Arsenite Oxidase Gene Research Articles

Novel insights into the co-selection of metal-driven antibiotic resistance in bacteria: a study of arsenic and antibiotic co-exposure.

The simultaneous development of antibiotic resistance in bacteria due to metal exposure poses a significant threat to the environment and human health. This study explored how exposure to both arsenic and antibiotics affects the ability of an arsenite oxidizer, Achromobacter xylosoxidans CAW4, to transform arsenite and its antibiotic resistance patterns. The bacterium was isolated from arsenic-contaminated groundwater in the Chandpur district of Bangladesh. We determined the minimum inhibitory concentration (MIC) of arsenite, cefotaxime, and tetracycline for A. xylosoxidans CAW4, demonstrating a multidrug resistance (MDR) trait. Following this determination, we aimed to mimic an environment where A. xylosoxidans CAW4 was exposed to both arsenite and antibiotics. We enabled the strain to grow in sub-MIC concentrations of 1mM arsenite, 40µg/mL cefotaxime, and 20µg/mL tetracycline. The expression dynamics of the arsenite oxidase (aioA) gene in the presence or absence of antibiotics were analyzed. The findings indicated that simultaneous exposure to arsenite and antibiotics adversely affected the bacteria's capacity to metabolize arsenic. However, when arsenite was present in antibiotics-containing media, it promoted bacterial growth. The study observed a global downregulation of the aioA gene in arsenic-antibiotic conditions, indicating the possibility of increased susceptibility through co-resistance across the entire bacterial population of the environment. This study interprets that bacterial arsenic-metabolizing ability can rescue the bacteria from antibiotic stress, further disseminating environmental cross-resistance. Therefore, the co-selection of metal-driven antibiotic resistance in bacteria highlights the need for effective measures to address this emerging threat to human health and the environment.

Metagenomic and culture-dependent approaches unveil active microbial community and novel functional genes involved in arsenic mobilization and detoxification in groundwater

BackgroundArsenic (As) and its species are major pollutants in ecological bodied including groundwater in Bangladesh rendering serious public health concern. Bacteria with arsenotrophic genes have been found in the aquifer, converting toxic arsenite [As (III)] to less toxic arsenate [As (V)] that is easily removed using chemical and biological trappers. In this study, genomic and metagenomic approaches parallel to culture-based assay (Graphical abstract) have made it possible to decipher phylogenetic diversity of groundwater arsenotrophic microbiomes along with elucidation of their genetic determinants.ResultsSeventy-two isolates were retrieved from six As-contaminated (average As concentration of 0.23 mg/L) groundwater samples from Munshiganj and Chandpur districts of Bangladesh. Twenty-three isolates harbored arsenite efflux pump (arsB) gene with high abundance, and ten isolates possessing arsenite oxidase (aioA) gene, with a wide range of minimum inhibitory concentration, MICAs (2 to 32 mM), confirming their role in arsenite metabolism. There was considerable heterogeneity in species richness and microbial community structure. Microbial taxa from Proteobacteria, Firmicutes and Acidobacteria dominated these diversities. Through these combinatorial approaches, we have identified potential candidates such as, Pseudomonas, Acinetobacter, Stenotrophomonas, Achromobacter, Paraburkholderia, Comamonas and Klebsiella and associated functional genes (arsB, acr3, arsD, arsH, arsR) that could significantly contribute to arsenite detoxification, accumulation, and immobilization.ConclusionsCulture-dependent and -independent shotgun metagenomic investigation elucidated arsenotrophic microbiomes and their functions in As biogeochemical transformation. These findings laid a foundation for further large-scale researches on the arsenotrophic microbiomes and their concurrent functions in As biogeochemical transformation in As-contaminated areas of Bangladesh and beyond.

