Comparison of the direct enzyme assay method with the membrane filtration technique in the quantification and monitoring of microbial indicator organisms – seasonal variations in the activities of coliforms and E. coli, temperature and pH

  • VC Wutor
  • CA Togo
  • BI Pletschke
Keywords: chlorophenol red â-D-galactopyranoside (CPRG), chlorophenol red (CPR), coliforms, E. coli, p-nitrophenyl-â-D-glucuronide (PNPG), p-nitrophenyl (PNP)


The aim of this project was to monitor variations and relationships between coliform and E. coli counts, the activities of their marker enzymes GAL and GUD, and temperature and pH over a period of 12 months in river samples obtained from the Eastern Cape, South Africa. Several polluted water samples were collected for direct coliform ƒÀ-D-galactosidase (B-GAL) and Escherichia coli ƒÀ-D-glucuronidase (B-GUD) assays and the membrane filtration technique. While all the samples showed enzyme activities, not all exhibited growth on CM1046 media. Variation in B-GAL activity (40%) was observed between November (highest activity month) and May (lowest activity month). The highest and lowest B-GUD activities were observed in the months of September and May/June, respectively. The sensitivity of the spectrophotometric assay method was indicated by a limit of detection (LOD) of 1 coliform forming unit (CFU)/100 m. and 2 CFU/100 m. for coliforms and E. coli, respectively. There was a significant (P < 0.05) positive correlation between E. coli counts and GUD activity (R2 = 0.8909). A correlation of R2 = 0.9151 was also observed between total coliforms and B-GAL activity, even though the CFUs were not evenly distributed. Direct enzyme assays were also shown to be more sensitive than the membrane filtration (MF) technique.

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eISSN: 0378-4738