Heterotrophic plate count vs. in situ bacterial 16S rRNA gene amplicon profiles from drinking water reveal completely different communities with distinct spatial and temporal allocations in a distribution net

Abstract

Heterotrophic plate count using ISO 6222 agar (HPC) vs. in situ bacterial (DF) community structure from corresponding samples of a drinking water distribution system were investigated by 16S rRNA gene-based polymerase chain reaction denaturing gradient gel electrophoresis (PCR DGGE) profiling. The investigation regime covered 10 different sampling locations and 2 points in time (t1, t2). In order to ensure accurate and reproducible 16S rRNA gene profile analysis, rigorous methodical evaluation and standardisation procedures were undertaken (DGGE optimisation, replication of PCR, multiplelane standardisation, representative sampling volume determination, application of multiple similarity coefficients). The reproducibility level of the profile analysis was determined to be ≥ 90% similarity. Two completely different communities were revealed from HPC vs. DF as indicated by DGGE analysis and sequencing.

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