Anaerobic Oxidation of Arsenite Linked to Chlorate Reduction

Abstract

Microorganisms play a significant role in the speciation and mobility of arsenic in the environment. In this study, the oxidation of arsenite [As(III)] to arsenate [As(V)] linked to chlorate (ClO₃⁻) reduction was shown to be catalyzed by sludge samples, enrichment cultures (ECs), and pure cultures incubated under anaerobic conditions. No activity was observed in treatments lacking inoculum or with heat-killed sludge, or in controls lacking ClO₃⁻. The As(III) oxidation was linked to the complete reduction of ClO₃⁻ to Cl⁻, and the molar ratio of As(V) formed to ClO₃⁻ consumed approached the theoretical value of 3:1 assuming the e⁻ equivalents from As(III) were used to completely reduce ClO₃⁻. In keeping with O₂ as a putative intermediate of ClO₃⁻ reduction, the ECs could also oxidize As(III) to As(V) with O₂ at low concentrations. Low levels of organic carbon were essential in heterotrophic ECs but not in autotrophic ECs. 16S rRNA gene clone libraries indicated that the ECs were dominated by clones of Rhodocyclaceae (including Dechloromonas, Azospira, and Azonexus phylotypes) and Stenotrophomonas under autotrophic conditions. Additional phylotypes (Alicycliphilus, Agrobacterium, and Pseudoxanthomonas) were identified in heterotrophic ECs. Two isolated autotrophic pure cultures, Dechloromonas sp. strain ECC1-pb1 and Azospira sp. strain ECC1-pb2, were able to grow by linking the oxidation of As(III) to As(V) with the reduction of ClO₃⁻. The presence of the arsenite oxidase subunit A (aroA) gene was demonstrated with PCR in the ECs and pure cultures. This study demonstrates that ClO₃⁻ is an alternative electron acceptor to support the microbial oxidation of As(III).