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Management of tailings at metal mine smelter sites can reduce the potential hazards associated with exposure to toxic metal(loid)s and residual organic flotation reagents. In addition, microbes in the tailings harboring multi-resistance genes (e.g., tolerance to multiple antimicrobial agents) can cause high rates of morbidity and global economic problems. The potential co-selection mechanisms of antibiotic resistance genes (ARGs) and metal(loid) resistance genes (MRGs) during tailings sulfate-reducing bacteria (SRB) treatment have been poorly investigated. Samples were collected from a nonferrous metal mine tailing site treated with an established SRB protocol and were analyzed for selected geochemical properties and high throughput sequencing of 16S rRNA gene barcoding. Based on the shotgun metagenomic analysis, the bacterial domain was dominant in nonferrous metal(loid)-rich tailings treated with SRB for 12 months. KEGGs related to ARGs and MRGs were detected. Thiobacillus and Sphingomonas were the main genera carrying the bacA and mexEF resistance operons, along with Sulfuricella which were also found as the main genera carrying MRGs. The SRB treatment may mediate the distribution of numerous resistance genes. KOs based on the metagenomic database indicated that ARGs (mexNW, merD, sul, and bla) and MRGs (czcABCR and copRS genes) were found on the same contig. The SRB strains (Desulfosporosinus and Desulfotomaculum), and the acidophilic strain Acidiphilium significantly contributed to the distribution of sul genes. The functional metabolic pathways related to siderophores metabolism were largely from anaerobic genera of Streptomyces and Microbacterium. The presence of arsenate reductase, metal efflux pump, and Fe transport genes indicated that SRB treatment plays a key role in the metal(loid)s transformation. Overall, our findings show that bio-treatment is an effective tool for managing ARGs/MRGs and metals in tailings that contain numerous metal(loid) contaminants., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

To uncover metal toxicity targets and defense mechanisms of the facultative anaerobe Pantoea sp. strain MT58 (MT58), we used a multiomic strategy combining two global techniques, random bar code transposon site sequencing (RB-TnSeq) and activity-based metabolomics. MT58 is a metal-tolerant Oak Ridge Reservation (ORR) environmental isolate that was enriched in the presence of metals at concentrations measured in contaminated groundwater at an ORR nuclear waste site. The effects of three chemically different metals found at elevated concentrations in the ORR contaminated environment were investigated: the cation Al 3+ , the oxyanion CrO 4 2- . Both global techniques were applied using all three metals under both aerobic and anaerobic conditions to elucidate metal interactions mediated through the activity of metabolites and key genes/proteins. These revealed that Al 2 2+ . Both global techniques were applied using all three metals under both aerobic and anaerobic conditions to elucidate metal interactions mediated through the activity of metabolites and key genes/proteins. These revealed that Al 3+ toxicity. In addition, the Tol outer membrane system contributed to the protection of cellular integrity from the toxic effects of all three metals. Likewise, we found evidence of regulation of lipid content in membranes under metal stress. Individually, RB-TnSeq and metabolomics are powerful tools to explore the impact various stresses have on biological systems. Here, we show that together they can be used synergistically to identify the molecular actors and mechanisms of these pertubations to an organism, furthering our understanding of how living systems interact with their environment. 4 2- sp. strain MT58, and metals contaminating a site where the organism can be located. A number of novel metal-microbe interactions were uncovered, including Al 2 2+ toxicity. In addition, the Tol outer membrane system contributed to the protection of cellular integrity from the toxic effects of all three metals. Likewise, we found evidence of regulation of lipid content in membranes under metal stress. Individually, RB-TnSeq and metabolomics are powerful tools to explore the impact various stresses have on biological systems. Here, we show that together they can be used synergistically to identify the molecular actors and mechanisms of these pertubations to an organism, furthering our understanding of how living systems interact with their environment. IMPORTANCE Studying microbial interactions with their environment can lead to a deeper understanding of biological molecular mechanisms. In this study, two global techniques, RB-TnSeq and activity metabolomics, were successfully used to probe the interactions between a metal-resistant microorganism, Pantoea sp. strain MT58, and metals contaminating a site where the organism can be located. A number of novel metal-microbe interactions were uncovered, including Al 3+ toxicity targeting arginine synthesis, which could lead to a deeper understanding of the impact Al 3+ contamination has on microbial communities as well as its impact on higher-level organisms, including plants for whom Al 3+ contamination is an issue. Using multiomic approaches like the one described here is a way to further our understanding of microbial interactions and their impacts on the environment overall.

