An Assessment of Variability of Pulp Mill Wastewater Treatment System Bacterial Communities using Molecular Methods

Abstract

Abstract The DNA fingerprinting techniques, 16S-restriction fragment length polymorphism (16S-RFLP), ribosomal intergenic spacer analysis (RISA) and repetitive extragenic palindrome PCR (Rep-PCR), were used for analyzing the bacterial communities of seven pulp and paper wastewater treatment systems. All three methods generate DNA fingerprints that can be compared using the computer-assisted program, Gelcompar©. Community similarity coefficients were based on quantitative determinations of both the positions of the DNA bands and the band intensities in order to compare the relative differences in the populations. Unique 16S-RFLP DNA fingerprints were observed for each mill suggesting that individual mills contained phylogenetically different communities. However this method was not sensitive enough to detect differences within a mill treatment system from different locations or from different sampling times. The RISA method, which generated more complex fingerprints than 16S-RFLP, could, for some mills, discern differences between samples. The Rep-PCR technique, however, showed the highest degree of resolution and produced not only distinct patterns for each mill but also distinct fingerprints for the temporal and spatial samples from some of the treatment systems. The sensitivity of this method might potentially be used to monitor the stability of the bacterial community within a secondary treatment system.