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Int J Syst Evol Microbiol 56 (2006), 1055-1058; DOI  10.1099/ijs.0.64171-0
© 2006 International Union of Microbiological Societies

Psychroserpens mesophilus sp. nov., a mesophilic marine bacterium belonging to the family Flavobacteriaceae isolated from a young biofilm

Kae Kyoung Kwon1, Soon Jae Lee1, Jae Hyun Park1,2, Tae-Young Ahn2 and Hong Kum Lee3

1 Marine Biotechnology Research Center, Korea Ocean Research and Development Institute (KORDI), Ansan PO Box 29, 425-600, Republic of Korea
2 Department of Microbiology, Dankook University, Cheonan 330-714, Republic of Korea
3 Polar BioCenter, Korea Polar Research Institute, Korea Ocean Research and Development Institute (KORDI), Ansan PO Box 29, 425-600, Republic of Korea

Correspondence
Hong Kum Lee
hklee{at}kopri.re.kr


   ABSTRACT
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A number of marine bacteria isolated from young biofilms were characterized as belonging to the family Flavobacteriaceae. The taxonomic characterization of strain KOPRI 13649T, which was isolated from an acrylic surface at the seashore at Gangneung, Korea, is reported here. The nearly complete 16S rRNA gene sequence of strain KOPRI 13649T was determined and was found to have a high level of similarity with that of Psychroserpens burtonensis (95.0–95.6 %). In addition, phylogenetic analysis and comparison with closely related strains confirmed that the strain represented a novel member of the genus Psychroserpens. The major respiratory quinone of strain KOPRI 13649T was MK-6 and the DNA G+C content was 29.8 mol%. The dominant fatty acid methyl esters were i-15 : 0, a-15 : 0, i-16 : 0, i-15 : 1{omega}10, 16 : 1{omega}7 and 15 : 0. Growth was observed at 10–34 °C (optimum 30 °C), at pH 6–9 (optimum 6.5–8.0) and with 0.5–4 % NaCl (optimum 1 %). On the basis of the polyphasic taxonomic evidence presented, strain KOPRI 13649T (=KCCM 42261T=JCM 13413T) should be classified as the type strain of a novel species in the genus Psychroserpens, for which the name Psychroserpens mesophilus sp. nov. is proposed.

The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain KOPRI 13649T is DQ001321.

A table detailing the cellular fatty acid content of strain KOPRI 13649T and closely related members of the Flavobacteriaceae is available as supplementary material in IJSEM Online.


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The genus Psychroserpens is a member of the family Flavobacteriaceae in the phylum Bacteroidetes. Psychroserpens burtonensis, isolated from the Antarctic ice at Lake Burton (Bowman et al., 1997Go), is the only currently recognized member of the genus. In the last decade, several bacterial isolates have been reported as putative members of the genus Psychroserpens (Brinkmeyer et al., 2003Go; DeLong et al., 1993Go; Li et al., 1999Go) but have yet to be formally taxonomically characterized. We have isolated several bacteria belonging to the family Flavobacteriaceae from a young marine biofilm and herein characterize one of these, designated strain KOPRI 13649T, as representing a novel member of the genus Psychroserpens.

Strain KOPRI 13649T was isolated from an acrylic surface after immersion in sea water for 3 days. The acrylic surface was withdrawn from the sea water, and the biofilm that had formed on it was detached, dispersed into sterilized sea water and spread on marine agar 2216 (MA; Difco). Among the colonies subsequently formed, a yellow-coloured, morphologically distinct colony was isolated and named KOPRI 13649T. This strain was cultivated on MA for morphological and biochemical characterization.

Unless otherwise stated, the physiological and morphological characterization was conducted according to the methods given by Sohn et al. (2004)Go and Kwon et al. (2005a)Go. Requirement for NaCl was tested using modified marine broth 2216 (5 g peptone, 1 g yeast extract, 0.01 g FePO4, 1.89 g MgCl2.6H2O, 0.36 g CaCl2.2H2O, per litre distilled water) supplemented with 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 10, 15 or 20 % (w/v) NaCl. Degradation of starch and casein and production of hydrogen sulfide were tested by using the procedures of Smibert & Krieg (1994)Go. The physiological, biochemical and morphological characteristics of strain KOPRI 13649T are given in the species description (see later) and in Table 1Go.


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  		Table 1. Phenotypic characteristics differentiating strain KOPRI 13649T from closely related members of the family Flavobacteriaceae

Taxa: 1, KOPRI 13649T; 2, Psychroserpens burtonensis; 3, Formosa algae KMM 3553T; 4, Algibacter lectus KMM 3914T; 5, Gelidibacter. Data were taken from Bowman et al. (1997)Go, Macián et al. (2002)Go, Ivanova et al. (2004)Go, Nedashkovskaya et al. (2004)Go and Bowman & Nichols (2005)Go. All strains have menaquinone MK-6 as the major respiratory quinone, produce catalase and do not produce H2S or indole. All are able to liquefy gelatin. +, Positive; –, negative; ND, not determined; V, variable.

 
Cells of strain KOPRI 13649T were Gram-negative, non-motile rods, 0.57–0.63 µm in length and 0.41–0.51 µm in diameter. During growth, the length of the cells increased up to 1.7 µm. Colonies were yellow to yellowish orange when grown on MA plates at 30 °C. Growth of strain KOPRI 13649T was observed at 10–34 °C; growth was poor at temperatures below 20 °C or above 34 °C, and no growth was observed at 8 or 37 °C.

