The extensive use of 3-nitro-4-hydroxybenzene arsonic acid (roxarsone) in the production of broiler chickens can lead to increased soil arsenic concentration and arsenic contaminated dust. While roxarsone is the dominant arsenic species in fresh litter, inorganic As (V) predominates in composted litter. Microbial activity has been implicated as the cause, but neither the specific processes nor the organisms have been identified. Here we demonstrate the rapid biotransformation of roxarsone under anaerobic conditions by Clostridium species in chicken litter enrichments and a pure culture of a fresh water arsenate respiring species (Clostridium sp. strain OhILAs). The main products were 3-amino-4-hydroxybenzene arsonic acid and inorganic arsenic. Growth experiments and genomic analysis indicate strain OhILAs may use roxarsone as a terminal electron acceptor for anaerobic respiration. Electronic structure analysis suggests that the reducing equivalents should go to the nitro group, while liberation of inorganic arsenic from the intact benzene ring by cleaving the C-As bond is unlikely. Clostridium and Lactobacillus species are common in the chicken cecum and litter. Thus, the organic-rich manure and anaerobic conditions typically associated with composting provide the conditions necessary for the native microbial populations to transform the roxarsone in the litter releasing the more toxic inorganic arsenic.
Alkaliphilus oremlandii sp. nov. strain OhILAs is a mesophilic, spore-forming, motile, low mole%GC gram positive. It was enriched from Ohio River sediments on a basal medium with 20 mM lactate and 5 mM arsenate and isolated through passage on medium with increased arsenic concentration (10 and 20 mM), tindalization, and serial dilution. The pH optimal for growth was 8.4 and 16S rRNA gene sequence analysis indicated it is most closely related to species in the genus Alkaliphilus (A. crotonoxidans 95%, A. auruminator 95%, A. metalliredigens, 94%). A strict anaerobe, it can ferment lactate via the acrylate pathway as well as fructose and glycerol. A. oremlandii also has respiratory capability, as it is able to use arsenate and thiosulfate as terminal electron acceptors with acetate, pyruvate, formate, lactate, fumarate, glycerol, or fructose as the electron donor. A respiratory arsenate reductase, which is constitutively expressed, has been identified through biochemical and Western blot analyses and confirmed by cloning and sequencing of the gene encoding the structural subunit arrA. The entire arr operon as well as the ars operon have also been identified in the fully annotated genome. A. oremlandii also transforms the organoarsenical 3-nitro-4-hydroxy benzene arsonic acid (roxarsone). Growth experiments and genomic analysis suggest that it couples the reduction of the nitro group of the organoarsenical to the oxidation of either lactate or fructose in a dissimilatory manner, generating ATP via a sodium dependent ATP synthase.
Syntrophobacter fumaroxidans strain MPOBT is the best-studied species of the genus Syntrophobacter. The species is of interest because of its anaerobic syntrophic lifestyle, its involvement in the conversion of propionate to acetate, H2 and CO2 during the overall degradation of organic matter, and its release of products that serve as substrates for other microorganisms. The strain is able to ferment fumarate in pure culture to CO2 and succinate, and is also able to grow as a sulfate reducer with propionate as an electron donor. This is the first complete genome sequence of a member of the genus Syntrophobacter and a member genus in the family Syntrophobacteraceae. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 4,990,251 bp long genome with its 4,098 protein-coding and 81 RNA genes is a part of the Microbial Genome Program (MGP) and the Genomes to Life (GTL) Program project.
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