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Microbiology 142 (1996), 1097-1106; DOI  10.1099/13500872-142-5-1097
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Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment

Werner Manz1,3,{dagger}, Rudolf Amann1, Wolfgang Ludwig1, Marc Vancanneyt2 and Karl-Heinz Schleifer1

Lehrstuhl für Mikrobiologie, Technische Universität München, D-80290 München, Germany
Laboratorium voor Microbiologie, Universiteit Gent, Belgium

3Author for correspondence: Werner Manz. Tel: +49 30 314 25831. Fax: +49 30 314 73461. e-mail: manz0654@mailszrz.zrz.tu-berlin.de

ABSTRACT

We designed a panel of four 16S rRNA-targeted oligonucleotide probes specific for bacteria of the phylum cytophaga-flavobacter-bacteroides (CFB). Probes CF319a and CF319b are targeted to members of the flavobacteria-cytophaga group and the genus Porphyromonas, whereas probe BAC303 has a target region characteristic for the genera Prevotella and Bacteroides within the bacteroides group. The probe FFE8b was developed for species-specific hybridizations with Flavobacterium ferrugineum. All probes were designed by computer-assisted sequence analysis and compared to all currently accessible 16S and 23S rRNA sequences. The oligonucleotides were further evaluated by whole-cell and non-radioactive dot-blot hybridization against reference strains of the CFB phylum and other major lineages of Bacteria. The newly developed probes were used together with other higher-order probes to analyse the structure and community composition in complex environments. In activated sludge samples, members of the flavobacteria-cytophaga group were revealed by in situ hybridization as important constituents of sludge flocs and characteristic colonizers of filamentous bacteria. By application of fluorescent probe BAC303, members of the genera Bacteroides and Prevotella could be visualized without prior cultivation as an important part of the human faecal microflora.


Keywords: 16S rRNA-targeted oligonucleotide probes, in situ hybridization, phylum cytophaga-flavobacter-bacteroides, flavobacteria-cytophaga group, bacteroides group

{dagger} Present address: Fachgebiet ökologie der Mikroorganismen, Technische Universität Berlin, D-10587 Berlin, Germany.




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Appl. Environ. Microbiol.Home page
T. Kenzaka, A. Ishidoshiro, N. Yamaguchi, K. Tani, and M. Nasu
rRNA Sequence-Based Scanning Electron Microscopic Detection of Bacteria
Appl. Envir. Microbiol., September 1, 2005; 71(9): 5523 - 5531.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
M. W. Hahn, M. Pockl, and Q. L. Wu
Low Intraspecific Diversity in a Polynucleobacter Subcluster Population Numerically Dominating Bacterioplankton of a Freshwater Pond
Appl. Envir. Microbiol., August 1, 2005; 71(8): 4539 - 4547.
[Abstract] [Full Text] [PDF]


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J. Nutr.Home page
K. Whelan, P. A. Judd, V. R. Preedy, R. Simmering, A. Jann, and M. A. Taylor
Fructooligosaccharides and Fiber Partially Prevent the Alterations in Fecal Microbiota and Short-Chain Fatty Acid Concentrations Caused by Standard Enteral Formula in Healthy Humans
J. Nutr., August 1, 2005; 135(8): 1896 - 1902.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
A. Loy, W. Beisker, and H. Meier
Diversity of Bacteria Growing in Natural Mineral Water after Bottling
Appl. Envir. Microbiol., July 1, 2005; 71(7): 3624 - 3632.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
A. W. Walker, S. H. Duncan, E. C. McWilliam Leitch, M. W. Child, and H. J. Flint
pH and Peptide Supply Can Radically Alter Bacterial Populations and Short-Chain Fatty Acid Ratios within Microbial Communities from the Human Colon
Appl. Envir. Microbiol., July 1, 2005; 71(7): 3692 - 3700.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
S. Okabe, T. Kindaichi, and T. Ito
Fate of 14C-Labeled Microbial Products Derived from Nitrifying Bacteria in Autotrophic Nitrifying Biofilms
Appl. Envir. Microbiol., July 1, 2005; 71(7): 3987 - 3994.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
N. Fierer, J. A. Jackson, R. Vilgalys, and R. B. Jackson
Assessment of Soil Microbial Community Structure by Use of Taxon-Specific Quantitative PCR Assays
Appl. Envir. Microbiol., July 1, 2005; 71(7): 4117 - 4120.
[Abstract] [Full Text] [PDF]


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J. Clin. Microbiol.Home page
A. Swidsinski, J. Weber, V. Loening-Baucke, L. P. Hale, and H. Lochs
Spatial Organization and Composition of the Mucosal Flora in Patients with Inflammatory Bowel Disease
J. Clin. Microbiol., July 1, 2005; 43(7): 3380 - 3389.
[Abstract] [Full Text] [PDF]