Alteromonas is a widely distributed genus of marine Gammaproteobacteria , with representatives shown to be key players in diverse processes, including biogeochemical cycling and biofouling of marine substrata. While Alteromonas spp. are early colonizers of copper-based antifouling paints on marine vessels, their mechanism of tolerance is poorly understood. PacBio whole-genome sequencing of Alteromonas macleodii strains CUKW and KCC02, isolated from Cu/Ni alloy test coupons submerged in oligotrophic coastal waters, indicated the presence of multiple megaplasmids (ca. 200 kb) in both. A pulsed-field gel electrophoresis method was developed and used to confirm the presence of multiple megaplasmids in these two strains; it was then used to screen additional Alteromonas strains for which little to no sequencing data exist. Plasmids were not detected in any of the other strains. Bioinformatic analysis of the CUKW and KCC02 plasmids identified numerous genes associated with metal resistance. Copper resistance orthologs from both the Escherichia coli Cue and Cus and Pseudomonas syringae Cop systems were present, at times as multiple copies. Metal growth assays in the presence of copper, cobalt, manganese, and zinc performed with 10 Alteromonas strains demonstrated the ability of CUKW and KCC02 to grow at metal concentrations inhibitory to all the other strains tested. This study reports multiple megaplasmids in Alteromonas strains. Bioinformatic analysis of the CUKW and KCC02 plasmids indicate that they harbor elements of the Tra system conjugation apparatus, although their type of mobility remains to be experimentally verified. IMPORTANCE Copper is commonly used as an antifouling agent on ship hulls. Alteromonas spp. are early colonizers of copper-based antifouling paint, but their mechanism of tolerance is poorly understood. Sequencing of A. macleodii strains isolated from copper test materials for marine ships indicated the presence of multiple megaplasmids. Plasmids serve as key vectors in horizontal gene transfer and confer traits such as metal resistance, detoxification, ecological interaction, and antibiotic resistance. Bioinformatic analysis identified many metal resistance genes and genes associated with mobility. Understanding the molecular mechanisms and capacity for gene transfer within marine biofilms provides a platform for the development of novel antifouling solutions targeting genes involved in copper tolerance and biofilm formation., (Copyright © 2020 American Society for Microbiology.)

Naturally occurring plasmids constitute a major category of mobile genetic elements responsible for harboring and transferring genes important in survival and fitness. A targeted evaluation of plasmidomes can reveal unique adaptations required by microbial communities. We developed a model system to optimize plasmid DNA isolation procedures targeted to groundwater samples which are typically characterized by low cell density (and likely variations in the plasmid size and copy numbers). The optimized method resulted in successful identification of several hundred circular plasmids, including some large plasmids (11 plasmids more than 50 kb in size, with the largest being 1.7 Mb in size). Several interesting observations were made from the analysis of plasmid DNA isolated in this study. The plasmid pool (plasmidome) was more conserved than the corresponding microbiome distribution (16S rRNA based). The circular plasmids were diverse as represented by the presence of seven plasmid incompatibility groups. The genes carried on these groundwater plasmids were highly enriched in metal resistance. Results from this study confirmed that traits such as metal, antibiotic, and phage resistance along with toxin-antitoxin systems are encoded on abundant circular plasmids, all of which could confer novel and advantageous traits to their hosts. This study confirms the ecological role of the plasmidome in maintaining the latent capacity of a microbiome, enabling rapid adaptation to environmental stresses. IMPORTANCE Plasmidomes have been typically studied in environments abundant in bacteria, and this is the first study to explore plasmids from an environment characterized by low cell density. We specifically target groundwater, a significant source of water for human/agriculture use. We used samples from a well-studied site and identified hundreds of circular plasmids, including one of the largest sizes reported in plasmidome studies. The striking similarity of the plasmid-borne ORFs in terms of taxonomical and functional classifications across several samples suggests a conserved plasmid pool, in contrast to the observed variability in the 16S rRNA-based microbiome distribution. Additionally, the stress response to environmental factors has stronger conservation via plasmid-borne genes as marked by abundance of metal resistance genes. Last, identification of novel and diverse plasmids enriches the existing plasmid database(s) and serves as a paradigm to increase the repertoire of biological parts that are available for modifying novel environmental strains., (Copyright © 2019 Kothari et al.)

Metal-tolerant thermophiles are inhabitants of a wide range of extreme habitats like solfatara fields, hot springs, mud holes, hydrothermal vents oozing out from metal-rich ores, hypersaline pools and soil crusts enriched with metals and other elements. The ability to withstand adverse environmental conditions, like high temperature, high metal concentration and sometimes high pH in their niche, makes them an interesting subject for understanding mechanisms behind their ability to deal with multiple duress simultaneously. Metals are essential for biological systems, as they participate in biochemistries that cannot be achieved only by organic molecules. However, the excess concentration of metals can disrupt natural biogeochemical processes and can impose toxicity. Thermophiles counteract metal toxicity via their unique cell wall, metabolic factors and enzymes that carry out metal-based redox transformations, metal sequestration by metallothioneins and metallochaperones as well as metal efflux. Thermophilic metal resistance is heterogeneous at both genetic and physiology levels and may be chromosomally, plasmid or transposon encoded with one or more genes being involved. These effective response mechanisms either individually or synergistically make proliferation of thermophiles in metal-rich habitats possibly. This article presents the state of the art and future perspectives of responses of thermophiles to metals at genetic as well as physiological levels.

