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Some sulfate-reducing bacteria (SRB), mainly belonging to the Desulfovibrionaceae family, have evolved the capability to conserve energy through microbial extracellular electron transfer (EET), suggesting that this process may be more widespread than previously believed. While previous evidence has shown that mobile genetic elements drive the plasticity and evolution of SRB and iron-reducing bacteria (FeRB), few have investigated the shared molecular mechanisms related to EET. To address this, we analyzed the prevalence and abundance of EET elements and how they contributed to their differentiation among 42 members of the Desulfovibrionaceae family and 23 and 59 members of Geobacteraceae and Shewanellaceae, respectively. Proteins involved in EET, such as the cytochromes PpcA and CymA, the outer membrane protein OmpJ, and the iron-sulfur cluster-binding CbcT, exhibited widespread distribution within Desulfovibrionaceae. Some of these showed modular diversification. Additional evidence revealed that horizontal gene transfer was involved in the acquiring and losing of critical genes, increasing the diversification and plasticity between the three families. The results suggest that specific EET genes were widely disseminated through horizontal transfer, where some changes reflected environmental adaptations. These findings enhance our comprehension of the evolution and distribution of proteins involved in EET processes, shedding light on their role in iron and sulfur biogeochemical cycling.

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Sugarcane vinasse exits the distillation process at high temperatures, which may differ from the optimal temperatures for dark fermentation and anaerobic digestion. A 15 °C temperature increase, for example, stops sugarcane vinasse methane generation, making distillery vinasse digestion complicated. Conversely, in other aspects, co-digesting vinasse and glycerol has been proven to stabilize methane production from vinasse because of sulfate dilution. However, glycerol has not been tested to stabilize vinasse digestion under temperature changes. Thus, this study compared the effects of different temperature settings on the co-digestion of 10 g COD L-1 of vinasse and glycerol (50 %:50 % on a COD basis) in anaerobic fluidized bed reactors (AFBR), i.e., an acidogenic and a methanogenic one-stage AFBRs operated at 55, 60, and 65 °C, and two methanogenic AFBRs fed both with acidogenic effluent (one operated at room temperature (25 °C) and the other at 55, 60, and 65 °C). The co-digestion provided steady methane generation at all AFBRs, with methane production rates ranging from 2.27 to 2.93 L CH4 d-1 L-1, whether in one or two stages. A feature of this research was to unravel the black box of the role of sulfate in the digestion of sugarcane vinasse, which was rarely studied. Desulfovibrio was the primary genus degrading 1,3-propanediol into 3-hydroxypropanoate after genome sequencing. Phosphate acetyltransferase (EC: 2.3.1.8, K00625) and acetate kinase (EC: 2.7.2.1, K00925) genes were also found, suggesting propionate was metabolized. In practical aspects, regarding the two-stage systems, the thermophilic-mesophilic (acidogenic-methanogenic) configuration is best for extracting additional value-added products because 1,3-propanediol may be recovered at high yields with steady methane production at reduced energy expenditure in a reactor operated at room temperature. However, the one-stage design is best for methane generation per system volume since it remained stable with rising temperatures, and all systems presented similar methane production rates.

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AIMS: The aim was to develop an electrochemical/optical set-up and correlate it (as validation) with other chemical and physical methods to obtain a simple and cost-effective system to study biofilm formation. METHODS AND RESULTS: A simple microfluidic cell and methods allowed continuous monitoring of the first, critical steps of microbial attachment. We monitored sulfate-reducing bacteria (SRB) at the early stages of biofilm formation. Herein, we studied the formation and adherence of SRB consortium biofilms over an indium tin oxide (ITO) conducting surface using microbiological and chemical methods, microscopic observations [scanning electron microscopy (SEM) and optical], and electrochemical impedance spectroscopy (EIS) measurements. The SRB biofilm formation was evaluated for 30 d by SEM and EIS. Charge transfer resistance decreased when the microbial population colonized the electrode. The monitoring of early-stage biofilm formation was performed using EIS at a single frequency of 1 Hz during the first 36 h. CONCLUSIONS: The simultaneous use of optical, analytical, and microbiological methods allowed us to connect the kinetics of the growth of the microbial consortium to the values obtained via the electrochemical technique. The simple setup we present here can help laboratories with limited resources to study biofilm attachment and facilitates the development of various strategies to control biofilm development in order to avoid damage to metallic structures (microbiologically influenced corrosion, MIC) or the colonization of other industrial structures and medical devices.

