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1 Institut für Angewandte Mikrobiologie, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 2632, D-35390 Giessen, Germany
2 Fachgebiet Hygiene und Umweltmikrobiologie, Technische Universität Berlin, D-13353 Berlin, Germany
3 Institut für Hygiene und Umweltmedizin, Rheinisch-Westfälische Technische Hochschule, D-52057 Aachen, Germany
4 Institut für Bakteriologie, Mykologie und Hygiene, Veterinärmedizinische Universität, A-1210 Vienna, Austria
5 Institut für Mikrobiologie und Genetik, Universität Wien, A-1030 Vienna, Austria
Correspondence
Peter Kämpfer
peter.kaempfer{at}agrar.uni-giessen.de
| ABSTRACT |
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| MAIN TEXT |
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During selective enrichment experiments of a phosphorus-removing defined mixed bacterial culture from activated sludge (Hollender et al., 2002
), four bacterial isolates were obtained, one of them showing yellow-pigmented colony morphology on nutrient agar. This strain (B2T) was isolated from an activated sludge enrichment on R2A agar (Oxoid). The isolate was subcultivated on this medium at 25 °C for 48 h for further analyses. The Gram reaction was tested as described by Gerhardt et al. (1994)
. Cell morphologies were observed under a light microscope (1000x times; magnification) (Zeiss) with cells grown for 3 days at 30 °C on tryptone-soy agar (TSA; Oxoid). On nutrient agar (Oxoid), strain B2T formed visible colonies (diameter of about 2 mm) within 24 h at 30 °C. No growth was observed within 14 days at 4 °C or at temperatures above 45 °C. At 15 °C, very small colonies were visible after 7 days incubation. The colonies were yellowish, translucent and shiny with entire edges. After prolonged incubation, colonies were not detectable as single entities because of the production of extracellular slimy substances. Oxidase activity was tested using Bactident-Oxidase test strips (Merck) according to the manufacturer's instructions. Strain B2T was oxidase-positive. Cells were non-motile, non-spore-forming rods (about 2 µm in length) and stained Gram-negative. Strain B2T was able to grow on various nutrient-rich media, including TSA (Oxoid), but was not able to grow on MacConkey agar. Physiological characterization was done as described previously (Kämpfer et al., 1991
), with the modification that all media were diluted threefold. Additional tests were performed using the Micronaut-E gallery (Merlin, formerly known as TTE-AS; Kämpfer, 1990
).
Strain B2T produced acid from D-glucose, D-maltose, trehalose and D-cellobiose (weak). No acid was produced from several other sugars and related compounds. In addition, only a few carbon sources were utilized. Most of the tested p-nitrophenyl derivatives were hydrolysed (see species description).
Fatty acid methyl esters were extracted and prepared by the standard protocol of the Microbial Identification System (MIDI; Microbial ID). Extracts were analysed using a Hewlett Packard model HP6890A GC equipped with an FID, an automatic sampler, an integrator and a computer, as described previously (Kämpfer & Kroppenstedt, 1996
). In strain B2T, the fatty acids 15 : 0 iso (58·5 %), 17 : 0 iso 3-OH (14·1 %) and summed feature 4 (16 : 1
7c/t and/or 15 : 0 iso 2-OH, 8·4 %) were predominant. The detailed fatty acid composition is shown in Table 1
. Because the results shown in this table are based on fatty acid analyses under identical conditions, these results can be compared directly. It is obvious that isolate B2T had larger relative amounts of 15 : 0 iso and 13 : 0 iso than all other Chryseobacterium species. Although representatives of the genera Riemerella and Bergeyella showed similar fatty acid profiles, differentiation on the basis of amounts of several other fatty acids is possible (e.g. iso 17 : 1
9c and 17 : 0 iso 3-OH).
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The detection of menaquinone MK-6 as the only quinone, sym-homospermidine in the polyamine pattern, the large amounts of 15 : 0 iso and the 16S rRNA sequence clearly placed strain B2T in the genus Chryseobacterium. However, strain B2T differs from all other previously described species belonging to this genus.
The main reason for creation of a novel species for strain B2T was the rather high degree of sequence divergence of the 16S rRNA gene in comparison with the corresponding sequences from different species of the genus Chryseobacterium. It was shown clearly by Yamaguchi & Yokoe (2000)
that Chryseobacterium species that share 16S rRNA gene sequence similarities of 94·996·0 % are distinct species on the basis of DNADNA pairing studies, with only 3·031·0 % DNADNA similarity. In addition, differences in physiological features were found in comparison with described Chryseobacterium species.