Microalgae are a source of numerous compounds that can be used in many branches of industry. Synthesis of such compounds in microalgal cells can be amplified under stress conditions. Exposure to various metals can be one of methods applied to induce cell stress and synthesis of target products in microalgae cultures. In this review, the potential of producing diverse biocompounds (pigments, lipids, exopolymers, peptides, phytohormones, arsenoorganics, nanoparticles) from microalgae cultures upon exposure to various metals, is evaluated. Additionally, different methods to alter microalgae response towards metals and metal stress are described. Finally, possibilities to sustain high growth rates and productivity of microalgal cultures in the presence of metals are discussed.

Metal-resistant bacteria are recommended for metal removal applications due to their rapid multiplication and growth rates. To ensure safety replenishment in contaminated areas frequently hampered by heavy metal toxicity, it is crucial to comprehend their coping mechanisms under heavy metal stress. This study primarily examines the role of EPS (exopolysaccharide) in Stenotrophomonas maltophilia, a Gram-negative, aerobic, and rod-shaped bacteria, in response to Cd, as well as the binding behavior and biosorption mechanism between EPS and Cd, using SEM and FTIR. The studies showed that Stenotrophomonas maltophilia, can resist up to 150 μM of Cd due to the binding of Cd to EPS. SEM analysis showed significant morphological changes and FTIR was to identify main structural groups like carboxyl and hydroxyl which confirms the presence of EPS. The study will also describe the mechanism of cross-reactivity between exopolysaccharide and siderophore production in metal-tolerant Stenotrophomonas maltophilia. This study proved that siderophore-mediated metal detoxification and effective absorption have been linked to metal chelation. [ABSTRACT FROM AUTHOR]

Contamination of environments with nitrate generated by industrial processes and the use of nitrogen-containing fertilizers is a growing problem worldwide. While nitrate can be removed from contaminated areas by microbial denitrification, nitrate frequently occurs with other contaminants, such as heavy metals, that have the potential to impede the process. Here, nitrate-reducing microorganisms were enriched and isolated from both groundwater and sediments at the Oak Ridge Reservation (ORR) using concentrations of nitrate and metals (Al, Mn, Fe, Co, Ni, Cu, Cd, and U) similar to those observed in a contaminated environment at ORR. Seven new metal-resistant, nitrate-reducing strains were characterized, and their distribution across both noncontaminated and contaminated areas at ORR was examined. While the seven strains have various pH ranges for growth, carbon source preferences, and degrees of resistance to individual and combinations of metals, all were able to reduce nitrate at similar rates both in the presence and absence of the mixture of metals found in the contaminated ORR environment. Four strains were identified in groundwater samples at different ORR locations by exact 16S RNA sequence variant analysis, and all four were found in both noncontaminated and contaminated areas. By using environmentally relevant metal concentrations, we successfully isolated multiple organisms from both ORR noncontaminated and contaminated environments that are capable of reducing nitrate in the presence of extreme mixed-metal contamination.IMPORTANCE Nitrate contamination is a global issue that affects groundwater quality. In some cases, cocontamination of groundwater with nitrate and mixtures of heavy metals could decrease microbially mediated nitrate removal, thereby increasing the duration of nitrate contamination. Here, we used metal and nitrate concentrations that are present in a contaminated site at the Oak Ridge Reservation to isolate seven metal-resistant strains. All were able to reduce nitrate in the presence of high concentrations of a mixture of heavy metals. Four of seven strains were located in pristine as well as contaminated sites at the Oak Ridge Reservation. Further study of these nitrate-reducing strains will uncover mechanisms of resistance to multiple metals that will increase our understanding of the effect of nitrate and metal contamination on groundwater microbial communities.

The metals present in freshwater have a toxic profile with bioaccumulation and are biomagnified along the aquatic food chain. The metals induce high sensitivity in most aquatic organisms, while others, such as some microalgae species, evolve towards resistance. Therefore, this research predicted through the Combination Index method the binary interaction exposed to divalent metals by inhibiting population growth in a Cd-resistant strain (DcRCd100) compared to the wild-type strain (Dc1Mwt) of Dictyosphaerium chlorelloides and evaluate the specific resistance level obtained by DcRCd100 to Cd relative to other divalent metals.The results showed that DcRCd100 presents resistance compared to Dc1Mwt in individual exposure in the order of Fe2+ > Ni2+ > Cd2+ > Co2+ > Zn2+ > Cu2+ > Hg2+ with 50% inhibitory concentration at 72 h of exposure (IC50(72)) values 1253, 644.4, 423, 162.7, 141.3, 35.1, and 9.9 µM, respectively. It induces cross-resistance with high antagonistic rates (Combination Index (CI); CI > > 1) in the Cd/Zn and Cd/Cu. Cd/Ni, its initial response, is antagonistic, and it ends in an additive (CI = 1). DcRCd100 showed a lower resistance in Co, and Cd/Fe resistance was reduced individually. The interaction with Hg increased its resistance ten times more than individually.This research highlights the use of the CI as a highly efficient prediction method of the binary metal interactions in wild-type and Cd-resistant strains of D.chlorelloides. It may have the potential for metal accumulation, allowing the development of new methods of bioremediation of metals in effluents, and to monitor the concentration of metals in wastewater, its relative availability, transport, and mechanisms on resistant strains of microalgae. [ABSTRACT FROM AUTHOR]

