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State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Zhong-Guan-Cun, Haidian, Beijing 100080, P. R. China
Correspondence
Shuang-Jiang Liu
shuangjiang{at}hotmail.com
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ABSTRACT |
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The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of Roseomonas lacus TH-G33T is AJ786000.
A table showing the cellular fatty acid profile of Roseomonas lacus TH-G33T is available as supplementary material in IJSEM Online.
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MAIN TEXT |
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TOPABSTRACTMAIN TEXTREFERENCES |
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) was created for pink coccoid isolates that caused bacteraemia and other human infections. Roseomonas species have been isolated frequently from blood, wounds, exudates, abscesses and genitourinary specimens (Rihs et al., 1993; Sandoe et al., 1997; Bibashi et al., 2000; Subudhi et al., 2001). Recently, bacterial strains that are phylogenetically related to members of the genus Roseomonas have also been isolated frequently from environmental samples such as drinking water distribution systems (September et al., 2004), oil wells (Roseomonas strain BON 1; GenBank accession no. AY219712), biological soil crust (Roseomonas strain CP4B2; AJ871456), agriculture drainage water (Roseomonas strain SA-2; AY730026), as well as from water (Rihs et al., 1993). Roseomonas-related 16S rRNA gene sequences have also been detected frequently during molecular surveys of microbial diversity in various environments, for example, in the freshwater bacterial communities associated with cyanobacterial blooms in four Swedish lakes (Eiler & Bertilsson, 2004). All these facts indicate that members of the genus Roseomonas could occur widely in nature. However, no names of environmental isolates (non-medical source) of Roseomonas have been validly published. Currently, all members of the genus Roseomonas with validly published names, including Roseomonas cervicalis, Roseomonas fauriae, Roseomonas gilardii subsp. gilardii (Rihs et al., 1993), Roseomonas gilardii subsp. rosea and Roseomonas mucosa (Han et al., 2003), were isolated from medical samples. Strain TH-G33T was isolated from sediment of the freshwater Taihu Lake (also called lake Tai; 120° 02' 16·8'' E 31° 27' 10·7'' N, Jiangsu Province, China), by plating 10-fold dilutions of samples on dilute nutrient medium (DNM) containing (l–1): 0·3 g beef extract, 1·0 g fish peptone, 0·1 g yeast extract, 0·5 g NaCl and 15 g agar. Sediment samples were obtained from 10 cm beneath the sediment surface. LB or nutrient medium (NM) agar or broth at 30 °C were used for routine cultivation.
Gram reactions were determined by staining cells grown on NM agar at 30 °C for 24 h according to the method described by Gerhardt et al. (1994). Endospore formation was determined after malachite green staining of cells grown on LB agar. Cell flagellation and morphology were examined by using transmission and scanning electron microscopy. For assimilation of carbon sources, the standard mineral base solution of Stanier et al. (1966) was used. Each compound was added at a concentration of 0·2 % (w/v) after the mineral base solution had been autoclaved. Growth was examined after incubation at 30 °C for 1, 3, 7 and 14 days. Aerobic and anaerobic production of acid (O–F reaction) from carbohydrates was determined using O–F basal medium (Hugh & Leifson, 1953). Carbohydrate solutions sterilized by filtration were added at a final concentration of 1 % (w/v) and acid production was recorded after 7 and 14 days of incubation. For cellular fatty acid analysis, strains were grown for 72 h at 30 °C on NM agar. Cells were harvested from the plates and saponified, and the liberated fatty acids were methylated and analysed by using the Sherlock system (Microbial ID Inc.), following the manufacturer's instructions. Isoprenoid quinones were extracted from freeze-dried cells (200 mg) with methyl chloride/methanol (2 : 1) and analysed by reversed-phase HPLC. The sensitivity to various antibiotics was tested by using the disc diffusion method (Kirby–Bauer method).
The DNA base composition was determined by thermal denaturation (Marmur & Doty, 1962) using DNA from Escherichia coli DH-5
as a control. The 16S rRNA gene was amplified as described previously (Zhang et al., 2003) and 16S rRNA gene sequence alignments were performed with the CLUSTAL_X program (version 1.64b; Thompson et al., 1997). The phylogenetic tree was constructed by using the neighbour-joining method (Saitou & Nei, 1987) with Kimura's two-parameter calculation model in TREECON W version 1.3b.
