Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Oct;62(10):3847–3857. doi: 10.1128/aem.62.10.3847-3857.1996

Competition and Coexistence of Sulfate-Reducing and Methanogenic Populations in Anaerobic Biofilms

L Raskin, B E Rittmann, D A Stahl
PMCID: PMC1388966  PMID: 16535428

Abstract

The microbial population structure and function of natural anaerobic communities maintained in laboratory fixed-bed biofilm reactors were tracked before and after a major perturbation, which involved the addition of sulfate to the influent of a reactor that had previously been fed only glucose (methanogenic), while sulfate was withheld from a reactor that had been fed both glucose and sulfate (sulfidogenic). The population structure, determined by using phylogenetically based oligonucleotide probes for methanogens and sulfate-reducing bacteria, was linked to the functional performance of the biofilm reactors. Before the perturbation, the methanogenic reactor contained up to 25% methanogens as well as 15% sulfate-reducing bacteria, even though sulfate was not present in the influent of this reactor. Methanobacteriales and Desulfovibrio spp. were the most abundant methanogens and sulfate-reducing bacteria, respectively. The presence of sulfate-reducing bacteria (primarily Desulfovibrio spp. and Desulfobacterium spp.) in the absence of sulfate may be explained by their ability to function as proton-reducing acetogens and/or fermenters. Sulfate reduction began immediately following the addition of sulfate consistent with the presence of significant levels of sulfate-reducing bacteria in the methanogenic reactor, and levels of sulfate-reducing bacteria increased to a new steady-state level of 30 to 40%; coincidentally, effluent acetate concentrations decreased. Notably, some sulfate-reducing bacteria (Desulfococcus/Desulfosarcina/Desulfobotulus group) were more competitive without sulfate. Methane production decreased immediately following the addition of sulfate; this was later followed by a decrease in the relative concentration of methanogens, which reached a new steady-state level of approximately 8%. The changeover to sulfate-free medium in the sulfidogenic reactor did not cause a rapid shift to methanogenesis. Methane production and a substantial increase in the levels of methanogens were observed only after approximately 50 days following the perturbation.

Full Text

The Full Text of this article is available as a PDF (369.7 KB).