Supramolecular cages have been constructed by anion‐coordination‐driven assembly (ACDA) in recent years and have shown unique host‐guest interactions. However, most of the reported cages are made of the phosphate ion (PO43−); other anions have rarely been explored. Here we show for the first time that the sulfate ion (SO42−) is also able to form the A4L4 tetrahedral motif with tris‐bis(urea) ligands, but this is dependent on the stoichiometry of the sulfate ion (in solution). Notably, the sulfate cages display enhanced resistance for both Brønsted (pH as low as 4.3 in acetone containing 15 % water) and Lewis acids (metal complexes) compared to the corresponding phosphate cages, and thus could find applications where an acidic (proton or metals) environment is required. [ABSTRACT FROM AUTHOR]

Sulfate-reducing bacteria (SRB) can be used to remove metals from wastewater, sewage, and contaminated areas. However, metals can be toxic to this group of bacteria. Sediments from port areas present abundance of SRB and also metal contamination. Their microbial community has been exposed to metals and can be a good inoculum for isolation of metal-resistant SRB. The objective of the study was to analyze how metals influence activity and composition of sulfate-reducing bacteria. Enrichment cultures were prepared with a different metal (Zn, Cr, Cu, and Cd) range concentration tracking activity of SRB and 16S rRNA sequencing in order to access the community. The SRB activity decreased when there was an increase in the concentration of the metals tested. The highest concentration of metals precipitated were 0.2 mM of Cd, 5.4 mM of Zn, 4.5 mM of Cu, and 9.6 mM of Cr. The more toxic metals were Cd and Cu and had a greater community similarity with less SRB and more fermenters (e.g., Citrobacter and Clostridium). Meanwhile, the enrichments with less toxic metals (Cr and Zn) had more sequences affiliated to SRB genera (mainly Desulfovibrio). A new Desulfovibrio species was isolated. This type of study can be useful to understand the effects of metals in SRB communities and help to optimize wastewater treatment processes contaminated by metals. The new Desulfovibrio species may be important in future studies on bioremediation of neutral pH effluents contaminated by metals. [ABSTRACT FROM AUTHOR]

Cupriavidus metallidurans , and in particular type strain CH34, became a model bacterium to study bacterial resistance to metals. Although nowadays the routine use of a wide variety of omics and molecular techniques allow refining, deepening and expanding our knowledge on adaptation and resistance to metals, these were not available at the onset of C. metallidurans research starting from its isolation in 1976. This minireview describes the early research and legacy tools used to study its metal resistance determinants, characteristic megaplasmids, ecological niches and environmental applications. [ABSTRACT FROM AUTHOR]

Five strains of Cupriavidus plantarum, a metal-resistant, plant-associated bacterium, were selected for genome sequencing through the Genomic Encyclopedia of Bacteria and Archaea (GEBA) Phase IV project at the Joint Genome Institute (JGI) of the U.S. Department of Energy (DOE). The genome of the strains was in the size range of 6.2–6.4 Mbp and encoded 5605–5834 proteins; 16.9–23.7% of these genes could not be assigned to a COG-associated functional category. The G + C content was 65.83–65.99%, and the genomes encoded 59–67 stable RNAs. The strains were resistant in vitro to arsenite, arsenate, cobalt, chromium, copper, nickel and zinc, and their genomes possessed the resistance genes for these metals. The genomes also encoded the biosynthesis of potential antimicrobial compounds, such as terpenes, phosphonates, bacteriocins, betalactones, nonribosomal peptides, phenazine and siderophores, as well as the biosynthesis of cellulose and enzymes such as chitinase and trehalase. The average nucleotide identity (ANI) and DNA-DNA in silico hybridization of the genomes confirmed that C. plantarum is a single species. Moreover, the strains cluster within a single group upon multilocus sequence analyses with eight genes and a phylogenomic analyses. Noteworthy, the ability of the species to tolerate high concentrations of different metals might prove useful for bioremediation of naturally contaminated environments. [ABSTRACT FROM AUTHOR]

