Nitrate-Driven Trophic Association of Sulfur-Cycling Microorganisms in Tsunami-Deposited Marine Sediment Revealed by High-Sensitivity 13C-Bicarbonate Probing.

Although denitrification-dependent chemolithotrophic sulfur oxidizers proliferated in tsunami-deposited marine sediment with nitrate amendment, their ecophysiological roles in biogeochemical carbon transfer are not addressed. We employed time-resolved high-sensitivity 13C-bicarbonate probing of rRNA to unveil the carbon fixation and resulting trophic relationship of the nitrate-amended sediment microorganisms. Nitrate reduction and sulfur oxidation co-occurred along with significant decreases in the 13CO2 and dissolved bicarbonate concentrations for the first 4 days of the incubation, during which the denitrification-dependent sulfur-oxidizing chemolithotrophs, i.e., the Sulfurimonas sp. HDS01 and Thioalkalispira sp. HDS22 relatives, and the sulfate-reducing heterotrophs, i.e., the Desulfobulbus spp. and Desulfofustis glycolicus relatives, actively incorporated 13C. These indicated that the sulfur oxidizers and sulfate reducers were tightly associated with each other through the direct carbon transfer. Relatives of the fermentative Thalassomonas sediminis and the hydrolytic Pararheinheimera aquatica, in addition to various sulfur-cycling microorganisms, significantly assimilated 13C at day 14. Although the incorporation of 13C was not detected, a syntrophic volatile-fatty-acid oxidizer and hydrogenotrophic methanogens significantly expressed their 16S rRNA molecules at day 21, indicating the metabolic activation of these final decomposers under the latter nutrient-limited conditions. The results demonstrated the nitrate-driven trophic association of sulfur-cycling microorganisms and the subsequent microbial activation and diversification, triggering the restoration of the marine ecosystem function.

Direct comparison of bacterial communities in soils contaminated with different levels of radioactive cesium from the first Fukushima nuclear power plant accident.

The Great East Japan Earthquake caused a serious accident at the first Fukushima nuclear power plant (NPP), which in turn released a large amount of radionuclides. Little attention has been paid to in-situ soil microorganisms exposed to radioactive contamination by the actual NPP accident. We herein investigated bacterial communities in the radioactive cesium (Cs)-contaminated and non-contaminated soils by high-throughput sequencing. The uppermost and ectorhizosphere soil samples were collected from the base of mugwort grown in the same soil type with the same soil-use history in order to compare the bacterial communities at geographically separated areas. The concentrations of radioactive Cs in the soils ranged from 10 to 563,000 Bq 137Cs/kg dry soil, with the highest concentration being detected at 1 km from the NPP. Alpha-diversity indices, i.e., Chao1, Shannon and Simpson reciprocal, of the sequence data showed the lower bacterial diversity in the most highly Cs-contaminated soil. Principal coordinate analysis with principle components 1 and 3 based on unweighted UniFrac distances indicated the significant difference in bacterial communities of the most contaminated area from those of the other areas. Operational taxonomic unit-based assay revealed higher abundance of the radio-resistant Geodermatophilus bullaregiensis relative in the most contaminated soil. Thus, it was strongly suggested that the radioactive accident facilitated the growth and/or survival of radio-resistant bacteria in the Cs-contaminated soils. The results of this study show that information on the soil type, vegetation and soil-use history enhances the direct comparison of geographically distant soil bacterial communities exposed to different levels of radioactive contamination.

Stratification of Sulfur Species and Microbial Community in Launched Marine Sediment by an Improved Sulfur-Fractionation Method and 16S rRNA Gene Sequencing.

