Response to vanadate exposure in Ochrobactrum tritici strains.

Vanadium is a transition metal that has been added recently to the EU list of Raw Critical Metals. The growing needs of vanadium primarily in the steel industry justify its increasing economic value. However, because mining of vanadium sources (i. e. ores, concentrates and vanadiferous slags) is expanding, so is vanadium environmental contamination. Bioleaching comes forth as smart strategy to deal with supply demand and environmental contamination. It requires organisms that are able to mobilize the metal and at the same time are resistant to the leachate generated. Here, we investigated the molecular mechanisms underlying vanadium resistance in Ochrobactrum tritici strains. The highly resistant strain 5bvl1 was able to grow at concentrations > 30 mM vanadate, while the O. tritici type strain only tolerated < 3 mM vanadate concentrations. Screening of O. tritici single mutants (chrA, chrC, chrF and recA) growth during vanadate exposure revealed that vanadate resistance was associated with chromate resistance mechanisms (in particular ChrA, an efflux pump and ChrC, a superoxide dismutase). We also showed that sensitivity to vanadate was correlated with increased accumulation of vanadate intracellularly, while in resistant cells this was not found. Other up-regulated proteins found during vanadate exposure were ABC transporters for methionine and iron, suggesting that cellular responses to vanadate toxicity may also induce changes in unspecific transport and chelation of vanadate.

Attenuation of bacterial cytotoxicity of carbon nanotubes by riverine suspended solids in water.

The impact of solid particles on ecotoxicity of nanomaterials in water environments is poorly understood. This study investigated the effect of natural riverine suspended solids (SPS) on the cytotoxicity of single-walled carbon nanotubes (SWCNTs) towards a bacterium, Ochrobactrum sp. in water. Compared with SWCNT suspension without SPS, the presence of SPS at different concentrations ranging from 20 to 400 mg L-1 markedly increased the survival rates of bacteria exposed to 50 mg L-1 SWCNTs and bacterial survival rates increased with SPS concentrations by a power law. Sedimentation experiments and field emission scanning electron microscopy revealed the occurrence of heteroaggregation between SWCNTs and SPS, probably responsible for the reduced SWCNT toxicity. Furthermore, the extended Derjaguin-Landau-Verwey-Overbeek (ExDLVO) calculation showed the mitigated toxicity might also result from the decreased SWCNT-bacterium interaction energy with the increased SPS concentrations and the stronger SPS-SWCNT interaction than the SWCNT-bacterium interaction. This work provides new insights into our understanding of environmental hazards of engineered nanomaterials in aquatic systems.

Identification and analysis of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from glyphosate-resistant Ochrobactrum intermedium Sq20.

BACKGROUND: Glyphosate is a herbicide that acts by inhibition of the enzyme, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), involved in the catalysis of an essential step in the biosynthesis of aromatic amino acids. The objective of this study was the isolation of glyphosate-resistant bacterial strains and subsequent characterization of the gene(s) encoding glyphosate resistance in these isolates. Using an enrichment culture technique, a glyphosate-resistant bacterium, Ochrobactrum intermedium Sq20 was isolated from glyphosate-contaminated indigenous soil and characterized. RESULTS: An open reading frame (ORF) comprising of 1353 bp potentially encoding aroAO. intermediumSq20 was amplified from O. intermedium Sq20. It showed 97% homology with aroA genes from other Ochrobactrum spp. Physicochemical characterization revealed that aroAO. intermediumSq20 encodes a polypeptide of 450 amino acids with a calculated molecular mass of 48.9782 kDa and an isoelectric point of 5.21. Secondary structure prediction of AroAO. intermediumSq20 demonstrated a high percentage of random coils and α helices. Methodical optimization and validation of the protein structure helped to build a reliable protein model indicating the presence of 91.8% amino acid residues in most favoured regions. In addition, strain Sq20 was found to be capable of complete degradation of glyphosate at 500 mg L-1 initial concentration as the sole carbon and energy source within 4 days. CONCLUSION: A glyphosate-resistant bacterial strain O. intermedium Sq20 was discovered. Sequence analysis and structure modelling demonstrated that AroAO. intermediumSq20 closely resembles class II EPSPS and possesses high glyphosate resistance. This provides a good foundation for functional analysis of experimentally derived crystal structures. The cloning and characterization of AroAO. intermediumSq20 will further help in understanding its role at the molecular level and its potential use in the production of glyphosate-resistant transgenic crops. © 2017 Society of Chemical Industry.