Selected References

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

  1. Alm E. W., Oerther D. B., Larsen N., Stahl D. A., Raskin L. The oligonucleotide probe database. Appl Environ Microbiol. 1996 Oct;62(10):3557–3559. doi: 10.1128/aem.62.10.3557-3559.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amann R. I., Binder B. J., Olson R. J., Chisholm S. W., Devereux R., Stahl D. A. Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol. 1990 Jun;56(6):1919–1925. doi: 10.1128/aem.56.6.1919-1925.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amann R. I., Ludwig W., Schleifer K. H. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev. 1995 Mar;59(1):143–169. doi: 10.1128/mr.59.1.143-169.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Amann R. I., Stromley J., Devereux R., Key R., Stahl D. A. Molecular and microscopic identification of sulfate-reducing bacteria in multispecies biofilms. Appl Environ Microbiol. 1992 Feb;58(2):614–623. doi: 10.1128/aem.58.2.614-623.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barlaz M. A., Schaefer D. M., Ham R. K. Bacterial population development and chemical characteristics of refuse decomposition in a simulated sanitary landfill. Appl Environ Microbiol. 1989 Jan;55(1):55–65. doi: 10.1128/aem.55.1.55-65.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bryant M. P., Campbell L. L., Reddy C. A., Crabill M. R. Growth of desulfovibrio in lactate or ethanol media low in sulfate in association with H2-utilizing methanogenic bacteria. Appl Environ Microbiol. 1977 May;33(5):1162–1169. doi: 10.1128/aem.33.5.1162-1169.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. DeLong E. F., Wu K. Y., Prézelin B. B., Jovine R. V. High abundance of Archaea in Antarctic marine picoplankton. Nature. 1994 Oct 20;371(6499):695–697. doi: 10.1038/371695a0. [DOI] [PubMed] [Google Scholar]
  9. Fuhrman J. A., McCallum K., Davis A. A. Novel major archaebacterial group from marine plankton. Nature. 1992 Mar 12;356(6365):148–149. doi: 10.1038/356148a0. [DOI] [PubMed] [Google Scholar]
  10. Fuhrman J. A., McCallum K., Davis A. A. Phylogenetic diversity of subsurface marine microbial communities from the Atlantic and Pacific Oceans. Appl Environ Microbiol. 1993 May;59(5):1294–1302. doi: 10.1128/aem.59.5.1294-1302.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fukuzaki S., Nishio N., Nagai S. Kinetics of the methanogenic fermentation of acetate. Appl Environ Microbiol. 1990 Oct;56(10):3158–3163. doi: 10.1128/aem.56.10.3158-3163.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gibson G. R., Macfarlane G. T., Cummings J. H. Occurrence of sulphate-reducing bacteria in human faeces and the relationship of dissimilatory sulphate reduction to methanogenesis in the large gut. J Appl Bacteriol. 1988 Aug;65(2):103–111. doi: 10.1111/j.1365-2672.1988.tb01498.x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Isa Z., Grusenmeyer S., Verstraete W. Sulfate reduction relative to methane production in high-rate anaerobic digestion: microbiological aspects. Appl Environ Microbiol. 1986 Mar;51(3):580–587. doi: 10.1128/aem.51.3.580-587.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones W. J., Guyot J. P., Wolfe R. S. Methanogenesis from sucrose by defined immobilized consortia. Appl Environ Microbiol. 1984 Jan;47(1):1–6. doi: 10.1128/aem.47.1.1-6.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kane M. D., Poulsen L. K., Stahl D. A. Monitoring the enrichment and isolation of sulfate-reducing bacteria by using oligonucleotide hybridization probes designed from environmentally derived 16S rRNA sequences. Appl Environ Microbiol. 1993 Mar;59(3):682–686. doi: 10.1128/aem.59.3.682-686.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lee Monica J., Zinder Stephen H. Isolation and Characterization of a Thermophilic Bacterium Which Oxidizes Acetate in Syntrophic Association with a Methanogen and Which Grows Acetogenically on H(2)-CO(2). Appl Environ Microbiol. 1988 Jan;54(1):124–129. doi: 10.1128/aem.54.1.124-129.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Min H., Zinder S. H. Kinetics of Acetate Utilization by Two Thermophilic Acetotrophic Methanogens: Methanosarcina sp. Strain CALS-1 and Methanothrix sp. Strain CALS-1. Appl Environ Microbiol. 1989 Feb;55(2):488–491. doi: 10.1128/aem.55.2.488-491.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nielsen P. H. Biofilm Dynamics and Kinetics during High-Rate Sulfate Reduction under Anaerobic Conditions. Appl Environ Microbiol. 1987 Jan;53(1):27–32. doi: 10.1128/aem.53.1.27-32.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ogram A., Sun W., Brockman F. J., Fredrickson J. K. Isolation and characterization of RNA from low-biomass deep-subsurface sediments. Appl Environ Microbiol. 1995 Feb;61(2):763–768. doi: 10.1128/aem.61.2.763-768.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Poulsen L. K., Ballard G., Stahl D. A. Use of rRNA fluorescence in situ hybridization for measuring the activity of single cells in young and established biofilms. Appl Environ Microbiol. 1993 May;59(5):1354–1360. doi: 10.1128/aem.59.5.1354-1360.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Powell M. R., Doebbler G. F., Hamilton R. W., Jr Serum enzyme level changes in pigs following decompression trauma. Aerosp Med. 1974 May;45(5):519–524. [PubMed] [Google Scholar]
  23. Raskin L., Capman W. C., Kane M. D., Rittmann B. E., Stahl D. A. Critical evaluation of membrane supports for use in quantitative hybridizations. Appl Environ Microbiol. 1996 Jan;62(1):300–303. doi: 10.1128/aem.62.1.300-303.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Raskin L., Poulsen L. K., Noguera D. R., Rittmann B. E., Stahl D. A. Quantification of methanogenic groups in anaerobic biological reactors by oligonucleotide probe hybridization. Appl Environ Microbiol. 1994 Apr;60(4):1241–1248. doi: 10.1128/aem.60.4.1241-1248.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Raskin L., Stromley J. M., Rittmann B. E., Stahl D. A. Group-specific 16S rRNA hybridization probes to describe natural communities of methanogens. Appl Environ Microbiol. 1994 Apr;60(4):1232–1240. doi: 10.1128/aem.60.4.1232-1240.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Raskin L., Zheng D., Griffin M. E., Stroot P. G., Misra P. Characterization of microbial communities in anaerobic bioreactors using molecular probes. Antonie Van Leeuwenhoek. 1995 Nov;68(4):297–308. doi: 10.1007/BF00874140. [DOI] [PubMed] [Google Scholar]
  27. Risatti J. B., Capman W. C., Stahl D. A. Community structure of a microbial mat: the phylogenetic dimension. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):10173–10177. doi: 10.1073/pnas.91.21.10173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith M. R., Mah R. A. Acetate as sole carbon and energy source for growth of methanosarcina strain 227. Appl Environ Microbiol. 1980 May;39(5):993–999. doi: 10.1128/aem.39.5.993-999.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stahl D. A., Flesher B., Mansfield H. R., Montgomery L. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology. Appl Environ Microbiol. 1988 May;54(5):1079–1084. doi: 10.1128/aem.54.5.1079-1084.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Thiele Jurgen H., Chartrain M., Zeikus J. Gregory. Control of Interspecies Electron Flow during Anaerobic Digestion: Role of Floc Formation in Syntrophic Methanogenesis. Appl Environ Microbiol. 1988 Jan;54(1):10–19. doi: 10.1128/aem.54.1.10-19.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thiele Jurgen H., Zeikus J. Gregory. Control of Interspecies Electron Flow during Anaerobic Digestion: Significance of Formate Transfer versus Hydrogen Transfer during Syntrophic Methanogenesis in Flocs. Appl Environ Microbiol. 1988 Jan;54(1):20–29. doi: 10.1128/aem.54.1.20-29.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Traore A. S., Fardeau M. L., Hatchikian C. E., Le Gall J., Belaich J. P. Energetics of Growth of a Defined Mixed Culture of Desulfovibrio vulgaris and Methanosarcina barkeri: Interspecies Hydrogen Transfer in Batch and Continuous Cultures. Appl Environ Microbiol. 1983 Nov;46(5):1152–1156. doi: 10.1128/aem.46.5.1152-1156.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zehnder A. J., Huser B. A., Brock T. D., Wuhrmann K. Characterization of an acetate-decarboxylating, non-hydrogen-oxidizing methane bacterium. Arch Microbiol. 1980 Jan;124(1):1–11. doi: 10.1007/BF00407022. [DOI] [PubMed] [Google Scholar]
  35. Zinder S. H., Mah R. A. Isolation and Characterization of a Thermophilic Strain of Methanosarcina Unable to Use H(2)-CO(2) for Methanogenesis. Appl Environ Microbiol. 1979 Nov;38(5):996–1008. doi: 10.1128/aem.38.5.996-1008.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES