Biodiversity study of Ambamata multimetal mine, Gujarat: Isolation and characterization of Sulphur and Iron utilizers
DOI:
https://doi.org/10.69980/jaz.v35i01.5403Keywords:
Iron oxidizers, Acidithiobacillus ferrooxidans, Pseudomonas stutzeri, metal sulphide, consortium, 16s rRNA, Heterotrophic thiosulphate- oxidizing bacteria, chemolithotrophs, chemolithoheterotrophs.Abstract
After screening of many ores available at department of Microbiology, Gujarat University, for their metal content and gangue material, pollymetallic sulphidic ore from the Ambamata multimetal mine, Ambaji, Gujarat, India was selected for study. The ore contained zinc, copper and lead along with Sulphur and pyrite. Chemical analysis of the ore showed the presence of 7.5% Zn, 6.7% Fe, 3.0% Pb, 0.545 % Cu, 0.064% Cd, and 0.002% Ag. Because of the lower concentration of the minerals, this ore could not be utilized for metal recovery by conventional metal extraction methods and hence heap bioleaching was tested for its feasibility. Three Iron oxidizers DA, DB and DC were isolated and tested for iron oxidation kinetics. Isolate DC was the fastest growing among them, so was used for further study after adaptation. Sulphur oxidizing isolate DS1 decreased the medium pH from 3.5 to 0.4, indicating it as potent acid generator at the site. Thiosulphate oxidizing isolate TH1 showing 78% thiosulphate oxidation within 11 days and decreased the medium pH from 4.5 to less than 3.0, proved to be the other potential acid generator at the site. Eleven strains of heterotrophic bacteria were isolated from various water samples collected from Ambamata multi metal mine site on the basis of their potential to utilize thiosulphate as an energy source under the aerobic conditions. All the isolates were analyzed for their growth pattern, sulphate production, medium pH change and thiosulphate utilization. Isolated organisms showed great diversity in colony morphology. Sequence analysis the 16S rRNA genes of the selected isolates showed their affiliation to genomovars of Pseudomonas stutzeri. The data suggested that thiosulphate utilizing bacteria, in particular those belonging to Pseudomonas stutzeri, can play a significant role in bioleaching. The isolates showed different substrate utilization profile when tested on Biolog® plates. A mixed consortium, predominantly Acidithiobacillus ferrooxidans isolated from Ambamata multi metal mine water sample was used as inoculum for the experiment. Repeated sub culturing was performed to ensure activation and the adaptation of the strain and reduction in lag phase. Subsequently, the bacterial lag phase was reduced and the log phase initiated immediately after inoculation of the strains in fresh media. After three sets of sub culturing, a steady state of the iron oxidation rate was achieved. This bacterial culture was then inoculated in 9k medium containing 2% pyrite and aerated continuously. Repeated transfer of the consortium in pyrite containing media reduced the time from 104 hours to 48 hours required for >98% pyrite oxidation. The cells were finally transferred to 9k medium having 2% metal sulphide.
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