Diversity of Ochrobactrum species in food animals, antibiotic resistance phenotypes and polymorphisms in the blaOCH gene.

Twenty-six lactose non-fermenting, oxidase, urease and citrate-positive Gram-negative rods, isolated from broiler chickens, pigs and cattle at slaughter, were subjected to the matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and 16S rDNA sequencing for identification. Susceptibility to 14 antimicrobials was determined by the disc diffusion method. Ochrobactrum isolates resistant to third-generation cephalosporins were PCR-screened for the presence of the Ochrobactrum anthropi ampC gene (blaOCH). A 547-bp internal segment of blaOCH in the Ochrobactrum spp isolates was amplified with a newly designed primer set, and a phylogenetic reconstruction based on the complete amino acid sequence of blaOCH obtained from nine Ochrobactrum strains in our collection and 20 O. anthropi available in the GenBank was undertaken. All the Ochrobactrum isolates were resistant to the expanded-spectrum beta-lactams and streptomycin. None of the isolates was resistant to imipenem while 41.7% to 50.0% of them were resistant to fluoroquinolones. The blaOCH gene was detected in 16 (66.7%) and 20 (83.3%) of the 24 Ochrobactrum isolates (O. intermedium/O. tritici species), using primers designed for O. anthropi and the newly designed primer set, respectively. Six blaOCH variants grouped into two divergent clusters were identified. This is the first report of the complete nucleotide sequence of the blaOCH gene in non-antropi Ochrobactrum species.

Characterization of Cu(II) and Cd(II) resistance mechanisms in Sphingobium sp. PHE-SPH and Ochrobactrum sp. PHE-OCH and their potential application in the bioremediation of heavy metal-phenanthrene co-contaminated sites.

Soil that is co-contaminated with heavy metals (HMs) and polycyclic aromatic hydrocarbons (PAHs) is difficult to bioremediate due to the ability of toxic metals to inhibit PAH degradation by bacteria. We demonstrated the resistance mechanisms to Cu(II) and Cd(II) of two newly isolated strains of Sphingobium sp. PHE-SPH and Ochrobactrum sp. PHE-OCH and further tested their potential application in the bioremediation of HM-phenanthrene (PhA) co-contaminated sites. The PHE-SPH and PHE-OCH strains tolerated 4.63 and 4.34 mM Cu(II) and also showed tolerance to 0.48 and 1.52 mM Cd(II), respectively. Diverse resistance patterns were detected between the two strains. In PHE-OCH cells, the maximum accumulation of Cu(II) occurred in the cell wall, while the maximum accumulation was in the cytoplasm of PHE-SPH cells. This resulted in a sudden suppression of growth in PHE-OCH and a gradual inhibition in PHE-SPH as the concentration of Cu(II) increased. Organic acid production was markedly higher in PHE-OCH than in PHE-SPH, which may also have a role in the resistance mechanisms, and contributes to the higher Cd(II) tolerance of PHE-OCH. The factors involved in the absorption of Cu(II) or Cd(II) in PHE-SPH and PHE-OCH were identified as proteins and carbohydrates by Fourier transform infrared (FT-IR) spectroscopy. Furthermore, both strains showed the ability to efficiently degrade PhA and maintained this high degradation efficiency under HM stress. The high tolerance to HMs and the PhA degradation capacity make Sphingobium sp. PHE-SPH and Ochrobactrum sp. PHE-OCH excellent candidate organisms for the bioremediation of HM-PhA co-contaminated sites.

First Case Report of Human Infection With Ochrobactrum tritici Causing Bacteremia and Cholecystitis

No abstract available.

Hyper Accumulation of Arsenic in Mutants of Ochrobactrum tritici Silenced for Arsenite Efflux Pumps.