Alteromonas is a widely distributed genus of marine Gammaproteobacteria with representatives shown to be key players in diverse processes including biogeochemical cycling and biofouling of marine substrata. While, Alteromonas spp. are early colonizers of copper-based antifouling paints on marine vessels, their mechanism of tolerance is poorly understood. PacBio whole genome sequencing of A. macleodii strains CUKW and KCC02, isolated from Cu/Ni alloy test coupons submerged in oligotrophic coastal waters indicated the presence of multiple mega-plasmids (ca. 200 KB) in both. A pulsed field gel electrophoresis method was developed and used to confirm the presence of multiple mega-plasmids in these two strains; it was then used to screen additional Alteromonas strains for which little to no sequencing data exist. Plasmids were not detected in any of the other strains. Bioinformatic analysis of the CUKW and KCC02 plasmids identified numerous genes associated with metal resistance. Copper resistance orthologs from both the Escherichia coli Cue and Cus and Pseudomonas syringae Cop systems were present, at times as multiple copies. Metal growth assays in the presence of copper, cobalt, manganese and zinc performed with 10 Alteromonas strains demonstrated the ability of CUKW and KCC02 to grow at metal concentrations inhibitory to all the other strains tested. This is the first report of multiple mega-plasmids in Alteromonas strains. Bioinformatic analysis of the CUKW and KCC02 plasmids indicate they harbor elements of the Tra system conjugation apparatus, although their type of mobility remains to be experimentally verified. Copper is commonly used as an antifouling agent on ship hulls. Alteromonas spp. are early colonizers of copper-based antifouling paint but their mechanism of tolerance is poorly understood. Sequencing of A. macleodii strains isolated from copper test materials for marine ships indicated the presence of multiple mega-plasmids. Plasmids serve as key vectors in horizontal gene transfer and confer traits such as metal resistance, detoxification, ecological interaction, and antibiotic resistance. Bioinformatic analysis identified many metal resistance genes and genes associated with mobility. Understanding the molecular mechanisms and capacity for gene transfer within marine biofilms provides a platform for the development of novel anti-fouling solutions targeting genes involved in copper tolerance and biofilm formation. [ABSTRACT FROM AUTHOR]

Spent catalysts represent an environmental concern, mainly due to their elevated metal content. Although conventional treatment methods for spent catalysts are available, they generate large volumes of potentially harmful wastes and gaseous emissions. To overcome the environmental impact, biotechnological approaches are currently being explored and developed. Thus, the current study assayed the capability of Bacillus megaterium strain MNSH1-9K-1 to remove Al, Ni, V and Ti contained in the spent catalyst coded as ECAT-TL-II. To this end, B. megaterium MNSH1-9K-1 growth and metal uptake abilities in the presence of ECAT-TL-II spent catalyst at 15% (wt/vol) pulp density were evaluated in modified Starkey medium at 37 °C and 200 rpm. The results presented here show B. megaterium resistance capability to the high-metal content residue, and its Al, V and Ni removal ability, in 1,059.15 ± 197.28 mg kg–1 of Al, 43.39 ± 24.13 mg kg–1 of V and 0.58 ± 0.00 mg kg–1 of Ni, corresponding to the 0.79%, 1.63% and 0.46% of each metal content, respectively, while no Ti removal was detected. Besides, it was observed that the sporulation process took place in B. megaterium cells in the presence of the spent catalyst. The results shown in this study suggest the potential of the strain MNSH1-9K-1 for the removal of metals contained in high-metal content residues, contributing also to the knowledge of the metal resistance and removal abilities of B. megaterium in the presence of a spent catalyst, and how morphological cell changes may be occurring while metal removal is taking place. [ABSTRACT FROM AUTHOR]

Chromosomal co-existence of metal and antibiotic resistance genes in bacteria offers a new perspective to the bacterial resistance proliferation in contaminated environment. In this study, an arsenotrophic bacterium Achromobacter xylosoxidans BHW-15, isolated from Arsenic (As) contaminated tubewell water in the Bogra district of Bangladesh, was analyzed using high throughput Ion Torrent Personal Genome Machine (PGM) complete genome sequencing scheme to reveal its adaptive potentiality. The assembled draft genome of A. xylosoxidans BHW-15 was 6.3 Mbp containing 5,782 functional genes, 1,845 pseudo genes, and three incomplete phage signature regions. Comparative genome study suggested the bacterium to be a novel strain of A. xylosoxidans showing significant dissimilarity with other relevant strains in metal resistance gene islands. A total of 35 metal resistance genes along with arsenite-oxidizing aioSXBA, arsenate reducing arsRCDAB, and mercury resistance merRTPADE operonic gene cluster and 20 broad range antibiotic resistance genes including β-lactams, aminoglycosides, and multiple multidrug resistance (MDR) efflux gene complex with a tripartite system OM-IM-MFP were found co-existed within the genome. Genomic synteny analysis with reported arsenotrophic bacteria revealed the characteristic genetic organization of ars and mer operonic genes, rarely described in β-Proteobacteria. A transposon Tn21 and mobile element protein genes were also detected to the end of mer (mercury) operonic genes, possibly a carrier for the gene transposition. In vitro antibiotic susceptibility assay showed a broad range of resistance against antibiotics belonging to β-lactams, aminoglycosides, cephalosporins (1st, 2nd, and 3rd generations), monobactams and even macrolides, some of the resistome determinants were predicted during in silico analysis. KEGG functional orthology analysis revealed the potential of the bacterium to utilize multiple carbon sources including one carbon pool by folate, innate defense mechanism against multiple stress conditions, motility, a proper developed cell signaling and processing unit and secondary metabolism-combination of all exhibiting a robust feature of the cell in multiple stressed conditions. The complete genome of the strain BHW-15 stands as a genetic basis for the evolutionary adaptation of metal and the antibiotic coexistence phenomenon in an aquatic environment. [ABSTRACT FROM AUTHOR]

Members of genus Acidithiobacillus which can adapt up to extremely high concentration of heavy metals are universally found at acid mine drainage (AMD) sites. We here perform comparative genomic analysis of 37 strains within the genus Acidithiobacillus to answer the untouched questions as to the mechanisms and the evolutionary history of metal resistance genes in Acidithiobacillus spp.. Results showed that the evolutionary history of metal resistance genes in Acidithiobacillus spp. involved a combination of gene gains and losses, HGT and gene duplication. Phylogenetic analyses revealed that metal resistance genes in Acidithiobacillus spp. were acquired by early horizontal gene transfer (HGT) events from species that shared habitats with Acidithiobacillus spp. such as Acidihalobacter, Thiobacillus, Acidiferrobacter and Thiomonas. Multi-copper oxidase genes involved in copper detoxification were lost in iron-oxidizing A. ferridurans, A. ferrivorans and A. ferrooxidans, and were replaced by rusticyanin during evolution. In addition, widespread purifying selection and predicted high expression levels emphasized the indispensable roles of metal resistance genes for Acidithiobacillus spp. to adapt harsh environments. Taking together, the results suggested that Acidithiobacillus spp. recruited and consolidated additional novel functionalities in adaption to challenging environments via HGT, gene duplication and purifying selection. This study shed light on the distribution, organization, functionality and the complex evolutionary history of metal resistance genes in Acidithiobacillus spp. [ABSTRACT FROM AUTHOR]

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Long-term contaminated environments have been recognized as potential hotspots for bacterial discovery in taxonomic and functional terms for bioremediation purposes. Here, bacterial diversity in waste sediment collected from a former industrial dumpsite and contaminated with petroleum hydrocarbon and heavy metals was investigated through the parallel application of culture-independent (16S rRNA gene amplicon sequencing) and -dependent (plate culturing followed by colony picking and identification of isolates by 16S rRNA gene Sanger sequencing) approaches. The bacterial diversities retrieved by both approaches greatly differed. Bacteroidetes and Proteobacteria were dominant in the culture-independent community, while Firmicutes and Actinobacteria were the main culturable groups. Only 2.7% of OTUs (operational taxonomic units) in the culture-independent dataset were cultured. Most of the culturable OTUs were absent or in very low abundances in the culture-independent dataset, revealing that culturing is a useful tool to study the rare bacterial biosphere. One culturable OTUs (comprising only the isolate SPR117) was identified as a potential new species in the genus Rhizorhapis (class Alphaproteobacteria) and was selected for further characterization. Phytopathogenicity tests showed that Rhizorhapis sp. strain SPR117 (ATCC TSD-228) is not pathogenic to lettuce, despite the only described species in this genus, Rhizorhapis suberifaciens , is causal agent of the lettuce corky root disease. The genome of the strain SPR117 was sequenced, assembled in 256 contigs, with a length of 4,419,522 bp and a GC content of 59.9%, and its further annotation revealed the presence of genes related to the resistance to arsenic, copper, iron, and mercury, among other metals. Therefore, the coupling of metataxonomics and culturing is a useful tool to obtain not only an improved description of bacterial communities in contaminated environments, but also to isolate microorganisms with bioremediation potential. • Metataxonomics and culturing were coupled to study bacteria from a waste sediment. • Culture-dependent and -independent diversities differed and were complementary. • Most of the isolated strains were members of the rare bacterial biosphere. • A rare biosphere member and new species within the genus Rhizorhapis was isolated. • Genome sequence of Rhizorhapis sp. SPR117 evidenced its metal resistance potential. [ABSTRACT FROM AUTHOR]

The present study first time reports the utilization of metal tolerant fungi as a potential source for efficient enzyme production. The purification and characterization of alkaline protease from an indigenous zinc-metal tolerant fungal isolate Aspergillus flavus NJP08 has been demonstrated. The specific activity of enzyme was determined as 89.1 U mg −1 which is found to be the highest among the reported Aspergillus flavus isolates so far. The protease was purified 55.87 fold up to homogeneity and identified as “alkaline protease” with a molecular mass of 33 kDa. The N-terminal sequence was GLTTQKSAPWGLG which showed high similarity with other reported proteases of genus Aspergillus . Physico-chemical characterization of enzyme revealed an estimated half-life of >20 h with aliphatic and GRAVY index values of 79.65 and −0.161 respectively, depicting high thermo-stability and secretory nature of protease. The protease was active within the temperature range of 25–50 °C with an optimum temperature of 50 °C and was stable in the pH range of 6.0–11.0. The enzyme was activated by Ca 2+ and Fe 2+ ions, partially inhibited by Cu 2+ ions and strongly inhibited by PMSF. High enzyme stability in presence of various detergents further strengthens enzyme applicability in industrial applications. [ABSTRACT FROM AUTHOR]