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J. Nutr.Home page
G. Tzortzis, A. K. Goulas, J. M. Gee, and G. R. Gibson
A Novel Galactooligosaccharide Mixture Increases the Bifidobacterial Population Numbers in a Continuous In Vitro Fermentation System and in the Proximal Colonic Contents of Pigs In Vivo
J. Nutr., July 1, 2005; 135(7): 1726 - 1731.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
M. Schauer and M. W. Hahn
Diversity and Phylogenetic Affiliations of Morphologically Conspicuous Large Filamentous Bacteria Occurring in the Pelagic Zones of a Broad Spectrum of Freshwater Habitats
Appl. Envir. Microbiol., April 1, 2005; 71(4): 1931 - 1940.
[Abstract] [Full Text] [PDF]


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GutHome page
A Swidsinski, P Schlien, A Pernthaler, U Gottschalk, E Barlehner, G Decker, S Swidsinski, J Strassburg, V Loening-Baucke, U Hoffmann, et al.
Bacterial biofilm within diseased pancreatic and biliary tracts
Gut, March 1, 2005; 54(3): 388 - 395.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
D. C. Gillan, B. Danis, P. Pernet, G. Joly, and P. Dubois
Structure of Sediment-Associated Microbial Communities along a Heavy-Metal Contamination Gradient in the Marine Environment
Appl. Envir. Microbiol., February 1, 2005; 71(2): 679 - 690.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
J. L. Nielsen, A. Schramm, A. E. Bernhard, G. J. van den Engh, and D. A. Stahl
Flow Cytometry-Assisted Cloning of Specific Sequence Motifs from Complex 16S rRNA Gene Libraries
Appl. Envir. Microbiol., December 1, 2004; 70(12): 7550 - 7554.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
M.-T. Wong, F. M. Tan, W. J. Ng, and W.-T. Liu
Identification and occurrence of tetrad-forming Alphaproteobacteria in anaerobic-aerobic activated sludge processes
Microbiology, November 1, 2004; 150(11): 3741 - 3748.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
R. Sekar, B. M. Fuchs, R. Amann, and J. Pernthaler
Flow Sorting of Marine Bacterioplankton after Fluorescence In Situ Hybridization
Appl. Envir. Microbiol., October 1, 2004; 70(10): 6210 - 6219.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
J. Pernthaler, E. Zollner, F. Warnecke, and K. Jurgens
Bloom of Filamentous Bacteria in a Mesotrophic Lake: Identity and Potential Controlling Mechanism
Appl. Envir. Microbiol., October 1, 2004; 70(10): 6272 - 6281.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
C.-L. Chen, H. Macarie, I. Ramirez, A. Olmos, S. L. Ong, O. Monroy, and W.-T. Liu
Microbial community structure in a thermophilic anaerobic hybrid reactor degrading terephthalate
Microbiology, October 1, 2004; 150(10): 3429 - 3440.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
J. Vulevic, A. L. McCartney, J. M. Gee, I. T. Johnson, and G. R. Gibson
Microbial Species Involved in Production of 1,2-sn-Diacylglycerol and Effects of Phosphatidylcholine on Human Fecal Microbiota
Appl. Envir. Microbiol., September 1, 2004; 70(9): 5659 - 5666.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
H. M. Probert, J. H. A. Apajalahti, N. Rautonen, J. Stowell, and G. R. Gibson
Polydextrose, Lactitol, and Fructo-Oligosaccharide Fermentation by Colonic Bacteria in a Three-Stage Continuous Culture System
Appl. Envir. Microbiol., August 1, 2004; 70(8): 4505 - 4511.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
U. Stingl, A. Maass, R. Radek, and A. Brune
Symbionts of the gut flagellate Staurojoenina sp. from Neotermes cubanus represent a novel, termite-associated lineage of Bacteroidales: description of 'Candidatus Vestibaculum illigatum'
Microbiology, July 1, 2004; 150(7): 2229 - 2235.
[Abstract] [Full Text] [PDF]


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Infect. Immun.Home page
M. Schuppler, K. Lotzsch, M. Waidmann, and I. B. Autenrieth
An Abundance of Escherichia coli Is Harbored by the Mucosa- Associated Bacterial Flora of Interleukin-2-Deficient Mice
Infect. Immun., April 1, 2004; 72(4): 1983 - 1990.
[Abstract] [Full Text] [PDF]


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GutHome page
A Fite, G T Macfarlane, J H Cummings, M J Hopkins, S C Kong, E Furrie, and S Macfarlane
Identification and quantitation of mucosal and faecal desulfovibrios using real time polymerase chain reaction
Gut, April 1, 2004; 53(4): 523 - 529.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
T. Kindaichi, T. Ito, and S. Okabe
Ecophysiological Interaction between Nitrifying Bacteria and Heterotrophic Bacteria in Autotrophic Nitrifying Biofilms as Determined by Microautoradiography-Fluorescence In Situ Hybridization
Appl. Envir. Microbiol., March 1, 2004; 70(3): 1641 - 1650.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
M. Sunamura, Y. Higashi, C. Miyako, J.-i. Ishibashi, and A. Maruyama
Two Bacteria Phylotypes Are Predominant in the Suiyo Seamount Hydrothermal Plume
Appl. Envir. Microbiol., February 1, 2004; 70(2): 1190 - 1198.
[Abstract] [Full Text] [PDF]


