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PubMed1996

Intersubjectivity, social microcosm, and the here-and-now in a support group for nurses.

Intersubjectivity, a concept from Watson's nursing model is interwoven with the group dynamics of social microcosm and the here-and-now. Examples from a graduate nursing student's clinical experience illustrate the application of a transpersonal dynamic, formerly associated with dyads, to interactions between a nurse and a support group. A psychiatric consultation liaison nurse specialist student facilitates a support group for registered nurses in a neurological intensive care unit, in response to a group-perceived need to improve communications between nurses, nurses and patients, and nurses and physicians. Discovery and processing of hidden anger and pain activates healing for these nurses.

Anger↗
PubMed2006

Effect of simulated rhizodeposition on the relative abundance of polynuclear aromatic hydrocarbon catabolic genes in a contaminated soil.

Microcosms were used to investigate whether soil exposure to mulberry root extracts (rich in phenolic compounds) select for bacteria that degrade polynuclear aromatic hydrocarbons (PAHs). Unlike previous studies with freshly spiked soil, the present experiments were conducted with soils aged for 518 d with [14C]phenanthrene to decrease bioavailability and avoid exaggerating the selective pressure exerted by PAHs relative to the rhizosphere effect. Microcosms simulating contaminated planted soil were exposed to carbon at 20 mg/L/week of mulberry root extract for 211 d to simulate rhizodeposition. Contaminated bulk soils microcosms were amended with a C-free mineral medium to discern the effect of rhizodeposition. Uncontaminated soil controls also were exposed to similar dose regimes. Real-time quantitative polymerase chain reaction was used to enumerate total bacteria and PAH degraders harboring the genes nahAc (coding for naphthalene dioxygenase), todC1 (coding for toluene/benzene/chlorobenzene dioxygenase), bmoA (coding for hydroxylating monooxygenases), and dmpN (coding for phenol hydroxylase). Exposure to root extracts enhanced the growth of total bacteria and PAH degraders in both contaminated and uncontaminated rhizosphere microcosms. The relative abundance of PAH-degrader gene copies (as a fraction of the total bacteria) was similar for different treatments, suggesting that the root extracts did not select for PAH degraders. Overall, these results suggest that rhizodeposition from phenolic releasers contributes to the fortuitous (but not selective) proliferation of PAH degraders, which may enhance phytoremediation.

Bacteria↗
PubMed1992

Microbial degradation of toluene under sulfate-reducing conditions and the influence of iron on the process.

Toluene degradation occurred concomitantly with sulfate reduction in anaerobic microcosms inoculated with contaminated subsurface soil from an aviation fuel storage facility near the Patuxent River (Md.). Similar results were obtained for enrichment cultures in which toluene was the sole carbon source. Several lines of evidence suggest that toluene degradation was directly coupled to sulfate reduction in Patuxent River microcosms and enrichment cultures: (i) the two processes were synchronous and highly correlated, (ii) the observed stoichiometric ratios of moles of sulfate consumed per mole of toluene consumed were consistent with the theoretical ratio for the oxidation of toluene to CO2 coupled with the reduction of sulfate to hydrogen sulfide, and (iii) toluene degradation ceased when sulfate was depleted, and conversely, sulfate reduction ceased when toluene was depleted. Mineralization of toluene was confirmed in experiments with [ring-U-14C]toluene. The addition of millimolar concentrations of amorphous Fe(OH)3 to Patuxent River microcosms and enrichment cultures either greatly facilitated the onset of toluene degradation or accelerated the rate once degradation had begun. In iron-amended microcosms and enrichment cultures, ferric iron reduction proceeded concurrently with toluene degradation and sulfate reduction. Stoichiometric data and other observations indicate that ferric iron reduction was not directly coupled to toluene oxidation but was a secondary, presumably abiotic, reaction between ferric iron and biogenic hydrogen sulfide.

