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Biomedical subjects

J B Herrick

Publications and source records attributed to J B Herrick.

At least 19 recordsLinked to original sources

Natural horizontal transfer of a naphthalene dioxygenase gene between bacteria native to a coal tar-contaminated field site.

Horizontal transfer of genes responsible for pollutant biodegradation may play a key role in the evolution of bacterial populations and the adaptation of microbial communities to environmental contaminants. However, field evidence for horizontal gene transfer between microorganisms has traditionally been very difficult to obtain. In this study, the sequences of the 16S rRNA and naphthalene dioxygenase iron-sulfur protein (nahAc) genes of nine naphthalene-degrading bacteria isolated from a coal tar waste-contaminated site, as well as a naphthalene-degrading bacterium from a contaminated site in Washington state and two archetypal naphthalene-degrading strains, were compared. Seven strains from the study site had a single nahAc allele, whereas the 16S rRNA gene sequences of the strains differed by as much as 7.9%. No nahAc alleles from the site were identical to those of the archetypal strains, although the predominant allele was closely related to that of Pseudomonas putida NCIB 9816-4, isolated in the British Isles. However, one site-derived nahAc allele was identical to that of the Washington state strain. Lack of phylogenetic congruence of the nahAc and 16S rRNA genes indicates that relatively recent in situ horizontal transfer of the nahAc gene has occurred, possibly as a direct or indirect consequence of pollutant contamination. Alkaline lysis plasmid preparations and pulsed-field gel electrophoresis have revealed the presence of plasmids ranging in size from 70 to 88 kb in all site isolates. Southern hybridizations with a 407-bp nahAc probe have suggested that the nahAc gene is plasmid borne in all the site isolates but one, a strain isolated from subsurface sediment 400 m upstream from the source of the other site isolates. In this strain and in the naphthalene-degrading strain from Washington state, nahAc appears to be chromosomally located. In addition, one site isolate may carry nahAc on both chromosome and plasmid. Within the group of bacteria with identical nahAc sequences the Southern hybridizations showed that the gene was distributed between plasmids of different sizes and a chromosome. This suggests that plasmid modification after transfer may have been effected by transposons. Horizontal transfer of catabolic genes may play a significant role in the acclimation of microbial communities to pollutants.

Alleles

Natural selection of PAH-degrading bacterial guilds at coal-tar disposal sites.

Microbial activity patterns at buried coal-tar disposal sites have been under investigation for several years to determine the response of naturally occurring microflora to polycyclic aromatic hydrocarbons (PAHs) at the sites. At one site in upstate New York, data have shown enrichment of PAH-degrading bacteria in subsurface contaminated zones but not in uncontaminated zones. Similar work at a midwestern site showed that the same trends existed in a heterogeneous disposal site except that a borehole outside the plume showed some PAH-mineralization activity. Polymerase chain reaction amplification of DNA extracted from sediment samples from the New York site indicated the presence of naphthalene metabolism genes nahAc and nahR, similar to those found on the NAH7 plasmid of Pseudomonas putida G7. Significant sequence polymorphism was observed in amplified nahAc products, indicating that divergent homologs of nahAc were present in the native community. Protozoan numbers were elevated in sediment samples displaying relatively high PAH-degrading activity, suggesting that a food chain was established based on PAH-degrading bacteria. Removal of the coal-tar source at the site occurred in 1991. In 1992, sampling of three key borehole stations revealed that mixing and backfilling operations had introduced soil microorganisms into the source area and introduced 14C-PAH-mineralization activity into the previously inactive pristine area. Thus removal of the source of the contaminants and restoration at the site have altered the microbial activity patterns outside the contaminant plume as well as in the source area.

Bacteria