[en] The U.S. Department of Energy is engaged in a suitability study for a potential geological repository at Yucca Mountain, Nevada, for the containment and storage of commercially generated spent fuel and defense high-level nuclear waste. There is growing recognition of the role that biotic factors could play in this repository, either directly through microbially induced corrosion (MIC), or indirectly by altering the chemical environment or contributing to the transport of radionuclides. As a first step toward describing and predicting these processes, a workshop was held on April 10-12, 1995, in Lafayette, California. The immediate aims of the workshop were: (1) To identify microbially related processes relevant to the design of a radioactive waste repository under conditions similar to those at Yucca Mountain. (2) To determine parameters that are critical to the evaluation of a disturbed subterranean environment. (3) To define the most effective means of investigating the factors thus identified

[en] The integrity of candidate waste package materials for the proposed Yucca Mountain (YM) repository may be compromised by the corrosive activities of microorganisms. Bacterial activities will be dependent on the abilities of deleterious bacteria to grow and multiply in the repository environment. Therefore, preliminary to assessing microbial induced corrosion, experiments were undertaken to determine the growth of native YM bacterial communities in modified YM pore water. Specifically, we sought to define nutrients that limit bacterial growth in Yucca Mountain and determine growth rates under aerobic, saturated conditions

[en] A promoterless chloramphenicol acetyltransferase gene (cat) was used to construct recA-cat operon fusions to quantitatively examine the transcriptional regulation of the Pseudomonas aeruginosa recA gene in P. aeruginosa PAO. Wild-type P. aeruginosa containing the recA8-cat fusion was treated with methyl methanesulfonate (MMS) and showed immediate induction of chloramphenicol acetyltransferase (CAT) specific activity, whereas a recA::Tn501 mutant of P. aeruginosa containing recA8-cat showed no induction with MMS. This indicated that a functional copy of recA was required for derepression of recA transcription and that P. aeruginosa recA protein was a positive regulatory factor promoting its own expression. Compared with that in the wild type, the uninduced level of CAT in recA8-cat-containing cells was reduced by approximately one-half in the recA::Tn501 mutant, indicating that recA+-dependent spontaneous induction contributes to the uninduced levels of recA expression in P. aeruginosa. MMS (0.012%) caused recA-directed CAT synthesis to increase almost immediately, with maximum CAT activity, fourfold higher than uninduced levels, attained at 60 min postinduction. The kinetics of recA8-cat fusion activity were shown to be directly related to the MMS doses used. Another fusion called recAa1-cat, where cat was located between the two transcriptional terminators of the P. aeruginosa recA gene, also showed dose-dependent induction by MMS, but the CAT activity from recAa1-cat was only one-half of that obtained with recA8-cat under the same conditions. Treatment of recA+ P. aeruginosa containing recA8-cat with UV irradiation produced an immediate effect on recA8-cat transcription and showed little UV dose dependency at doses of 5 J/m2 or greater

[en] The U.S. Department of Energy has been charged with assessing the suitability of a geologic nuclear waste repository at Yucca Mountain (YM), NV. Microorganisms, both those endogenous to the repository site and those introduced as a result of construction and operational activities, may contribute to the corrosion of metal nuclear waste packaging and thereby decrease their useful lifetime as barrier materials. Evaluation of potential Microbiological Influenced Corrosion (MIC) on candidate waste package materials was undertaken reactor systems incorporating the primary elements of the repository: YM rock (either non-sterile or presterilized), material coupons, and a continual feed of simulated YM groundwater. Periodically, both aqueous reactor efflux and material coupons were analyzed for chemical and surfacial characterization. Alloy 22 coupons exposed for a year at room temperature in reactors containing non-sterile YM rock demonstrated accretion of chromium oxide and silaceous scales, with what appear to be underlying areas of corrosion

[en] We present time-of-flight measurements of biological material ejected from bacterial spores following laser irradiation. Ion impacts are registered on a microchannel plate detector and on a Superconducting Tunnel Junction (STJ) detector. We compare mass spectra obtained with the two detectors. The STJ has better sensitivity to massive ions and also measures the energy of each ion. We show evidence that spores of different bacillus species produce distinctive mass spectra and associate the observed mass peaks with coat proteins

[en] The impacts of the native and introduced bacteria on the performance of geologic nuclear waste disposal facilities should be evaluated because these bacteria could promote corrosion of repository components and alteration of chemical and hydrological properties of the surrounding engineered and rock barriers. As a first step towards investigating these potentialities, native and introduced bacteria obtained from post-construction Yucca Mountain (YM) rock were isolated under varying conditions, including elevated temperature, low nutrient availability, and the absence of available oxygen. Individual isolates are being screened for activities associated with microbially induced corrosion of metals (MIC). Preliminary determination of growth rates of whole YM microbial communities under varying conditions was also undertaken