Transcriptional Activity of Arsenic-Reducing Bacteria and Genes Regulated by Lactate and Biochar during Arsenic Transformation in Flooded Paddy Soil.

Organic substrates and biochar are important in controlling arsenic release from sediments and soils; however, little is known about their impact on arsenic-reducing bacteria and genes during arsenic transformation in flooded paddy soils. In this study, microcosm experiments were established to profile transcriptional activity of As(V)-respiring gene (arrA) and arsenic resistance gene (arsC) as well as the associated bacteria regulated by lactate and/or biochar in anaerobic arsenic-contaminated paddy soils. Chemical analyses revealed that lactate as the organic substrate stimulated microbial reduction of As(V) and Fe(III), which was simultaneously promoted by lactate+biochar, due to biochar's electron shuttle function that facilitates electron transfer from bacteria to As(V)/Fe(III). Sequencing and phylogenetic analyses demonstrated that both arrA closely associated with Geobacter (>60%, number of identical sequences/number of the total sequences) and arsC related to Enterobacteriaceae (>99%) were selected by lactate and lactate+biochar. Compared with the lactate microcosms, transcriptions of the bacterial 16S rRNA gene, Geobacter spp., and Geobacter arrA and arsC genes were increased in the lactate+biochar microcosms, where transcript abundances of Geobacter and Geobacter arrA closely tracked with dissolved As(V) concentrations. Our findings indicated that lactate and biochar in flooded paddy soils can stimulate the active As(V)-respiring bacteria Geobacter species for arsenic reduction and release, which probably increases arsenic bioavailability to rice plants.

Simultaneous alleviation of cadmium and arsenic accumulation in rice by applying zero-valent iron and biochar to contaminated paddy soils.

The fates of cadmium (Cd) and arsenic (As) in paddy fields are generally opposite; thus, the inconsistent transformation of Cd and As poses large challenges for their remediation. In this study, the impacts of zero valent iron (ZVI) and/or biochar amendments on Cd and As bioavailability were examined in pot trials with rice. Comparison with the untreated soil, both Cd and As accumulation in different rice tissues decreased significantly in the ZVI-biochar amendments and the Cd and As accumulation in rice decreased with increasing ZVI contents. In particular, the concentrations of Cd (0.15 ± 0.01 mg kg-1) and As (0.17 ± 0.01 mg kg-1) in rice grains were decreased by 93% and 61% relative to the untreated soil, respectively. A sequential extraction analysis indicated that with increasing Fe ratios in the ZVI-biochar mixtures, bioavailable Cd and As decreased, and the immobilized Cd and As increased. Furthermore, high levels of Fe, Cd, and As were detected in Fe plaque of the ZVI-biochar amendments in comparison with the single biochar or single ZVI amendments. The ZVI-biochar mixture may have a synergistic effect that simultaneously reduces Cd and As bioavailability by increasing the formation of amorphous Fe and Fe plaque for Cd and As immobilization. The single ZVI amendment significantly decreased As bioavailability, while the single biochar amendment significantly reduced the bioavailability of Cd compared with the combined amendments. Hence, using a ZVI-biochar mixture as a soil amendment could be a promising strategy for safely-utilizing Cd and As co-contaminated sites in the future.

Roles of different active metal-reducing bacteria in arsenic release from arsenic-contaminated paddy soil amended with biochar.

Although biochar has great potential for heavy metal removal from sediments or soils, its impact on arsenic biogeochemistry in contaminated paddy fields remains poorly characterized. In this study, anaerobic microcosms were established with arsenic-contaminated paddy soil to investigate arsenic transformation as well as the potentially active microbial community and their transcriptional activities in the presence of biochar. The results demonstrated that biochar can simultaneously stimulate microbial reduction of As(V) and Fe(III), releasing high levels of As(III) into the soil solution relative to the control. Total RNAs were extracted to profile the potentially active microbial communities, which suggested that biochar increased the abundance of arsenic- and iron-related bacteria, such as Geobacter, Anaeromyxobacter and Clostridium compared to the control. Reverse transcription, quantitative PCR (RT-qPCR) showed that the abundance of Geobacter transcripts were significantly stimulated by biochar throughout the incubation. Furthermore, significant positive correlations were observed between the abundance of Geobacter transcripts and As(V) concentrations, and between that of Clostridium transcripts and Fe(III) concentrations in biochar-amended microcosms. Our findings suggest that biochar can stimulate the activity of metal-reducing bacteria to promote arsenic mobility. The Geobacter may contribute to As(V) reduction in the presence of biochar, while Clostridium has a role in Fe(III) reduction.

Bioaugmentation with an acetate-type fermentation bacterium Acetobacteroides hydrogenigenes improves methane production from corn straw.