Bayesian network highlights the contributing factors for efficient arsenic phytoextraction by Pteris vittata in a contaminated field

Phytoextraction is a low-cost and eco-friendly method for removing pollutants, such as arsenic (As), from contaminated soil. One of the most studied As hyperaccumulators for soil remediation include Pteris vittata. Although phytoextraction using plant-assisted microbes has been considered a promising soil remediation method, microbial harnessing has not been achieved due to the complex and difficult to understand interactions between microbes and plants. This problem can possibly be addressed with a multi-omics approach using a Bayesian network. However, limited studies have used Bayesian networks to analyze plant–microbe interactions. Therefore, to understand this complex interaction and to facilitate efficient As phytoextraction using microbial inoculants, we conducted field cultivation experiments at two sites with different total As contents (62 and 8.9 mg/kg). Metabolome and microbiome data were obtained from rhizosphere soil samples using nuclear magnetic resonance and high-throughput sequencing, respectively, and a Bayesian network was applied to the obtained multi-omics data. In a highly As-contaminated site, inoculation with Pseudomonas sp. strain m307, which is an arsenite-oxidizing microbe having multiple copies of the arsenite oxidase gene, increased As concentration in the shoots of P. vittata to 157.5 mg/kg under this treatment; this was 1.5-fold higher than that of the other treatments. Bayesian network demonstrated that strain m307 contributed to As accumulation in P. vittata. Furthermore, the network showed that microbes belonging to the MND1 order positively contributed to As accumulation in P. vittata. Based on the ecological characteristics of MND1, it was suggested that the rhizosphere of P. vittata inoculated with strain m307 was under low-nitrogen conditions. Strain m307 may have induced low-nitrogen conditions via arsenite oxidation accompanied by nitrate reduction, potentially resulting in microbial iron reduction or the prevention of microbial iron oxidation. These conditions may have enhanced the bioavailability of arsenate, leading to increased As accumulation in P. vittata.

Cloning and functional characterization of arsenite oxidase (aoxB) gene associated with arsenic transformation in Pseudomonas sp. strain AK9

Arsenic transforming bacterial strains belong to genus Pseudomonas sp.AK9 (KY569424), were isolated from the middle Gangetic plains of Bihar, India. The Pseudomonas sp. AK9 strains were able to transform toxic arsenite to a less toxic arsenate. In the present work, the presence of different arsenic resistance genes (aoxB, arsB, acr3 and aoxAB) were observed in isolated strain. Furthermore, the aoxB gene was amplified from genomic DNA of AK9, cloned in E.coli/DH5αcells, and sequenced. The BLASTn results and phylogenetic study of the aoxB gene showed 95.32 % and 90.07 % identity with the large subunit of aoxB gene of previous reported Thiomonas arsenivorans strain DSM16361 and Thiomonas arsenivorans strain b6, respectively. Further overhang primers were designed for amplifications of full length aoxB gene (∼1200 bp), and cloned in to the expression vector and host E.coli/BL21 cells. The GST-aoxB gene was expressed in BL21 cells, and a profound expression product of ∼ 72 kDa was observed in SDS PAGE. The detection of a large subunit (aoxB) of arsenate oxidase protein in western blotting assay affirmed the expression of aoxB gene in recombinant E.coli/BL21 clone. Further, the recombinant E.coli/BL21cells showed increased growth than the normal E.coli/BL21 cells against As (III). Thus, this study showed the presence of aoxB gene in Pseudomonas sp. AK9 genome which regulates the resistant ability to arsenic toxicity.

Empirical Evidence of Arsenite Oxidase Gene as an Indicator Accounting for Arsenic Phytoextraction by Pteris vittata.