Biodegradation, Environmental↗
PubMed2005

Removal of microcystins by phototrophic biofilms. A microcosm study.

BACKGROUND, AIMS AND SCOPE: Microcystins (MCs) are a family of natural toxins produced by cyanobacteria (blue-green algae). As a result of eutrophication, massive cyanobacterial blooms occur more frequently and MCs represent important contaminants of freshwater ecosystems. Bacterial biodegradation is considered a main mechanism for MC breakdown in environmental conditions. While existing studies were mostly focused on MC biodegradation by planktonic bacteria, our experiments examined the fate and kinetics of MC degradation in river-originated phototrophic biofilms and investigated factors influencing the rate of MC removal. METHODS: The fate of dissolved MCs was studied in laboratory microcosms with different composition (containing water only, water with phytoplankton and/or phototrophic biofilms). Biofilms originated from river ecosystem were pre-incubated under various conditions (with/without presence of cyanobacterial biomass or model organic substrates: glucose and protein--casein). Changes in MC concentration (0-14 days) in water columns were measured by HPLC-DAD after external additions of purified MCs (160 microg L(-1), MC-LR and MC-YR), and halftimes (t1/2) of MC removal were estimated. RESULTS AND DISCUSSION: The slow degradation of MC was revealed in tap water (t1/2 approximately 14 days) and river water without cyanobacteria (t1/2 approximately 8 days). Enhanced removal occurred in the presence of natural planktonic cyanobacteria (t1/2 approximately 44 h), most probably due to microorganisms associated with the biomass of cyanobacterial bloom. More rapid MC elimination occurred in the variants containing phototrophic biofilms, and was particularly pronounced at those biofilms pre-cultivated in the presence of cyanobacterial blooms (t1/2 approximately 20 h). Much slower removal was observed in the variants simulating possible substrate-dependent induction of microorganism metabolism (biofilms pre-incubated with glucose: t1/2 approximately 35 h, and casein: t1/2 approximately 80 h). After termination of experiments, total amounts of MCs accumulated in the biofilms were below 5% of the initial toxin level revealing significant biodegradation processes. CONCLUSION: The microcosm studies contributed to understanding of the environmental fate of MCs and revealed a rapid biodegradation by phototrophic biofilms. The rate of MC elimination depends on history of biofilm community, previous contact with cyanobacteria seems to be a selective factor improving the biodegradation potential. RECOMMENDATION AND OUTLOOK: Our results experimentally showed a positive role of biofilms in MC elimination during water treatment processes such as bank filtration or slow sand filtration, and could eventually serve for further research of biofilm-based technological applications for MCs removal in small-scale drinking water treatment facilities.

Bacterial Toxins↗
PubMed2006

Effects of temperature and sand on E. coil survival in a northern lake water microcosm.

A concern for public health officials is the presence of Escherichia coli (E. coli), an indicator of fecal contamination, in monitoring recreational waters. While E. coli is unlikely to cause disease in humans, its presence may indicate other more pathogenic microorganisms. Many factors can lead to changes in the survival of E. coli outside of the animal intestine and may affect the probability of colonizing a new host. Survival of bacteria in recreational water has been linked to water temperature, and most recently to the presence of sand on the beach. This project looked at the survival of an environmental E. coli isolate in lake water. Lake water microcosms were placed at 4, 10, 14, or 25 degrees C for up to 36 d and an enzyme-substrate test (Colisure, IDEXX Corp.) was used to determine the most probable number (MPN) of E. coli/100 ml water. E. coli numbers at all temperatures declined over the duration of the experiment. The decline was most pronounced at 14 degrees C and was slowest at 4 degrees C. The presence of sand in the microcosm increased the time that E. coli survived, regardless of temperature. From a beach management standpoint, these findings indicate that E.coli may persist in the environment in cooler water longer than in the warmer water encountered in late summer.

