Molecular characterization of dechlorination potential in kraft pulp mill effluent treatment systems

Many low molecular weight chlorinated organic compounds produced by the pulp and paper industry during kraft bleaching of the wood pulp are toxic. Mass balance studies suggest that mineralization of chlorinated organics is occurring in pulp and paper mill wastewater treatment systems. To understand the nature of dechlorination activity, molecular tools such as oligonucleotide primers and corresponding DNA probes were developed to monitor the presence of microorganisms possessing key genes (dehalogenases) responsible for the degradation of chloroaliphatic organics in kraft pulp mill effluent treatment systems. Oligonucleotide primers designed from the coding sequence of known dehalogenases and a methane monooxygenase gene, which is known to catalyze dehalogenation reactions, were used for polymerase chain reaction (PCR) analysis, using genomic DNA extracted from dehalogenating bacterial isolates and total community DNA extracted from water and sediments of lagoon treatment systems. PCR amplification with dhlB oligonucleotide primers, designed from the haloacid dehalogenase of Xanthobacter autotrophicus, revealed the presence of dehalogenase genes in both aerated lagoons and stabilization basins. Similar results were obtained with mmoX primers designed from the methane monooxygenase gene of Methylococcus capsulatus. DNA sequence analysis of several PCR fragments showed significant similarity to known dehalogenase genes. The molecular tools developed in this study revealed the presence of different types of microorganisms with dechlorination potential in the microbial community of pulp mill effluents