Arsenic (As) is a toxic semi-metallic element that is ubiquitous in the environment and poses serious human health risks. Phytoextraction by Pteris vittata is considered a low-cost and environmentally friendly approach to treat As-contaminated soil. P. vittata mainly absorbs arsenate thus the bioavailability of As to P. vittata depends on the chemical form of As. Microbial redox of As contributes to the biogeochemical cycling of As, and rhizobacterium-assisted phytoextraction by P. vittata was proposed. In this study, this microbe-assisted phytoextraction was applied to two fields, and the effectiveness of phytoextraction was evaluated. The results revealed that P. vittata was able to grow in temperate and subarctic climate zones. The biomass was influenced by the weather, and the As concentration in plants was dependent on the As content in the soil. The ratio of arsenite oxidase genes (aioA-like genes) to 16S rRNA genes was employed to evaluate the effect of As phytoextraction, and the results exhibited that the ratio was related to the As concentration in P. vittata. Our results showed that arsenite oxidation in the rhizosphere might not be achieved by single-strain inoculation, while this study provided empirical evidence that the rhizospheric aioA-like genes could be an indicator for evaluating the effectiveness of As phytoextraction.

Bioaccumulation and detoxification of trivalent arsenic by Achromobacter xylosoxidans BHW-15 and electrochemical detection of its transformation efficiency

Arsenotrophic bacteria play an essential role in lowering arsenic contamination by converting toxic arsenite [As (III)] to less toxic and less bio-accumulative arsenate [As (V)]. The current study focused on the qualitative and electrocatalytic detection of the arsenite oxidation potential of an arsenite-oxidizing bacteria A. xylosoxidans BHW-15 (retrieved from As-contaminated tube well water), which could significantly contribute to arsenic detoxification, accumulation, and immobilization while also providing a scientific foundation for future electrochemical sensor development. The minimum inhibitory concentration (MIC) value for the bacteria was 15 mM As (III). Scanning Electron Microscopy (SEM) investigation validated its intracellular As uptake capacity and demonstrated a substantial association with the MIC value. During the stationary phase, the strain’s As (III) transformation efficiency was 0.0224 mM/h. Molecular analysis by real-time qPCR showed arsenite oxidase (aioA) gene expression increased 1.6-fold in the presence of As (III) compared to the untreated cells. The immobilized whole-cell also showed As (III) conversion up to 18 days. To analyze the electrochemical oxidation in water, we developed a modified GCE/P-Arg/ErGO-AuNPs electrode, which successfully sensed and quantified conversion of As (III) into As (V) by accepting electrons; implying a functional As oxidase enzyme activity in the cells. To the best of our knowledge, this is the first report on the electrochemical observation of the As-transformation mechanism with Achromobactersp. Furthermore, the current work highlighted that our isolate might be employed as a promising candidate for arsenic bioremediation, and information acquired from this study may be helpful to open a new window for the development of a cost-effective, eco-friendly biosensor for arsenic species detection in the future.

New Arsenite Oxidase Gene (aioA) PCR Primers for Assessing Arsenite-Oxidizer Diversity in the Environment Using High-Throughput Sequencing.

Gene encoding the large subunit of As(III) oxidase (AioA), an important component of the microbial As(III) oxidation system, is a widely used biomarker to characterize As(III)-oxidizing communities in the environment. However, many studies were restricted to a few sequences generated by clone libraries and Sanger sequencing, which may have underestimated the diversity of As(III)-oxidizers in natural environments. In this study, we designed a primer pair, 1109F (5′-ATC TGG GGB AAY RAC AAY TA−3′) and 1548R (5′-TTC ATB GAS GTS AGR TTC AT−3′), targeting gene sequence encoding for the conserved molybdopterin center of the AioA protein, yielding amplicons approximately 450 bp in size that are feasible for highly parallel amplicon sequencing. By utilizing in silico analyses and the experimental construction of clone libraries using Sanger sequencing, the specificity and resolution of 1109F/1548R are approximated with two other previously published and commonly used primers, i.e., M1-2F/M3-2R and deg1F/deg1R. With the use of the 1109F/1548R primer pair, the taxonomic composition of the aioA genes was similar both according to the Sanger and next-generation sequencing (NGS) platforms. Furthermore, high-throughput amplicon sequencing using the primer pair, 1109F/1548R, successfully identified the well-known As(III)-oxidizers in paddy soils and sediments, and they also revealed the differences in the community structure and composition of As(III)-oxidizers in above two biotopes. The random forest analysis showed that the dissolved As(III) had the highest relative influence on the Chao1 index of the aioA genes. These observations demonstrate that the newly designed PCR primers enhanced the ability to detect the diversity of aioA-encoding microorganisms in environments using highly parallel short amplicon sequencing.