Cell Proliferation↗
PubMed1994

Changes in Ester-Linked Phospholipid Fatty Acid Profiles of Subsurface Bacteria during Starvation and Desiccation in a Porous Medium.

Ester-linked phospholipid fatty acid (PLFA) profiles of a Pseudomonas aureofaciens strain and an Arthrobacter protophormiae strain, each isolated from a subsurface sediment, were quantified in a starvation experiment in a silica sand porous medium under moist and dry conditions. Washed cells were added to sand microcosms and maintained under saturated conditions or subjected to desiccation by slow drying over a period of 16 days to final water potentials of approximately - 7.5 MPa for the P. aureofaciens and - 15 MPa for the A. protophormiae. In a third treatment, cells were added to saturated microcosms along with organic nutrients and maintained under saturated conditions. The numbers of culturable cells of both bacterial strains declined to below detection level within 16 days in both the moist and dried nutrient-deprived conditions, while direct counts and total PLFAs remained relatively constant. Both strains of bacteria maintained culturability in the nutrient-amended microcosms. The dried P. aureofaciens cells showed changes in PLFA profiles that are typically associated with stressed gram-negative cells, i.e., increased ratios of saturated to unsaturated fatty acids, increased ratios of trans- to cis-monoenoic fatty acids, and increased ratios of cyclopropyl fatty acids to their monoenoic precursors. P. aureofaciens starved under moist conditions showed few changes in PLFA profiles during the 16-day incubation, whereas cells incubated in the presence of nutrients showed decreases in the ratios of both saturated fatty acids to unsaturated fatty acids and cyclopropyl fatty acids to their monoenoic precursors. The PLFA profiles of A. protophormiae changed very little in response to either nutrient deprivation or desiccation. Diglyceride fatty acids, which have been proposed to be indicators of dead or lysed cells, remained relatively constant throughout the experiment. Only the A. protophormiae desiccated for 16 days showed an increase in the ratio of diglyceride fatty acids to PLFAs. The results of this laboratory experiment can be useful for interpreting PLFA profiles of subsurface communities of microorganisms for the purpose of determining their physiological status.

Journal Article↗
PubMed2003

Intraspecific litter diversity and nitrogen deposition affect nutrient dynamics and soil respiration.

Anthropogenic forces are concurrently reducing biodiversity and altering terrestrial nutrient cycles. As natural populations decline, genetic diversity within single species also declines. The consequences of intraspecific genetic loss for ecosystem functions are poorly understood, and interactions among intraspecific diversity, nitrogen deposition, and nutrient cycling are unknown. We present results from an experiment that simulated both a decline in biodiversity and an increase in nitrogen deposition. In soil microcosms, we tested effects of variation in intraspecific litter diversity and nitrogen deposition on soil respiration and nitrogen leaching. Increases in intraspecific litter diversity increased soil respiration overall, with the greatest increases in respiration occurring under high nitrogen deposition. Nitrogen deposition increased the amount of inorganic nitrogen leached, while the amount of dissolved organic nitrogen leached was correlated with initial litter chemistry (lignin concentration) and remained independent of litter diversity and nitrogen deposition treatments. Our results demonstrate the potential for losses in genetic diversity to interact with other global environmental changes to influence terrestrial nutrient cycles.

Ecosystem↗
PubMed2005

Cadmium bioaccumulation in Tubificidae from the overlying water source and effects on bioturbation.

Cadmium bioaccumulation in tubificid oligochaetes in relation to metal vertical distribution in sediment and bioturbation intensity was studied during a 56-day experiment with a constant contamination source in the overlying water (20 microg L(-1)). The indoor microcosms simulate a two-compartment biotope with three experimental treatments based on metal exposure and faunal composition: contaminated water column with or without worms and uncontaminated water column with worms. Cadmium bioaccumulation in worms was studied after 7, 14, 21, 28, and 56 days. Bioturbation was analyzed as a functional parameter representative of organisms' activity and using conservative particulate tracers: luminophores (phi = 63-100 microm and 100-315 microm) and microspheres (phi = 1 microm). The results show no significant effects of cadmium exposure on bioturbation, despite high bioaccumulation levels in worms (50 microg g(-1) dry wt.), suggesting the existence of detoxification/sequestration processes.