On the basis of these results, strain B2T is proposed as a representative of a novel species of the genus Chryseobacterium, Chryseobacterium defluvii sp. nov.
Description of Chryseobacterium defluvii sp. nov.
Chryseobacterium defluvii (de.flu'vi.i. L. gen. n. defluvii of sewage).
Cells are non-motile, non-spore-forming rods (approx. 2 µm in length). Gram-negative, oxidase-positive, showing an oxidative metabolism. Good growth is observed on R2A agar, TSA and nutrient agar at 2530 °C, but not on MacConkey agar. Colonies are yellowish, translucent and shiny with entire edges. Menaquinone MK-6 is the predominant quinone and sym-homospermidine is the major polyamine. Phosphatidylethanolamine is the major lipid; several unknown polar lipids are also present. The fatty acid profile is composed largely of 15 : 0 iso (58·5 %), 17 : 0 iso 3-OH (14·1 %) and summed feature 4 (16 : 1
7c/t and/or iso 15 : 0 2-OH, 8·4 %). Produces acid from D-glucose, D-maltose, trehalose and D-cellobiose (weak). No acid produced from adonitol, L-arabinose, D-arabitol, dulcitol, erythritol, i-inositol, lactose, D-mannitol, D-melibiose, methyl
-D-glucoside, raffinose, L-rhamnose, salicin, D-sorbitol, sucrose or D-xylose. The following compounds are utilized as sole sources of carbon: D-glucose, D-maltose, D-mannose, D-trehalose, acetate (weak) and propionate (weak). The following carbon sources are not utilized as sole sources of carbon: N-acetylgalactosamine, N-acetylglucosamine, L-arabinose, L-arbutin, D-cellobiose, D-galactose, gluconate, glycerol, D-fructose, D-mannitol, maltitol,
-D-melibiose, L-rhamnose, D-ribose, D-sucrose, salicin, D-trehalose, D-xylose, adonitol, i-inositol, D-sorbitol, putrescine, cis-aconitate, trans-aconitate, 4-aminobutyrate, adipate, azelate, fumarate, glutarate, DL-3-hydroxybutyrate, itaconate, DL-lactate, 2-oxoglutarate, pyruvate, suberate, citrate, mesaconate, L-alanine,
-alanine, L-ornithine, L-phenylalanine, L-serine, L-aspartate, L-histidine, L-leucine, L-proline, L-tryptophan, 3-hydroxybenzoate, 4-hydroxybenzoate and phenylacetate. The chromogenic substrates p-nitrophenyl
-D-glucopyranoside, p-nitrophenyl
-D-glucopyranoside, bis-p-nitrophenyl phosphate, bis-p-nitrophenyl phenylphosphonate, bis-p-nitrophenyl phosphorylcholine, 2-deoxythymidine-2'-p-nitrophenyl phosphate, L-alanine p-nitroanilide,
-L-glutamate p-nitroanilide and L-proline p-nitroanilide are hydrolysed. The following compounds are not hydrolysed: p-nitrophenyl
-D-galactopyranoside, p-nitrophenyl
-D-glucuronide and p-nitrophenyl
-D-xylopyranoside.
The type strain is strain B2T (=DSM 14219T =CIP 107207T), isolated from sewage sludge (for details see Hollender et al., 2002
).