Ochrobactrum tritici SCII24T is a highly As-resistant bacterium, with two previously described arsenic resistance operons, ars1 and ars2. Among a large number of genes, these operons contain the arsB and Acr3 genes that encode the arsenite efflux pumps responsible for arsenic resistance. Exploring the genome of O. tritici SCII24T, an additional putative operon (ars3) was identified and revealed the presence of the Acr3_2 gene that encodes for an arsenite efflux protein but which came to prove to not be required for full As resistance. The genes encoding for arsenite efflux pumps, identified in this strain, were inactivated to develop microbial accumulators of arsenic as new tools for bioremediation. Six different mutants were produced, studied and three were more useful as biotools. O. tritici wild type and the Acr3-mutants showed the highest resistance to As(III), being able to grow up to 50 mM of arsenite. On the other hand, arsB-mutants were not able to grow at concentrations higher than 1 mM As(III), and were the most As(III) sensitive mutants. In the presence of 1 mM As(III), the strain with arsB and Acr3_1 mutated showed the highest intracellular arsenic concentration (up to 17 ng(As)/mg protein), while in assays with 5 mM As(III), the single arsB-mutant was able to accumulate the highest concentration of arsenic (up to 10 ng(As)/mg protein). Therefore, arsB is the main gene responsible for arsenite resistance in O. tritici. However, both genes arsB and Acr3_1 play a crucial role in the resistance mechanism, depending on the arsenite concentration in the medium. In conclusion, at moderate arsenite concentrations, the double arsB- and Acr3_1-mutant exhibited a great ability to accumulate arsenite and can be seen as a promising bioremediation tool for environmental arsenic detoxification.

Evaluation of bacterial biosensors to determine chromate bioavailability and to assess ecotoxicity of soils.

Chromate can be considered a potent environmental contaminant and consequently, an understanding of chromate availability and toxicity to soil biology is essential for effective ecological assessment of metal impact in soils. This study shows the response of two bacterial bioreporters, pCHRGFP1 Escherichiacoli and pCHRGFP2 Ochrobactrumtritici, to increasing concentrations of chromate in two different soils. The bioreporters, carrying the regulatory gene chrB transcriptionally fused to the gfp reporter system, exhibited different features. In both, the fluorescence signal and the chromate concentration could be linearly correlated but E. coli biosensor functioned within the range of 0.5-2 µM and O. tritici biosensor within 2-10 µM chromate. The bioreporters were validated through comparative measurements using the chemical chromate methods of diphenylcarbazide and ionic chromatography. The bacterial sensors were used for the estimation of bioavailable fraction of chromate in a natural soil and OECD artificial soil, both spiked with chromate in increasing concentrations of 0-120 mg Cr(VI) kg(-1) of soil. OECD soil showed a faster chromate decrease comparing to the natural soil. The toxicity of soils amended with chromate was also evaluated by ecotoxicological tests through collembolan reproduction tests using Folsomia candida as test organism. Significant correlations were found between collembolans reproduction and chromate concentration in soil (lower at high chromate concentrations) measured by biosensors. Data obtained showed that the biosensors tested are sensitive to chromate presence in soil and may constitute a rapid and efficient method to measure chromate availability in soils.

Glutamate decarboxylase-dependent acid resistance in Brucella spp.: distribution and contribution to fitness under extremely acidic conditions.

Brucella is an expanding genus of major zoonotic pathogens, including at least 10 genetically very close species occupying a wide range of niches from soil to wildlife, livestock, and humans. Recently, we have shown that in the new species Brucella microti, the glutamate decarboxylase (Gad)-dependent system (GAD system) contributes to survival at a pH of 2.5 and also to infection in mice by the oral route. In order to study the functionality of the GAD system in the genus Brucella, 47 isolates, representative of all known species and strains of this genus, and 16 strains of the closest neighbor genus, Ochrobactrum, were studied using microbiological, biochemical, and genetic approaches. In agreement with the genome sequences, the GAD system of classical species was not functional, unlike that of most strains of Brucella ceti, Brucella pinnipedialis, and newly described species (B. microti, Brucella inopinata BO1, B. inopinata-like BO2, and Brucella sp. isolated from bullfrogs). In the presence of glutamate, these species were more acid resistant in vitro than classical terrestrial brucellae. Expression in trans of the gad locus from representative Brucella species in the Escherichia coli MG1655 mutant strain lacking the GAD system restored the acid-resistant phenotype. The highly conserved GAD system of the newly described or atypical Brucella species may play an important role in their adaptation to acidic external and host environments. Furthermore, the GAD phenotype was shown to be a useful diagnostic tool to distinguish these latter Brucella strains from Ochrobactrum and from classical terrestrial pathogenic Brucella species, which are GAD negative.

Impact of environmental stress on biochemical parameters of bacteria reducing chromium.