Adaptation, Physiological↗
PubMed2006

Reoxidation behavior of technetium, iron, and sulfur in estuarine sediments.

Technetium is a redox active radionuclide, which is present as a contaminant at a number of sites where nuclear fuel cycle operations have been carried out. Recent studies suggest that Tc(VII), which is soluble under oxic conditions, will be retained in sediments as Fe(III)-reducing conditions develop, due to reductive scavenging as hydrous TcO2. However, the behavior of technetium during subsequent reoxidation of sediments remains poorly characterized. Here, we describe a microcosm-based approach to investigate the reoxidation behavior of reduced, technetium-contaminated sediments. In reoxidation experiments, the behavior of Tc was strongly dependent on the nature of the oxidant. With air, reoxidation of Fe(II) and, in sulfate-reducing sediments, sulfide occurred accompanied by approximately 50% remobilization of Tc to solution as TcO4-. With nitrate, reoxidation of Fe(II) and, in sulfate-reducing sediments, sulfide only occurred in microbially active experiments where Fe(II) and sulfide oxidation coupled to nitrate reduction was occurring. Here, Tc was recalcitrant to remobilization with <10% Tc remobilized to solution even when extensive Fe(II) and sulfide reoxidation had occurred. X-ray absorption spectroscopy on reoxidized sediments suggested that 15-50% of Tc bound to sediments was present as Tc(VII). Overall, these results suggest that Tc reoxidation behavior is not directly coupled to Fe or S oxidation and that the extent of Tc remobilization is dependent on the nature of the oxidant.

Ferric Compounds↗
PubMed2002

Escherichia coli survival in groundwater and effluent measured using a combination of propidium iodide and the green fluorescent protein.

AIMS: The aim of this study was to deterimine the survival of an enteric bacterium in anaerobic groundwater and effluent microcosms using the green fluorescent protein (GFP) marker gene in combination with the viability indicator propidium iodide (PI). METHODS AND RESULTS: The pEGFP vector (Clontech) was transformed into Escherichia coli DH5alpha and was stable for at least 100 generations of growth in nonselective medium at 28 degrees C and 37 degrees C. Using an epifluorescent microscope, GFP cells could be detected under blue light (450-490 nm) and the numbers of PI-positive GFPs could be detected under green light (530-560 nm). GFP-tagged E. coli could be detected for at least 132 d in sterilized water microcosms. GFP fluorescence was not lost from the culturable cell population for the duration of the experiment. However, a slow decline in the number of GFP-fluorescent cells in sterilized groundwater was observed. Escherichia coli die-off and loss of green fluorescence was more rapid in nonsterilized waters than in sterilized. Viable numbers of the GFP-tagged E. coli determined by PI counterstaining were compatible with numbers of colony-forming units. CONCLUSIONS: The long-term survival of E. coli and maintainance of GFP-conferred fluorescence in these cells was demonstrated in both groundwater and effluent, under sterilized conditions. However, severe starvation and/or the presence of indigenous microorganisms were found to be factors affecting the maintenance of fluorescence in dead or dying cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the successful application of PI with GFP-tagging to monitor long-term bacterial survival in nutrient-limited conditions and mixed microbial populations.

Anaerobiosis↗
PubMed1994

Getting dressed in the early morning: styles of staff/patient interaction on rehabilitation hospital wards for elderly people.

This paper presents findings from a study of the nursing role in rehabilitation hospital wards for elderly people and focuses on early morning dressing; this is a microcosm of service delivery but it illustrates the way in which institutional practices impinge on patient experiences and highlights areas which warrant attention for the future development of rehabilitation services for elderly people in hospital. The reported findings are drawn from a non-participant structured observational study of 'early morning rising' on rehabilitation wards for elderly people.

