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1 Institut für Angewandte Mikrobiologie, Justus-Liebig-Universität Giessen, D-35392 Giessen, Germany
2 Culture Collection University Göteborg, Dept. of Clinical Bacteriology, S-41346 Göteborg, Sweden
3 Institut für Bakteriologie, Mykologie und Hygiene, Veterinärmedizinische Universität, A-1210 Wien, Austria
Correspondence
Peter Kämpfer
peter.kaempfer{at}agrar.uni-giessen.de
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The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain CCUG 2513T is AM748811.
A two-dimensional TLC of the polar lipids of P. mephitica CCUG 2513T is available as supplementary material with the online version of this paper.
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). The species description was based on morphological characteristics, i.e. a Gram-negative bacillus with polar flagellation, as well as physiological traits. The species name was included in the Approved Lists of Bacterial Names (Skerman et al., 1980).
Previously, it was recognized that P. mephitica is phylogenetically related to Janthinobacterium and related organisms (Anzai et al., 2000). The current definition of the genus Janthinobacterium is based on the descriptions of De Ley et al. (1978) and Sneath (1984) with the emendations of Lincoln et al. (1999) and Gillis & Logan (2005). Most Janthinobacterium lividum strains produce violacein, but non-pigmented colonies are often encountered (Gillis & Logan, 2005). P. mephitica did not produce pigmented colonies. Lincoln et al. (1999) pointed out that the fatty acid pattern serves to differentiate the genus Janthinobacterium from members of other genera showing a high degree (95 %) of 16S rRNA gene sequence similarity. Q-8 is the major respiratory lipoquinone, as in all members of the Betaproteobacteria studied to date (Yokota et al., 1992). The major phospholipids are phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The fatty acid composition comprises C10 : 0 3-OH, C12 : 0, C12 : 0 2-OH, C14 : 0, C16 : 0, C16 : 1
7c, C17 : 0 cyclo and C18 : 17c. The polyamine pattern, with the major compounds 2-hydroxyputrescine and putrescine, is in agreement with the characteristics of the Betaproteobacteria (Busse & Auling, 1988).
In this study, P. mephitica CCUG 2513T was studied for its exact taxonomic position along with J. lividum CCUG 2344T. The 16S rRNA gene sequences (both 1388 bp) were studied as described by Kämpfer et al. (2003) and shared 99.8 % similarity.
For polar lipid, quinone and polyamine analyses, cells were grown on PYE medium (0.3 % peptone from casein, 0.3 % yeast extract, pH 7.2). Analyses were carried out as described previously (Busse & Auling, 1988; Tindall, 1990a, b; Altenburger et al., 1996; Stolz et al., 2007). The major phospholipids were detected to be phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. In addition, moderate amounts of an unknown aminolipid and an unknown phospholipid and minor to trace amounts of another unknown aminolipid and five unknown polar lipids were found (Supplementary Fig. S1, available in IJSEM Online). J. lividum CCUG 2344T exhibited the same profile (results not shown), and both profiles were in perfect agreement with those reported for the type strains of J. lividum and Janthinobacterium agaricidamnosum (Lincoln et al., 1999), although these authors did not detect minor components. The quinone system of P. mephitica CCUG 2513T consisted of 2 % ubiquinone Q-7 and 98 % Q-8 and the polyamine pattern also exhibited the characteristics of members of the Betaproteobacteria [µmol (g dry weight)–1: 2-hydroxyputrescine, 19.4; putrescine, 71.7; cadaverine, 0.3; spermidine, 1.0; spermine, 0.6].
Fatty acid methyl esters were prepared, separated and identified according to the instructions of the Microbial Identification System (MIDI, Microbial ID; Kämpfer & Kroppenstedt, 1996). The fatty acid profiles of P. mephitica CCUG 2513T and J. lividum CCUG 2344T are shown in Table 1. No significant differences were found between the fatty acid profiles of the two strains.
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) and a polyamine pattern with the predominant compounds putrescine and spermidine (Busse & Auling, 1988), and are in excellent agreement with the characteristics of J. lividum.