Chromium pollution is produced in connection with industrial processes like in tanneries. It has been suggested that bioremediation could be a good option for clean up. The stress effect of variable chromate levels, pHs and growth temperatures on biochemical parameters of two Cr(VI) reducing bacterial strains Pseudomonas aeruginosa Rb-1 and Ochrobactrum intermedium Rb-2 was investigated. Transmission electrone microscopy (TEM) was performed to study the intracellular distribution of Cr(VI). It was observed that initial stress of 1000 µgmL(-1) caused significant enhancement of all studied biochemical parameters at pH 7.0 and growth temperature of 37 °C showing great bioremediation potential of the strains. Transmission electron microscopy revealed that the distribution of chromium precipitates was not uniform as they were distributed in the cytoplasm as well as found associated with the periplasm and outer membrane. Fourier transform infrared spectroscopy showed the possible involvement of carboxyl, amino, sulpohonate and hydroxyl groups present on the bacterial cell surface for the binding of Cr(VI) ions. Cr(VI) stress brought about changes in the distridution of these functional groups. It can be concluded that the investigated bacterial strains adjust well to Cr(VI) stress in terms of biochemical parameters and along that exhibited alteration in morphology.

Impact of environmental stress on biochemical parameters of bacteria reducing chromium

Chromium pollution is produced in connection with industrial processes like in tanneries. It has been suggested that bioremediation could be a good option for clean up. The stress effect of variable chromate levels, pHs and growth temperatures on biochemical parameters of two Cr(VI) reducing bacterial strains Pseudomonas aeruginosa Rb-1 and Ochrobactrum intermedium Rb-2 was investigated. Transmission electrone microscopy (TEM) was performed to study the intracellular distribution of Cr(VI). It was observed that initial stress of 1000 µgmL-1 caused significant enhancement of all studied biochemical parameters at pH 7.0 and growth temperature of 37 °C showing great bioremediation potential of the strains. Transmission electron microscopy revealed that the distribution of chromium precipitates was not uniform as they were distributed in the cytoplasm as well as found associated with the periplasm and outer membrane. Fourier transform infrared spectroscopy showed the possible involvement of carboxyl, amino, sulpohonate and hydroxyl groups present on the bacterial cell surface for the binding of Cr(VI) ions. Cr(VI) stress brought about changes in the distridution of these functional groups. It can be concluded that the investigated bacterial strains adjust well to Cr(VI) stress in terms of biochemical parameters and along that exhibited alteration in morphology.

Modification of Fatty acids in membranes of bacteria: implication for an adaptive mechanism to the toxicity of carbon nanotubes.

We explored whether bacteria could respond adaptively to the presence of carbon nanotubes (CNTs) by investigating the influence of CNTs on the viability, composition of fatty acids, and cytoplasmic membrane fluidity of bacteria in aqueous medium for 24 h exposure. The CNTs included long single-walled carbon nanotubes (L-SWCNTs), short single-walled carbon nanotubes (S-SWCNTs), short carboxyl single-walled carbon nanotubes (S-SWCNT-COOH), and aligned multiwalled carbon nanotubes (A-MWCNTs). The bacteria included three common model bacteria, Staphyloccocus aureus (Gram-positive), Bacillus subtilis (Gram-positive), and Escherichia coli (Gram-negative), and one polybrominated diphenyl ether degrading strain, Ochrobactrum sp. (Gram-negative). Generally, L-SWCNTs were the most toxic to bacteria, whereas S-SWCNT-COOH showed the mildest bacterial toxicity. Ochrobactrum sp. was more susceptible to the toxic effect of CNTs than E. coli. Compared to the control in the absence of CNTs, the viability of Ochrobactrum sp. decreased from 71.6-81.4% to 41.8-70.2%, and E. coli from 93.7-104.0% to 67.7-91.0% when CNT concentration increased from 10 to 50 mg L(-1). The cytoplasmic membrane fluidity of bacteria increased with CNT concentration, and a significant negative correlation existed between the bacterial viabilities and membrane fluidity for E. coli and Ochrobactrum sp. (p < 0.05), indicating that the increase in membrane fluidity induced by CNTs was an important factor causing the inactivation of bacteria. In the presence of CNTs, E. coli and Ochrobactrum sp. showed elevation in the level of saturated fatty acids accompanied with reduction in unsaturated fatty acids, compensating for the fluidizing effect of CNTs. This demonstrated that bacteria could modify their composition of fatty acids to adapt to the toxicity of CNTs. In contrast, S. aureus and B. subtilis exposed to CNTs increased the proportion of branched-chain fatty acids and decreased the level of straight-chain fatty acids, which was also favorable to counteract the toxic effect of CNTs. This study suggests that the bacterial tolerances to CNTs are associated with both the adaptive modification of fatty acids in the membrane and the physicochemical properties of CNTs. This is the first report about the physiologically adaptive response of bacteria to CNTs, and may help to further understand the ecotoxicological effects of CNTs.