Drivers and ecological consequences of arsenite detoxification in aged semi-aerobic landfill

Microbial populations responsible for arsenite [As(III)] detoxification were examined in aged refuse treated with 75 μM As(III) under semi-aerobic conditions. As(III) was rapidly oxidized to As(V) via microbial activity, and substantial As was fixed in the solid phase. The abundance of arsenite oxidase genes (aioA) was about four times higher in the moderate As(III) stressed treatment than in the untreated control. Network analysis of microbial community 16S rRNA genes based on MRT (random matrix theory) further illuminated details about microbe-microbe interactions, and showed six ecological clusters. A total of 166 “core” taxa were identified by within-module connectivity and among-module connectivity values. When compared with the control treatment without As(III), 12 putative keystone operational taxonomic units were positively correlated with As(III) oxidation, of which 10 of these were annotated to genera level. Eight genera were associated with As(III) detoxification: Pseudomonas, Paenalcaligenes, Proteiniphilum, Moheibacter, Mobilitalea, Anaerosporobacter, Syntrophomonas and Pusillimonas. Most of those putative keystone taxa were rare species in landfill, which suggests that low-abundance taxa might significantly contribute to As(III) oxidation.

Bacteria responsible for nitrate-dependent antimonite oxidation in antimony-contaminated paddy soil revealed by the combination of DNA-SIP and metagenomics

Antimonite (Sb(III)) oxidation (SbO) can decrease the toxicity of antimony (Sb) and its uptake into rice, thus serving an ecological role in bioremediation of Sb contamination in rice paddies and decreasing the translocation and accumulation of Sb in rice. Nitrate as the electron acceptor can be coupled to SbO in anoxic environments, which, however, has not been reported in paddy soils. Here we investigate the potential for nitrate-dependent SbO in Sb-contaminated rice paddies and identify nitrate-dependent Sb(III)-oxidizing bacteria (SbOB) using stable isotope probing (SIP) coupled with amplicon and shotgun metagenomic sequencing. Anaerobic SbO was exclusively observed in the paddy soil amended with both Sb(III) and NO3−, whereas no apparent SbO was detected in the soil amended with Sb(III) only. The increasing abundance of the arsenite oxidase gene (aioA) suggests that nitrate-dependent SbO was catalysed by microorganisms harbouring the aioA gene. After 60-day DNA-SIP incubation, an obvious shift in the relative abundance of aioA gene to heavy DNA fractions occurred only in the treatment amended with 13C–NaHCO3, Sb(III) and NO3−, suggesting the incorporation of 13C by nitrate-dependent SbOB. Accordingly, a number of putative nitrate-dependent SbOB were identified in the paddy soil, including Azoarcus, Azospira and Chelativorans. Metagenomic analysis further revealed that they contained aioA genes and genes involved in denitrification and carbon fixation, supporting their capability for nitrate-dependent SbO. These observations suggested the occurrence of nitrate-dependent SbO in paddy soils. A number of putative nitrate-dependent SbOB (i.e., Azoarcus, Azospira and Chelativorans) were reported here, which expands our current knowledge regarding the diversity of nitrate-dependent SbOB. In addition, this study provides a proof of concept using DNA-SIP to identify nitrate-dependent SbOB.

