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1 Pacific Institute of Bioorganic Chemistry of the Far-Eastern Branch of the Russian Academy of Sciences, Pr. 100 Let Vladivostoku 159, 690022, Vladivostok, Russia
2 Korean Collection for Type Cultures, Genetic Resources Center, Korea Institute of Bioscience and Biotechnology, Yusong, Daejon 305-333, Republic of Korea
3 Institute of Microbiology of the Russian Academy of Sciences, Pr. 60 Let October 7/2, 117811, Moscow, Russia
4 Bereich Mikrobiologie, Abt. Mikrobielle Pathogenitat und Impfstoffforschung, GBF – Gesellschaft für Biotechnologische Forschung, Mascheroder Weg 1, D-38124 Braunschweig, Germany
5 Institute of Marine Biology of the Far-Eastern Branch of the Russian Academy of Sciences, Pal'chevskogo St 17, 690032, Vladivostok, Russia
6 Culture Collection, Department of Clinical Bacteriology, University of Göteborg, Guldhedsgatan 10, S-413 46 Göteborg, Sweden
Correspondence
Olga I. Nedashkovskaya
olganedashkovska{at}piboc.dvo.ru
or olganedashkovska{at}yahoo.com
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Published online ahead of print on 23 May 2003 as DOI 10.1099/ijs.0.02626-0.
The GenBank/EMBL/DDBJ accession numbers for the 16S rDNA sequences of Mesonia algae KMM 3909T, KMM 3910, KMM 3936 and KMM 3937 are AF536383, AF536384, AF536385 and AF536386, respectively.
A micrograph showing rod-shaped cells of Mesonia algae is available as supplementary material in IJSEM Online.
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; Fuhrman et al., 1993; Suzuki et al., 1997; Brambilla et al., 2001). Bacteria that belong to the phylum Cytophaga–Flavobacterium–Bacteroides (CFB) are often found in natural microbial communities of marine environments (Glöckner et al., 1999; Bano & Hollibaugh, 2002; Kirchman, 2002). In recent years, isolation procedures supported by phylogenetic methods have led to the description of the novel genera Gelidibacter, Psychroserpens, Polaribacter, Cellulophaga, Zobellia, Tenacibaculum, Arenibacter, Muricauda, Aequorivita and Reichenbachia (Bowman et al., 1997; Gosink et al., 1998; Johansen et al., 1999; Barbeyron et al., 2001; Bruns et al., 2001; Ivanova et al., 2001; Suzuki et al., 2001; Bowman & Nichols, 2002; Nedashkovskaya et al., 2003). Among recently described marine bacteria of the family Flavobacteriaceae that have been isolated from unique Antarctic saline lakes are representatives of the genera Psychroflexus and Salegentibacter, which are characterized by the ability to grow in nutrient media that contain up to 15–20 % NaCl (Dobson et al., 1993; Bowman et al., 1998; McCammon & Bowman, 2000). In the course of the study of the microbial community associated with the common green alga Acrosiphonia sonderi, which inhabits coastal waters of the Sea of Japan, we isolated another novel, moderately halophilic bacterium that forms a phylogenetic cluster with species of the genera Salegentibacter and Psychroflexus. Phenotypic and genotypic characteristics and fatty acid and menaquinone compositions, in combination with phylogenetic findings, support the establishment a novel genus, for which the name Mesonia gen. nov. is proposed. In the present work, we describe a novel member of the family Flavobacteriaceae, which is designated Mesonia algae gen. nov., sp. nov. Strains KMM 3909T, KMM 3910, KMM 3936 and KMM 3937 were isolated from samples of the green alga Acrosiphonia sonderi that were collected in Troitsa Bay, Gulf of Peter the Great, Sea of Japan, during June 2000. For strain isolation, 0·1 ml homogenates of algal fronds were transferred onto plates of marine agar 2216 (Difco). After primary isolation and purification, strains were cultivated at 28 °C on the same medium and stored at -80 °C in marine broth (Difco) supplemented with 20 % (v/v) glycerol.