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This study aimed to compare microscopic counting, culture, and quantitative or real-time PCR (qPCR) to quantify sulfate-reducing bacteria in environmental and engineered sludge samples. Four sets of primers that amplified the dsrA and apsA gene encoding the two key enzymes of the sulfate-reduction pathway were initially tested. qPCR standard curves were constructed using genomic DNA from an SRB suspension and dilutions of an enriched sulfate-reducing sludge. According to specificity and reproducibility, the DSR1F/RH3-dsr-R primer set ensured a good quantification based on dsrA gene amplification; however, it exhibited inconsistencies at low and high levels of SRB concentrations in environmental and sulfate-reducing sludge samples. Ultimately, we conducted a qPCR method normalized to dsrA gene copies, using a synthetic double-stranded DNA fragment as a calibrator. This method fulfilled all validation criteria and proved to be specific, accurate, and precise. The enumeration of metabolically active SRB populations through culture methods differed from dsrA gene copies but showed a plausible positive correlation. Conversely, microscopic counting had limitations due to distinguishing densely clustered organisms, impacting precision. Hence, this study proves that a qPCR-based method optimized with dsrA gene copies as a calibrator is a sensitive molecular tool for the absolute enumeration of SRB populations in engineered and environmental sludge samples.

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Sulfate-rich effluents have been successfully treated in anaerobic reactors using sulfate-reducing bacteria (SRB). Many authors have demonstrated that these systems require nitrogen and phosphorous supplementation to achieve high sulfate removal rates. However, the resource ratio theory assumes that some species can be dominant according to the nutritional relations used or even without external nutrient supplementation. Thus, this study evaluated the SRB communities in batch reactors without external nitrogen and phosphorus sources based on most probable number (MPN) quantification, denaturing gradient gel electrophoresis (DGGE) analyses and sequencing. The sulfate and chemical oxygen demand (COD) removal and kinetic parameters were also determined. After 100 days of operation, the sulfate and COD removal achieved 71.8 ± 10% and 86.5 ± 10%, respectively. The SRB population increased from 8.106 to 4 × 1012 MPN 100 mL-1, and the richness of SRB bands was much higher at the end of the experiment compared to the inoculum. In addition, the sequenced bands from SRB-DGGE showed similarities to Desulfacinum infernum, Desulfobulbus sp, Syntrophobacter and Desulfomicrobium aestuarii-related sequences. Therefore, biological treatment of acid mine drainage wastewater was effective in the absence of nutrients, lowering costs and providing high sulfate removal efficiency.

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A novel interdomain consortium composed of a methanogenic Archaeon and a sulfate-reducing bacterium was isolated from a microbial biofilm in an oil well in Cahuita National Park, Costa Rica. Both organisms can be grown in pure culture or as stable co-culture. The methanogenic cells were non-motile rods producing CH4 exclusively from H2/CO2. Cells of the sulfate-reducing partner were motile rods forming cell aggregates. They utilized hydrogen, lactate, formate, and pyruvate as electron donors. Electron acceptors were sulfate, thiosulfate, and sulfite. 16S rRNA sequencing revealed 99% gene sequence similarity of strain CaP3V-M-L2AT to Methanobacterium subterraneum and 98.5% of strain CaP3V-S-L1AT to Desulfomicrobium baculatum. Both strains grew from 20 to 42 °C, pH 5.0-7.5, and 0-4% NaCl. Based on our data, type strains CaP3V-M-L2AT (= DSM 113354 T = JCM 39174 T) and CaP3V-S-L1AT (= DSM 113299 T = JCM 39179 T) represent novel species which we name Methanobacterium cahuitense sp. nov. and Desulfomicrobium aggregans sp. nov.