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P. Kampfer, C.-C. Young, K. R. Sridhar, A. B. Arun, W. A. Lai, F. T. Shen, and P. D. Rekha Transfer of [Flexibacter] sancti, [Flexibacter] filiformis, [Flexibacter] japonensis and [Cytophaga] arvensicola to the genus Chitinophaga and description of Chitinophaga skermanii sp. nov. Int J Syst Evol Microbiol, September 1, 2006; 56(Pt 9): 2223 - 2228. [Abstract] [Full Text] [PDF] |
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H.-Y. Weon, B.-Y. Kim, S.-H. Yoo, S.-W. Kwon, Y.-H. Cho, S.-J. Go, and E. Stackebrandt Chryseobacterium wanjuense sp. nov., isolated from greenhouse soil in Korea. Int J Syst Evol Microbiol, July 1, 2006; 56(Pt 7): 1501 - 1504. [Abstract] [Full Text] [PDF] |
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V. Gallego, M. T. Garcia, and A. Ventosa Chryseobacterium hispanicum sp. nov., isolated from the drinking water distribution system of Sevilla, Spain. Int J Syst Evol Microbiol, July 1, 2006; 56(Pt 7): 1589 - 1592. [Abstract] [Full Text] [PDF] |
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T. D. Allen, P. A. Lawson, M. D. Collins, E. Falsen, and R. S. Tanner Cloacibacterium normanense gen. nov., sp. nov., a novel bacterium in the family Flavobacteriaceae isolated from municipal wastewater Int J Syst Evol Microbiol, June 1, 2006; 56(6): 1311 - 1316. [Abstract] [Full Text] [PDF] |
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H. de Beer, C. J. Hugo, P. J. Jooste, M. Vancanneyt, T. Coenye, and P. Vandamme Chryseobacterium piscium sp. nov., isolated from fish of the South Atlantic Ocean off South Africa Int J Syst Evol Microbiol, June 1, 2006; 56(6): 1317 - 1322. [Abstract] [Full Text] [PDF] |
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P. Kampfer, R. Rossello-Mora, E. Falsen, H.-J. Busse, and B. J. Tindall Cohnella thermotolerans gen. nov., sp. nov., and classification of 'Paenibacillus hongkongensis' as Cohnella hongkongensis sp. nov. Int J Syst Evol Microbiol, April 1, 2006; 56(Pt 4): 781 - 786. [Abstract] [Full Text] [PDF] |
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W.-A. Lai, P. Kampfer, A. B. Arun, F.-T. Shen, B. Huber, P. D. Rekha, and C.-C. Young Deinococcus ficus sp. nov., isolated from the rhizosphere of Ficus religiosa L. Int J Syst Evol Microbiol, April 1, 2006; 56(Pt 4): 787 - 791. [Abstract] [Full Text] [PDF] |
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P. Kampfer, K. Denger, A. M. Cook, S.-T. Lee, U. Jackel, E. B. M. Denner, and H.-J. Busse Castellaniella gen. nov., to accommodate the phylogenetic lineage of Alcaligenes defragrans, and proposal of Castellaniella defragrans gen. nov., comb. nov. and Castellaniella denitrificans sp. nov. Int J Syst Evol Microbiol, April 1, 2006; 56(Pt 4): 815 - 819. [Abstract] [Full Text] [PDF] |
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P. Kampfer, H.-J. Busse, and E. Falsen Polaromonas aquatica sp. nov., isolated from tap water. Int J Syst Evol Microbiol, March 1, 2006; 56(Pt 3): 605 - 608. [Abstract] [Full Text] [PDF] |
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P. Kampfer, J. M. Lindh, O. Terenius, S. Haghdoost, E. Falsen, H.-J. Busse, and I. Faye Thorsellia anophelis gen. nov., sp. nov., a new member of the Gammaproteobacteria Int J Syst Evol Microbiol, February 1, 2006; 56(2): 335 - 338. [Abstract] [Full Text] [PDF] |
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P. Kampfer, O. Terenius, J. M. Lindh, and I. Faye Janibacter anophelis sp. nov., isolated from the midgut of Anopheles arabiensis Int J Syst Evol Microbiol, February 1, 2006; 56(2): 389 - 392. [Abstract] [Full Text] [PDF] |
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M. S. Park, S. R. Jung, K. H. Lee, M.-S. Lee, J. O. Do, S. B. Kim, and K. S. Bae Chryseobacterium soldanellicola sp. nov. and Chryseobacterium taeanense sp. nov., isolated from roots of sand-dune plants Int J Syst Evol Microbiol, February 1, 2006; 56(2): 433 - 438. [Abstract] [Full Text] [PDF] |
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L. A. O'Sullivan, J. Rinna, G. Humphreys, A. J. Weightman, and J. C. Fry Culturable phylogenetic diversity of the phylum 'Bacteroidetes' from river epilithon and coastal water and description of novel members of the family Flavobacteriaceae: Epilithonimonas tenax gen. nov., sp. nov. and Persicivirga xylanidelens gen. nov., sp. nov. Int J Syst Evol Microbiol, January 1, 2006; 56(1): 169 - 180. [Abstract] [Full Text] [PDF] |
