, Volume 17, Issue 2, pp 57–65 | Cite as

Sulfidogenesis in Low pH (3.8–4.2) Media by a Mixed Population of Acidophilic Bacteria

  • Sakurako Kimura
  • Kevin B. Hallberg
  • D. Barrie Johnson


A defined mixed bacterial culture was established which catalyzed dissimilatory sulfate reduction, using glycerol as electron donor, at pH 3.8–4.2. The bacterial consortium comprised a endospore-forming sulfate reducing bacterium (isolate M1) that had been isolated from acidic sediment in a geothermal area of Montserrat (West Indies) and which had 94% sequence identity (of its 16S rRNA gene) to the Gram-positive neutrophile Desulfosporosinus orientis, and a Gram-negative (non sulfate-reducing) acidophile (isolate PFBC) that shared 99% gene identity with Acidocella aromatica. Whilst M1 was an obligate anaerobe, isolate PFBC, as other Acidocella spp., only grew in pure culture in aerobic media. Analysis of microbial communities, using a combination of total bacterial counts and fluorescent in situ hybridization, confirmed that concurrent growth of both bacteria occurred during sulfidogenesis under strictly anoxic conditions in a pH-controlled fermenter. In pure culture, M1 oxidized glycerol incompletely, producing stoichiometric amounts of acetic acid. In mixed culture with PFBC, however, acetic acid was present only in small concentrations and its occurrence was transient. Since M1 did not oxidize acetic acid, it was inferred that this metabolite was catabolized by Acidocella PFBC which, unlike glycerol, was shown to support the growth of this acidophile under aerobic conditions. In fermenter cultures maintained at pH 3.8–4.2, sulfidogenesis resulted in the removal of soluble zinc (as solid phase ZnS) whilst ferrous iron remained in solution. Potential syntrophic interactions, involving hydrogen transfer between M1 and PFBC, are discussed, as is the potential of sulfidogenesis in acidic liquors for the selective recovery of heavy metals from wastewaters.

Key words

Acidocella acid mine drainage Desulfosporosinus metal recovery sulfate-reducing bacteria syntrophy 