With a focus on marine sediment launched by the tsunami accompanying the Great East Japan Earthquake, we examined the vertical (i.e., depths of 0-2, 2-10, and 10-20 mm) profiles of reduced inorganic sulfur species and microbial community using a newly improved sulfur-fractionation method and 16S rRNA gene sequencing. S0 accumulated at the largest quantities at a depth of 2-10 mm, while the reduced forms of sulfur, such as iron(II) sulfide and pyrite, were abundant below 2 mm of the sediment. Operational taxonomic units (OTUs) related to chemolithotrophically sulfur-oxidizing Sulfurimonas denitrificans and Sulfurimonas autotrophica were only predominant at 2-10 mm, suggesting the involvement of these OTUs in the oxidation of sulfide to S0. In addition, Desulfocapsa sulfexigens, which is capable of chemolithotrophically disproportionating S0, prevailed at the same depth, indicating that accumulated S0 was converted to sulfate and sulfide. Although no significant differences were observed in sulfate concentrations across the depths examined, specific species of chemoorganotrophic sulfate reducers, i.e., Desulfotignum toluenicum and Desulfosalsimonas propionicica, showed significantly higher abundance at a depth of 2-10 mm than at the other depths examined. Organic matter potentially generated from sulfur oxidation and disproportionation may have served as the carbon source for the growth of these sulfate reducers. The present results demonstrated that sulfur oxidizers, a sulfur disproportionator, and sulfate reducers played vital roles in sulfur cycling with S0 as the key inorganic sulfur species in the oxic-anoxic boundary layer of the launched marine sediment.

Sulfur-Oxidizing Bacteria Mediate Microbial Community Succession and Element Cycling in Launched Marine Sediment.

A large amount of marine sediment was launched on land by the Great East Japan earthquake. Here, we employed both on-site and laboratory studies on the launched marine sediment to investigate the succession of microbial communities and its effects on geochemical properties of the sediment. Twenty-two-month on-site survey showed that microbial communities at the uppermost layer (0-2 mm depth) of the sediment changed significantly with time, whereas those at the deeper layer (20-40 mm depth) remained nearly unchanged and kept anaerobic microbial communities. Nine months after the incidence, various sulfur-oxidizing bacteria (SOB) prevailed in the uppermost layer, in which afterwards diverse chemoorganotrophic bacteria predominated. Geochemical analyses indicated that the concentration of metals other than Fe was lower in the uppermost layer than that in the deeper layer. Laboratory study was carried out by incubating the sediment for 57 days, and clearly indicated the dynamic transition of microbial communities in the uppermost layer exposed to atmosphere. SOB affiliated in the class Epsilonproteobacteria rapidly proliferated and dominated at the uppermost layer during the first 3 days, after that Fe(II)-oxidizing bacteria and chemoorganotrophic bacteria were sequentially dominant. Furthermore, the concentration of sulfate ion increased and the pH decreased. Consequently, SOB may have influenced the mobilization of heavy metals in the sediment by metal-bound sulfide oxidation and/or sediment acidification. These results demonstrate that SOB initiated the dynamic shift from the anaerobic to aerobic microbial communities, thereby playing a critical role in element cycling in the marine sediment.

Dynamic transition of chemolithotrophic sulfur-oxidizing bacteria in response to amendment with nitrate in deposited marine sediments.

Although environmental stimuli are known to affect the structure and function of microbial communities, their impact on the metabolic network of microorganisms has not been well investigated. Here, geochemical analyses, high-throughput sequencing of 16S rRNA genes and transcripts, and isolation of potentially relevant bacteria were carried out to elucidate the anaerobic respiration processes stimulated by nitrate (20 mM) amendment of marine sediments. Marine sediments deposited by the Great East Japan Earthquake in 2011 were incubated anaerobically in the dark at 25∘C for 5 days. Nitrate in slurry water decreased gradually for 2 days, then more rapidly until its complete depletion at day 5; production of N2O followed the same pattern. From day 2 to 5, the sulfate concentration significantly increased and the sulfur content in solid-phase sediments significantly decreased. These results indicated that denitrification and sulfur oxidation occurred simultaneously. Illumina sequencing revealed the proliferation of known sulfur oxidizers, i.e., Sulfurimonas sp. and Chromatiales bacteria, which accounted for approximately 43.5% and 14.8% of the total population at day 5, respectively. These oxidizers also expressed 16S rRNA to a considerable extent, whereas the other microorganisms, e.g., iron(III) reducers and methanogens, became metabolically active at the end of the incubation. Extinction dilution culture in a basal-salts medium supplemented with sulfur compounds and nitrate successfully isolated the predominant sulfur oxidizers: Sulfurimonas sp. strain HDS01 and Thioalkalispira sp. strain HDS22. Their 16S rRNA genes showed 95.2-96.7% sequence similarity to the closest cultured relatives and they grew chemolithotrophically on nitrate and sulfur. Novel sulfur-oxidizing bacteria were thus directly involved in carbon fixation under nitrate-reducing conditions, activating anaerobic respiration processes and the reorganization of microbial communities in the deposited marine sediments.