Effects of Arsenic and Iron on the Community and Abundance of Arsenite-Oxidizing Bacteria in an Arsenic-Affected Groundwater Aquifer

Arsenic (As) contamination of groundwater aquifers is a global environmental problem, especially in South and Southeast Asian regions, and poses a risk to human health. Arsenite-oxidizing bacteria that transform As(III) to less toxic As(V) can be potentially used as a groundwater As remediation strategy. This study aimed to examine the community and abundance of arsenite-oxidizing bacteria in groundwater with various As concentrations from Rayong Province, Thailand using PCR-cloning-sequencing and quantitative PCR (qPCR) of catalytic subunit of arsenite oxidase gene (aioA). Key factors influencing their community and abundance were also identified. The results demonstrated that arsenite-oxidizing bacteria retrieved from groundwater were phylogenetically related to Betaproteobacteria and Alphaproteobacteria. The aioA gene abundances ranged from 8.6 × 101 to 1.1 × 104 copies per ng of genomic DNA, accounting for 0.16-1.37% of the total 16S rRNA bacterial gene copies. Although the abundance of arsenite-oxidizing bacteria in groundwater was low, groundwater with As(III) dominance likely promoted their abundance which possibly played an important role in chemolithoautotrophic oxidation of As(III) to As(V). Fe and As(III) were the major environmental factors influencing the community and abundance of arsenite-oxidizing bacteria. The knowledge gained from this study can be used to further contribute to the development of bioremediation strategies for As removal from groundwater resources.

Survival strategies and dominant phylotypes of maize-rhizosphere microorganisms under metal(loid)s contamination

Profound understanding of metabolic capabilities of microorganisms is prerequisite for effective bioremediation. However, microorganisms in neutral-alkaline soils affected by metal(loid)s contamination around mining area remains under-studied. Herein, microbial community and survival strategies of microorganisms in rhizospheric soils highly contaminated by multiple metal(loid)s (such as lead (Pb), zinc (Zn), cadmium (Cd), arsenic (As), and antimony (Sb)) from a carbonate-hosted PbZn deposit in South China were profiled via 16S ribosomal RNA (rRNA) gene sequence and shotgun metagenomic analysis. The results indicated that enriched levels of toxic metal(loid)s had significant impacts on the diversity and abundance of rhizosphere microorganisms. The siderophores produced by Sphingomonas might enhance maize uptake of toxic metal(loid)s. Variovorax and Terrimonas may be potential remediation microorganisms for metal(loid)s contamination. Shotgun metagenomics indicated that metal(loid)s related genes (such as zntA and arsC) may enhance the tolerance of rhizosphere microorganisms to toxic metal(loid)s. The lack of arsenite oxidase gene (aoxAB) may inhibit the biogeochemical cycle of As in the studied area. The findings are critical for facilitating microbiome bioremediation for soil affected by neutral-alkaline mining.

Microbial community structure in aquifers associated with arsenic: analysis of 16S rRNA and arsenite oxidase genes.

The microbiomes of deep and shallow aquifers located in an agricultural area, impacted by an old tin mine, were explored to understand spatial variation in microbial community structures and identify environmental factors influencing microbial distribution patterns through the analysis of 16S rRNA and aioA genes. Although Proteobacteria, Cyanobacteria, Actinobacteria, Patescibacteria, Bacteroidetes, and Epsilonbacteraeota were widespread across the analyzed aquifers, the dominant taxa found in each aquifer were unique. The co-dominance of Burkholderiaceae and Gallionellaceae potentially controlled arsenic immobilization in the aquifers. Analysis of the aioA gene suggested that arsenite-oxidizing bacteria phylogenetically associated with Alpha-, Beta-, and Gamma proteobacteria were present at low abundance (0.85 to 37.13%) and were more prevalent in shallow aquifers and surface water. The concentrations of dissolved oxygen and total phosphorus significantly governed the microbiomes analyzed in this study, while the combination of NO3--N concentration and oxidation-reduction potential significantly influenced the diversity and abundance of arsenite-oxidizing bacteria in the aquifers. The knowledge of microbial community structures and functions in relation to deep and shallow aquifers is required for further development of sustainable aquifer management.

Microaerophilic Oxidation of Fe(II) Coupled with Simultaneous Carbon Fixation and As(III) Oxidation and Sequestration in Karstic Paddy Soil.