Reduction of hexavalent chromium by a novel Ochrobactrum sp. - microbial characteristics and reduction kinetics.

A Gram negative hexavalent chromium (Cr(VI)) reducing bacteria, Ochrobactrum sp. Cr-B4 (genbank accession number: JF824998) was isolated from the aerator water of an activated sludge process of a wastewater treatment facility of a dye and pigment based specialty chemical industry. It showed a resistance for 1000 mg L(-1) Cr(VI). It exhibited resistance against other heavy metal ions like Ni(2+) (900 mg L(-1) ), Cu(2+) (500 mg L(-1) ), Pb(2+) (800 mg L(-1) ), and Cd(2+) (250 mg L(-1) ), Zn(2+) (700 mg L(-1) ), Fe(3+) (800 mg L(-1) ), and against selected antibiotics. Cr-B4 could efficiently reduce 200 mg L(-1) Cr(VI) completely in nutrient and LB media and could convert Cr(VI) to Cr(III) efficiently. Cr(VI) reduction in nutrient media followed allosteric enzyme kinetics with Km values of 59.39 mg L(-1) and Vmax values of 47.03 mg L(-1) h(-1) . The reduction in LB media followed Michaelis-Menten kinetics with Km values of 99.52 mg L(-1) and Vmax of 77.63 mg L(-1) h(-1) . Scanning electron micrograms revealed the presence of extracellular polymeric secretions.

Identification and characterization of the transcriptional regulator ChrB in the chromate resistance determinant of Ochrobactrum tritici 5bvl1.

Ochrobactrum tritici 5bvl1 is able to resist to high concentrations of chromate through the expression of an inducible chromate-resistant determinant, found in a mobile element (TnOtChr), which carries the genes, chrB, chrA, chrC and chrF. The regulation of chr operon present in TnOtChr, which is controlled by a transcriptional regulator, ChrB, was characterized in the current work. Fusions of chr promoter, or chr promoter and chrB gene, upstream of a gfp reporter gene, identified the most probable promoter sequence within the tnpR-chrB intergenic region. This region contains an AT-rich imperfect inverted repeat sequence, which overlaps a part of the -10 sequence. The results of the in vitro DNA-binding assays with purified ChrB (His- or no-tagged) showed that the protein binds directly to the chr promoter region. In order to identify the ChrB functional domain for sensing chromate stress and for DNA-binding, site-directed mutagenesis of ChrB was performed. Among several single amino acid mutants, three mutants (R180; R187 and H229) prevented chromate induction without any modification to the protein's stability. Interestingly, two ChrB mutants (R18 and R23) were constitutively active, regardless of chromate stress conditions, indicating that the residues most probably belong to the protein-DNA binding site. As such, the ChrB was classified as a transcriptional regulator that recognizes a specific DNA sequence, regulating the expression of a chromate resistance determinant.

Acquired genetic mechanisms of a multiresistant bacterium isolated from a treatment plant receiving wastewater from antibiotic production.

The external environment, particularly wastewater treatment plants (WWTPs), where environmental bacteria meet human commensals and pathogens in large numbers, has been highlighted as a potential breeding ground for antibiotic resistance. We have isolated the extensively drug-resistant Ochrobactrum intermedium CCUG 57381 from an Indian WWTP receiving industrial wastewater from pharmaceutical production contaminated with high levels of quinolones. Antibiotic susceptibility testing against 47 antibiotics showed that the strain was 4 to >500 times more resistant to sulfonamides, quinolones, tetracyclines, macrolides, and the aminoglycoside streptomycin than the type strain O. intermedium LMG 3301T. Whole-genome sequencing identified mutations in the Indian strain causing amino acid substitutions in the target enzymes of quinolones. We also characterized three acquired regions containing resistance genes to sulfonamides (sul1), tetracyclines [tet(G) and tetR], and chloramphenicol/florfenicol (floR). Furthermore, the Indian strain harbored acquired mechanisms for horizontal gene transfer, including a type I mating pair-forming system (MPFI), a MOBP relaxase, and insertion sequence transposons. Our results highlight that WWTPs serving antibiotic manufacturing may provide nearly ideal conditions for the recruitment of resistance genes into human commensal and pathogenic bacteria.

