An acidophilic,rod-shaped Gram-negative sulfur oxidizing strain BY-05 was isolated from an acid mine drainage of copper ore in Baiyin area,Gansu Province,China.Ultrastructural studies show that the isolate has a tuft of polar flagella and possesses sulfur granules with clear membrane adhering to the cell innermembrane.Physiological study shows that this isolate grows autotrophically and aerobically by oxidizing S0and reduced inorganic sulfur compounds(SO, 2 23-SO, 2 24- S2 -and ZnS)with the optimum growth at pH 3.5-4.0 and at the temperature range of 25-30℃.The 16S rRNA gene sequence(DQ 423683)of strain BY-05 has 100%sequence similarity to that of Acidithiobacillus albertensis(DSM 14366).So it is identified and named as A. albertensis BY-05.Bioleaching experiments with this new strain show that it can play an important role in recovery of metals from chalcopyrite and sphalerite.
Eleven acid mine drainage （AMD） samples were obtained from southeast of China for the analysis of the microbial communities diversity, and the relationship with geochemical variables and spatial distance by using a culture-independent 16S rDNA gene phylogenetic analysis approach and multivariate analysis respectively. The principle component analysis （PCA） of geochemical variables shows that eleven AMDs can be clustered into two groups, relative high and low metal rich （RHMR and RLMR） AMDs. Total 1691 clone sequences are obtained and the detrended correspondence analysis （DCA） of operational taxonomic units （OTUs） shows that, ~,-Proteobacteria, Acidobacteria, Actinobacteria, Cyanobacteria, Firmicutes and Nitrospirae are dominant species in RHMR AMDs. In contrast, a-Proteobacteria, fl-Proteobacteria, Planctomycetes and Bacteriodetes are dominant species in RLMR AMD. Results also show that high-abundance putative iron-oxidizing and only putative sulfur-oxidizing microorganisms are found in RHMR AMD. Multivariate analysis shows that both geochemical variables （r=0.429 3, P=-0.037 7） and spatial distance （r=0.321 3, P=-0.018 1） are significantly positively correlated with microbial community and pH, Mg, Fe, S, Cu and Ca are key geochemistry factors in shaping microbial community. Variance partitioning analysis shows that geochemical variables and spatial distance can explain most （92%） of the variation.
According to physiological and biochemical characteristics of Leptospirillum ferriphilum, a strain of object bacteria was isolated successfully. Bacteria were enriched by selective liquid medium and plated on designed single-layered agar solid medium. Colony was cultured and bacteria were collected. The morphologies of the object bacteria were observed using crystal violet staining, scanning electron microscope（SEM） and transmission electron microscope （TEM）. The result of 16S rDNA identification shows that this bacterium belongs to Leptospirillum ferriphilum and it is named as Leptospirillum ferriphilum strain D1. These results indicate that this new single-layered agar solid medium is efficient and physiological-biochemical characteristics show that the optimum simple for isolation of Leptospirillum ferriphilum. Additionally, initial pH value and its growth temperature are 1.68 and 40℃.
Mixed microorganisms with elevated activity of chalcocite-leaching were screened by mutation methods. The original microorganisms collected from acid mine drainage of different sites were mixed and then treated with mutagens NO2^- , diethyl sulfate （DES）, UV and their combinations, respectively. Five groups of mixed microorganisms with much stronger ore-leaching ability were obtained by screening on the leaching media. Among them, group E of mixed microorganisms （treated with 1% DES for 60 min） with the best perfonnance on chalcocite-leaching, increases the content of Cu^2＋ by 101.4% in 20 d of leaching compared with the control culture. In addition, group E is more tolerant to Cu^2＋ in media than the control without mutation treatment. Analysis for the diversity of microbial clones indicates that half of operational taxonomic units （OTUs） in group E are Acidithiobacillus ferrooxidans. These observations suggest that group E might have potentials for industrial application.
The effect of mineral particle size, pulp potential and category of oxidant on pyrite leaching was studied. The results show that a smaller mineral particle size leads to a higher leaching rate of pyrite, and the optimum result with pyrite leaching rate of 2.92% is obtained when mineral particle size is less than 0.037 mm. The pulp potential reflects the leaching process. The increase of pulp potential can improve pyrite leaching. The leaching rate and velocity of pyrite can be enhanced rapidly by adding strong oxidant. The kind and the method of adding oxidant have important effect on the pyrite leaching. Appropriate concentration of Fe3+ can enhance pyrite leaching but the precipitation generated by high concentration of ferric ion covers the surface of pyrites and prevents the leaching process. The leaching rate increases with the constant addition of H2O2.