Microaerophilic Fe(II)-oxidizing bacteria are often chemolithoautotrophs, and the Fe(III) (oxyhydr)oxides they form could immobilize arsenic (As). If such microbes are active in karstic paddy soils, their activity would help increase soil organic carbon and mitigate As contamination. We therefore used gel-stabilized gradient systems to cultivate microaerophilic Fe(II)-oxidizing bacteria from karstic paddy soil to investigate their capacity for Fe(II) oxidation, carbon fixation, and As sequestration. Stable isotope probing demonstrated the assimilation of inorganic carbon at a maximum rate of 8.02 mmol C m-2 d-1. Sequencing revealed that Bradyrhizobium, Cupriavidus, Hyphomicrobium, Kaistobacter, Mesorhizobium, Rhizobium, unclassified Phycisphaerales, and unclassified Opitutaceas were fixing carbon. Fe(II) oxidation produced Fe(III) (oxyhydr)oxides, which can absorb and/or coprecipitate As. Adding As(III) decreased the diversity of functional bacteria involved in carbon fixation, the relative abundance of predicted carbon fixation genes, and the amount of carbon fixed. Although the rate of Fe(II) oxidation was also lower in the presence of As(III), over 90% of the As(III) was sequestered after oxidation. The potential for microbially mediated As(III) oxidation was revealed by the presence of arsenite oxidase gene (aioA), denoting the potential of the Fe(II)-oxidizing and autotrophic microbial community to also oxidize As(III). Thisstudy demonstrates that carbon fixation coupled to Fe(II) oxidation can increase the carbon content in soils by microaerophilic Fe(II)-oxidizing bacteria, as well as accelerate As(III) oxidation and sequester it in association with Fe(III) (oxyhydr)oxides.

Potential of versatile bacteria isolated from activated sludge for the bioremediation of arsenic and antimony

Two bacterial strains belonging to the genus Shinella were isolated from the activated sludge of a domestic wastewater treatment plant and could autotrophically oxidize As(III) to As(V) under aerobic conditions. Kinetic studies indicated that the autotrophic As(III) oxidation rate of both strains matched the Monod model well, and strain VKA3 exhibited higher kinetic parameters compared to strain VKA4 (μmax = 124.16 μM/h and Ks = 4407.19 μM vs. μmax = 59.00 μM/h and Ks = 1787.03 μM). The versatility of both strains was expressed through their flexible growth under heterotrophic conditions in the presence of yeast extract and anaerobic As(III) coupled with denitrification. Both strains can oxidize Sb(III) to Sb(V) under aerobic conditions. The arsenite oxidase (aioA) gene, which might be involved in As(III) and Sb(III) oxidation, was detected in the isolated strains. This study demonstrated that the versatile bacteria with potential applications for bioremediation of As and Sb could be obtained from an easily accessible microbiological source.

Long-term effectiveness of microbe-assisted arsenic phytoremediation by Pteris vittata in field trials

Phytoremediation is a promising inexpensive method of detoxifying arsenic (As) contaminated soils using plants and associated soil microorganisms. The potential of Pteris vittata to hyperaccumulate As contamination has been investigated widely. Since As(V) is efficiently taken up by P. vittata than As(III), As speciation by associated rhizobacteria could offer enormous possibility to enhance As phytoremediation. Specifically, increased rhizobacteria mediated As(III) to As(V) conversion appeared to be a crucial step in As mobilization and translocation. In this study, Pseudomonasvancouverensis strain m318 with the potential to improve As phytoremediation was inoculated to P. vittata in a field trial for three years to evaluate its long-term efficacy and stability for enhancing As phytoextraction. The biomass, As concentration, and As accumulation of ferns showed to be increased by inoculation treatment. Although this trend occasionally declined which may be accounted to lower As concentration in soil and amount of precipitation during experiments, the potential of inoculation was observed in increased enrichment coefficients. Further, the arsenite oxidase (aioA-like) genes in the rhizosphere were detected to evaluate the influence of inoculation on As phytoremediation. The findings of this study suggested the potential application of rhizosphere regulation to improve phytoremediation technologies for As contaminated soils. However, the conditions which set the efficacy of this method could be further optimized.