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J. Nutr.Home page
V. Mai, H. A. Katki, H. Harmsen, D. Gallaher, A. Schatzkin, D. J. Baer, and B. Clevidence
Effects of a Controlled Diet and Black Tea Drinking on the Fecal Microflora Composition and the Fecal Bile Acid Profile of Human Volunteers in a Double-Blinded Randomized Feeding Study
J. Nutr., February 1, 2004; 134(2): 473 - 478.
[Abstract] [Full Text] [PDF]


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GutHome page
F Hoentjen, H J M Harmsen, H Braat, C D Torrice, B A Mann, R B Sartor, and L A Dieleman
Antibiotics with a selective aerobic or anaerobic spectrum have different therapeutic activities in various regions of the colon in interleukin 10 gene deficient mice
Gut, December 1, 2003; 52(12): 1721 - 1727.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
R. Brinkmeyer, K. Knittel, J. Jurgens, H. Weyland, R. Amann, and E. Helmke
Diversity and Structure of Bacterial Communities in Arctic versus Antarctic Pack Ice
Appl. Envir. Microbiol., November 1, 2003; 69(11): 6610 - 6619.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
D. Schmitt-Wagner, M. W. Friedrich, B. Wagner, and A. Brune
Phylogenetic Diversity, Abundance, and Axial Distribution of Bacteria in the Intestinal Tract of Two Soil-Feeding Termites (Cubitermes spp.)
Appl. Envir. Microbiol., October 1, 2003; 69(10): 6007 - 6017.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
A. S. Engel, N. Lee, M. L. Porter, L. A. Stern, P. C. Bennett, and M. Wagner
Filamentous "Epsilonproteobacteria" Dominate Microbial Mats from Sulfidic Cave Springs
Appl. Envir. Microbiol., September 1, 2003; 69(9): 5503 - 5511.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
J. Bertaux, M. Schmid, N. C. Prevost-Boure, J. L. Churin, A. Hartmann, J. Garbaye, and P. Frey-Klett
In Situ Identification of Intracellular Bacteria Related to Paenibacillus spp. in the Mycelium of the Ectomycorrhizal Fungus Laccaria bicolor S238N
Appl. Envir. Microbiol., July 1, 2003; 69(7): 4243 - 4248.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
J. Peplies, F. O. Glockner, and R. Amann
Optimization Strategies for DNA Microarray-Based Detection of Bacteria with 16S rRNA-Targeting Oligonucleotide Probes
Appl. Envir. Microbiol., March 1, 2003; 69(3): 1397 - 1407.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
S. H. Duncan, K. P. Scott, A. G. Ramsay, H. J. M. Harmsen, G. W. Welling, C. S. Stewart, and H. J. Flint
Effects of Alternative Dietary Substrates on Competition between Human Colonic Bacteria in an Anaerobic Fermentor System
Appl. Envir. Microbiol., February 1, 2003; 69(2): 1136 - 1142.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
G. R. Crocetti, J. F. Banfield, J. Keller, P. L. Bond, and L. L. Blackall
Glycogen-accumulating organisms in laboratory-scale and full-scale wastewater treatment processes
Microbiology, November 1, 2002; 148(11): 3353 - 3364.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
N. H. Salzman, H. de Jong, Y. Paterson, H. J. M. Harmsen, G. W. Welling, and N. A. Bos
Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria
Microbiology, November 1, 2002; 148(11): 3651 - 3660.
[Abstract] [Full Text] [PDF]


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J PLANKTON RESHome page
J.-P. Gattuso, S. Peduzzi, M.-D. Pizay, and M. Tonolla
Changes in freshwater bacterial community composition during measurements of microbial and community respiration
J. Plankton Res., November 1, 2002; 24(11): 1197 - 1206.
[Abstract] [Full Text] [PDF]


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Nucleic Acids ResHome page
K. E. Ashelford, A. J. Weightman, and J. C. Fry
PRIMROSE: a computer program for generating and estimating the phylogenetic range of 16S rRNA oligonucleotide probes and primers in conjunction with the RDP-II database
Nucleic Acids Res., August 1, 2002; 30(15): 3481 - 3489.
[Abstract] [Full Text] [PDF]


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GutHome page
P V Kirjavainen, T Arvola, S J Salminen, and E Isolauri
Aberrant composition of gut microbiota of allergic infants: a target of bifidobacterial therapy at weaning?
Gut, July 1, 2002; 51(1): 51 - 55.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
H. J. M. Harmsen, G. C. Raangs, T. He, J. E. Degener, and G. W. Welling
Extensive Set of 16S rRNA-Based Probes for Detection of Bacteria in Human Feces
Appl. Envir. Microbiol., June 1, 2002; 68(6): 2982 - 2990.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
E. O. Casamayor, C. Pedros-Alio, G. Muyzer, and R. Amann
Microheterogeneity in 16S Ribosomal DNA-Defined Bacterial Populations from a Stratified Planktonic Environment Is Related to Temporal Changes and to Ecological Adaptations
Appl. Envir. Microbiol., April 1, 2002; 68(4): 1706 - 1714.
[Abstract] [Full Text] [PDF]




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