A defined substrate technology for the enumeration of microbial indicators of environmental pollution.

The examination of water and other environmental sources for microbial pollution is a major public health undertaking. Currently, there are two accepted methods in use: the multiple-tube fermentation (MTF) and the membrane filtration (MF) tests. Both methods are designed to enumerate the secondary indicator group, total coliforms. Both tests suffer several inherent limitations, including a time delay of three to seven days to obtain a definitive result, the subjective nature of the test interpretation, and the inability to provide directly useful public health information. A defined substrate technology, originally used to enumerate specific bacterial species from mixtures in clinical urine specimens, was applied to water testing; the technology was constituted to enumerate simultaneously both total coliforms and the primary indicator bacterium E. coli. Examination of environmental isolates of these two classes of target microbes showed sensitivity equal to available methods, with potentially greater specificity. It was not subject to inhibition by bacteria other than the targets, grew injured coliforms, did not require confirmatory tests, and the maximum time to a positive was 24 hours. The defined substrate technology provides both regulatory and directly useful public health information.