Diverse respiratory capacity among Thermus strains from US Great Basin hot springs.

Thermus species are thermophilic heterotrophs, with most capable of using a variety of organic and inorganic electron donors for respiration. Here, a combined cultivation-independent and -dependent approach was used to explore the diversity of Thermus in Great Boiling Spring (GBS) and Little Hot Creek (LHC) in the US Great Basin. A cultivation-independent 16S rRNA gene survey of ten LHC sites showed that Thermus made up 0-3.5% of sequences and were predominately Thermus thermophilus. 189 Thermus isolates from GBS and LHC were affiliated with T. aquaticus (73.0%), T. oshimai (25.4%), T. sediminis (1.1%), and T. thermophilus (0.5%), with T. aquaticus and T. oshimai forming biogeographic clusters. 22 strains were selected for characterization, including chemolithotrophic oxidation of thiosulfate and arsenite, and reduction of ferric iron, polysulfide, and nitrate, revealing phenotypic diversity and broad respiratory capability within each species. PCR demonstrated the wide distribution of aerobic arsenite oxidase genes. A GBS sediment metaproteome contained sulfite oxidase and Fe3+ ABC transporter permease peptides, suggesting sulfur and iron transformations in situ. This study expands our knowledge of the physiological diversity of Thermus, suggesting widespread chemolithotrophic and anaerobic respiration phenotypes, and providing a foundation for better understanding the ecology of this genus in thermal ecosystems.

Biological Fe(II) and As(III) oxidation immobilizes arsenic in micro-oxic environments

Fe(III) oxyhydroxides play critical roles in arsenic immobilization due to their strong surface affinity for arsenic. However, the role of bacteria in Fe(II) oxidation and the subsequent immobilization of arsenic has not been thoroughly investigated to date, especially under the micro-oxic conditions present in soils and sediments where these microorganisms thrive. In the present study, we used gel-stabilized gradient systems to investigate arsenic immobilization during microaerophilic microbial Fe(II) oxidation and Fe(III) oxyhydroxide formation. The removal and immobilization of dissolved As(III) and As(V) proceeded via the formation of biogenic Fe(III) oxyhydroxides through microbial Fe(II) oxidation. After 30 days of incubation, the concentration of dissolved arsenic decreased from 600 to 4.8 μg L−1. When an Fe(III) oxyhydroxide formed in the presence of As(III), most of the arsenic ultimately was found as As(V), indicating that As(III) oxidation accompanied arsenic immobilization. The structure of the microbial community in As(III) incubations was highly differentiated with respect to the As(V)-bearing ending incubations. The As(III)-containing incubations contained the arsenite oxidase gene, suggesting the potential for microbially mediated As(III) oxidation. The findings of the present study suggest that As(III) immobilization can occur in micro-oxic environments after microbial Fe(II) oxidation and biogenic Fe(III) oxyhydroxide formation via the direct microbial oxidation of As(III) to As(V). This study demonstrates that microbial Fe(II) and As(III) oxidation are important geochemical processes for arsenic immobilization in micro-oxic soils and sediments.

Ample Arsenite Bio-Oxidation Activity in Bangladesh Drinking Water Wells: A Bonanza for Bioremediation?