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K. Shimomura, S. Kaji, and A. Hiraishi Chryseobacterium shigense sp. nov., a yellow-pigmented, aerobic bacterium isolated from a lactic acid beverage Int J Syst Evol Microbiol, September 1, 2005; 55(5): 1903 - 1906. [Abstract] [Full Text] [PDF] |
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H. de Beer, C. J. Hugo, P. J. Jooste, A. Willems, M. Vancanneyt, T. Coenye, and P. A. R. Vandamme Chryseobacterium vrystaatense sp. nov., isolated from raw chicken in a chicken-processing plant Int J Syst Evol Microbiol, September 1, 2005; 55(5): 2149 - 2153. [Abstract] [Full Text] [PDF] |
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P. Kampfer, S. Engelhart, M. Rolke, and J. Sennekamp Extrinsic Allergic Alveolitis (Hypersensitivity Pneumonitis) Caused by Sphingobacterium spiritivorum from the Water Reservoir of a Steam Iron J. Clin. Microbiol., September 1, 2005; 43(9): 4908 - 4910. [Abstract] [Full Text] [PDF] |
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K. K. Kim, M. K. Kim, J. H. Lim, H. Y. Park, and S.-T. Lee Transfer of Chryseobacterium meningosepticum and Chryseobacterium miricola to Elizabethkingia gen. nov. as Elizabethkingia meningoseptica comb. nov. and Elizabethkingia miricola comb. nov. Int J Syst Evol Microbiol, May 1, 2005; 55(3): 1287 - 1293. [Abstract] [Full Text] [PDF] |
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F.-T. Shen, P. Kampfer, C.-C. Young, W.-A. Lai, and A. B. Arun Chryseobacterium taichungense sp. nov., isolated from contaminated soil Int J Syst Evol Microbiol, May 1, 2005; 55(3): 1301 - 1304. [Abstract] [Full Text] [PDF] |
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P. Kampfer, R. M. Kroppenstedt, and I. Grun-Wollny Nonomuraea kuesteri sp. nov. Int J Syst Evol Microbiol, March 1, 2005; 55(2): 847 - 851. [Abstract] [Full Text] [PDF] |
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K. K. Kim, H.-S. Bae, P. Schumann, and S.-T. Lee Chryseobacterium daecheongense sp. nov., isolated from freshwater lake sediment Int J Syst Evol Microbiol, January 1, 2005; 55(1): 133 - 138. [Abstract] [Full Text] [PDF] |
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P. Kampfer, R. Schulze, U. Jackel, K. A. Malik, R. Amann, and S. Spring Hydrogenophaga defluvii sp. nov. and Hydrogenophaga atypica sp. nov., isolated from activated sludge Int J Syst Evol Microbiol, January 1, 2005; 55(1): 341 - 344. [Abstract] [Full Text] [PDF] |
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H. Yi, H. I. Yoon, and J. Chun Sejongia antarctica gen. nov., sp. nov. and Sejongia jeonii sp. nov., isolated from the Antarctic Int J Syst Evol Microbiol, January 1, 2005; 55(1): 409 - 416. [Abstract] [Full Text] [PDF] |
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C.-C. Young, P. Kampfer, F.-T. Shen, W.-A. Lai, and A. B. Arun Chryseobacterium formosense sp. nov., isolated from the rhizosphere of Lactuca sativa L. (garden lettuce) Int J Syst Evol Microbiol, January 1, 2005; 55(1): 423 - 426. [Abstract] [Full Text] [PDF] |
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M. K. Kim, W.-T. Im, Y. K. Shin, J. H. Lim, S.-H. Kim, B. C. Lee, M.-Y. Park, K. Y. Lee, and S.-T. Lee Kaistella koreensis gen. nov., sp. nov., a novel member of the Chryseobacterium-Bergeyella-Riemerella branch Int J Syst Evol Microbiol, November 1, 2004; 54(6): 2319 - 2324. [Abstract] [Full Text] [PDF] |
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N. Lee, C. M. Cellamare, C. Bastianutti, R. Rossello-Mora, P. Kampfer, W. Ludwig, K. H. Schleifer, and L. Stante Emended description of the species Lampropedia hyalina Int J Syst Evol Microbiol, September 1, 2004; 54(5): 1709 - 1715. [Abstract] [Full Text] [PDF] |
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P. Kampfer and R. M. Kroppenstedt Pseudonocardia benzenivorans sp. nov. Int J Syst Evol Microbiol, May 1, 2004; 54(3): 749 - 751. [Abstract] [Full Text] [PDF] |
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P. Kampfer, S. Buczolits, U. Jackel, I. Grun-Wollny, and H.-J. Busse Nocardia tenerifensis sp. nov. Int J Syst Evol Microbiol, March 1, 2004; 54(2): 381 - 383. [Abstract] [Full Text] [PDF] |
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P. Kampfer, S. Buczolits, A. Albrecht, H.-J. Busse, and E. Stackebrandt Towards a standardized format for the description of a novel species (of an established genus): Ochrobactrum gallinifaecis sp. nov. Int J Syst Evol Microbiol, May 1, 2003; 53(3): 893 - 896. [Abstract] [Full Text] [PDF] |
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