Wilfred Fm Röling

Subsurface removal of arsenic by injection with oxygenated groundwater has been proposed as a viable technology for obtaining ‘safe’ drinking water in Bangladesh. While the oxidation of ferrous iron to solid ferric iron minerals, to which arsenic adsorbs, is assumed to be driven by abiotic reactions, metal-cycling microorganisms may potentially affect arsenic removal. A cultivation-independent survey covering 24 drinking water wells in several geographical regions in Bangladesh was conducted to obtain information on microbial community structure and diversity in general, and on specific functional groups capable of the oxidation or reduction of arsenic or iron. Each functional group, targeted by either group-specific 16S rRNA or functional gene amplification, occurred in at least 79% of investigated samples. Putative arsenate reducers and iron-oxidizing Gallionellaceae were present at low diversity, while more variation in potentially arsenite-oxidizing microorganisms and iron-reducing Desulfuromonadales was revealed within and between samples. Relations between community composition on the one hand and hydrochemistry on the other hand were in general not evident, apart from an impact of salinity on iron-cycling microorganisms. Our data suggest widespread potential for a positive contribution of arsenite and iron oxidizers to arsenic removal upon injection with oxygenated water, but also indicate a potential risk for arsenic re-mobilization by anaerobic arsenate and iron reducers once injection is halted.

Summary: Whole genome amplification methods facilitate the detection and characterization of microbial communities in low biomass environments. We examined the extent to which the actual community structure is reliably revealed and factors contributing to bias. One widely used [multiple displacement amplification ( MDA )] and one new primer‐free method [primase‐based whole genome amplification ( pWGA )] were compared using a polymerase chain reaction ( PCR )‐based method as control. Pyrosequencing of an environmental sample and principal component analysis revealed that MDA impacted community profiles more strongly than pWGA and indicated that this related to species GC content, although an influence of DNA integrity could not be excluded. Subsequently, biases by species GC content, DNA integrity and fragment size were separately analysed using defined mixtures of DNA from various species. We found significantly less amplification of species with the highest GC content for MDA ‐based templates and, to a lesser extent, for pWGA . DNA fragmentation also interfered severely: species with more fragmented DNA were less amplified with MDA and pWGA . pWGA was unable to amplify low molecular weight DNA (< 1.5 kb), whereas MDA was inefficient. We conclude that pWGA is the most promising method for characterization of microbial communities in low‐biomass environments and for currently planned astrobiological missions to M ars.

The adsorption of nucleic acids to mineral matrixes can result in low extraction yields and negatively influences molecular microbial ecology studies,

Humankind's innate curiosity makes us wonder whether life is or was present on other planetary bodies such as Mars. The EuroGeoMars 2009 campaign was organized at the Mars Desert Research Station (MDRS) to perform multidisciplinary astrobiology research. MDRS in southeast Utah is situated in a cold arid desert with mineralogy and erosion processes comparable to those on Mars. Insight into the microbial community composition of this terrestrial Mars analogue provides essential information for the search for life on Mars: including sampling and life detection methodology optimization and what kind of organisms to expect. Soil samples were collected from different locations. Culture-independent molecular analyses directed at ribosomal RNA genes revealed the presence of all three domains of life (Archaea, Bacteria and Eukarya), but these were not detected in all samples. Spiking experiments revealed that this appears to relate to low DNA recovery, due to adsorption or degradation. Bacteria were most frequently detected and showed high alpha- and beta-diversity. Members of theActinobacteria, Proteobacteria, BacteroidetesandGemmatimonadetesphyla were found in the majority of samples. Archaea alpha- and beta-diversity was very low. For Eukarya, a diverse range of organisms was identified, such as fungi, green algae and several phyla of Protozoa. Phylogenetic analysis revealed an extraordinary variety of putative extremophiles, mainly Bacteria but also Archaea and Eukarya. These comprised radioresistant, endolithic, chasmolithic, xerophilic, hypolithic, thermophilic, thermoacidophilic, psychrophilic, halophilic, haloalkaliphilic and alkaliphilic micro-organisms. Overall, our data revealed large difference in occurrence and diversity over short distances, indicating the need for high-sampling frequency at similar sites. DNA extraction methods need to be optimized to improve extraction efficiencies.

Summary: Natural attenuation of the mono‐aromates benzene, toluene, ethylbenzene and xylene occurs under iron‐reducing conditions in a leachate‐contaminated aquifer near the Banisveld landfill, the Netherlands. The diversity of mono‐aromate‐degrading microorganisms was studied by targeting functional genes encoding benzylsuccinate synthase α‐subunit ( bssA ) and 6‐oxocyclohex‐1‐ene‐1‐carbonyl‐CoA hydrolase ( bamA ). Sixty‐four bssA and 188 bamA variants were sequenced from groundwater sampled along the pollution plume in 1999 and 2004. Species containing bssA sequences closest affiliated (> 91%) with the betaprotebacterium Georgfuchsia toluolica were the dominant alkylbenzene degraders (89% of bssA sequences). bssA genes were found at more than 10‐fold lower copy numbers than bamA genes, of which only a small fraction (< 2%) was closely related to the genes of Georgfuchsia . bamA gene diversity was high and bamA ‐based community composition was primarily affected by dissolved organic carbon (DOC) and ferrous iron concentrations. bamA sequences closest related to Geobacteraceae were dominantly (43.2%) observed and the presence of Geobacteraceae ‐related bamA sequences was associated with DOC. Our results indicate a key role for specialized Georgfuchsia spp. in the degradation of alkylbenzenes, whereas Geobacteraceae are involved in degradation of aromatics other than toluene and xylene.

Summary: Geobacteraceae dominate many iron‐reducing subsurface environments and are associated with biodegradation of organic pollutants. In order to enhance the understanding of the environmental role played by Geobacteraceae , the physiology of Geobacter metallireducens was investigated at the low growth rates found in its subsurface habitat. Cultivation in retentostats (a continuous culturing device with biomass retention) under electron acceptor and electron donor limitation enabled growth rates as low as 0.0008 h ⁻¹ . The maximum growth yield was between 0.05 and 0.09 C‐mol biomass per C‐mol acetate and comparable to that observed in batch experiments. Maintenance energy demand is among the lowest reported for heterotrophic bacteria, under both acetate and AQDS limitation. The cells were able to use alternative electron acceptors directly, without requiring de novo protein synthesis. We discuss how the extremely low maintenance energy demand and the ability to readily use alternative electron acceptors may help Geobacter species to become ubiquitous and dominant microorganisms in many iron‐reducing subsurface settings.