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Appl Environ Microbiol. Apr 1996; 62(4): 1405–1415.
PMCID: PMC167907

Distribution of sulfate-reducing bacteria in a stratified fjord (Mariager Fjord, Denmark) as evaluated by most-probable-number counts and denaturing gradient gel electrophoresis of PCR-amplified ribosomal DNA fragments.

Abstract

The sulfate-reducing bacterial populations of a stratified marine water column, Mariager Fjord, Denmark, were investigated by molecular and culture-dependent approaches in parallel. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA and DNA encoding rRNA (rDNA) isolated from the water column indicated specific bacterial populations in different water column layers and revealed a highly differentiated pattern of rRNA- and rDNA-derived PCR amplificates, probably reflecting active and resting bacterial populations. Hybridization of DGGE patterns with rRNA probes indicated the increased presence and activity (by at least 1 order of magnitude) of sulfate-reducing bacteria within and below the chemocline. Parallel to this molecular approach, an approach involving most-probable-number (MPN) counts was used, and it found a similar distribution of cultivable sulfate-reducing bacteria in the water column of Mariager Fjord, Approximately 25 cells and 250 cells per ml above and below the chemocline, respectively, were found. Desulfovibrio- and Desulfobulbus-related strains occurred in the oxic zone. DGGE bands from MPN cultures were sequenced and compared with those obtained from nucleic acids extracted from water column samples. The MPN isolates were phylogenetically affiliated with sulfate-reducing delta subdivision proteobacteria (members of the genera Desulfovibrio, Desulfobulbus, and Desulfobacter), whereas the molecular isolates constituted an independent lineage of the delta subdivision proteobacteria. DGGE of PCR-amplified nucleic acids with general eubacterial PCR primers conceptually revealed the general bacterial population, whereas the use of culture media allowed cultivable sulfate-reducing bacteria to be selected. A parallel study of Mariager Fjord biogeochemistry, bacterial activity, and bacterial counts complementing this investigation has been presented elsewhere (N.B. Ramsing, H. Fossing, T. G. Ferdelman, F. Andersen, and B. Thamdrup, Appl. Environ.

