The Role of Microorganisms in the Biogeochemical Cycle of Arsenic in the Environment

Abstract

Arsenic (As) is a highly toxic chemical that is widely distributed in groundwater around the world. As-bearing sulfide minerals (ASM) are known to contribute to high background concentrations of As in groundwater in regions where the geochemistry of the parent material is dominated by sulfide minerals. The fate of As in groundwater depends on the activity of microorganisms which can oxidize arsenite (Asᴵᴵᴵ), or reduce arsenate (Asᵛ). In oxidizing environments, Asᵛ is the predominant species, and the accumulation of As is limited by the sorption of As onto iron (Fe) oxides and hydroxides. Under reducing environments, Asᴵᴵᴵ is the predominant specie, and while the sorption strength of Asᴵᴵᴵ on the Fe-surface of Fe (oxy)hydroxides is weaker, the accumulation of As in water can be limited by the precipitation of As as part of an ASM. The main aim of this research is to study the impact of microbial activity on the mobilization and immobilization of As in the environment. The first objective of this research was to characterize the metabolic activity of three Asᴵᴵᴵ-oxidizing bacteria, Azoarcus sp. pb-1 strain EC1, Azoarcus sp. pb-1 strain EC3 and Diaphorobacter sp. pb-1 strain MC, isolated from a non-contaminated, pristine environment. These Asᴵᴵᴵ-oxidizing bacteria demonstrated a great metabolic flexibility to use oxygen and nitrate to oxidize Asᴵᴵᴵ as well as organic and inorganic substrates as alternative electron donors (e-donors) explains their presence in non-As-contaminated environments. The findings suggest that at least some Asᴵᴵᴵ-oxidizing bacteria are flexible with respect to electron-acceptors and e-donors and that they are potentially widespread in low As concentration environments. The second objective of this research was to investigate the stability of orpiment (As₂S₃) and arsenopyrite (FeAsS), at circumneutral pH and 30°C, under aerobic- and or anoxic conditions (nitrate amended as electron acceptor (e-acceptor)), in order to assess the feasibility of immobilizing As by formation of ASM as a long-term option for the bioremediation of As contamination. The percentage of As released from the minerals ranged from zero when FeAsS was biologically incubated to 87% for As₂S₃(s) under anoxic abiotic conditions. While the dissolution of ASM was greater in biological conditions, the presence of inoculum provided as sludge served as a sink for As, limiting the mobilization of As into aqueous phase. Thus, the mobilization of As from ASM can be controlled by altering the environmental conditions such as the redox conditions or by stimulating microbial activity. Further research investigated the formation of ASM catalyzed by biological reduction of Asᵛ and sulfate (SO₄²⁻). In particular, the third objective of this research was to study the effect of the pH on the removal of As due to the biological-mediated formation of ASM in an iron-poor system. A series of batch experiments were performed to study the reduction of SO₄²⁻ and Asᵛ by an anaerobic mixed culture in a range of pH conditions (6.1-7.2), using ethanol as the e-donor. A marked decrease of the total aqueous concentrations of As and S and the formation of a yellow precipitate was observed in the inoculated treatments amended with ethanol, but not in the non-inoculated controls, indicating that the As-removal was biologically mediated. The pH dramatically affected the extent and rate of As removal, as well as the stoichiometric composition of the precipitate. The precipitate was composed of a mixture of orpiment and realgar, and the proportion of orpiment in the sample increased with increasing pH. The results suggest that ASM formation is greatly enhanced at mildly acidic pH conditions. The fourth objective was to investigate the biomineralization of As through simultaneous Asᵛ and SO₄²⁻ reduction in a minimal iron environment for the As-contaminated groundwater bioremediation. A continuous bioreactor, inoculated with an anaerobic sludge was maintained at circumneutral pH (6.25-6.50) and fed with Asᵛ and SO₄²⁻, utilizing ethanol as an e-donor for over 250 d. A second bioreactor running under the same conditions but lacking SO₄²⁻ was operated as a control to study the fate of As removal. The reactor fed with both Asᵛ and SO₄²⁻ removed on the average 91.2% of the total soluble As, while less than 5% removal was observed in the control bioreactor without S. The biomineralization of As in the bioreactor was also evident from the formation of a yellow precipitate made of a mixture of As₂S₃ and AsS minerals. These results taken as a whole indicate that a bioremediation process relying on the addition of a simple, low-cost e-donor offers potential to promote the removal of As from groundwater by precipitation of ASM. The fifth objective was to evaluate the toxic impact that the exposure to soluble As or the formation of ASM could have on the anaerobic mixed culture used as inocula. The methanogenic community on the reactors was impacted by addition of As. The biogenic ASM inhibited the acetoclastic methanogens causing an accumulation of acetate. In the SO₄²⁻-free bioreactor, the methanogens were initially highly sensitive to Asᴵᴵᴵ (formed from Asᵛ reduction) but quickly adapted to its toxicity. Consequently, the formation of ASM would impact the methanogenic activity of an anaerobic biofilm, while the exposure to Asᴵᴵᴵ would not have a negative impact if the biofilm undergoes adaptation. The sixth and final objective was to study the stability of a biogenic ASM at two different pH values (6.5 and 7.5) and under different redox conditions. The long-term stability was evaluated in three different bioreactors that operated for 145 d: aerobic (R1), anoxic (nitrate as alternative e-acceptor (R2) and anaerobic (R3). The dissolution of ASM was greatly affected by the pH, and slightly by the presence and nature of the e-acceptor. The ASM was very stable at pH 6.5, however, the As mobilization rate was up to 7-fold higher at pH 7.5, likely due to the formation of thioarsenic species. The stability of ASM was also impacted by the e-acceptor present. The As mobilization rate was 77% higher under anaerobic conditions than under aerobic conditions, most likely due to the formation of secondary As-bearing minerals. Therefore, the stability of ASM depends on the conditions of the operation, and it can be controlled by altering the environmental conditions, such as the pH or the presence of the e-acceptor.