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Xichú River is a Mexican river located in an environmental preservation area called Sierra Gorda Biosphere Reserve. Around it, there are tons of abandoned mine residues that represent a serious environmental issue. Sediment samples of Xichú River, visibly contaminated by flows of an acid mine drainage, were collected to study their prokaryotic diversity. The study was based on both cultural and non-cultural approaches. The analysis of total 16S rRNA gene by MiSEQ sequencing allowed to identify 182 Operational Taxonomic Units. The community was dominated by Pseudomonadota, Bacteroidota, "Desulfobacterota" and Acidobacteriota (27, 21, 19 and 16%, respectively). Different culture conditions were used focusing on the isolation of anaerobic bacteria, including sulfate-reducing bacteria (SRB) and arsenate-reducing bacteria (ARB). Finally, 16 strains were isolated. Among them, 12 were phylogenetically identified, with two strains being SRB, belonging to the genus Solidesulfovibrio ("Desulfobacterota"), while ten are ARB belonging to the genera Azospira (Pseudomonadota), Peribacillus (Bacillota), Raineyella and Propionicimonas (Actinomycetota). The isolate representative of Raineyella genus probably corresponds to a new species, which, besides arsenate, also reduces nitrate, nitrite, and fumarate.

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Molecular dynamics (MD) simulations were used to evaluate some chelating agents as potential candidates to inhibitors for dissimilatory adenosine-5'-phosphosulfate reductase (APSrAB). Molecular docking methods were used to evaluate the best binding modes of these molecules to the enzyme at two binding sites: of the substrate (enzyme active site) by mean the redocking protocol of substrate; and of one of the [Fe4S4]2+ groups by mean of the clusterization protocol. The best docking poses were selected by criteria such as low energy and RMSD (redocking) and the cluster with the higher number of similar poses (clusterization), which were submitted to MD simulations. RMSD, RDF, and hydrogen bonds results revelated that all ligands left the cube site, while in the active site, some ligands remained in their docking region, pointing to the enzyme active site as the best target for the selected ligands. The binding energy results of ligands hydroxamic acid (HXA) and catechol (CAT) showed that they bonded favorably to the enzyme and key residues of the active site contributed significantly to the protein-ligand bind, indicating HAX and CAT may compete with the substrate for interactions with these residues and displaying potential as candidates for experimental studies about APSrAB inhibitors.Communicated by Ramaswamy H. Sarma.

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Sulfate-reducing bioreactors are a biotechnological alternative for the treatment of acid mine drainage (AMD). In this study, two separate bioreactors with pH and temperature-controlled (Bio I and II) were operated with two different acidophilic microbial consortia to determine their efficiencies in sulfate removal from a synthetic acidic mine water. The bioreactors were operated for 302 days in continuous flow mode under the same parameters: fed with a sulfate solution of ∼30 mM with a pH of 2.5, the temperature at 30°C, stirred gently at 40 rpm and using a continuous stream of nitrogen to help remove the H2S produced in the bioreactor. The glycerol consumption, acetate production, and sulfate removal were monitored throughout the course of the experiment. The community composition and potential metabolic functional groups were analyzed via 16S rRNA partial gene sequencing. Bio I consortium reduced the sulfate, achieving a range of sulfate concentration from 4.7 to 19 mM in the effluent liquor. The removal of sulfate in Bio II was between 5.6 and 18 mM. Both bioreactors' communities showed the presence of the genus De sulfosporosinus as the main sulfate-reducing bacteria (SRB). Despite differences in microbial composition, both bioreactors have similar potential metabolism, with a higher percentage of microorganisms that can use sulfate in respiration. Overall, both bioreactors showed similar performance in treating acidic mine water containing mostly sulfate using two different acidophilic sulfidogenic consortia obtained from different global locations.

