The use of a thermophilic acidophilic iron-oxidizing archaeon, Acidianus brierleyi, was investigated for oxidation and immobilization of As(III) from acidic refinery waste water. Some As(III) oxidation was measured in all Ac. brierleyi cultures independently of the presence or concentration of Fe(II) in bulk solution; the exception was at initial Fe(II) concentration ([Fe(II)](ini)) of 1000 mg l(-1) where As(III) oxidation became markedly facilitated and consequently approximately 70% of As was immobilized as amorphous ferric arsenate. Providing 1000 mg l(-1) Fe(III) instead of Fe(II) did not show the same effect, implying the importance of Fe(III) be microbially-produced and complexed in the archaeal EPS (extracellular polymeric substances) region for effective As(III) oxidation. The reaction towards secondary mineral formation shifted from ferric arsenate to jarosite at [Fe(II)](ini) of >1000 mg l(-1). Furthermore addition of jarosite seed crystals retarded the As(III) oxidation rate at [Fe(II)](ini) of 1000 mg l(-1). The observations indicate that by setting the appropriate bulk Fe(II)/As(III) ratio in Ac. brierleyi culture to achieve a certain concentration of Fe(III) within the EPS region, but at the same time to avoid jarosite formation, it is possible to maximize the As(III) oxidation rate and thus As immobilization efficiency. This study describes for the first time microbially-mediated simultaneous oxidation and immobilization of As(III) as ferric arsenate, using a thermoacidophilic iron-oxidizing archaeon, Ac. brierleyi. (C) 2012 Elsevier Ltd. All rights reserved.