Physiological/biochemical tests were performed with methods described previously (Kämpfer et al., 1991). Strains CCUG 2344T and CCUG 2513T shared the following biochemical characteristics. L-Alanine p-nitroanilide (pNA), bis-p-nitrophenyl (pNP) phosphate, bis-pNP phenylphosphonate, pNP β-D-glucopyranoside and L-proline pNA are hydrolysed on the basis of the method described by Kämpfer et al. (1991). The following compounds are not hydrolysed: pNP β-D-galactopyranoside, pNP β-D-glucuronide, pNP 
-D-glucopyranoside, pNP β-D-xylopyranoside, bis-pNP phosphorylcholine and 
-L-glutamate pNA. The following compounds are used as sole sources of carbon: N-acetylgalactosamine, L-arabinose, L-arbutin, D-fructose, D-galactose, D-glucose, maltose, D-ribose, sucrose, D-xylose (weakly), D-mannitol (weakly), D-sorbitol (weakly), citrate, fumarate, glutarate, DL-3-hydroxybutyrate, DL-lactate, L-malate, 2-oxoglutarate, pyruvate (weakly), L-alanine (weakly), L-proline (weakly) and L-serine (weakly). The following compounds are not utilized on the basis of the method described by Kämpfer et al. (1991): D-gluconate, acetate, propionate, cis- and trans-aconitate, 4-aminobutyrate, itaconate, mesaconate, β-alanine, L-aspartate, L-leucine, L-ornithine, L-serine, N-acetylglucosamine, D-cellobiose, D-mannose, -D-melibiose, L-rhamnose, salicin, trehalose, adonitol, inositol, maltitol, putrescine, adipate, azelate, suberate, L-histidine, L-phenylalanine and L-tryptophan. Acids are produced from D-glucose, sucrose (weakly) and L-arabinose. No acids are produced from lactose, D-mannitol, dulcitol, salicin, adonitol, inositol, sorbitol, raffinose, rhamnose, maltose, D-xylose, trehalose, cellobiose, methyl D-glucoside, erythritol, melibiose, D-arabitol or D-mannose.
On the basis of these results, it is clear that P. mephitica CCUG 2513T is not a member of the genus Pseudomonas sensu stricto and, hence, we propose that the name Pseudomonas mephitica Claydon and Hammer 1939 is a later heterotypic synonym of Janthinobacterium lividum (Eisenberg 1891) De Ley et al. 1978 and that P. mephitica CCUG 2513T should be assigned to J. lividum based on: (i) 99.8 % 16S rRNA gene sequence similarity between J. lividum CCUG 2344T (=DSM 1522T) and P. mephitica CCUG 2513T (our confirmation: GenBank accession numbers Y08846 and AM748811, respectively), (ii) identical quinone systems, (iii) identical polar lipid profiles, (iv) highly similar polyamine patterns, (v) similar fatty acid profiles (Table 1
) and (vi) identical biochemical test results.
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REFERENCES |
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Anzai, Y., Kim, H., Park, J. Y., Wakabayashi, H. & Oyaizu, H. (2000). Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence. Int J Syst Evol Microbiol 50, 1563–1589.[Abstract]
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Claydon, T. J. & Hammer, B. W. (1939). A skunk-like odor of bacterial origin in butter. J Bacteriol 37, 251–258.
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Kämpfer, P. & Kroppenstedt, R. M. (1996). Numerical analysis of fatty acid patterns of coryneform bacteria and related taxa. Can J Microbiol 42, 989–1005.
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Lincoln, S. P., Fermor, T. R. & Tindall, B. J. (1999). Janthinobacterium agaricidamnosum sp. nov., a soft rot pathogen of Agaricus bisporus. Int J Syst Bacteriol 49, 1577–1589.
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Stolz, A., Busse, H.-J. & Kämpfer, P. (2007). Pseudomonas knackmussii sp. nov. Int J Syst Evol Microbiol 57, 572–576.
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This article has been cited by other articles:
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  P. Kampfer, H.-J. Busse, and H. C. Scholz Chromobacterium piscinae sp. nov. and Chromobacterium pseudoviolaceum sp. nov., from environmental samples Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2486 - 2490. [Abstract] [Full Text] [PDF]
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