Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Apr;62(4):1323–1328. doi: 10.1128/aem.62.4.1323-1328.1996

Bacterial populations in samples of bioleached copper ore as revealed by analysis of DNA obtained before and after cultivation.

J Pizarro 1, E Jedlicki 1, O Orellana 1, J Romero 1, R T Espejo 1
PMCID: PMC167897  PMID: 8919792

Abstract

The composition of bacterial populations in copper bioleaching systems was investigated by analysis of DNA obtained either directly from ores or leaching solutions or after laboratory cultures. This analysis consisted of the characterization of the spacer regions between the 16 and 23S genes in the bacterial rRNA genetic loci after PCR amplification. The sizes of the spacer regions, amplified from DNAs obtained from samples, were compared with the sizes of those obtained from cultures of the main bacterial species isolated from bioleaching systems. This allowed a preliminary assessment of the bacterial species present in the samples. Identification of the bacteria was achieved by partial sequencing of the 16S rRNA genes adjacent to the spacer regions. The spacer regions observed in DNA from columns leached at different iron concentrations indicated the presence of a mixture of different bacteria. The spacer region corresponding to Thiobacillus ferrooxidans was the main product observed at high ferrous iron concentration. At low ferrous iron concentration, spacer regions of different lengths, corresponding to Thiobacillus thiooxidans and "Leptospirillum ferrooxidans" were observed. However, T. ferrooxidans appeared to predominate after culture of these samples in medium containing ferrous iron as energy source. Although some of these strains contained singular spacer regions, they belonged within previously described groups of T. ferrooxidans according to the nucleotide sequence of the neighbor 16S rRNA. These results illustrate the bacterial diversity in bioleaching systems and the selective pressure generated by different growth conditions.

Full Text

The Full Text of this article is available as a PDF (250.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Apel W. A., Dugan P. R., Filppi J. A., Rheins M. S. Detection of Thiobacillus ferrooxidans in acid mine environments by indirect fluorescent antibody staining. Appl Environ Microbiol. 1976 Jul;32(1):159–165. doi: 10.1128/aem.32.1.159-165.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. DeLong E. F. Archaea in coastal marine environments. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5685–5689. doi: 10.1073/pnas.89.12.5685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Espejo R. T., Escanilla D. Detection of HIV1 DNA by a simple procedure of polymerase chain reaction, using "primer-dimer" formation as an internal control of amplification. Res Virol. 1993 May-Jun;144(3):243–246. doi: 10.1016/s0923-2516(06)80035-x. [DOI] [PubMed] [Google Scholar]
  4. Giovannoni S. J., Britschgi T. B., Moyer C. L., Field K. G. Genetic diversity in Sargasso Sea bacterioplankton. Nature. 1990 May 3;345(6270):60–63. doi: 10.1038/345060a0. [DOI] [PubMed] [Google Scholar]
  5. Goebel B. M., Stackebrandt E. Cultural and phylogenetic analysis of mixed microbial populations found in natural and commercial bioleaching environments. Appl Environ Microbiol. 1994 May;60(5):1614–1621. doi: 10.1128/aem.60.5.1614-1621.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Harrison A. P., Jr The acidophilic thiobacilli and other acidophilic bacteria that share their habitat. Annu Rev Microbiol. 1984;38:265–292. doi: 10.1146/annurev.mi.38.100184.001405. [DOI] [PubMed] [Google Scholar]
  7. Hutchins S. R., Davidson M. S., Brierley J. A., Brierley C. L. Microorganisms in reclamation of metals. Annu Rev Microbiol. 1986;40:311–336. doi: 10.1146/annurev.mi.40.100186.001523. [DOI] [PubMed] [Google Scholar]
  8. Jensen M. A., Webster J. A., Straus N. Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol. 1993 Apr;59(4):945–952. doi: 10.1128/aem.59.4.945-952.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kostman J. R., Edlind T. D., LiPuma J. J., Stull T. L. Molecular epidemiology of Pseudomonas cepacia determined by polymerase chain reaction ribotyping. J Clin Microbiol. 1992 Aug;30(8):2084–2087. doi: 10.1128/jcm.30.8.2084-2087.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lane D. J., Harrison A. P., Jr, Stahl D., Pace B., Giovannoni S. J., Olsen G. J., Pace N. R. Evolutionary relationships among sulfur- and iron-oxidizing eubacteria. J Bacteriol. 1992 Jan;174(1):269–278. doi: 10.1128/jb.174.1.269-278.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lane D. J., Stahl D. A., Olsen G. J., Heller D. J., Pace N. R. Phylogenetic analysis of the genera Thiobacillus and Thiomicrospira by 5S rRNA sequences. J Bacteriol. 1985 Jul;163(1):75–81. doi: 10.1128/jb.163.1.75-81.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Liesack W., Stackebrandt E. Occurrence of novel groups of the domain Bacteria as revealed by analysis of genetic material isolated from an Australian terrestrial environment. J Bacteriol. 1992 Aug;174(15):5072–5078. doi: 10.1128/jb.174.15.5072-5078.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Muyzer G., de Bruyn A. C., Schmedding D. J., Bos P., Westbroek P., Kuenen G. J. A Combined Immunofluorescence-DNA-Fluorescence Staining Technique for Enumeration of Thiobacillus ferrooxidans in a Population of Acidophilic Bacteria. Appl Environ Microbiol. 1987 Apr;53(4):660–664. doi: 10.1128/aem.53.4.660-664.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Salazar O., Takamiya M., Orellana O. Characterization of the two rRNA gene operons present in Thiobacillus ferrooxidans. FEBS Lett. 1989 Jan 2;242(2):439–443. doi: 10.1016/0014-5793(89)80518-6. [DOI] [PubMed] [Google Scholar]
  15. Sand W., Rohde K., Sobotke B., Zenneck C. Evaluation of Leptospirillum ferrooxidans for Leaching. Appl Environ Microbiol. 1992 Jan;58(1):85–92. doi: 10.1128/aem.58.1.85-92.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schmidt T. M., DeLong E. F., Pace N. R. Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. J Bacteriol. 1991 Jul;173(14):4371–4378. doi: 10.1128/jb.173.14.4371-4378.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Venegas A., Hevia E., Sánchez H. Sequence of two tRNA genes from a Thiobacillus ferrooxidans ribosomal operon. Nucleic Acids Res. 1988 Aug 25;16(16):8179–8179. doi: 10.1093/nar/16.16.8179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ward D. M., Weller R., Bateson M. M. 16S rRNA sequences reveal numerous uncultured microorganisms in a natural community. Nature. 1990 May 3;345(6270):63–65. doi: 10.1038/345063a0. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES