A previously unknown chemolithoautotrophic arsenite-oxidizing bacterium has been isolated from a gold mine in the Northern Territory of Australia. The organism, designated NT-26, was found to be a gram-negative motile rod with two subterminal flagella. In a minimal medium containing only arsenite as the electron donor (5 mM), oxygen as the electron acceptor, and carbon dioxide-bicarbonate as the carbon source, the doubling time for chemolithoautotrophic growth was 7.6 h. Arsenite oxidation was found to be catalyzed by a periplasmic arsenite oxidase (optimum pH, 5.5). Based upon 16S rDNA phylogenetic sequence analysis, NT-26 belongs to the Agrobacterium/Rhizobium branch of the ␣-Proteobacteria and may represent a new species. This recently discovered organism is the most rapidly growing chemolithoautotrophic arsenite oxidizer known.Arsenic is found in many environments and is toxic to life in some forms. When present in insoluble forms, which are nontoxic, arsenic is frequently found as a mineral in combination with sulfur (e.g., orpiment [As 2 S 3 ] and realgar [AsS]) and especially with iron and sulfur (e.g., arsenopyrite [FeAsS]). The oxidation of these insoluble forms, chemically and/or microbiologically, results in the formation of arsenite (As III [H 3 AsO 3 ]). In environments such as acid mine drainage, arsenite concentrations can be extremely high (2 to 13 mg/liter) (22). The arsenite formed in various environments can then be oxidized to arsenate (As V [H 2 AsO 4 Ϫ ϩ H ϩ ]). Both of these soluble forms of arsenic, arsenite and arsenate, are toxic to living organisms, with arsenite considered more toxic than arsenate (5, 22).Interestingly, the chemical oxidation of arsenite to arsenate is slow. Most arsenite is, therefore, oxidized to arsenate microbiologically (22). Arsenite-oxidizing bacteria were first described in 1918 (7). These organisms, as well as a number of others that have been isolated more recently, are almost all heterotrophic arsenite-oxidizing bacteria, as they require the presence of organic matter for growth (17,18,23,24). The most common organism found has been Alcaligenes faecalis (7,17,18,23,24). For these heterotrophic bacteria, the oxidation (equation 1) is considered a detoxification mechanism rather than one that can support growth, despite the fact that the reaction is exergonic.Only one bacterium has been described that is able to use the energy gained from this reaction for growth. This organism, Pseudomonas arsenitoxidans, was found to grow chemolithoautotrophically with arsenite, oxygen, and carbon dioxide. The fastest doubling time reported for growth with arsenite was, however, in the order of 2 days (8). This report describes a new bacterium, isolated from a gold mine in the Northern Territory of Australia, that can also grow chemolithoautotrophically with arsenite as the electron donor, oxygen as the electron acceptor, and carbon dioxide (CO 2 ) or bicarbonate (HCO 3 Ϫ ) as the carbon source. Growth was rapid, with a doubling time of 7.6 h for chemolithoautotrophic grow...