Isolation of Leptospirillum ferriphilum by single-layered solid medium
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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 simple for isolation of Leptospirillum ferriphilum. Additionally, physiological-biochemical characteristics show that the optimum initial pH value and its growth temperature are 1.68 and 40 °C, respectively. The culture of it is used to leach a complex concentrate chalocpyrite, the leaching efficiencies of copper and iron are 1.93% and 13.74%, respectively, and it is more effective than the A. ferrooxidans culture in the leaching of the complex concentrate chalcopyrite.
Key wordsLeptospirillum ferriphilum isolation purification solid medium bioleaching concentrate chalcopyrite
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- PIVOVAROVA T A, MARKOSYAN G E, KARAVAIKO G I. The auxotrophic growth of Leptospirillum ferrooxidans[J]. Microbiology, 1981, 50(1): 339–344.Google Scholar
- HIPPE H. Leptospirillum gen. nov. (ex Markosyan 1972), nom. rev., including Leptospirillum ferrooxidans sp. nov. (ex Markosyan 1972), nom. rev. and Leptospirillum thermoferrooxidans sp. nov. (Golovacheva et al. 1992) [J]. Int J Syst Evol Microbiol, 2000, 50(5): 501–503.Google Scholar
- NICOLETTE J C, RAWLINGS D E. Molecular relationship between two groups of the genus Leptospirillum and the finding that Leptospirillum ferriphilum sp. nov. dominates south african commercial biooxidation tanks that operate at 40 °C [J]. Appl Environ Microbiol, 2002, 68(1): 838–845.Google Scholar
- RAWLINGS D E. Restriction enzyme analysis of 16S rRNA genes for the rapid identification of Thiobacillus ferrooxidans, Thiobacillus thiooxidans and Leptospirillum ferrooxidans strains in leaching environments[C]// JEREZ C A, VARGAS T, TOLEDO H, et al. Biohydrometallurgical Processing, vol. II. Santiago: University of Chile Press, 1995: 9–17Google Scholar
- RAWLINGS D E, CORAM N J, GARDNER M N, et al. Thiobacillus caldus and Leptospirillum ferrooxidans are widely distributed in continuous flow biooxidation tanks used to treat a variety of metal containing ores and concentrates[C]// AMILS R, BALLESTER A. Biohydrometallurgy and the Environment: toward the Mining of the 21st Century, Part A. Amsterdam: Elsevier, 1999: 777–786.CrossRefGoogle Scholar
- LIU Ying, LIU Xiang-mei, TIAN Ke-li. Growth and morphology of Leptospirillum ferrooxidans on solid medium[J]. Microbiology, 2003, 30(6): 70–72. (in Chinese)Google Scholar
- ZHOU J, BRUNS M A, TIEDJE J M. DNA recovery from soils of diverse composition[J]. Appl Environ Microbiol, 1996, 62(2): 316–322.Google Scholar
- LANE D J. 16S/23S rRNA sequencing[C]// STACKEBRANDT E, GOODFELLOW M. Nucleic Acid Techniques in Bacterial Systematics. Chichester: John Wiley & Sons, 1991: 115–175.Google Scholar
- ZHANG Zai-hai, QIU Guan-zhou, HU Yue-hua, et al. The investigation of the colony isolation of thiobacillus fenooxidans[J]. Multipurpose Utilization of Mineral Resources, 2001, (1): 19–23. (in Chinese)Google Scholar
- SAND W, ROHDE K, SOBOTKE B, et al. Evaluation of Leptospirillum ferrooxidans for leaching[J]. Appl Environ Microbiol, 1992, 58(1): 85–92.Google Scholar