acid mine drainage


energy dispersive analysis of X-rays


fluorescent in situ hybridization


oxygen-free nitrogen


sulfate reducing bacteria


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  1. EW Alm, DB Oerther, N Larsen, DA Stahl and L Raskin, The oligonucleotide probe database. Appl. Environ. Microbiol. 62 (1996) 3557-3559Google Scholar
  2. RI Amann, L Krumholz and DA Stahl, Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology. J. Bacteriol. 172 (1990) 762-770Google Scholar
  3. PL Bond and JF Banfield, Design and performance of rRNA targeted oligonucleotide probes for in situ detection and phylogenetic identification of microorganisms inhabiting acid mine drainage environments. Microbial. Ecol. 41 (2001) 149-161Google Scholar
  4. Boonstra J, van Lier R, Janssen G, Dijkman H & Buisman CJN (1999) Biological treatment of acid mine drainage. In: Amils R & Ballester A (Ed) Biohydrometallurgy and the Environment Toward the Mining of the 21st Century. Process Metallurgy, Vol 9B (pp 559–567). Elsevier, AmsterdamGoogle Scholar
  5. HF Castro, NH Williams and A Ogram, Phylogeny of sulfate-reducing bacteria. FEMS Microbiol. Ecol. 31 (2000) 1-9Google Scholar
  6. BM Fuchs, FO Glockner, J Wulf and R Amann, Unlabeled helper oligonucleotides increase the in situ accessibility to 16S rRNA of fluorescently labeled oligonucleotide probes. Appl. Environ. Microbiol. 66 (2000) 3603-3607CrossRefGoogle Scholar
  7. RT Gemmell and CJ Knowles, Utilisation of aliphatic compounds by acidophilic heterotrophic bacteria. The potential for bioremediation of acidic wastewaters contaminated with toxic organic compounds and heavy metals. FEMS Microbiol. Lett. 192 (2000) 185-190CrossRefGoogle Scholar
  8. RA Gyure, A Konopka, A Brooks and W Doemel, Microbial sulfate reduction in acidic (pH 3) strip-mine lakes. FEMS Microbiol. Ecol. 73 (1990) 193-202CrossRefGoogle Scholar
  9. KB Hallberg and DB Johnson, Biodiversity of acidophilic microorganisms. Adv. Appl. Microbiol. 49 (2001) 37-84CrossRefGoogle Scholar
  10. KB Hallberg, Å Kolmert, DB Johnson and PA Williams, A novel metabolic phenotype among acidophilic bacteria: aromatic degradation and the potential use of these microorganisms for the treatment of wastewater containing organic and inorganic pollutants. In: R Amils and A Ballester (eds.) Biohydrometallurgy and the Environment Toward the Mining of the 21st Century. Process Metallurgy, Vol 9A. Amsterdam: Elsevier (1999) pp. 719-728CrossRefGoogle Scholar
  11. BC Hard, S Friedrich and W Babel, Bioremediation of acid mine water using facultatively methylotrophic metal-tolerant sulfate-reducing bacteria. Microbiol. Res. 152 (1997) 65-73Google Scholar
  12. DB Johnson, Selective solid media for isolating and enumerating acidophilic bacteria. J. Microbiol. Meth. 23 (1995) 205-218CrossRefGoogle Scholar
  13. DB Johnson, Chemical and microbiological characteristics of mineral spoils and drainage waters at abandoned coal and metal mines. Water Air Soil Pollut.:Focus 3 (2003) 47-66Google Scholar
  14. Johnson DB, Roberto FF (1997) Biodiversity of acidophilic bacteria in mineral leaching and related environments. IBS Biomine ‘97 Conference Proceedings. (pp. P3.1–10). Australian Mineral Foundation, Glenside, AustraliaGoogle Scholar
  15. KA Küsel, U Roth, T Trinkwalter and S Peiffer, Effect of pH on the anaerobic microbial cycling of sulfur in mining-impacted freshwater lake sediments. Environ. Exp. Bot. 46 (2001) 213-223CrossRefGoogle Scholar
  16. DR Lovley and EJP Phillips, Rapid assay for microbially reduced ferric iron in aquatic sediments. Appl. Environ. Microbiol. 53 (1987) 1536-1540Google Scholar
  17. MT Madigan, JM Martinko and J Parker, Biology of Microorganisms. Upper Saddle River: Prentice Hall International, Inc. (2003).Google Scholar
  18. PR Norris and WJ Ingledew, Acidophilic bacteria: adaptations and applications. In: RA Herbert and RJ Sharp (eds.) Molecular Biology and Biotechnology of Extremophiles. Cambridge: Royal Society for Chemistry (1992) pp. 121-131Google Scholar
  19. JR Postgate, The Sulphate-Reducing Bacteria. London: Cambridge University Press (1979).Google Scholar
  20. B Pott and B Mattiasson, Separations of heavy metals from water solutions at the laboratory scale. Biotechnol. Lett. 26 (2004) 421-456CrossRefGoogle Scholar
  21. Sen AM (2001) Acidophilic sulphate reducing bacteria: candidates for bioremediation of acid mine drainage pollution. Ph.D. Thesis, University of Wales, Bangor, United KingdomGoogle Scholar
  22. AM Sen and DB Johnson, Acidophilic sulphate-reducing bacteria: candidates for bioremediation of acid mine drainage. In: R Amils and A Ballester (eds.) Biohydrometallurgy and the Environment Toward the Mining of the 21st Century. Process Metallurgy Vol, 9A. Amsterdam: Elsevier (1999) pp. 709-718CrossRefGoogle Scholar
  23. HH Tabak and R Govind, Advances in biotreatment of acid mine drainage and biorecovery of metals: 2. Membrane bioreactor system for sulfate reduction. Biodegradation 14 (2003) 437-452CrossRefGoogle Scholar
  24. HH Tabak, R Scharp, J Burckle, FK Kawahara and R Govind, Advances in biotreatment of acid mine drainage and biorecovery of metals: 1. Metal precipitation for recovery and recycle. Biodegradation 14 (2003) 423-436CrossRefGoogle Scholar
  25. JH Tuttle, PR Dugan, CB Macmillan and CI Randles, Microbial dissimilatory sulfur cycle in acid mine water. J. Bacteriol. 97 (1969) 594-602Google Scholar
  26. F Widdel and N Pfennig, Studies on dissimilatory sulfate-reducing bacteria that decompose fatty-acids. I. Isolation of new sulfate-reducing bacteria enriched with acetate from saline environments – description of Desulfobacter postgatei gen. nov., sp. nov.. Arch. Microbiol. 129 (1981) 395-400CrossRefGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • Sakurako Kimura
    • 1
  • Kevin B. Hallberg
    • 1
  • D. Barrie Johnson
    • 1
  1. 1.School of Biological SciencesUniversity of WalesBangorU.K.

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