The effect of bioaugmentation with an acetate-type fermentation bacterium in the phylum Bacteroidetes on the anaerobic digestion of corn straw was evaluated by batch experiments. Acetobacteroides hydrogenigenes is a promising strain for bioaugmentation with relatively high growth rate, hydrogen yields and acetate tolerance, which ferments a broad spectrum of pentoses, hexoses and polyoses mainly into acetate and hydrogen. During corn straw digestion, bioaugmentation with A. hydrogenigenes led to 19-23% increase of the methane yield, with maximum of 258.1 mL/g-corn straw achieved by 10% inoculation (control, 209.3 mL/g-corn straw). Analysis of lignocellulosic composition indicated that A. hydrogenigenes could increase removal rates of cellulose and hemicelluloses in corn straw residue by 12% and 5%, respectively. Further experiment verified that the addition of A. hydrogenigenes could improve the methane yields of methyl cellulose and xylan (models for cellulose and hemicelluloses, respectively) by 16.8% and 7.0%.

Hydrogenispora ethanolica gen. nov., sp. nov., an anaerobic carbohydrate-fermenting bacterium from anaerobic sludge.

An anaerobic, spore-forming, ethanol-hydrogen-coproducing bacterium, designated LX-BT, was isolated from an anaerobic sludge treating herbicide wastewater. Cells of strain LX-BT were non-motile rods (0.3-0.5×3.0-18.0 µm). Spores were terminal with a bulged sporangium. Growth occurred at 20-50 °C (optimum 37-45 °C), pH 5.0-8.0 (optimum pH 6.0-7.7) and 0-2.5% (w/v) NaCl. The strain could grow fermentatively on glucose, maltose, arabinose, fructose, xylose, ribose, galactose, mannose, raffinose, sucrose, pectin, starch, glycerol, fumarate, tryptone and yeast extract. The major end-products of glucose fermentation were acetate, ethanol and hydrogen. Yeast extract was not required but stimulated growth. Nitrate, sulfate, thiosulfate, elemental sulfur, sulfite, anthraquinone-2,6-disulfonate, fumarate and Fe (III) nitrilotriacetate were not used as terminal electron acceptors. The G+C content of the genomic DNA was 56.1 mol%. The major cellular fatty acids were anteiso-C15:0, iso-C14:0 and C16:0. The most abundant polar lipids of strain LX-BT were diphosphatidylglycerol and phosphatidylglycerol. 16S rRNA gene sequence analysis revealed that it belongs to an as-yet-unidentified taxon at the order- or class-level (OPB54) within the phylum Firmicutes, showing 86.5% sequence similarity to previously described species of the Desulfotomaculum cluster. The name Hydrogenispora ethanolica gen. nov., sp. nov. is proposed to accommodate strain LX-BT (=DSM 25471T=JCM 18117T=CGMCC 1.5175T) as the type strain.

Molecular characterization of bacterial and archaeal communities in a full-scale anaerobic reactor treating corn straw.

A 16S rRNA gene-based method was used to characterize the structure of bacterial and archaeal communities in a full-scale, anaerobic reactor treating corn straw. Degradability experiment indicated biogas slurry had high microbial activity, the TS removal rate was 53% and the specific methanogenic activity was 86 mL CH4 g VSS(-1) d(-1). During anaerobic degradation of corn straw, volatile acids and aromatic compounds (p-cresol, phenylpropionate, phenol and benzoate) were detected as transient intermediates. Phylogenetic analysis revealed bacterial community exhibited high diversity, 69 bacterial phylotypes in 13 phyla were identified. Firmicutes (48.3%), Chloroflexi (20.1%), Actinobacteria (9.1%), Bacteroidetes (7.7%), and Proteobacteria (7.2%) represented the most abundant bacterial phyla. Hydrolytic and fermentative bacteria were major bacterial populations. Moreover, a relatively high proportion of syntrophic propionate and aromatic compounds degrading bacteria were detected. In the archaeal clone library, 11 archaeal phylotypes affiliated with two phyla of Crenarchaeota (10%) and Euryarchaeota (90%) were identified.

[Phylogenetic analysis of methanogenic corn stalk degrading microbial communities].

Methanogenic corn stalk degrading enrichment cultures were constructed using corn stalk as the sole carbon source and eight types of environmental samples as inocula. All the cultures could degrade corn stalk within 30-50 days and the total solids (TS) removal rates were in the range of 30%-40%. In six out of eight cultures, the cumulative methane yields per gram TS were 62.1-118.4 mL x g(-1), with acetate, propionate and butyrate as the major volatile fatty acids (100-500 mg x L(-1)), and the final pH were 6.5-6.7. In the other two cultures, the cumulative methane yields per gram TS were 8.5-9.7 mL xg(-1), while the concentrations of acetate were high (1200 mg x L(-1)), and the final pH were low (5.6-5.9). The bacterial and archaeal structures in eight enrichments were investigated with a 16S rRNA genes-based clone library method. Clones belonging to the bacterial phyla Firmicutes, Bacteroidetes, Synergistetes and Thermotogae were observed in abundance within the bacterial clone libraries, which accounted for 37.8%, 34.3%, 11.6% and 6.4% of the total number of bacterial clones, respectively. Within the domain Archaea, clones affiliated with the classes Methanomicrobia and Methanobacteria were found to be abundant in the archaeal clone libraries, which accounted for 61.1% and 38.9% of the total number of archaeal clones, respectively.