Oxidative or fermentative utilization of glucose was determined by using Hugh–Leifson medium modified for marine bacteria (Lemos et al., 1985). Degradation of agar, starch, casein, gelatin, cellulose (filter paper and CM-cellulose), chitin, DNA, urea and alginic acids, flexirubin production, growth at different pH values, production of acid from carbohydrates and susceptibility to antibiotics were tested as described previously (Nedashkovskaya et al., 2003). Gram-staining reaction, hydrolysis of Tweens 20, 40 and 80, nitrate reduction, production of hydrogen sulphide, indole and acetoin (Voges–Proskauer reaction) and activities of 
-galactosidase, oxidase, catalase and alkaline phosphatase were tested according to the methods of Gerhardt et al. (1994). To examine carbon source utilization, commercial API 20NE identification strips (bioMérieux), following the instructions of the manufacturer, and a medium that contained 0·2 g NaNO3, 0·2 g NH4Cl, 0·05 g yeast extract (Difco) and 0·4 % (w/v) carbon source in 1000 ml artificial sea water were used. In order to study the temperature range for growth, bacteria were cultivated on medium A, which consisted of (l-1): 5 g Bacto peptone (Difco), 2 g Bacto yeast extract (Difco), 1 g glucose, 0·02 g KH2PO4 and 0·05 g MgSO4.7H2O in 50 % (v/v) natural sea water and 50 % (v/v) distilled water. Bacterial growth at different concentrations of NaCl was checked on medium A that was prepared with distilled water and contained 0, 1, 2, 3, 5, 6, 8, 10, 12, 13, 15, 16, 17 or 18 % (w/v) NaCl. Spreading growth was observed by cultivation on medium B, which contained (l-1): 1 g Bacto peptone (Difco), 1 g yeast extract (Difco), 15 g agar and half-strength natural sea water under high-moisture conditions. Gliding motility was determined as described by Bowman (2000). On marine agar, colonies of strains KMM 3909T, KMM 3936 and KMM 3937 were yellow-pigmented and those of strain KMM 3910 were whitish. Growth of strains KMM 3936 and KMM 3937 occurred in media that contained 1–12 % NaCl; strains KMM 3909T and KMM 3910 grew in the presence of 1–15 % NaCl. Other physiological and biochemical characteristics of the strains studied are listed in the species description and Table 1. Cell morphology was observed by scanning electron microscopy (SEM) as described by Bruns et al. (2001). The rod-shaped cells ranged from 0·4 to 0·5 µm in width and from 1·6 to 2·3 µm in length (see Supplementary Fig. A in IJSEM Online). Analysis of fatty acid methyl esters was carried out according to the standard protocol of the Microbial Identification system (Microbial ID). The predominant cellular fatty acids of KMM 3909T were straight- and branched-chain unsaturated fatty acids, namely i-C15 : 0, i-C15 : 1, i-C16 : 0, i-C16 : 0 3-OH and i-C17 : 0 3-OH (Table 2). Isoprenoid quinones were extracted from lyophilized cells and analysed as described by Akagawa-Matsushita et al. (1992). Menaquinones were detected by monitoring at 270 nm and were identified by comparison with known quinones from the reference strain Salegentibacter salegens DSM 5424T. The main isoprenoid quinone of the strains studied was MK-6. DNA was isolated following the method of Marmur (1961) and the G+C content of the DNA was determined by the thermal denaturation method (Marmur & Doty, 1962). DNA–DNA hybridization was performed spectrophotometrically and initial renaturation rates were recorded as described by De Ley et al. (1970). The DNA G+C content of the strains studied ranged from 32·7 to 34·0 mol%, as determined by the thermal denaturation method. The level of DNA–DNA binding between KMM 3909T, KMM 3910, KMM 3936 and KMM 3937 was 81–95 %.
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). Obtained sequence data were aligned with those of representative members of selected genera that belong to the family Flavobacteriaceae by using PHYDIT version 3.2 (http://plaza.snu.ac.kr/
jchun/phydit/). Phylogenetic trees were inferred by using suitable programs of the PHYLIP package (Felsenstein, 1993). Phylogenetic distances were calculated from the model of Jukes & Cantor (1969) and trees were constructed on the basis of the neighbour-joining (Saitou & Nei, 1987), least-squares (Fitch & Margoliash, 1967) and maximum-likelihood (Felsenstein, 1993) algorithms. Bootstrap analysis was performed with 1000 resampled datasets by using the SEQBOOT and CONSENSE programs of the PHYLIP package.
Phylogenetic analysis of almost-complete 16S rDNA sequences (1385 nucleotide positions) revealed that the strains studied formed a distinct lineage within the family Flavobacteriaceae (Bernardet et al., 2002), whilst belonging to a cluster group with [Cytophaga] latercula, S. salegens and Psychroflexus species (Fig. 1
). 16S rDNA sequence similarity values of the strains studied and their close relatives, [C.] latercula, Psychroflexus torquis and Psychroflexus gondwanensis, were 91·1, 90·4 and 90·6 %, respectively. The nearest neighbour, S. salegens DSM 5424T, had 92·6 % 16S rDNA sequence similarity (102 nucleotide differences) with KMM 3909T and KMM 3910 without significant bootstrap support (61 %). In addition, S. salegens DSM 5424T was rather closer to [C.] latercula ATCC 23177T (16S rDNA similarity between them was 92·8 %), whereas the latter shared around 91·2 % similarity with the strains studied. The overall topology of the unrooted tree based on 16S rDNA was not changed in the trees by using other algorithms, except that KMM 3909T clustered with P. torquis ACAM 623T in the first place and then with S. salegens DSM 5424T in the maximum-likelihood tree (data not shown).
It is therefore evident from phylogenetic analysis that the strains studied can be considered to form a genus separate from Salegentibacter. Low sequence similarities of the strains tested with other members of the CFB phylum described to date (87·2–91·1 %) demonstrate clearly that the bacteria isolated in this study represent a novel genus. Significant differences in the whole-cell fatty acid compositions of strains KMM 3909T and S. salegens DSM 5424T should be noted (Table 2). For example, fatty acid a-C15 : 1 presents in significant quantity in the cell extract of S. salegens DSM 5424T (3·5 %) but only in minor quantity in that of strain KMM 3909T (0·4 %). The hydroxy fatty acid C15 : 0 3-OH is represented in S. salegens DSM 5424T (3·9 %) but is absent in strain KMM 3909T. Fatty acid a-C17 : 1
9c was found in the cell extract of strain KMM 3909T (1·9 %), but not in that of S. salegens DSM 5424T. Phenotypic examination of the strains studied (including their inability to grow without Na+ ions and to utilize carbohydrates, lack of DNase activity, absence of starch hydrolysis, presence of casein hydrolysis and negative nitrate reduction), in combination with phylogenetic differences and distinctiveness of cellular fatty acid composition, allows the differentiation of strains KMM 3909T, KMM 3910, KMM 3936 and KMM 3937 from their closest relative, S. salegens. Phenotypic features that separate the strains studied from other relatives of the family Flavobacteriaceae are listed in Table 1.
Thus, the polyphasic data presented in this paper support the conclusion that the bacteria studied could not be affiliated to any taxa currently included in the family Flavobacteriaceae. Consequently, we propose that strains KMM 3909T, KMM 3910, KMM 3936 and KMM 3937 should be placed in a novel genus as Mesonia algae gen. nov., sp. nov.
Description of Mesonia gen. nov.
Mesonia [Me.so'ni.a. N.L. fem. n. Mesonia arbitrary name derived from the abbreviation MES (Marine Experimental Station of the Pacific Institute of Bioorganic Chemistry, FEB RAS) near the site where the bacteria were first isolated].
Rod-shaped, non-motile, Gram-negative cells. Endospores are not formed. Na+ ions are required for growth. Strictly aerobic. Non-diffusible yellow pigments are produced. No flexirubins are formed. Chemo-organotrophic. Cytochrome oxidase-, catalase- and alkaline phosphatase-positive. Major respiratory quinone is MK-6. Main cellular fatty acids are the straight- and branched-chain unsaturated fatty acids i-C15 : 0, i-C15 : 1, i-C16 : 0, i-C16 : 0 3-OH and i-C17 : 0 3-OH. As determined by 16S rDNA sequence analysis, the genus Mesonia is a member of the family Flavobacteriaceae in the phylum Cytophaga–Flavobacterium–Bacteroides. The type species is Mesonia algae.
Description of Mesonia algae sp. nov.
Mesonia algae (al'gae. L. gen. n. algae of alga, seaweed; bacterium isolated from alga).
