberg R. W. Evaluation of the Autoanalysis Colilert test for detection and enumeration of total coliforms. Appl Environ Microbiol. 1989 Oct;55(10):2443–2447. doi: 10.1128/aem.55.10.2443-2447.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dalbow D. G., Bremer H. Metabolic regulation of beta-galactosidase synthesis in Escherichia coli. A test for constitutive ribosome synthesis. Biochem J. 1975 Jul;150(1):1–8. doi: 10.1042/bj1500001b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Delisle G. J., Ley A. Rapid detection of Escherichia coli in urine samples by a new chromogenic beta-glucuronidase assay. J Clin Microbiol. 1989 Apr;27(4):778–779. doi: 10.1128/jcm.27.4.778-779.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dufour A. P., Strickland E. R., Cabelli V. J. Membrane filter method for enumerating Escherichia coli. Appl Environ Microbiol. 1981 May;41(5):1152–1158. doi: 10.1128/aem.41.5.1152-1158.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edberg S. C., Allen M. J., Smith D. B. 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. Appl Environ Microbiol. 1988 Jun;54(6):1595–1601. doi: 10.1128/aem.54.6.1595-1601.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Entis P. Hydrophobic grid membrane filter/MUG method for total coliform and Escherichia coli enumeration in foods: collaborative study. J Assoc Off Anal Chem. 1989 Nov-Dec;72(6):936–950. [PubMed] [Google Scholar]
  10. Feng P. C., Hartman P. A. Fluorogenic assays for immediate confirmation of Escherichia coli. Appl Environ Microbiol. 1982 Jun;43(6):1320–1329. doi: 10.1128/aem.43.6.1320-1329.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Freier T. A., Hartman P. A. Improved membrane filtration media for enumeration of total coliforms and Escherichia coli from sewage and surface waters. Appl Environ Microbiol. 1987 Jun;53(6):1246–1250. doi: 10.1128/aem.53.6.1246-1250.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iritani B., Inzana T. J. Evaluation of a rapid tube assay for presumptive identification of Escherichia coli from veterinary specimens. J Clin Microbiol. 1988 Mar;26(3):564–566. doi: 10.1128/jcm.26.3.564-566.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kilian M., Bülow P. Rapid identification of Enterobacteriaceae. II. Use of a beta-glucuronidase detecting agar medium (PGUA agar) for the identification of E. coli in primary cultures of urine samples. Acta Pathol Microbiol Scand B. 1979 Oct;87(5):271–276. [PubMed] [Google Scholar]
  14. Ley A. N., Bowers R. J., Wolfe S. Indoxyl-beta-D-glucuronide, a novel chromogenic reagent for the specific detection and enumeration of Escherichia coli in environmental samples. Can J Microbiol. 1988 May;34(5):690–693. doi: 10.1139/m88-115. [DOI] [PubMed] [Google Scholar]
  15. Manafi M., Kneifel W., Bascomb S. Fluorogenic and chromogenic substrates used in bacterial diagnostics. Microbiol Rev. 1991 Sep;55(3):335–348. doi: 10.1128/mr.55.3.335-348.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Manafi M., Kneifel W. Ein kombiniertes Chromogen-Fluorogen-Medium zum simultanen Nachweis der Coliformengruppe und von E. coli in Wasser. Zentralbl Hyg Umweltmed. 1989 Dec;189(3):225–234. [PubMed] [Google Scholar]
  17. Mates A., Shaffer M. Membrane filtration differentiation of E. coli from coliforms in the examination of water. J Appl Bacteriol. 1989 Sep;67(3):343–346. doi: 10.1111/j.1365-2672.1989.tb02503.x. [DOI] [PubMed] [Google Scholar]
  18. Petzel J. P., Hartman P. A. Monensin-based medium for determination of total gram-negative bacteria and Escherichia coli. Appl Environ Microbiol. 1985 Apr;49(4):925–933. doi: 10.1128/aem.49.4.925-933.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Thaller M. C., Berlutti F., Dainelli B., Pezzi R. New plate medium for screening and presumptive identification of gram-negative urinary tract pathogens. J Clin Microbiol. 1988 Apr;26(4):791–793. doi: 10.1128/jcm.26.4.791-793.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thompson J. S., Hodge D. S., Borczyk A. A. Rapid biochemical test to identify verocytotoxin-positive strains of Escherichia coli serotype O157. J Clin Microbiol. 1990 Oct;28(10):2165–2168. doi: 10.1128/jcm.28.10.2165-2168.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tuveson R. W., Larson R. A., Kagan J. Role of cloned carotenoid genes expressed in Escherichia coli in protecting against inactivation by near-UV light and specific phototoxic molecules. J Bacteriol. 1988 Oct;170(10):4675–4680. doi: 10.1128/jb.170.10.4675-4680.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Watkins W. D., Rippey S. R., Clavet C. R., Kelley-Reitz D. J., Burkhardt W., 3rd Novel compound for identifying Escherichia coli. Appl Environ Microbiol. 1988 Jul;54(7):1874–1875. doi: 10.1128/aem.54.7.1874-1875.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES