Membrane filter procedure for enumerating the component genera of the coliform group in seawater

A facile, quantitative, membrane filter procedure (mC) for defining the distribution of coliform populations in seawater according to the component genera was developed. The procedure, which utilizes a series of in situ substrate tests to obviate the picking of colonies for identification, also provides an estimate of the total coliform density. When pure cultures of Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae were suspended in seawater and held at 4 C for 24 h, between 56 and 100% of the cells which grew on nutrient agar spread plates at 35 C could be recovered by the mC procedure. Confirmation as coliforms of typical colonies from natural samples was about 95%. Assay variability was found to be insignificant. The recovery of coliforms from marine waters by the mC procedure was comparable to those obtainable by current methods. Klebsiella was differentiated by the urease reaction and E. coli by its ability to form indole. The confirmation frequencies for colonies designated as Klebsiella and E. coli by the in situ tests approached 95% for the former and 98% for the latter.