Main characteristics are otherwise the same as those given for the genus. In addition, cells range from 0·4 to 0·5 µm in width and from 1·6 to 2·3 µm in length. On marine agar, colonies are 2–4 mm in diameter, circular, shiny with entire edges and yellow in colour (whitish strains occur). Growth is observed at 4–34 °C. Optimal temperature for growth is 21–23 °C. Growth occurs at 1–15 % NaCl. Gelatin, casein and Tweens 20, 40 and 80 are decomposed. No hydrolysis of agar, alginate, starch, cellulose (CM-cellulose or filter paper) or chitin occurs. No acid is formed from arabinose, cellobiose, fucose, galactose, glucose, lactose, maltose, mannose, melibiose, raffinose, rhamnose, sucrose, xylose, citrate, adonitol, dulcitol, glycerol, inositol or mannitol. Arabinose, glucose, lactose, mannose, sucrose, mannitol, inositol, sorbitol, N-acetylglucosamine, gluconate, caprate, adipate, malate, malonate, phenylacetate and citrate are not utilized. Nitrate is not reduced. H2S is produced. Indole and acetoin (Voges–Proskauer reaction) production is negative. G+C content of the DNA is 32·7–34·0 mol%.
Type strain is KMM 3909T (=KCTC 12089T=CCUG 47092T). Isolated from the green alga Acrosyphonia sonderi, collected in Troitsa Bay of the Gulf of Peter the Great of the Sea of Japan.
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ACKNOWLEDGEMENTS |
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J.-H. Yoon, S.-J. Kang, S.-Y. Jung, H. W. Oh, and T.-K. Oh Gaetbulimicrobium brevivitae gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from a tidal flat of the Yellow Sea in Korea Int J Syst Evol Microbiol, January 1, 2006; 56(1): 115 - 119. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, M. Vancanneyt, C. Snauwaert, A. M. Lysenko, M. Rohde, G. M. Frolova, N. V. Zhukova, V. V. Mikhailov, K. S. Bae, et al. Formosa agariphila sp. nov., a budding bacterium of the family Flavobacteriaceae isolated from marine environments, and emended description of the genus Formosa Int J Syst Evol Microbiol, January 1, 2006; 56(1): 161 - 167. [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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S. C. K. Lau, M. M. Y. Tsoi, X. Li, I. Plakhotnikova, S. Dobretsov, M. Wu, P.-K. Wong, J. R. Pawlik, and P.-Y. Qian Stenothermobacter spongiae gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from a marine sponge in the Bahamas, and emended description of Nonlabens tegetincola Int J Syst Evol Microbiol, January 1, 2006; 56(1): 181 - 185. [Abstract] [Full Text] [PDF]
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J.-H. Yoon, S.-J. Kang, C.-H. Lee, and T.-K. Oh Donghaeana dokdonensis gen. nov., sp. nov., isolated from sea water Int J Syst Evol Microbiol, January 1, 2006; 56(1): 187 - 191. [Abstract] [Full Text] [PDF]
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S. C. K. Lau, M. M. Y. Tsoi, X. Li, I. Plakhotnikova, S. Dobretsov, P.-K. Wong, and P.-Y. Qian Gramella portivictoriae sp. nov., a novel member of the family Flavobacteriaceae isolated from marine sediment Int J Syst Evol Microbiol, November 1, 2005; 55(6): 2497 - 2500. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, A. M. Lysenko, M. S. Park, V. V. Mikhailov, K. S. Bae, and H. Y. Park Roseivirga echinicomitans sp. nov., a novel marine bacterium isolated from the sea urchin Strongylocentrotus intermedius, and emended description of the genus Roseivirga Int J Syst Evol Microbiol, September 1, 2005; 55(5): 1797 - 1800. [Abstract] [Full Text] [PDF]
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L. A. O'Sullivan, J. Rinna, G. Humphreys, A. J. Weightman, and J. C. Fry Fluviicola taffensis gen. nov., sp. nov., a novel freshwater bacterium of the family Cryomorphaceae in the phylum 'Bacteroidetes' Int J Syst Evol Microbiol, September 1, 2005; 55(5): 2189 - 2194. [Abstract] [Full Text] [PDF]
