Effects of diesel exhaust on the microbiota within a tuffaceous tunnel system

The abundance and distribution of microbiota that may be impacted by diesel and diesel exhaust were investigated from three depths into the walls and invert (floor) of U12n tunnel at Rainier Mesa, Nevada Test Site, a potential geological analog of Yucca Mountain. Enumerations included total cell counts, and numbers of aerobic heterotrophic, sulfate-reducing, nitrate-reducing, and diesel-degrading bacteria. Additionally, the disappearance of total petroleum hydrocarbons was determined in microcosms containing subsurface materials that were amended with diesel fuel. Results revealed that microbes capable of utilizing diesel and diesel combustion products were present in the subsurface in both the walls and the invert of the tunnel. The abundance of specific bacterial types in the tunnel invert, a perturbed environment, was greater than that observed in the tunnel wall. Few trends of microbial distribution either into the tunnel wall or the invert were noted with the exception of aerobic heterotrophic abundance which increased with depth into the wall and decreased with depth into the invert. No correlation between microbiota and a specific introduced chemical species have yet been determined. The potential for microbial contamination of the tunnel wall during sampling was determined to be negligible by the use of fluorescently labeled latex spheres (1{mu}m in dia.) as tracers. Results indicate that additional investigations might be needed to examine the microbiota and their possible impacts on the geology and geochemistry of the subsurface, both indigenous microbiota and those microorganisms that will likely be introduced by anthropogenic activity associated with the construction of a high-level waste repository