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Perchlorate is a widespread drinking water contaminant with regulatory standards ranging from 2 to 18 µg/L. The hydrogen-based membrane-biofilm reactor (MBfR) can effectively reduce perchlorate, but it is challenging to achieve low-µg/L levels. We explored chlorate addition to increase the abundance of perchlorate-reducing bacteria (PRB) and improve removals. MBfR reactors were operated with and without chlorate addition. Results show that chlorate doubled the abundance of putative PRB (e.g., Rhodocyclales) and improved perchlorate reduction to 23 ± 17 µg/L, compared to 53 ± 37 µg/L in the control. Sulfate reduction was substantially inhibited during chlorate addition, but quickly recovered once suspended. Our results suggest that chlorate addition can enhance perchlorate reduction by providing a selective pressure for PRB. It also decreases net sulfate reduction. KEY POINTS: • Chlorate increased the abundance of perchlorate-reducing bacteria • Chlorate addition improved perchlorate removal • Chlorate appeared to suppress sulfate reduction.

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It is anticipated that copper mining output will significantly increase over the next 20 years because of the more intensive use of copper in electricity-related technologies such as for transport and clean power generation, leading to a significant increase in the impacts on water resources if stricter regulations and as a result cleaner mining and processing technologies are not implemented. A key concern of discarded copper production process water is sulfate. In this study we aim to transform sulfate into sulfur in real mining process water. For that, we operate a sequential 2-step membrane biofilm reactor (MBfR) system. We coupled a hydrogenotrophic MBfR (H2-MBfR) for sulfate reduction to an oxidizing MBfR (O2-MBfR) for oxidation of sulfide to elemental sulfur. A key process improvement of the H2-MBfR was online pH control, which led to stable high-rate sulfate removal not limited by biomass accumulation and with H2 supply that was on demand. The H2-MBfR easily adapted to increasing sulfate loads, but the O2-MBfR was difficult to adjust to the varying H2-MBfR outputs, requiring better coupling control. The H2-MBfR achieved high average volumetric sulfate reduction performances of 1.7-3.74 g S/m3-d at 92-97% efficiencies, comparable to current high-rate technologies, but without requiring gas recycling and recompression and by minimizing the H2 off-gassing risk. On the other hand, the O2-MBfR reached average volumetric sulfur production rates of 0.7-2.66 g S/m3-d at efficiencies of 48-78%. The O2-MBfR needs further optimization by automatizing the gas feed, evaluating the controlled removal of excess biomass and S0 particles accumulating in the biofilm, and achieving better coupling control between both reactors. Finally, an economic/sustainability evaluation shows that MBfR technology can benefit from the green production of H2 and O2 at operating costs which compare favorably with membrane filtration, without generating residual streams, and with the recovery of valuable elemental sulfur.

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The competition between sulfate-reducing bacteria and methane-producing archaea has a major influence on organic matter removal, as well as the success of sulfidogenic systems. This study investigated the performance of six batch sulfidogenic reactors in response to different COD/sulfate ratios (1.0 and 2.0) and electron donors (cheese whey, ethanol, and sodium lactate) by evaluating the biochemical mechanisms of sulfate reduction, organic matter oxidation, and microbial structure modification. A COD/sulfate ratio of 1.0 resulted in high sulfidogenic activity for all electron donors, thereby achieving a nearly 80% sulfate removal. Lactate provided high sulfate removal rates at COD/sulfate ratios of 1.0 (80%) and 2.0 (90%). A COD/sulfate ratio of 2.0 decreased the sulfate removal rates by 25 and 28% when ethanol and cheese whey were used as substrates. The sulfate-reducing bacteria populations increased using ethanol and lactate at a COD/sulfate ratio of 1.0. Particularly, Desulfovibrio, Clostridium, and Syntrophobacter were predominant. Influent composition and COD/sulfate ratio influenced the relative abundance of the microbial communities. Therefore, controlling these parameters may facilitate the wastewater treatment with high sulfate levels through bacterial activity.