Activities of Daily Living↗
PubMed2003

The effects of temperature, water activity and pH on the growth of Aeromonas hydrophila and on its subsequent survival in microcosm water.

AIMS: The influence of temperature, water activity and pH on the growth of Aeromonas hydrophila, and on its survival after transfer in nutrient-poor water were assessed. METHODS AND RESULTS: Experiments were carried out according to a Box-Behnken matrix at 10-30 degrees C, 0.95-0.99 water activity (aw) and pH 5-9. The effect of each factor on the kinetic parameters of growth (i.e. the maximal specific growth rate, mumax, and the lag time, lambda) and on the decline of the bacteria in microcosm water (time to obtain a reduction of 5 log, T5 log) were studied by applying central composite design. CONCLUSIONS: The major effect of temperature and water activity on the growth of A. hydrophila was highlighted, whereas the effect of pH in these experimental conditions was not significant. Models describing the effect of environmental parameters on the growth of A. hydrophila were proposed. The effect of the growth environment, and particularly the incubation temperature, have an influence on the survival ability of the bacteria in nutrient-poor water. SIGNIFICANCE AND IMPACT OF THE STUDY: The Box-Behnken design was well suited to determine the influence of environmental factors on the growth of A. hydrophila and to investigate the effect of previous growth conditions on its survival in microcosm water.

Aeromonas hydrophila↗
PubMed2001

Effects of atrazine and nicosulfuron on freshwater microalgae.

Growth modifications caused by various concentrations of atrazine and nicosulfuron were monitored in closed and continuous culture of Chlorella vulgaris (chlorophyta), Navicula accommoda (diatomophyta), and Oscillatoria limnetica (cyanophyta). The concentration at which algal growth rate was reduced twofold (EC50) was determined in the three species for both herbicides. Comparatively, the two toxicants were applied at 10 microg/l level in microcosms inoculated with natural phytoplankton from Lake Geneva. The relative abundances of major phytoplanktonic species were measured by algal cell count at the beginning and at the end of each experiment. Atrazine and nicosulfuron have different targets in plant metabolism, respectively, photosystem II (PSII) and acetolactate synthase (ALS), and the expected effects were different. Generally, the cultured phytoplankton exhibited various sensitivities, depending on species or herbicide. In the microcosms, the major taxa of natural phytoplanktonic samples exhibited various patterns, from acute toxicity to growth enhancement. For example, the diatoms inside the community were not affected by atrazine and nicosulfuron, except for Stephanodiscus minutulus that was sensitive to both, and Asterionella/formosa that was sensitive only to nicosulfuron. The specific physiology and the relationships among the phytoplanktonic communities have to be carefully considered when one would try to predict the extent of herbicide action on natural phytoplankton using in vitro tests. There is a need to test the toxic effect on various cultured strains, representative of most of the taxonomic composition of natural communities, to take into account the wide range of sensitivities and reaction to herbicide contamination. But this is not enough to give a solid frame when transposing the results to the field, and the use of more ecologically relevant systems is recommended.

Animals↗
PubMed2003

Biofilm structure and function and possible implications for riverine DOC dynamics.

