Investigations on the “Extreme” Microbial Arsenic Cycle within the Sediments of an Acidic Impoundment of the Former Sulfur Bank Mercury Mine: Herman Pit, Clear Lake, California.

Abstract:

The involvement of prokaryotes in the redox reactions of arsenic occurring between this element’s +5 [arsenate; As(V)] and + 3 [arsenite; As(III)] oxidation states has been well established. Most research has focused upon circum-neutral pH environments, such as freshwater lake and aquifer sediments, and extreme environments like hot springs and hypersaline soda lakes have also been well investigated. In contrast, little work has been conducted on acidic environments. The azure-hued, clear waters of the Herman Pit are acidic (pH 2-4), and overlie oxidized sediments that have a distinctive red/orange coloration indicative of the presence of ferrihydrites and other Fe(III) minerals. There is extensive ebullitive release of geothermal gases from the lake bottom in the form of numerous continuous-flow seeps which are composed primarily of mixtures of CO₂, CH₄, and H₂S. We collected near-shore surface sediments with an Eckman grab, and stored the “soupy” material in filled mason jars kept at 4˚C. Initial experiments were conducted using 3:1 mixtures of lake water: sediment so as to generate dilute slurries which were amended with mM levels of electron acceptors (arsenate, nitrate, oxygen), electron donors (arsenite, acetate, lactate, hydrogen), and incubated under N₂, air, or H₂. Owing to the large adsorptive capacity of the Fe(III)-rich slurries, we were unable to detect As(V) or As(III) in the aqueous phase of either live or autoclaved controls, although the former consumed lactate, acetate, nitrate, or hydrogen, while the latter did not. This prompted us to conduct a series of further diluted slurry experiments using the live materials from the first as a 10 % addition to lakewater. In these experiments we observed reduction of As(V) to As(III) in anoxic slurries and that rates were enhanced by addition of electron donors (H₂, acetate, or lactate). We also observed oxidation of As(III) to As(V) in oxic slurries and in anoxic slurries amended with nitrate. These results indicate the presence of a viable microbial As(V)/As(III) redox cycle in the sediments of this extreme environment. Further investigations using culture-independent protocols to identify participant prokaryotes and their functional arsenic genes (e.g., aioA, arrA, arxA) are underway at this time.