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Selected References

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  • Amann RI, Binder BJ, Olson RJ, Chisholm SW, Devereux R, Stahl DA. Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol. 1990 Jun;56(6):1919–1925. [PMC free article] [PubMed]
  • Canfield DE, Des Marais DJ. Aerobic sulfate reduction in microbial mats. Science. 1991 Mar 22;251:1471–1473. [PubMed]
  • Delong EF, Frankel RB, Bazylinski DA. Multiple evolutionary origins of magnetotaxis in bacteria. Science. 1993 Feb 5;259(5096):803–806. [PubMed]
  • Devereux R, Mundfrom GW. A phylogenetic tree of 16S rRNA sequences from sulfate-reducing bacteria in a sandy marine sediment. Appl Environ Microbiol. 1994 Sep;60(9):3437–3439. [PMC free article] [PubMed]
  • Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS. 'Touchdown' PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res. 1991 Jul 25;19(14):4008–4008. [PMC free article] [PubMed]
  • Ferguson RL, Buckley EN, Palumbo AV. Response of marine bacterioplankton to differential filtration and confinement. Appl Environ Microbiol. 1984 Jan;47(1):49–55. [PMC free article] [PubMed]
  • Fischer SG, Lerman LS. DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1579–1583. [PMC free article] [PubMed]
  • Fründ C, Cohen Y. Diurnal Cycles of Sulfate Reduction under Oxic Conditions in Cyanobacterial Mats. Appl Environ Microbiol. 1992 Jan;58(1):70–77. [PMC free article] [PubMed]
  • Fuhrman JA, Comeau DE, Hagström A, Chan AM. Extraction from natural planktonic microorganisms of DNA suitable for molecular biological studies. Appl Environ Microbiol. 1988 Jun;54(6):1426–1429. [PMC free article] [PubMed]
  • Jørgensen BB, Bak F. Pathways and microbiology of thiosulfate transformations and sulfate reduction in a marine sediment (kattegat, denmark). Appl Environ Microbiol. 1991 Mar;57(3):847–856. [PMC free article] [PubMed]
  • Kane MD, Poulsen LK, Stahl DA. 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. [PMC free article] [PubMed]
  • Kogure K, Simidu U, Taga N. A tentative direct microscopic method for counting living marine bacteria. Can J Microbiol. 1979 Mar;25(3):415–420. [PubMed]
  • Kogure K, Simidu U, Taga N. Distribution of viable marine bacteria in neritic seawater around Japan. Can J Microbiol. 1980 Mar;26(3):318–323. [PubMed]
  • Lovley DR, Phillips EJ. Novel processes for anaerobic sulfate production from elemental sulfur by sulfate-reducing bacteria. Appl Environ Microbiol. 1994 Jul;60(7):2394–2399. [PMC free article] [PubMed]
  • Maidak BL, Larsen N, McCaughey MJ, Overbeek R, Olsen GJ, Fogel K, Blandy J, Woese CR. The Ribosomal Database Project. Nucleic Acids Res. 1994 Sep;22(17):3485–3487. [PMC free article] [PubMed]
  • Muyzer G, de Waal EC, Uitterlinden AG. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol. 1993 Mar;59(3):695–700. [PMC free article] [PubMed]
  • Muyzer G, Teske A, Wirsen CO, Jannasch HW. Phylogenetic relationships of Thiomicrospira species and their identification in deep-sea hydrothermal vent samples by denaturing gradient gel electrophoresis of 16S rDNA fragments. Arch Microbiol. 1995 Sep;164(3):165–172. [PubMed]
  • Myers RM, Maniatis T, Lerman LS. Detection and localization of single base changes by denaturing gradient gel electrophoresis. Methods Enzymol. 1987;155:501–527. [PubMed]
  • Poulsen LK, Ballard G, Stahl DA. 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. [PMC free article] [PubMed]
  • Ramsing NB, Fossing H, Ferdelman TG, Andersen F, Thamdrup B. Distribution of bacterial populations in a stratified fjord (Mariager Fjord, Denmark) quantified by in situ hybridization and related to chemical gradients in the water column. Appl Environ Microbiol. 1996 Apr;62(4):1391–1404. [PMC free article] [PubMed]
  • Risatti JB, Capman WC, Stahl DA. Community structure of a microbial mat: the phylogenetic dimension. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):10173–10177. [PMC free article] [PubMed]
  • Roden EE, Lovley DR. Dissimilatory Fe(III) Reduction by the Marine Microorganism Desulfuromonas acetoxidans. Appl Environ Microbiol. 1993 Mar;59(3):734–742. [PMC free article] [PubMed]
  • Rosset R, Julien J, Monier R. Ribonucleic acid composition of bacteria as a function of growth rate. J Mol Biol. 1966 Jul;18(2):308–320. [PubMed]
  • Staley JT, Konopka A. Measurement of in situ activities of nonphotosynthetic microorganisms in aquatic and terrestrial habitats. Annu Rev Microbiol. 1985;39:321–346. [PubMed]
  • Teske A, Alm E, Regan JM, Toze S, Rittmann BE, Stahl DA. Evolutionary relationships among ammonia- and nitrite-oxidizing bacteria. J Bacteriol. 1994 Nov;176(21):6623–6630. [PMC free article] [PubMed]
  • Thamdrup B, Finster K, Hansen JW, Bak F. Bacterial disproportionation of elemental sulfur coupled to chemical reduction of iron or manganese. Appl Environ Microbiol. 1993 Jan;59(1):101–108. [PMC free article] [PubMed]
  • Wagner M, Amann R, Lemmer H, Schleifer KH. Probing activated sludge with oligonucleotides specific for proteobacteria: inadequacy of culture-dependent methods for describing microbial community structure. Appl Environ Microbiol. 1993 May;59(5):1520–1525. [PMC free article] [PubMed]
  • Weller R, Weller JW, Ward DM. 16S rRNA sequences of uncultivated hot spring cyanobacterial mat inhabitants retrieved as randomly primed cDNA. Appl Environ Microbiol. 1991 Apr;57(4):1146–1151. [PMC free article] [PubMed]
  • Woese CR. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. [PMC free article] [PubMed]

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