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Acid mine drainage (AMD) is the major effluent generated from metal and coal mines, causing serious ecological risks and degradation of aquatic habitats and surrounding soil quality. Biochemical passive reactors (BPRs) are an option for improving AMD affected water. This study investigates the effect of the size and concentration of zerovalent iron nanoparticles (nZVI) on the efficiency of batch BPRs during AMD remediation. Atomic force microscopy (AFM) and transmission electron microscopy (TEM) were also used as complementary techniques for the investigation of the changes in microbial cells and nZVI properties after the AMD remediation. The results from the batch experiment showed that the concentration of nZVI increases the pH and decreases ORP during AMD treatment, thus favouring the removal of metals. The results also suggest that metal sulfide precipitation occurred in all the batch with reactive mixture but was greater in reactors amended with nZVI of larger size. This study revealed that the presence of nZVI in the BPR leads to metal removal as well as the inhibition of sulfate-reducing bacteria (SRB) activity. Microscopy study indicated that the addition of nZVI creates a morphological change on certain microorganisms in which the cellular membrane was fully covered with nZVI, inducing cell lysis process. These results show that nZVI is a promising reactive material for the treatment of AMD in BPR systems.

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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.

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Successive years of gold mining in French Guiana has resulted in soil degradation and deforestation leading to the pollution and erosion of mining plots. Due to erosion and topography, gold panning sites are submitted to hydromorphy during rainfall and groundwater increases. This original study focused on characterizing the impact of hydromorphic anaerobic periods on bio-geochemical cycles. We sampled soil from five rehabilitated sites in French Guiana, including sites with herbaceous vegetation and sites restored with fabaceous plants, Clitoria racemosa (Cli) mon-oculture, Acacia mangium (Aca) monoculture, Clitoria racemosa and Acacia mangium (Mix) bi-culture. We conducted mesocosm experiments where soil samples were incubated in anaerobic conditions for 35 days. To evaluate the effect of anaerobic conditions on biogeochemical cycles, we measured the following parameters related to iron-reducing bacteria and sulfate-reducing bacteria metabolism throughout the experiment: CO2 release, carbon dissolution, sulphide production and sulphate mobilization. We also monitored the solubilization of iron oxyhydroxides, manganese oxides, aluminum oxides and mercury in the culture medium. Iron-reducing bacteria (IRB) and sulfate-reducing bacteria (SRB) are described as the major players in the dynamics of iron, sulfur and metal elements including mercury in tropical environments. The results revealed two trends in these rehabilitated sites. In the Aca and Mix sites, bacterial iron-reducing activity coupled with manganese solubilization was detected with no mercury solubilization. In herbaceous sites, a low anaerobic activity coupled with sulphide production and mercury solubilization were detected. These results are the first that report the presence and activity of iron- and sulfate-reductive communities at rehabilitated mining sites and their interactions with the dynamics of metallic elements and mercury. These results report, however, the positive impact of ecological restoration of mining sites in French Guiana by reducing IRB and SRB activities, the potential mobility of mercury and its risk of transfer and methylation.

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An iron reducing enrichment was obtained from sulfate reducing sludge and was evaluated on the capability of reducing Fe3+ coupled to acetate oxidation in a microbial fuel cell (MFC). Three molar ratios for acetate/Fe3+ were evaluated (2/16, 3.4/27 and 6.9/55 mM). The percentages of Fe3+ reduction were in a range of 80-90, 60-70 and 40-50% for the MFCs at closed circuit for the molar ratios of 2/16, 3.4/27 and 6.9/55 mM, respectively. Acetate consumption was in a range of 80-90% in all cases. The results obtained at closed circuit for current density were: 11.37 mA/m2, 4.5 mA/m2 and 7.37 mA/m2 for the molar ratios of 2/16, 3.4/27 and 6.9/55 mM, respectively. Some microorganisms that were isolated and identified in the MFCs were Azospira oryzae, Cupriavidus metallidurans CH34, Enterobacter bugandensis 247BMC, Citrobacter freundii ATCC8090 and Citrobacter murliniae CDC2970-59, these bacteria have been reported as exoelectrogens in MFC and in MFC involving metals removal but not all of them have been reported to utilize acetate as preferred substrate. The results demonstrate that the isolates can utilize acetate as the sole source of carbon and suggest that Fe3+ reduction was carried out by a combination of different mechanisms (direct contact and redox mediators) utilized by the bacteria identified in the MFC. Storage of the energy generated from the 2/16 mM MFC system arranged in a series of three demonstrated that it is possible to utilize the energy to charge a battery.

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RESUMO Rochas contendo sulfetos metálicos podem ser oxidadas em um processo catalisado por procariotos quimiolitoautotróficos ou Fe3+. A atividade mineradora acelera esse processo ao gerar resíduos contendo sulfetos metálicos com grande superfície de contato. O lixiviado resultante, conhecido como drenagem de mina (DM), é rico em sulfato, íons hidrogênio e contaminantes químicos inorgânicos como ferro (Fe), zinco (Zn), cádmio (Cd), manganês (Mn), níquel (Ni), arsênio (As) e alumínio (Al). Para remover tais poluentes, atualmente, o principal tratamento utilizado é a adição de reagentes alcalinos. Entretanto, esse método tem limitada eficiência, alto custo e gera grandes volumes de resíduos sólidos tóxicos de relativa solubilidade. Bactérias redutoras de sulfato (BRS) podem oxidar matéria orgânica com geração de sulfeto. Algumas vias metabólicas do processo consomem H+neutralizando o pH. O sulfeto produzido pode reagir com contaminantes inorgânicos e precipitá-los, permitindo sua recuperação da fase líquida. O uso de subprodutos industriais e urbanos contendo diferentes fontes de carbono como doadores de elétrons no tratamento de DM tem sido investigado. Este artigo sumariza dados sobre as variáveis relevantes para a atividade microbiana durante o tratamento biológico de DM, analisando o atual cenário de pesquisas com fontes alternativas de carbono. Discute-se ainda novas fontes de matéria orgânica ainda não aplicadas para tratamento biológico de efluentes e que, sob aspectos de sustentabilidade, dos pontos de vista sustentável e econômico, podem ser usadas no tratamento de resíduos.

ABSTRACT Rocks containing metal sulfides be can oxidized biologically or chemically. Chemolithoautotrophics prokaryotes and Fe3+ catalyze this process. The mining activities also accelerate the process for creates metal sulphides tailings with a big contact surface. The leached formed is called Mine Drainage (MD) whose composition is rich in sulphate, hydrogen ions and inorganic chemical contaminants such as Fe, Zn, Mn, Cd, Ni, As e Al. Currently, in order to remove these pollutants, the main treatment used is the addition of alkaline reagents. However, the method has limited efficiency, high cost with input reagents and generates wide amounts of toxic solid residues with high solubility. The sulphide reducing bacterias (RSB) can oxidize organic matter generating sulphide. Some metabolic pathways consume H+ neutralizing the pH. The sulphide formed can react and precipitate inorganic pollutants, allowing their recuperation from the liquid phase. The use of industrial and urban by-products containing different carbon sources have been investigated as an electron donor in the MD treatment. The diverse microbial consortia synergic acting can present bigger efficiency in the presence of mixed carbon sources, besides lower cost in relation to the pure matter. Here will be detailed the biological treatment about which and how the variables of the system can influence the microbial activity and relevant molecules to the treatment. After is described the current situation of the research about alternative carbon sources. New carbon sources whose are a by-product of the expanding industry presenting good feature to anaerobic degrading are suggested. The by-product potential is described from the point of view of sustainability, and waste management.

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A paramour factor limiting metal-microorganism interaction is the metal ion concentration, and the metal precipitation efficiency driven by microorganisms is sensitive to metal ion concentration. The aim of the work was to determine the tolerance of the sulfidogenic sludge generated from hydrothermal vent sediments at microcosms level to different concentrations of Fe, Cu and Zn and the effect on the microbial community. In this study the chemical oxygen demand (COD) removal, sulfate-reducing activity (SRA) determination, inhibition effect through the determination of IC50, and the characterization of the bacterial community´s diversity were conducted. The IC50 on SRA was 34 and 81 mg/L for Zn and Cu, respectively. The highest sulfide concentration (H2S mg/L) and % of sulfate reduction obtained were: 511.30 ± 0.75 and 35.34 ± 0.51 for 50 mg/L of Fe, 482.48 ± 6.40 and 33.35 ± 0.44 for 10 mg/L of Cu, 442.26 ± 17.1 and 30.57 ± 1.18 for 10 mg/L of Zn, respectively. The COD removal rates were of 71.81 ± 7.6, 53.92 ± 1.07 and 57.68 ± 10.2 mg COD/ L d for Fe (50 mg/L), Cu (40 mg/L) and Zn (20 mg/L), respectively. Proteobacteria, Firmicutes, Chloroflexi and Actinobacteria were common phyla to four microcosms (stabilized sulfidogenic and added with Fe, Cu or Zn). The dsrA genes of Desulfotomaculum acetoxidans, Desulfotomaculum gibsoniae and Desulfovibrio desulfuricans were expressed in the microcosms supporting the SRA results. The consortia could be explored for ex-situ bioremediation purposes in the presence of the metals tested in this work.

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The acid mine drainage (AMD) can causes negative impacts to the environment. Physico-chemical methods to treat AMD can have high operational costs. Through passive biological methods, such as anaerobic reactors, sulfate reduction, and recovery of metals are promoted. This study evaluated the performance of a horizontal anaerobic immobilized biomass (HAIB) reactor for the treatment of synthetic AMD using polyurethane foam as support material, and anaerobic sludge as inoculum. Ethanol was used as an electron donor for sulfate reduction, resulting in an influent chemical oxygen demand (COD) in the range of 500-1,500 mg/L and COD/sulfate ratio at 1. A gradual increase of sulfate and COD concentration was applied that resulted in COD removal efficiencies higher than 78%, and sulfate removal efficiencies of 80%. Higher sulfate and COD concentrations associated with higher hydraulic retention times (36 h) proved to be a better strategy for sulfate removal. The HAIB reactor was able to accommodate an increase in the SLR up to 2.25 g SO42-/L d-1 which achieved the greatest performance on the entire process. Moreover, the reactor proved a suitable alternative for reaching high levels of metal removal (86.95 for Zn, 98.79% for Fe, and 99.59% for Cu).

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Biomats that flourished in a fumarole located on the geothermal site Los Azufres (Mexico) were used as inocula to select aerobic and sulfate-reducing bacteria consortia for studying their capacity to reduce hexavalent chromium [Cr(VI)], aiming to use these consortia in biotransformation technologies. The sample site is characterized by slightly warm (nearly 27 [Formula: see text]C), acid (pH 3) and about hypoxic (1.8 mg L[Formula: see text] of dissolved oxygen) conditions. Four culture systems (2 aerobic and 2 anaerobic) were investigated, including their enzymatic activity, capacity to produce biofilms, and an analysis of the total bacterial populations. For the anaerobic condition (using sulfate and sulfur as electron acceptors), four pH values (from 2 to 8) and four carbon sources (pyruvate, glycerol, Na-lactate and Na-acetate) were probed. Significant biological Cr(VI) removal was observed for all the pH values probed, particularly during the first 12 h, being more effective at the most acid conditions. At a pH value of 4 and using pyruvate as carbon source, 100 mg L[Formula: see text] of Cr(VI) were completely depleted in less than 12 h, while the use of Na-lactate was less effective but still reasonable. These results indicate that sulfate-reducing bacteria consortia from geothermal sites like the one studied here are capable of biotransforming Cr(VI) and have the potential to provide metal bioremediation technologies.

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