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Microbial electroactive biofilms dominated by Geoalkalibacter spp. from a highly saline-alkaline environment.

ABSTRACT: Understanding of the extreme microorganisms that possess extracellular electron transfer (EET) capabilities is pivotal to advance electromicrobiology discipline and to develop niche-specific microbial electrochemistry-driven biotechnologies. Here, we report on the microbial electroactive biofilms (EABs) possessing the outward EET capabilities from a haloalkaline environment of the Lonar lake. We used the electrochemical cultivation approach to enrich haloalkaliphilic EABs under 9.5?pH and 20?g/L salinity conditions. The electrodes controlled at 0.2?V vs. Ag/AgCl yielded the best-performing biofilms in terms of maximum bioelectrocatalytic current densities of 548?±?23 and 437?±?17?µA/cm2 with acetate and lactate substrates, respectively. Electrochemical characterization of biofilms revealed the presence of two putative redox-active moieties with the mean formal potentials of 0.183 and 0.333?V vs. Ag/AgCl, which represent the highest values reported to date for the EABs. 16S-rRNA amplicon sequencing of EABs revealed the dominance of unknown Geoalkalibacter sp. at ~80% abundance. Further investigations on the haloalkaliphilic EABs possessing EET components with high formal potentials might offer interesting research prospects in electromicrobiology.

SUBMITTER: Yadav S

PROVIDER: S-EPMC7555509 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Microbial electroactive biofilms dominated by Geoalkalibacter spp. from a highly saline-alkaline environment.

Yadav Sukrampal S Patil Sunil A SA

NPJ biofilms and microbiomes 20201013 1

Understanding of the extreme microorganisms that possess extracellular electron transfer (EET) capabilities is pivotal to advance electromicrobiology discipline and to develop niche-specific microbial electrochemistry-driven biotechnologies. Here, we report on the microbial electroactive biofilms (EABs) possessing the outward EET capabilities from a haloalkaline environment of the Lonar lake. We used the electrochemical cultivation approach to enrich haloalkaliphilic EABs under 9.5 pH and 20 g/L ...[more]

PMID: 33051461

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Project description:UNLABELLED: Anode-respiring bacteria (ARB) generate electric current in microbial electrochemical cells (MXCs) by channeling electrons from the oxidation of organic substrates to an electrode. Production of high current densities by monocultures in MXCs has resulted almost exclusively from the activity of Geobacter sulfurreducens, a neutrophilic freshwater Fe(III)-reducing bacterium and the highest-current-producing member documented for the Geobacteraceae family of the Deltaproteobacteria. Here we report high current densities generated by haloalkaliphilic Geoalkalibacter spp., thus broadening the capability for high anode respiration rates by including other genera within the Geobacteraceae. In this study, acetate-fed pure cultures of two related Geoalkalibacter spp. produced current densities of 5.0 to 8.3 and 2.4 to 3.3 A m(-2) under alkaline (pH 9.3) and saline (1.7% NaCl) conditions, respectively. Chronoamperometric studies of halophilic Glk. subterraneus DSM 23483 and alkaliphilic Glk. ferrihydriticus DSM 17813 suggested that cells performed long-range electron transfer through electrode-attached biofilms and not through soluble electron shuttles. Glk. ferrihydriticus also oxidized ethanol directly to produce current, with maximum current densities of 5.7 to 7.1 A m(-2) and coulombic efficiencies of 84 to 95%. Cyclic voltammetry (CV) elicited a sigmoidal response with characteristic onset, midpoint, and saturation potentials, while CV performed in the absence of an electron donor suggested the involvement of redox molecules in the biofilm that were limited by diffusion. These results matched those previously reported for actively respiring Gb. sulfurreducens biofilms producing similar current densities (~5 to 9 A m(-2)). IMPORTANCE: This study establishes the highest current densities ever achieved by pure cultures of anode-respiring bacteria (ARB) under alkaline and saline conditions in microbial electrochemical cells (MXCs) and provides the first electrochemical characterization of the genus Geoalkalibacter. Production of high current densities among the Geobacteraceae is no longer exclusive to Geobacter sulfurreducens, suggesting greater versatility for this family in fundamental and applied microbial electrochemical cell (MXC) research than previously considered. Additionally, this work raises the possibility that different members of the Geobacteraceae have conserved molecular mechanisms governing respiratory extracellular electron transfer to electrodes. Thus, the capacity for high current generation may exist in other uncultivated members of this family. Advancement of MXC technology for practical uses must rely on an expanded suite of ARB capable of using different electron donors and producing high current densities under various conditions. Geoalkalibacter spp. can potentially broaden the practical capabilities of MXCs to include energy generation and waste treatment under expanded ranges of salinity and pH.