Biofilms are major sites of carbon cycling in streams and rivers. Here we elucidate the relationship between biofilm structure and function and river DOC dynamics. Metabolism (extracellular enzymatic activity) and structure (algae, bacteria, C/N content) of light-grown (in an open channel) and dark-grown (in a dark pipe) biofilms were studied over a year, and variations in dissolved organic carbon (DOC) and biodegradable DOC (BDOC) were also recorded. A laboratory experiment on 14C-glucose uptake and DOC dynamics was also performed by incubating natural biofilms in microcosms. On the basis of our field (annual DOC budget) and laboratory results, we conclude that light-grown biofilm is, on annual average, a net DOC consumer. This biofilm showed a high monthly variability in DOC uptake/release rates, but, on average, the annual uptake rate was greater than that of the dark-grown biofilm. The higher algal biomass and greater structure of the light-grown biofilm may enhance the development of the bacterial community (bacterial biomass and activity) and microbial heterotrophic activity. In addition, the light-grown biofilm may promote abiotic adsorption because of the development of a polysaccharide matrix. In contrast, the dark-grown biofilm is highly dependent on the amount and quality of organic matter that enters the system and is more efficient in the uptake of labile molecules (higher 14C-glucose uptake rate per mgC). The positive relationships between the extracellular enzymatic activity of biofilm and DOC and BDOC content in flowing water indicate that biofilm metabolism contributes to DOC dynamics in fluvial systems. Our results show that short-term fluvial DOC dynamics is mainly due to the use and recycling of the more labile molecules. At the river ecosystem level, the potential surface area for biofilm formation and the quantity and quality of available organic carbon might determine the effects of biofilm function on DOC dynamics.

Analysis of Variance↗
PubMed2005

Long-term recovery of PCB-contaminated sediments at the Lake Hartwell superfund site: PCB dechlorination. 2. Rates and extent.

This paper reports on extensive polychlorinated biphenyl (PCB) dechlorination measured in Lake Hartwell (Pickens County, SC) sediments. Vertical sediment cores were collected from 18 locations in Lake Hartwell (Pickens County, SC) and analyzed in 5-cm increments for PCB congeners. The preferential loss of meta and para chlorines with sediment depth demonstrated that PCBs in the sediments underwent reductive dechlorination after burial. Notably, ortho chlorines were highly conserved for more than 5 decades; since the first appearance of PCBs, ca. 1950-1955. These dechlorination characteristics resulted in the accumulation of lower chlorinated congeners dominated by ortho chlorine substituents. Dechlorination rates were determined by plotting the numbers of meta plus para chlorines per biphenyl molecule (mol of chlorine/mol of PCB) with sediment age. Regression analyses showed linear correlations between meta plus para chlorine concentrations with time. The average dechlorination rate was 0.094 +/- 0.063 mol of Cl/mol of PCB/yr. The rates measured using the 2001 cores were approximately twice those measured using the 2000 cores, most likely because the 2001 cores were collected only at transects O, L, and I, which had the highest rates measured in 2000. An inverse of the dechlorination rates indicated that 16.4 +/- 11.6 yr was required per meta plus para chlorine removal (ranging from 4.3 to 43.5 yr per chlorine removal). The rates determined from this study were 1-2 orders of magnitude lower than rates reported from laboratory microcosm studies using Hudson River and St. Lawrence River sediments, suggesting that dechlorination rates reported for laboratory experiments are much higher than those occurring in situ.

Bacteria, Aerobic↗
PubMed1999

Effects of salinity and temperature on long-term survival of the eel pathogen Vibrio vulnificus biotype 2 (serovar E).

Vibrio vulnificus biotype 2 (serovar E) is a primary eel pathogen. In this study, we performed long-term survival experiments to investigate whether the aquatic ecosystem can be a reservoir for this bacterium. We have used microcosms containing water of different salinities (ranging from 0.3 to 3.8%) maintained at three temperatures (12, 25, and 30 degrees C). Temperature and salinity significantly affected long-term survival: (i) the optimal salinity for survival was 1.5%; (ii) lower salinities reduced survival, although they were nonlethal; and (ii) the optimal temperature for survival was dependent on the salinity (25 degrees C for microcosms at 0.3 and 0.5% and 12 degrees C for microcosms at 1.5 to 3.8%). In the absence of salts, culturability dropped to zero in a few days, without evidence of cellular lysis. Under optimal conditions of salinity and temperature, the bacterium was able to survive in the free-living form for at least 3 years. The presence of a capsule on the bacterial cell seemed to confer an advantage, since the long-term survival rate of opaque variants was significantly higher than that of translucent ones. Long-term-starved cells maintained their infectivity for eels (as determined by both intraperitoneal and immersion challenges) and mice. Examination under the microscope showed that (i) the capsule was maintained, (ii) the cell size decreased, (iii) the rod shape changed to coccuslike along the time of starvation, and (iv) membrane vesicles and extracellular material were occasionally produced. In conclusion, V. vulnificus biotype 2 follows a survival strategy similar to that of biotype 1 of this species in response to starvation conditions in water. Moreover, the aquatic ecosystem is one of its reservoirs.