Millions of people worldwide are at risk of arsenic poisoning from their drinking water. In Bangladesh the problem extends to rural drinking water wells, where non-biological solutions are not feasible. In serial enrichment cultures of water from various Bangladesh drinking water wells, we found transfer-persistent arsenite oxidation activity under four conditions (aerobic/anaerobic; heterotrophic/autotrophic). This suggests that biological decontamination may help ameliorate the problem. The enriched microbial communities were phylogenetically at least as diverse as the unenriched communities: they contained a bonanza of 16S rRNA gene sequences. These related to Hydrogenophaga, Acinetobacter, Dechloromonas, Comamonas, and Rhizobium/Agrobacterium species. In addition, the enriched microbiomes contained genes highly similar to the arsenite oxidase (aioA) gene of chemolithoautotrophic (e.g., Paracoccus sp. SY) and heterotrophic arsenite-oxidizing strains. The enriched cultures also contained aioA phylotypes not detected in the previous survey of uncultivated samples from the same wells. Anaerobic enrichments disclosed a wider diversity of arsenite oxidizing aioA phylotypes than did aerobic enrichments. The cultivatable chemolithoautotrophic and heterotrophic arsenite oxidizers are of great interest for future in or ex-situ arsenic bioremediation technologies for the detoxification of drinking water by oxidizing arsenite to arsenate that should then precipitates with iron oxides. The microbial activities required for such a technology seem present, amplifiable, diverse and hence robust.

Potential of biological arsenite oxidation in sand filtration units at arsenic\u2013iron removal plants (AIRPs) in Bangladesh

This study is aimed to investigate biological oxidation of arsenic [As(III)] in sand component of arsenic–iron removal plant (AIRP). Water and sand samples were collected from the municipal and household AIRP units (MAIRP and HAIRPs) at Manikganj district, Bangladesh. These samples were analyzed using arsenic speciation cartridge with inductively coupled plasma-mass spectrometry for arsenic and iron detection and phylogenetic analysis based on aoxB gene sequencing. It was observed that arsenic concentration in the raw water, before and after sand filtration, ranged from 13.0–81.0 to 4.0–21.0 μg/L, respectively, which majorly contained reduced arsenic [As(III)] in the inlet and oxidized arsenic [As(V)] in the outlet. The results of our batch experiments showed that indeed sand unit of MAIRP oxidized As(III) with 36 g/h estimated biological oxidation potential which was considered enough to oxidize all the As(III) loading (6.3 g/h) in the influent. Additionally, bacterial arsenite oxidase gene (aoxB) was detected in the sand sample and has 80% sequence similarity with Polymorphum gilvum, an alphaproteobacteria on the phylogenetic tree. This study, therefore, revealed that AIRP sand units have enough potential of biological activity to ensure overall arsenic removal through As(III) oxidation. However, future research is necessary to unfold the basic mechanistic approach explored in this plant for further modification if need arises.

Arsenite biotransformation and bioaccumulation by Klebsiella pneumoniae strain SSSW7 possessing arsenite oxidase (aioA) gene.

Arsenite oxidizing Klebsiella pneumoniae strain SSSW7 isolated from shipyard waste Goa, India showed a minimum inhibitory concentration of 21mM in mineral salts medium. The strain possessed a small supercoiled plasmid and PCR amplification of arsenite oxidase gene (aioA) was observed on plasmid as well as chromosomal DNA. It was confirmed that arsenite oxidase enzyme was a periplasmic protein with a 47% increase in arsenite oxidase activity at 1mM sodium arsenite. Scanning electron microscopy coupled with electron dispersive X-ray spectroscopic (SEM-EDS) analysis of 15mM arsenite exposed cells revealed long chains of cells with no surface adsorption of arsenic. Transmission electron microscopy combined with electron dispersive X-ray spectroscopic (TEM-EDS) analysis demonstrated plasma membrane disruption, cytoplasmic condensation and periplasmic accumulation of arsenic. The bacterial strain oxidized 10mM of highly toxic arsenite to less toxic arsenate after 24h of incubation. Fourier transformed infrared (FTIR) spectroscopy confirmed the interaction of arsenite with functional groups present on the bacterial cell surface. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of 5mM arsenite exposed cells demonstrated over-expression of 87kDa and 14kDa proteins of two subunits aioA and aioB of heterodimer arsenite oxidase enzyme as compared to control cells. Therefore, this bacterial strain might be employed as a potential candidate for bioremediation of arsenite contaminated environmental sites.