The metal-exporting systems CusCFBA of Escherichia coli and GesABC of Salmonella are resistance-nodulation-division (RND)-type multiprotein systems responsible for detoxification during metal stress. In this study, the substrate range was determined for each metal transport system and possible amino acid residues important in substrate specificity were identified. The Ges system, previously identified as a gold-efflux system, conferred resistance to the greatest number and variety of organic chemicals including chloramphenicol, not recognized previously as a substrate. Phylogenetic analysis showed that GesB is most closely related to a class of RND transporters including MexF that have been shown to be responsible for exporting fluoroquinolones, chloramphenicol, and biocides. However, many of the closest homologs of GesABC appear to play a role in metal resistance judging from the genetic context. In contrast, CusCFBA belongs to a distinct family of RND-type monovalent metal-exporter systems containing a number of essential metal-binding methionines, resulting in a much narrower substrate range. [ABSTRACT FROM AUTHOR]

The article presents the study conducted regarding the behavior of microorganisms on the metallomorphic environments. The study of the impact of microbial behavior on minerals and mining areas geologically significant compounds has resulted to the understanding of the metallic elements cycle, stability and formation of biotopes and altered compounds. The microorganisms metabolic process is due to the dissolution of minerals which add to the formation of mineral deposits over geogical period. On the other hand, the effect of metals on microorganisms can influence habitat development.

Abstract: Agricultural soil irrigated with industrial wastewater (more than two decades) analysed for heavy metals revealed high levels of Fe, Cr, Cu, Zn, Ni and Cd. Out of a total of 40 bacterial isolates obtained from these soils, 17 belonged to the family enterobacteriaceae and 10 were Pseudomonas spp. A maximum MIC of 200 for Cd, 400 for Zn and Cu, 800 for Ni, and 1600μg/ml for Pb was observed. Biosorption of Ni and Cd studies over a range of metal ion concentrations with Escherichia coli WS11 both in single and bi-metal systems showed that the adsorption of Cd and Ni was dependent on the concentrations and followed the Freundlich adsorption isotherm. The biosorption of Ni increased from 6.96 to 55.31mg/g of cells, and Cd from 4.96 to 45.37mg/g of cells at a concentration ranging from 50 to 400μg/ml after 2h of incubation in a single metal solution. A further increase in incubation time had no significant effect on the biosorption of metals. [Copyright &y& Elsevier]

We examined cadmium and lead resistance in Pseudomonas sp. S8A, an isolate obtained from mine tailings- contaminated soil. Resistant to soluble metal concentrations up to 200 mg l-1 cadmium and 300 mg l-1 lead, S8A produced both exopolymer and biosurfactant. Upon growth, this pseudomonad diverged into two morphologically distinct colony subtypes; small and round or large and fiat. In the presence of lead and in the no metal control the large morphotype appeared only in late stationary phase. With cadmium the large morphotype appeared immediately following exposure. Results show that the large morphotype produced greater amounts of surfactant than the small morphotype, suggesting a unique subpopulation response to cadmium toxicity. Results also indicate that an unidentified 28 kDa protein was expressed following exposure to > 10 mg l-1 cadmium. This study demonstrates new links between surfactant production, differential subpopulation response and metal exposure. [ABSTRACT FROM AUTHOR]

The transition metal Ni is an essential cofactor for a number of enzymatic reactions in both prokaryotes and eukaryotes. Molecular analyses have revealed the existence of two major types of high-affinity Ni2+ transporters in bacteria. The Nik system of Escherichia coli is a member of the ABC transporter family and provides Ni2+ ion for the anaerobic biosynthesis of hydrogenases. The periplasmic binding protein of the transporter, NikA, is likely to play a dual role. It acts as the primary binder in the uptake process and is also involved in negative chemotaxis to escape Ni overload. Expression of the nik operon is controlled by the Ni-responsive repressor NikR, which shows functional similarity to the ferric ion uptake regulator Fur. The second type of Ni2+ transporter is represented by HoxN of Ralstonia eutropha, the prototype of a novel family of transition metal permeases. Members of this family have been identified in gram-negative and gram-positive bacteria and recently also in a fission yeast. They transport Ni2+ with very high affinity, but differ with regard to specificity. Site-directed mutagenesis experiments have identified residues that are essential for transport. Besides these uptake systems, different types of metal export systems, which prevent microorganisms from the toxic effects of Ni2+ at elevated intracellular concentrations, have also been described. [ABSTRACT FROM AUTHOR]