Animals↗
PubMed1999

Bioremediation of atrazine-contaminated soil by repeated applications of atrazine-degrading bacteria.

Bioaugmentation has previously been unreliable for the in situ clean-up of contaminated soils because of problems with poor survival and the rapid decline in activity of the bacterial inoculum. In an attempt to solve these problems, a 500-l batch fermenter was investigated for its ability to deliver inoculum repeatedly to contaminated soils via irrigation lines. In a field experiment, mesocosms were filled with 350 kg soil containing 100 mg kg-1 atrazine, and inoculated one, four or eight times with an atrazine-degrading bacterial consortium that was produced in the fermenter. After 12 weeks, no significant degradation of atrazine had occurred in soil that was inoculated only once; whereas, mesocosms inoculated four and eight times mineralized 38% and 72% of the atrazine respectively. Similar results were obtained in a laboratory experiment using soil contaminated with 100 mg kg-1 [14C]atrazine. After 35 days, soil that was inoculated once with 10(8) cfu ml-1 of the consortium or with the atrazine-degrading bacterium, Pseudomonas sp. strain ADP, mineralized 17% and 35% of the atrazine respectively. In comparison, microcosms inoculated every 3 days with the consortium or with Pseudomonas sp. (ADP) mineralized 64% or 90% of the atrazine over this same period. Results of these experiments suggest that repeated inoculation from an automated fermenter may provide a strategy for bioaugmentation of contaminated soil with xenobiotic-degrading bacteria.

Atrazine↗
PubMed1999

Effect of gamma radiation on native endolithic microorganisms from a radioactive waste deposit site.

A time-course experiment was conducted to evaluate the effects of gamma radiation on the indigenous microbiota present in rock obtained from Yucca Mountain, Nevada Test Site. Microcosms were constructed by placing pulverized Yucca Mountain rock in polystyrene cylinders. Continuous exposure (96 h) at a dose rate of 1.63 Gy/min was used to mimic the near-field environment surrounding waste canisters. The expected maximum surface dose rate from one unbreached canister designed to contain spent nuclear fuels is 0.06 Gy/min. Considering the current repository packing design, multiple canisters within one vault, the cumulative dose rate may well approach that used in this experiment. The microbial communities were characterized after receiving cumulative doses of 0, 0.098, 0. 58, 2.33, 4.67, 7.01 and 9.34 kGy. Radiation-resistant microorganisms in the pulverized rock became viable but nonculturable (VBNC) after a cumulative dose of 2.33 kGy. VBNC microorganisms lose the ability to grow on media on which they have routinely been cultured in response to the environmental stress imposed (i.e. radiation) but can be detected throughout the time course using direct fluorescence microscopy techniques. Two representative exopolysaccharide-producing isolates from Yucca Mountain were exposed to the same radiation regimen in sand microcosms. One isolate was much more radiation-resistant than the other, but both had greater resistance than the general microbial community based on culturable counts. However, when respiring cell counts (VBNC) were compared after irradiation, the results would indicate much more radiation resistance of the individual isolates and the microbial community in general. These results have significant implications for underground storage of nuclear waste as they indicate that indigenous microorganisms are capable of surviving gamma irradiation in a VBNC state.

Bacteria↗