Phylogenetic analysis using the 16S rRNA gene sequence of strain KOPRI 13649T was conducted according to the methods of Sohn et al. (2004)Go. The sequence was compared against the 16S rRNA gene sequences of strains belonging to P. burtonensis and of members of the genera Winogradskyella (Lau et al., 2005Go; Nedashkovskaya et al., 2005aGo), Gelidibacter (Bowman et al., 1997Go; Bowman & Nichols, 2005Go; Macián et al., 2002Go) and Subsaximicrobium (Bowman & Nichols, 2005Go), Subsaxibacter broadyi (Bowman & Nichols, 2005Go), Yeosuana aromativorans (Kwon et al., 2005bGo), Bizionia paragorgiae (Nedashkovskaya et al., 2005bGo), Formosa algae (Ivanova et al., 2004Go), Algibacter lectus (Nedashkovskaya et al., 2004Go) and Lacinutrix copepodicola (Bowman & Nichols, 2005Go). Closest sequence similarity was to P. burtonensis (95.0–95.6 %). Phylogenetic analysis based on 16S rRNA gene sequences placed strain KOPRI 13649T within the outer area of strains belonging to P. burtonensis, but in the same phylogenetic line with the genus Psychroserpens (Fig. 1Go).


Figure 1
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  		Fig. 1. Phylogenetic tree based on nearly complete 16S rRNA gene sequences (1326 unambiguously aligned base pairs) showing the relationship between strain KOPRI 13649T and other members of the family Flavobacteriaceae. The tree is based on the maximum-likelihood distances model and the neighbour-joining method. Bootstrap values of >50 % (from 1000 resampled datasets) are shown. Bar, 0.01 nucleotide substitutions per nucleotide position.

 
The cellular fatty acid methyl ester profile of strain KOPRI 13649T was determined according to the methods given by Sohn et al. (2004)Go. The dominant fatty acid methyl esters were i-15 : 0 (31.4 %), a-15 : 0 (10.9 %), i-16 : 0 (7.3 %), i-15 : 1{omega}10 (7.1 %), 16 : 1{omega}7 (5.6 %) and 15 : 0 (5.6 %). The isolate also contained small amounts of i-14 : 0 (4.1 %), 17 : 0 cyclic (3.5 %), 10-methyl 16 : 0 (3.2 %), a-17 : 0 (3.0 %), i-17 : 0 (2.7 %), 16 : 0 (2.2 %), 15 : 1 (1.6 %) and 14 : 0 (1.4 %). This profile differed from that of P. burtonensis (Bowman et al., 1997Go) as follows: relatively large amounts of i-15 : 0, presence of i-14 : 0, i-16 : 0, a-17 : 0, 10-methyl-16 : 0 and 17 : 0 cyclic, and insignificant amounts of 15 : 1, a-15 : 1 and br-16 : 1 (for more complete details see Supplementary Table S1 available in IJSEM Online). The difference in optimal growth temperature might explain these differences.

The DNA G+C content of strain KOPRI 13649T, as determined using the thermal denaturation method of Kim et al. (2000)Go, was 29.8 mol%, a value similar to that of P. burtonensis. The major respiratory quinone was determined to be menaquinone according to the reversed-phase TLC method described by Kim et al. (2000)Go and was confirmed to be MK-6 following the HPLC method of Collins (1985)Go.

The results of our phylogenetic analysis combined with the DNA G+C determination provide strong support that strain KOPRI 13649T should be classified as a member of the genus Psychroserpens. However, the isolate showed strict mesophilic growth characteristics and had a cellular fatty acid profile distinct from that of P. burtonensis. Thus, on the basis of the polyphasic taxonomic evidence presented, strain KOPRI 13649T is considered to represent a novel species in the genus Psychroserpens, for which the name Psychroserpens mesophilus sp. nov. is proposed.

Description of Psychroserpens mesophilus sp. nov.
Psychroserpens mesophilus [me.so.phi'lus. Gr. adj. mesos middle; Gr. adj. philus loving; N.L. adj. mesophilus middle (temperature)-loving, mesophilic].

Cells are Gram-negative, non-motile rods, 0.41–0.51x0.57–1.67 µm. Gliding motility is absent. Colonies on solid MA are yellow to yellowish orange. Oxidase and catalase activities are positive. Positive for lysine decarboxylase, tryptophan deaminase, nitrate reduction, Voges–Proskauer reaction, and assimilation of glucose, mannose, mannitol, sorbitol, rhamnose, melibiose, amylose and arabinose (according to the API 20E kit). Oxidizes {alpha}-cyclodextrin, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, L-arabinose, D-fructose, {alpha}-D-glucose, maltose, D-trehalose, sucrose, methyl pyruvate, monomethyl succinate, {alpha}- and beta-hydroxybutyric acid, {alpha}-ketobutyric acid, {alpha}-ketoglutaric acid, {alpha}-ketovaleric acid, DL-lactic acid, propionic acid, D-alanine, L-leucine, L-ornithine, L-proline, L-serine, L-threonine, uridine, inosine and DL-{alpha}-glycerol phosphate (Microlog GN2 MicroPlate). Growth is observed at 10–34 °C (optimum 30 °C), at pH 6–9 (optimum pH 6.5–8.0) and with 0.5–4 % NaCl (optimum 1 %). Requires either 0.18 % (w/v) CaCl2 or 0.59 % (w/v) MgCl2 for growth in addition to NaCl. The major respiratory quinone is MK-6. The DNA G+C content is 29.8 mol%. The dominant fatty acids are i-15 : 0, a-15 : 0, i-16 : 0, i-15 : 1{omega}10, 16 : 1{omega}7 and 15 : 0.

The type strain, KOPRI 13649T (=KCCM 42261T=JCM 13413T), was isolated from a young biofilm formed on an acrylic surface in Gangneung, Korea.


   ACKNOWLEDGEMENTS
 
This work was supported by the National Research Laboratory program of the Ministry of Science and Technology, Korea, with funding to H. K. L.


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