Gate resistance, middle of line resistance, and back end of line resistance in modern metal-gate CMOS increase drastically as the dimensions of the gates, interconnects and vias scale down close to or below the bulk electron mean free paths (MFPs) of the metal materials. These resistances, especially the gate resistance, impose more and more significant RC delay to CMOS circuits and become significant concerns in sub-22-nm CMOS. In order to optimize the metal-gate materials and structures for low resistance, accurate metal resistance model is needed. In this letter, we propose an analytical metal resistance model applicable for metal wires and films even with sub-MFP sizes. Our model includes scattering effects from surfaces, interfaces, and grain boundaries, and has been successfully verified on $W$ metal gates with the feature sizes ranging from 20 to 70 nm. [ABSTRACT FROM AUTHOR]

Cueva de la Mora (CM) is an acidic, meromictic pit lake in the Iberian Pyrite Belt characterized by extremely high metal(loid) concentrations and strong gradients in oxygen, metal, and nutrient concentrations. We hypothesized that geochemical variations with depth would result in differences in community composition and in metal resistance strategies among active microbial populations. We also hypothesized that metal resistance gene (MRG) expression would correlate with toxicity levels for dissolved metal species in the lake. Water samples were collected in the upper oxic layer, chemocline, and deep anoxic layer of the lake for shotgun metagenomic and metatranscriptomic sequencing. Metagenomic analyses revealed dramatic differences in the composition of the microbial communities with depth, consistent with changing geochemistry. Based on relative abundance of taxa identified in each metagenome, Eukaryotes (predominantly Coccomyxa) dominated the upper layer, while Archaea (predominantly Thermoplasmatales) dominated the deep layer, and a combination of Bacteria and Eukaryotes were abundant at the chemocline. We compared metal resistance across communities using a curated list of protein-coding MRGs with KEGG Orthology identifiers (KOs) and found that there were broad differences in the metal resistance strategies (e.g., intracellular metal accumulation) expressed by Eukaryotes, Bacteria, and Archaea. Although normalized abundances of MRG and MRG expression were generally higher in the deep layer, expression of metal-specific genes was not strongly related to variations in specific metal concentrations, especially for Cu and As. We also compared MRG potential and expression in metagenome assembled genomes (MAGs) from the deep layer, where metal concentrations are highest. Consistent with previous work showing differences in metal resistance mechanisms even at the strain level, MRG expression patterns varied strongly among MAG populations from the same depth. Some MAG populations expressed very few MRG known to date, suggesting that novel metal resistance strategies remain to be discovered in uncultivated acidophiles. [ABSTRACT FROM AUTHOR]

(Ni) is a common cocontaminant at many waste sites where the soils and sediments often are acidic, thereby influencing metal availability. Growth of Burkholderia cepacia PR1301 was not affected at 3.41 mM Ni at pH 5, but was inhibited by 73.2% at pH 6 and inhibited completely at pH 7 compared to growth without Ni. This pH effect was not observed in the Ni-resistant strains, Ralstonia metallidurans CH34 and 31A. Predicted Ni speciation did not explain the observed toxicity trends. Sorption of Ni to PR1 increased with increasing pH (1.49, 1.12, and 3.88 mg Ni/g dry weight at pH 5, 6, and 7, respectively), but was low at all three pH values, and most likely does not explain the observed pH effect. Growth inhibition of PR1 with increasing pH also was observed for other divalent cations, with growth observed at 4.24 mM Co, 2.22 mM Cd, and 3.82 mM Zn at pH 5 and 6, but totally inhibited at pH 7. These studies suggest that, at circumneutral pH, PR1 would be considered sensitive to Ni and other divalent cations, in spite of the ability to grow in higher concentrations at lower pH values. [ABSTRACT FROM AUTHOR]

(Ni) is a common cocontaminant at many waste sites where the soils and sediments often are acidic, thereby influencing metal availability. Growth of Burkholderia cepacia PR1301 was not affected at 3.41 mM Ni at pH 5, but was inhibited by 73.2% at pH 6 and inhibited completely at pH 7 compared to growth without Ni. This pH effect was not observed in the Ni-resistant strains, Ralstonia metallidurans CH34 and 31A. Predicted Ni speciation did not explain the observed toxicity trends. Sorption of Ni to PR1 increased with increasing pH (1.49, 1.12, and 3.88 mg Ni/g dry weight at pH 5, 6, and 7, respectively), but was low at all three pH values, and most likely does not explain the observed pH effect. Growth inhibition of PR1 with increasing pH also was observed for other divalent cations, with growth observed at 4.24 mM Co, 2.22 mM Cd, and 3.82 mM Zn at pH 5 and 6, but totally inhibited at pH 7. These studies suggest that, at circumneutral pH, PR1 would be considered sensitive to Ni and other divalent cations, in spite of the ability to grow in higher concentrations at lower pH values. [ABSTRACT FROM AUTHOR]