Abstract

O. A. Ogunseitan

Perennial wildfires in Africa and other continents contribute an estimated 8 x 105 kg of mercury to the global atmosphere with a residence time of approximately one year. This phenomenon changes the flux of biologically available mercury in natural microbial communities where enzymatic actions, including mercuric reductase and organomercurial lyase activities, underpin the biogeochemical cycling of mercury with repercussions for human exposure to toxic forms of the element. To elucidate the impact of episodic mercury bioavailability on the response of microbial communities, the expression of microbial proteins and nucleic acids in environmental strains of Pseudomonas species were evaluated under various concentrations of mercury ranging from 0 to 500 µM. Routine cultivation of Pseudomonas aeruginosa PU21 containing the 142.5 kb plasmid Rip64 in medium containing 100 µg of Hg++/ml (500 µM) exhibited a prolonged lag phase survived by hyper-resistant cells able to grow in medium containing 200 µg of Hg++/ml. Nucleic acid analyses showed a distinct mutation in the merA gene encoding for mercuric reductase activity in cells able to grow at elevated mercury concentrations. A similar mutation was detected in the merR locus which serves as the regulator of the mer operon. Mutations were not detected in merC which encodes for a hydrophobic membrane-associated protein implicated in active mercury transport. Protein profiles of cells grown with elevated mercury concentrations were associated with a stable increase in the production of specific polypeptides. In addition, the survival and genetic response of naturally-occurring mercury resistant bacteria inoculated into contaminated environmental samples were monitored in microcosm experiments over a 30 day period. The results suggest that sudden exposure to high concentrations of mercury either decimates the bacterial population or selects for hyper-resistant strains with high level of constitutive expression of active proteins, including mercuric reductase. Methyl mercury was observed to cause a higher level of induction for mercuric reductase than the specific substrate, inorganic mercury. The selection of hyper-resistant strains is potentially useful for biotechnological strategies to control the bioavailability of mercury, and thereby potentially reducing the re-uptake of mercury into vegetation in regions frequently subjected to wildfires.