Geochemical diversity in S processes mediated by culture-adapted and environmental-enrichments of Acidithiobacillus spp.

Coupled S speciation and acid generation resulting from S processing associated with five different microbial treatments, all primarily Acidithiobacillus spp. (i.e. autotrophic S-oxidizers) were evaluated in batch laboratory experiments. Microbial treatments included two culture-adapted strains, Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, their consortia and two environmental enrichments from a mine tailings lake that were determined to be >95% Acidithiobacillus spp., by whole-cell fluorescent hybridization. Using batch experiments simulating acidic mine waters with no carbon amendments, acid generation, and S speciation associated with the oxidation of three S substrates (thiosulfate, tetrathionate, and elemental S) were evaluated. Aseptic controls showed no observable pH decrease over the experimental time course (1 month) for all three S compounds examined. In contrast, pH decreased in all microbial treatments from starting pH values of 4 to 2 or less for all three S substrates. Results show a non-linear relationship between the pH dynamics of the batch cultures and their corresponding sulfate concentrations, and indicate how known microbial S processing pathways have opposite impacts, ultimately on pH dynamics. Associated geochemical modeling indicated negligible abiogenic processes contributing to the observed results, indicating strong microbial control of acid generation extending over pH ranges from 4 to less than 2. However, the observed acid generation rates and associated S speciation were both microbial treatment and substrate-specific. Results reveal a number of novel insights regarding microbial catalysis of S oxidation: (1) metabolic diversity in S processing, as evidenced by the observed geochemical signatures in S chemical speciation and rates of acid generation amongst phylogenetically similar organisms (to the genus level); (2) consortial impacts differ from those of individual strain members; (3) environmental enrichments of Acidithiobacillus spp. catalyze different S reaction arrays than pure strain Acidithiobacillus spp.; and (4) microbial catalysis of S reactions involves significant disproportionation tied to substantial H⁺ consumption, with the formation of as yet, poorly characterized intermediate S species, most likely polythionates and polysulfane monosulfonic acids that are thought to be involved in microbial S storage mechanisms.