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J. P. Bowman and D. S. Nichols Novel members of the family Flavobacteriaceae from Antarctic maritime habitats including Subsaximicrobium wynnwilliamsii gen. nov., sp. nov., Subsaximicrobium saxinquilinus sp. nov., Subsaxibacter broadyi gen. nov., sp. nov., Lacinutrix copepodicola gen. nov., sp. nov., and novel species of the genera Bizionia, Gelidibacter and Gillisia Int J Syst Evol Microbiol, July 1, 2005; 55(4): 1471 - 1486. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, M. Vancanneyt, P. Dawyndt, K. Engelbeen, K. Vandemeulebroecke, I. Cleenwerck, B. Hoste, J. Mergaert, T.-L. Tan, G. M. Frolova, et al. Reclassification of [Cytophaga] marinoflava Reichenbach 1989 as Leeuwenhoekiella marinoflava gen. nov., comb. nov. and description of Leeuwenhoekiella aequorea sp. nov. Int J Syst Evol Microbiol, May 1, 2005; 55(3): 1033 - 1038. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, S. K. Han, C. Snauwaert, M. Vancanneyt, J. Swings, K.-O. Kim, A. M. Lysenko, M. Rohde, G. M. Frolova, et al. Winogradskyella thalassocola gen. nov., sp. nov., Winogradskyella epiphytica sp. nov. and Winogradskyella eximia sp. nov., marine bacteria of the family Flavobacteriaceae Int J Syst Evol Microbiol, January 1, 2005; 55(1): 49 - 55. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, A. M. Lysenko, G. M. Frolova, V. V. Mikhailov, K. H. Lee, and K. S. Bae Description of Aquimarina muelleri gen. nov., sp. nov., and proposal of the reclassification of [Cytophaga] latercula Lewin 1969 as Stanierella latercula gen. nov., comb. nov. Int J Syst Evol Microbiol, January 1, 2005; 55(1): 225 - 229. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, D. H. Lee, A. M. Lysenko, L. S. Shevchenko, G. M. Frolova, V. V. Mikhailov, K. H. Lee, and K. S. Bae Roseivirga ehrenbergii gen. nov., sp. nov., a novel marine bacterium of the phylum 'Bacteroidetes', isolated from the green alga Ulva fenestrata Int J Syst Evol Microbiol, January 1, 2005; 55(1): 231 - 234. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, A. M. Lysenko, G. M. Frolova, V. V. Mikhailov, and K. S. Bae Bizionia paragorgiae gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from the soft coral Paragorgia arborea Int J Syst Evol Microbiol, January 1, 2005; 55(1): 375 - 378. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, A. M. Lysenko, G. M. Frolova, V. V. Mikhailov, K. S. Bae, D. H. Lee, and I. S. Kim Gramella echinicola gen. nov., sp. nov., a novel halophilic bacterium of the family Flavobacteriaceae isolated from the sea urchin Strongylocentrotus intermedius Int J Syst Evol Microbiol, January 1, 2005; 55(1): 391 - 394. [Abstract] [Full Text] [PDF]
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J.-C. Cho and S. J. Giovannoni Robiginitalea biformata gen. nov., sp. nov., a novel marine bacterium in the family Flavobacteriaceae with a higher G+C content Int J Syst Evol Microbiol, July 1, 2004; 54(4): 1101 - 1106. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, M. Suzuki, M. Vancanneyt, I. Cleenwerck, N. V. Zhukova, M. V. Vysotskii, V. V. Mikhailov, and J. Swings Salegentibacter holothuriorum sp. nov., isolated from the edible holothurian Apostichopus japonicus Int J Syst Evol Microbiol, July 1, 2004; 54(4): 1107 - 1110. [Abstract] [Full Text] [PDF]
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O. I. Nedashkovskaya, S. B. Kim, S. K. Han, M.-S. Rhee, A. M. Lysenko, M. Rohde, N. V. Zhukova, G. M. Frolova, V. V. Mikhailov, and K. S. Bae Algibacter lectus gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from green algae Int J Syst Evol Microbiol, July 1, 2004; 54(4): 1257 - 1261. [Abstract] [Full Text] [PDF]
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S. Van Trappen, I. Vandecandelaere, J. Mergaert, and J. Swings Gillisia limnaea gen. nov., sp. nov., a new member of the family Flavobacteriaceae isolated from a microbial mat in Lake Fryxell, Antarctica Int J Syst Evol Microbiol, March 1, 2004; 54(2): 445 - 448. [Abstract] [Full Text] [PDF]
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