Radioactive waste microbiology: predicting microbial survival and activity in changing extreme environments.

The potential for microbial activity to occur within the engineered barrier system (EBS) of a geological disposal facility (GDF) for radioactive waste is acknowledged by waste management organizations as it could affect many aspects of the safety functions of a GDF. Microorganisms within an EBS will be exposed to changing temperature, pH, radiation, salinity, saturation, and availability of nutrient and energy sources, which can limit microbial survival and activity. Some of the limiting conditions are incorporated into GDF designs for safety reasons, including the high pH of cementitious repositories, the limited pore space of bentonite-based repositories, or the high salinity of GDFs in evaporitic geologies. Other environmental conditions such as elevated radiation, temperature, and desiccation, arise as a result of the presence of high heat generating waste (HHGW). Here, we present a comprehensive review of how environmental conditions in the EBS may limit microbial activity, covering HHGW and lower heat generating waste (LHGW) in a range of geological environments. We present data from the literature on the currently recognized limits to life for each of the environmental conditions described above, and nutrient availability to establish the potential for life in these environments. Using examples where each variable has been modelled for a particular GDF, we outline the times and locations when that variable can be expected to limit microbial activity. Finally, we show how this information for multiple variables can be used to improve our understanding of the potential for microbial activity to occur within the EBS of a GDF and, more broadly, to understand microbial life in changing environments exposed to multiple extreme conditions.

Seasonality of enteric viruses in groundwater-derived public water sources.

We investigated the seasonal prevalence of seven enteric viruses in groundwater-derived public water sources distributed across the dominant aquifers of England. Sampling targeted four periods in the hydrological cycle with typically varying microbial risks, as indicated using a decade of Escherichia coli prevalence data. Viruses were concentrated onsite by filtration of raw groundwater, and extracted nucleic acid (NA) was amplified by qPCR or RT-qPCR. Seven out of eight sources, all aquifers, and 31% of samples were positive for viral NA. The most frequently detected viral NA targets were Hepatitis A virus (17% samples, 63% sites), norovirus GI (14% samples, 38% sites), and Hepatitis E virus (7% samples, 25% sites). Viral NA presence was episodic, being most prevalent and at its highest concentration during November and January, the main groundwater recharge season, with 89% of all positive detects occurring during a rising water table. Seasonal norovirus NA detections matched its seasonal incidence within the population. Viral NA is arriving with groundwater recharge, as opposed to persisting for long-periods within the saturated zone. Neither total coliforms nor E. coli were significant predictors of viral NA presence-absence, and there was limited co-occurrence between viruses. Nevertheless, a source with an absence of E. coli in regularly collected historical data is unlikely to be at risk of viral contamination. To manage potential groundwater viral contamination via risk assessment, larger scale studies are required to understand key risk factors, with the evidence here suggesting viral NA is widespread across a range of typical microbial risk settings.

Subsurface Microbial Hydrogen Cycling: Natural Occurrence and Implications for Industry.

Hydrogen is a key energy source for subsurface microbial processes, particularly in subsurface environments with limited alternative electron donors, and environments that are not well connected to the surface. In addition to consumption of hydrogen, microbial processes such as fermentation and nitrogen fixation produce hydrogen. Hydrogen is also produced by a number of abiotic processes including radiolysis, serpentinization, graphitization, and cataclasis of silicate minerals. Both biotic and abiotically generated hydrogen may become available for consumption by microorganisms, but biotic production and consumption are usually tightly coupled. Understanding the microbiology of hydrogen cycling is relevant to subsurface engineered environments where hydrogen-cycling microorganisms are implicated in gas consumption and production and corrosion in a number of industries including carbon capture and storage, energy gas storage, and radioactive waste disposal. The same hydrogen-cycling microorganisms and processes are important in natural sites with elevated hydrogen and can provide insights into early life on Earth and life on other planets. This review draws together what is known about microbiology in natural environments with elevated hydrogen, and highlights where similar microbial populations could be of relevance to subsurface industry.

Advances in the in-field detection of microorganisms in ice.

The historic view of ice-bound ecosystems has been one of a predominantly lifeless environment, where microorganisms certainly exist but are assumed to be either completely inactive or in a state of long-term dormancy. However, this standpoint has been progressively overturned in the past 20years as studies have started to reveal the importance of microbial life in the functioning of these environments. Our present knowledge of the distribution, taxonomy, and metabolic activity of such microbial life has been derived primarily from laboratory-based analyses of collected field samples. To date, only a restricted range of life detection and characterization techniques have been applied in the field. Specific examples include direct observation and DNA-based techniques (microscopy, specific stains, and community profiling based on PCR amplification), the detection of biomarkers (such as adenosine triphosphate), and measurements of metabolism [through the uptake and incorporation of radiolabeled isotopes or chemical alteration of fluorescent substrates (umbelliferones are also useful here)]. On-going improvements in technology mean that smaller and more robust life detection and characterization systems are continually being designed, manufactured, and adapted for in-field use. Adapting technology designed for other applications is the main source of new methodology, and the range of techniques is currently increasing rapidly. Here we review the current use of technology and techniques to detect and characterize microbial life within icy environments and specifically its deployment to in-field situations. We discuss the necessary considerations, limitations, and adaptations, review emerging technologies, and highlight the future potential. Successful application of these new techniques to in-field studies will certainly generate new insights into the way ice bound ecosystems function.

In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments.

The discovery over the past two decades of viable microbial communities within glaciers has promoted interest in the role of glaciers and ice sheets (the cryosphere) as contributors to subglacial erosion, global biodiversity, and in regulating global biogeochemical cycles. In situ or in-field detection and characterisation of microbial communities is becoming recognised as an important approach to improve our understanding of such communities. Within this context we demonstrate, for the first time, the ability to detect Gram-negative bacteria in glacial field-environments (including subglacial environments) via the detection of lipopolysaccharide (LPS); an important component of Gram-negative bacterial cell walls. In-field measurements were performed using the recently commercialised PyroGene® recombinant Factor C (rFC) endotoxin detection system and used in conjunction with a handheld fluorometer to measure the fluorescent endpoint of the assay. Twenty-seven glacial samples were collected from the surface, bed and terminus of a low-biomass Arctic valley glacier (Engabreen, Northern Norway), and were analysed in a field laboratory using the rFC assay. Sixteen of these samples returned positive LPS detection. This work demonstrates that LPS detection via rFC assay is a viable in-field method and is expected to be a useful proxy for microbial cell concentrations in low biomass environments.