Prior exposure of microbial communities to seawater reduces resilience but increases compositional and functional resistance to flooding events.

Storm surges, flooding, and the encroachment of seawater onto agricultural land are predicted to increase with climate change. These flooding events fundamentally alter many soil properties and have knock-on effects on the microbial community composition and its functioning. The hypotheses tested in this study were (1) that the extent of change (resistance) of microbial community functioning and structure during seawater flooding is a factor of pre-adaptation to the stress, and (2) if structure and function are altered, the pre-adaptation will result in communities returning to previous state prior to flooding (resilience) faster than unexposed communities. We chose a naturally occurring saltmarsh-terrestrial pasture gradient from which three elevations were selected to create mesocosms. By selecting these sites, we were able to incorporate the legacy of differing levels of seawater ingress and exposure. Mesocosms were submerged in seawater for 0, 1, 96- and 192-h, with half of the mesocosms sacrificed immediately after flooding, and the other half taken after a 14 day "recovery" period. The following parameters were monitored: 1) changes in soil environmental parameters, 2) prokaryotic community composition, and 3) microbial functioning. Our results indicated that any length of seawater inundation significantly altered the physicochemical properties of all the soils, although a greater change is observed in the pasture site compared to the saltmarsh sites. These changes remained following a recovery period. Interestingly, our results indicated that for community composition, there was a high degree of resistance for the Saltmarsh mesocosms, with the Pasture mesocosm displaying higher resilience. Further, we observed a functional shift in the enzyme activities with labile hemicellulose being preferentially utilised over cellulose, with the effect increasing with longer floods. These results suggest that changing bacterial physiology is more critical to understanding the impact of storm surges on agricultural systems than bulk community change.

Identification of a Stable Hydrogen-Driven Microbiome in a Highly Radioactive Storage Facility on the Sellafield Site.

The use of nuclear power has been a significant part of the United Kingdom's energy portfolio with the Sellafield site being used for power production and more recently reprocessing and decommissioning of spent nuclear fuel activities. Before being reprocessed, spent nuclear fuel is stored in water ponds with significant levels of background radioactivity and in high alkalinity (to minimize fuel corrosion). Despite these challenging conditions, the presence of microbial communities has been detected. To gain further insight into the microbial communities present in extreme environments, an indoor, hyper-alkaline, oligotrophic, and radioactive spent fuel storage pond (INP) located on the Sellafield site was analyzed. Water samples were collected from sample points within the INP complex, and also the purge water feeding tank (FT) that supplies water to the pond, and were screened for the presence of the 16S and 18S rRNA genes to inform sequencing requirements over a period of 30 months. Only 16S rRNA genes were successfully amplified for sequencing, suggesting that the microbial communities in the INP were dominated by prokaryotes. Quantitative Polymerase Chain Reaction (qPCR) analysis targeting 16S rRNA genes suggested that bacterial cells in the order of 104-106 mL-1 were present in the samples, with loadings rising with time. Next generation Illumina MiSeq sequencing was performed to identify the dominant microorganisms at eight sampling times. The 16S rRNA gene sequence analysis suggested that 70% and 91% from of the OTUs samples, from the FT and INP respectively, belonged to the phylum Proteobacteria, mainly from the alpha and beta subclasses. The remaining OTUs were assigned primarily to the phyla Acidobacteria, Bacteroidetes, and, Cyanobacteria. Overall the most abundant genera identified were Hydrogenophaga, Curvibacter, Porphyrobacter, Rhodoferax, Polaromonas, Sediminibacterium, Roseococcus, and Sphingomonas. The presence of organisms most closely related to Hydrogenophaga species in the INP areas, suggests the metabolism of hydrogen as an energy source, most likely linked to hydrolysis of water caused by the stored fuel. Isolation of axenic cultures using a range of minimal and rich media was also attempted, but only relatively minor components (from the phylum Bacteroidetes) of the pond water communities were obtained, emphasizing the importance of DNA-based, not culture-dependent techniques, for assessing the microbiome of nuclear facilities.

Microbial bloom formation in a high pH spent nuclear fuel pond.

Microorganisms are able to colonise a wide range of extreme environments, including nuclear facilities. In this study, the First Generation Magnox Storage Pond (FGMSP) a high pH, legacy spent nuclear fuel pond (SNFP) situated at Sellafield, Cumbria, UK, was studied. Despite the inhospitable conditions in the FGMSP, microorganisms can cause "blooms" within the facility which to date have not been studied. These microbial blooms significantly reduce visibility in the engineered facility, disrupting fuel retrieval operations and slowing decommissioning. The microbial community colonising the pond during two microbial bloom periods was determined by using physiological measurements and high throughput next generation sequencing techniques. In situ probes within the ponds targeting photosynthetic pigments indicated a cyanobacterial bloom event. Analysis of the 16S rRNA gene data suggested that a single cyanobacterial genus was dominant during the bloom events, which was most closely related to Pseudanabaena sp. Comparisons between the microbial community of FGMSP and an adjacent SNFP that is periodically purged into the FGMSP, showed different community profiles. Data confirm the onset of the microbial blooms occurred when the pond purge rate was reduced, and blooms could be controlled by re-establishing the purging regime. The presence of Pseudanabaena sp. that can colonise the pond and dominate during bloom periods is notable since they have received little attention for their role in cyanobacterial bloom formation. This work also informs bioremediation efforts to treat waters contaminated with radionuclides, which could benefit from the use of cyanobacteria able to tolerate extreme environments and accumulate priority radionuclides.