Role of heavy metal resistant Ochrobactrum sp. and Bacillus spp. strains in bioremediation of a rice cultivar and their PGPR like activities.

The present study demonstrates the metal toxicity ameliorating and growth promoting abilities of three different bacterial isolates when applied to rice as host plant. The three bacterial strains included a cadmium resistant Ochrobactrum sp., a lead resistant Bacillus sp. and an arsenic resistant Bacillus sp. designated as CdSP9, PbSP6, and AsSP9, respectively. When these isolates were used as inocula applied to metal-treated rice plants of variety Satabdi, the germination percentage, relative root elongation (RRE), amylase and protease activities were increased. The toxic effect of metal was reduced in presence of these bacteria. The overall biomass and root/shoot ratio were also enhanced by bacterial inoculation. Hydroponic studies showed that the superoxide dismutase (SOD) activity and malondialdehyde (MDA) level, which had been increased in the presence of metal stress in rice roots, were lowered by the bacterial inoculation. In addition, all three strains were 1-aminocyclopropane-1-carboxylate (ACC) deaminase and catalase positive, whereas siderophore producing ability was lacking in PbSP6. However, both PbSP6 and AsSP9 were protease positive and could hydrolyse starch. The data indicate that these bacteria have promise for bioremediation as well as for plant growth promotion.

Influence of heavy metals on the activity of antioxidant enzymes in the metal resistant strains of Ochrobactrum and Bacillus sp.

Three bacterial strains, a cadmium resistant Ochrobactrum sp. designated as CdSP9 and two strains of Bacillus sp. named PbSP6 and AsSP9 resistant to lead and arsenate, respectively were characterized here with respect to their oxidative enzyme activities. The bacterial strains were grown in basal medium supplemented with 50 microg ml(-1) of respective elements to determine the changes in the level of oxidative enzymes. The superoxide dismutase activity increased in all three isolates, but the catalase activity and malondialdehyde concentration were relatively more in CdSP9 than PbSP6 and AsSP9. The glutathione peroxidase, however, remained almost uninduced in CdSP9 but was enhanced in PbSP6 and AsSP9. A possible role of these enzymes in metal tolerance is evident from these results.

Characterization of arsenic resistant bacteria from arsenic rich groundwater of West Bengal, India.

Sixty-four arsenic (As) resistant bacteria isolated from an arsenic rich groundwater sample of West Bengal were characterized to investigate their potential role in subsurface arsenic mobilization. Among the isolated strains predominance of genera Agrobacterium/Rhizobium, Ochrobactrum and Achromobacter which could grow chemolitrophically and utilize arsenic as electron donor were detected. Higher tolerance to As(3+) [maximum tolerable concentration (MTC): ≥10 mM], As(5+) (MTC: ≥100 mM) and other heavy metals like Cu(2+), Cr(2+), Ni(2+) etc. (MTC: ≥10 mM), presence of arsenate reductase and siderophore was frequently observed among the isolates. Ability to produce arsenite oxidase and phosphatase enzyme was detected in 50 and 34 % of the isolates, respectively. Although no direct correlation among taxonomic identity of bacterial strains and their metabolic abilities as mentioned above was apparent, several isolates affiliated to genera Ochrobactrum, Achromobacter and unclassified Rhizobiaceae members were found to be highly resistant to As(3+) and As(5+) and positive for all the test properties. Arsenate reductase activity was found to be conferred by arsC gene, which in many strains was coupled with arsenite efflux gene arsB as well. Phylogenetic incongruence between the 16S rRNA and ars genes lineages indicated possible incidence of horizontal gene transfer for ars genes. Based on the results we propose that under the prevailing low nutrient condition inhabitant bacteria capable of using inorganic electron donors play a synergistic role wherein siderophores and phosphatase activities facilitate the release of sediment bound As(5+), which is subsequently reduced by arsenate reductase resulting into the mobilization of As(3+) in groundwater.