Cells of strain TH-G33T were Gram-negative, aerobic, non-spore-forming coccoid rods or rods (Fig. 1). Flagella were not observed. Growth occurred at 15–40 °C and pH 6·0–9·0, with optima at 30 °C and pH 7·0. Cultures grown in NM broth were colourless. Colonies on NM agar were colourless to pale-pink. Colonies were pinpoint, raised, entire, runny and mucoid, with a diameter of 0·5–1·0 mm after incubation for 3–4 days. Catalase and oxidase reactions were positive. 16S rRNA gene sequence analysis showed that strain TH-G33T was phylogenetically related to members of the genus Roseomonas (similarities 93·2–94·8 %), with the greatest similarity to R. gilardii subsp. rosea (94·8 %) followed by R. mucosa (94·4 %). A neighbour-joining tree based on these 16S rRNA gene sequences was constructed (Fig. 2), which showed that strain TH-G33T and other species of the genus Roseomonas except R. fauriae, which has been suggested not to be a member of the genus Roseomonas (Cohen et al., 2004; Han et al., 2003), were grouped together. Thus, strain TH-G33T represents a novel species of the genus Roseomonas.
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. The major cellular fatty acids of strain TH-G33T were iso-C15 : 0 (19·9 %), iso-C16 : 0 (38 %) and iso-C17 : 1
9c (12·1 %). The cellular fatty acid profile of strain TH-G33T (Supplementary Table in IJSEM Online) was also significantly different from those reported for R. mucosa, R. gilardii, R. cervicalis and R. fauriae. Strain TH-G33T was susceptible to ampicillin, piperacillin, amoxicillin, piperacillin/tazobactam, cefazolin, cephalothin, cefuroxime, cefotaxime, cefaclor, cefoperazone, ceftazidime, amikacin, gentamicin, tobramycin, kanamycin, streptomycin, tetracycline, doxycycline, minocycline, ciprofloxacin, levofloxacin, norfloxacin, lomefloxacin, fleroxacin, erythromycin, nitrofurantoin, polymyxin and chloramphenicol, but not to trimethoprim/sulfamethoxazole or enoxacin.
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Description of Roseomonas lacus sp. nov.
Roseomonas lacus (la'cus. L. masc. gen. n. lacus of a lake, indicating the site of isolation of this organism).
Cells are coccoid rods or rods, 0·5–0·8x0·8–1·5 µm. Non-motile, without flagella. Gram-negative. Colonies on NM or DNM agar are circular, raised, entire, runny and mucoid, colourless to pale-pink and 0·5–1·0 mm in diameter after incubation for 3–4 days. Grows at 15–40 °C and pH 6·0–9·0, with optima at 30 °C and pH 7·0. Oxidase- and catalase-positive. Grows in NaCl at concentrations up to 6·5 %. Nitrate is reduced. Casein, gelatin, starch, Tween 80 and aesculin are not hydrolysed. Voges–Proskauer test and methyl red reaction are negative. Oxidizes ribose, L-rhamnose, D-galactose, L-arabinose, DL-sorbose, fucose, sucrose, D-cellobiose, D-melibiose and D-xylose. Does not utilize D-glucose, fructose, DL-sorbitol, DL-raffinose, DL-lactose, L-lysine, D-mannitol, acetate, propionate, glycerol or citrate. The major quinone is ubiquinone 10 (Q-10). The major cellular fatty acids (>1 %) are iso-C14 : 0 (5·8 %), iso-C15 : 0 (19·9 %), iso-C15 : 1 (1·6 %), anteiso-C15 : 0 (4·3 %), C15 : 0 (1·5 %), iso-C16 : 0 (38 %), iso-C16 : 1 (5·0 %), C16 : 0 (1·9 %), iso-C17 : 0 (4 %), anteiso-C17 : 0 (2·1 %) and iso-C17 : 19c (12·1 %). Sensitive to a range of antibiotics but not to trimethoprim/sulfamethoxazole or enoxacin.
The type strain, TH-G33T (=CGMCC 1.3617T=JCM 13283T), was isolated from freshwater lake sediment. The G+C content of the type strain is 71·9 mol%.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Cohen, M. F., Han, X. Y. & Mazzola, M. (2004). Molecular and physiological comparison of Azospirillum spp. isolated from Rhizoctonia solani mycelia, wheat rhizosphere, and human skin wounds. Can J Microbiol 50, 291–297.[CrossRef][Medline]
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