Project description:Three kinds of alkaliphilic bacteria able to utilize thiocyanate (CNS-) at pH 10 were found in highly alkaline soda lake sediments and soda soils. The first group included obligate heterotrophs that utilized thiocyanate as a nitrogen source while growing at pH 10 with acetate as carbon and energy sources. Most of the heterotrophic strains were able to oxidize sulfide and thiosulfate to tetrathionate. The second group included obligately autotrophic sulfur-oxidizing alkaliphiles which utilized thiocyanate nitrogen during growth with thiosulfate as the energy source. Genetic analysis demonstrated that both the heterotrophic and autotrophic alkaliphiles that utilized thiocyanate as a nitrogen source were related to the previously described sulfur-oxidizing alkaliphiles belonging to the gamma subdivision of the division Proteobacteria (the Halomonas group for the heterotrophs and the genus Thioalkalivibrio for autotrophs). The third group included obligately autotrophic sulfur-oxidizing alkaliphilic bacteria able to utilize thiocyanate as a sole source of energy. These bacteria could be enriched on mineral medium with thiocyanate at pH 10. Growth with thiocyanate was usually much slower than growth with thiosulfate, although the biomass yield on thiocyanate was higher. Of the four strains isolated, the three vibrio-shaped strains were genetically closely related to the previously described sulfur-oxidizing alkaliphiles belonging to the genus Thioalkalivibrio. The rod-shaped isolate differed from the other isolates by its ability to accumulate large amounts of elemental sulfur inside its cells and by its ability to oxidize carbon disulfide. Despite its low DNA homology with and substantial phenotypic differences from the vibrio-shaped strains, this isolate also belonged to the genus Thioalkalivibrio according to a phylogenetic analysis. The heterotrophic and autotrophic alkaliphiles that grew with thiocyanate as an N source possessed a relatively high level of cyanase activity which converted cyanate (CNO-) to ammonia and CO2. On the other hand, cyanase activity either was absent or was present at very low levels in the autotrophic strains grown on thiocyanate as the sole energy and N source. As a result, large amounts of cyanate were found to accumulate in the media during utilization of thiocyanate at pH 10 in batch and thiocyanate-limited continuous cultures. This is a first direct proof of a "cyanate pathway" in pure cultures of thiocyanate-degrading bacteria. Since it is relatively stable under alkaline conditions, cyanate is likely to play a role as an N buffer that keeps the alkaliphilic bacteria safe from inhibition by free ammonia, which otherwise would reach toxic levels during dissimilatory degradation of thiocyanate.

Project description:The application of organic amendments to saline-alkaline soil has been recommended as an agricultural strategy to improve crop productivity and soil health. However, there has been limited research on how organic soil amendment strategies affect the health of oats and their associated rhizosphere fungal communities in saline-alkaline conditions. Thus, the objectives of this study were to understand the effects of oat cultivars with contrasting saline-alkaline tolerances and different amendments on plant morphologies, root exudates (soluble sugars and organic acids), and rhizosphere fungal communities in a saline-alkaline environment. Experiments were conducted on a saline-alkaline tolerant cultivar, Baiyan2, and a saline-alkaline sensitive cultivar, Caoyou1, under four different organic amendment strategies: 1. control (no amendment application), 2. bio-fertilizer application, 3. rotten straw application, and 4. a co-application of bio-fertilizer and rotten straw. Results showed that plant morphological characters of Baiyan2 were better than Caoyou1, and that soluble sugar and organic acid levels in the rhizosphere of Baiyan2 were significantly lower than Caoyou1. Compared to the control, oat root and plant development was significantly improved by the combined bio-fertilizer and rotten straw amendment. Bio-fertilizer application promoted malic and citric acid levels, contributing to a higher total organic acid level, and significantly increased the abundance of Rhizopus arrhizus and decreased the abundance of the fungal pathogens Alternaria, Cladosporium, Sarocladium and Heydenia of Ascomycota in both oat cultivars. All amendment treatments containing rotten straw, except the combined amendment in Baiyan2, significantly increased the relative abundance of Ascomycota (specifically Gibberella, Talaromyces, Fusarium, and Bipolaris) and decreased the relative abundance of R. arrhizus by reducing soluble sugar and organic acid levels. For the combined amendment in Baiyan2, there were no significant changes in Gibberella and Rhizopus between the control and amendment treatment. Our results suggest that co-application of bio-fertilizer and rotten straw, combined with a tolerant oat cultivar, is an effective method to increase crop productivity and enhance soil health in a saline-alkaline environment.

Dataset Information

Microbial electroactive biofilms dominated by Geoalkalibacter spp. from a highly saline-alkaline environment.

Publications

Microbial electroactive biofilms dominated by Geoalkalibacter spp. from a highly saline-alkaline environment.

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