A total of 301 strains of fluorescent pseudomonads previously characterized by conventional phenotypic and/or genomic taxonomic methods were analyzed through siderotyping, i.e., by the isoelectrophoretic characterization of their main siderophores and pyoverdines and determination of the pyoverdine-mediated iron uptake specificity of the strains. As a general rule, strains within a well-circumscribed taxonomic group, namely the species Pseudomonas brassicacearum, Pseudomonas fuscovaginae, Pseudomonas jessenii, Pseudomonas mandelii, Pseudomonas monteilii, "Pseudomonas mosselii," "Pseudomonas palleronii," Pseudomonas rhodesiae, "Pseudomonas salomonii," Pseudomonas syringae, Pseudomonas thivervalensis, Pseudomonas tolaasii, and Pseudomonas veronii and the genomospecies FP1, FP2, and FP3 produced an identical pyoverdine which, in addition, was characteristic of the group, since it was structurally different from the pyoverdines produced by the other groups. In contrast, 28 strains belonging to the notoriously heterogeneous Pseudomonas fluorescens species were characterized by great heterogeneity at the pyoverdine level. The study of 23 partially characterized phenotypic clusters demonstrated that siderotyping is very useful in suggesting correlations between clusters and welldefined species and in detecting misclassified individual strains, as verified by DNA-DNA hybridization. The usefulness of siderotyping as a determinative tool was extended to the nonfluorescent species Pseudomonas corrugata, Pseudomonas frederiksbergensis, Pseudomonas graminis, and Pseudomonas plecoglossicida, which were seen to have an identical species-specific siderophore system and thus were easily differentiated from one another. Thus, the fast, accurate, and easy-to-perform siderotyping method compares favorably with the usual phenotypic and genomic methods presently necessary for accurate identification of pseudomonads at the species level.

Twenty-f ive non-ident if ied fluorescent Pseudomonas strains isolated from natural mineral waters were previously clustered into three phenotypic subclusters, Xlllb, XVa and XVc. These strains were characterized genotypically in the present study. DNA-DNA hybridization results and DNA base composition analysis revealed that these strains were members of two new species, for which the names Pseudomonas gessardii sp. nov. (type strain CIP 1054693 and Pseudomonas migulae sp. nov. (type strain CIP 1054703 are proposed. P. gessardii included 13 strains from phenotypic subclusters XVa and XVc. P. migulae included 10 strains from phenotypic subcluster Xlllb. The levels of DNA-DNA relatedness ranged from 71 to 100% for P. gessardii and from 74 to 100% for P. migulae. The G+C content of the DNA of each type strain was 58 mol%. DNA similarity levels, measured with 67 reference strains of Pseudomonas species, were below 55%, with ATm values of 13 "C or more. The two new species presented basic morphological characteristics common to all pseudomonads. Various phenotypic features were found to differentiate them: P. gessardii strains utilized L-arabitol, myo-inositol, adonitol, xylitol and meso-erythritol as carbon sources, whereas P. migulae strains assimilated Larabinose, D-xylose, D-saccharate, meso-tartrate, tricarballylate, D-glucuronate, D-galacturonate, phenylacetate and histamine. The complete 16s rRNA sequences of each type strain were determined and compared with those of the type strains of Pseudomonas species. Finally, a phylogenetic tree was inferred from sequence analysis and demonstrated that the two new species fell into the 'Pseudomonas fluorescens intrageneric cluster'. To date, their clinical significance is unknown.

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The taxonomic position of eight fluorescent Pseudomonas isolates, from two Lebanese spring waters, which were previously recognized by numerical analysis as members of a new subcluster (subcluster Vb) was examined. Except for one strain, the new subcluster exhibited internal DNA hybridization values of 76-100%, and 9-53% hybridization was measured with the type or reference strains of other Pseudomonas species. The highest DNA binding value was found with Pseudomonas marginalis strains (37-53%). The G+C content of the DNA of the type strain was 58 mol%. A comparison of 1322 nt of the 16S rRNA gene sequence of the strain representing subcluster Vb (CFML 96-195T) with the sequence of other strains of the genus Pseudomonas revealed that strain CFML 96-195T was part of the 'Pseudomonas fluorescens intrageneric cluster'. On the basis of the results of phenotypic, DNA-DNA and phylogenetic analyses, a new Pseudomonas species, Pseudomonas libanensis sp. nov., is proposed for the seven strains of subcluster Vb. The type strain is P. libanensis CFML 96-195T and has been deposited in the Collection de l'Institut Pasteur (Paris, France) as CIP 105460T. The P. libanensis strains are phenotypically and genotypically homogeneous and can be differentiated from most other fluorescent species by several phenotypic features. Differentiation of P. libanensis and Pseudomonas aeruginosa is based mainly on pyocyanin production; P. libanensis can be differentiated from P. fluorescens (all biovars) by alpha-aminobutyrate assimilation. The clinical significance of P. libanensis is unknown.

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