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Int J Syst Evol Microbiol 58 (2008), 1268-1270; DOI  10.1099/ijs.0.2008/001388-0
© 2008 International Union of Microbiological Societies

Request for an Opinion

Bacillus aeolius DSM 15084T (=CIP 107628T) is a strain of Bacillus licheniformis.

R. Pukall1, P. Schumann1, D. Clermont2 and C. Bizet2

1 DSMZ – Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstrasse 7b, 38124 Braunschweig, Germany
2 Collection de l'Institut Pasteur, 25–28 rue du Docteur Roux, 75724 Paris Cedex 15, France

Correspondence
C. Bizet
bizet{at}pasteur.fr

ABSTRACT

Based on phenotypic properties, 16S rRNA gene sequence and MALDI-TOF analysis, strains DSM 15084T and CIP 107628T, deposited as the type strain of Bacillus aeolius, do not represent the original type strain, strain 4-1T. It is therefore proposed that the Judicial Commission of the International Committee on Systematics of Prokaryotes places the name Bacillus aeolius on the list of rejected specific and subspecific epithets in names of species and subspecies of bacteria if a suitable replacement for the type strain or a neotype cannot be found within 2 years of publication of this Request.

A 16S rRNA gene sequence alignment, a dendrogram based on Riboprinting results and results of MALDI-TOF analysis are available as supplementary material with the online version of this paper.

In 2003, strain 4-1T (=DSM 15084T =CIP 107628T) was proposed to represent a novel species within the genus Bacillus, Bacillus aeolius, on the basis of differences between its chemotaxonomic and phenotypic properties and those of related moderately thermophilic species, e.g. Bacillus smithii, B. fumarioli, B. oleronis, B. sporothermodurans and B. infernus (Gugliandolo et al., 2003aGo, bGo). The G+C content of the DNA of strain 4-1T was 40.8 mol%, falling within range of values determined for species of the genus Bacillus rRNA group 1 as defined by Ash et al. (1991)Go. Phylogenetic analysis based on analysis of the nearly complete 16S rRNA gene sequence placed strain 4-1T close to the type strain of B. smithii (96.7 %), and binary similarity values determined with other type strains ranged between 95 and 96 % (Bacillus methanolicus, 96.7 %; B. fumarioli, 95.9 %; B. infernus, 95.3 %).

Strain 4-1T was originally isolated by Maugeri et al. (2001)Go from a water sample collected at a depth of 15 m during an oceanographic cruise around the Eolian islands (Italy) at the site Vulcano – Punta Congigliara. These authors published a polyphasic taxonomic study of 18 thermophilic bacilli including strain 4-1T, whose partial 16S rRNA gene sequence was found to be distantly related to that of the type strain of B. methanolicus.

Phenotypic properties of the strain published in 2001 compared with the data published within the species description in 2003 are summarized in Table 1Go. Further results obtained from API 50CHB analysis of the strain deposited as DSM 15084T and Bacillus licheniformis DSM 13T are given for comparison. Although the phenotypic characteristics of strains 4-1T (as reported in 2003) and DSM 15084T were not identical with respect to some of the substrates tested, the profiles differ significantly from that published in 2001 for strain 4-1T. In addition, the two species B. aeolius and B. licheniformis are similar in many of the properties detected: aerobic growth, positive result in an oxidase test and acid production from glycerol, D-xylose, ribose, mannose, glucose, fructose, inositol, mannitol, salicin, cellobiose, maltose, sucrose and turanose.


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  		Table 1. Acid production from different substrates as determined by the use of API 50CH

Data for strain DSM 15084T and B. licheniformis DSM 13T were obtained in this study at 55 and 37 °C, respectively, after 5 and 24 h. +, Change of indicator dye to yellow; (+), positive at 5 h time point only; W, weak reaction. Glycerol gave a positive result in all assays. The following substrates gave a negative result in all assays: erythritol, D-arabinose, L-xylose, adonitol, methyl β-D-xylopyranoside, L-sorbose, dulcitol, inulin, xylitol, gentiobiose, D-lyxose, D- and L-fucose, D- and L-arabitol and gluconate.

 
In order to verify the identity of the strain, the DSMZ and CIP independently determined the 16S rRNA gene sequence of the strains deposited as the type strain of B. aeolius and, in both cases, sequence analyses revealed B. licheniformis as the nearest phylogenetic neighbour. The sequence deposited for B. aeolius 4-1T (GenBank accession no. AJ504797) does not match the sequences obtained at the DSMZ and CIP, suggesting that the wrong strain has been deposited in the culture collections. Within a stretch of more than 600 bp, the sequence submitted for strain B. aeolius 4-1T differs at 18 positions when compared with the sequence obtained at the DSMZ for strain DSM 15084T (Supplementary Fig. S1, available in IJSEM Online). Recently, seven rRNA genes were found to be present in the whole genome of B. licheniformis DSM 13T. One 16S rRNA gene sequence consists of 1549 bp, and all seven sequences differed at only six positions. It is therefore unlikely that the differences detected for B. aeolius 4-1T could be caused by rrn operon heterogeneities.

Strain DSM 15084T was analysed using the Qualicon Riboprinter system (DuPont Qualicon) for further characterization. Cluster analysis of the riboprint pattern obtained for strain DSM 15084T allocated the strain to the B. licheniformis cluster, even though the fingerprints obtained for the different strains of the species B. licheniformis vary in the intensity and number of bands (Supplementary Fig. S2).

MALDI-TOF analysis (Schumann, 2007Go) was applied in order to confirm that the strains deposited at the DSMZ (DSM 15084T) and the CIP (CIP 107628T) are identical. Spectra recorded of both strains in five replicated measurements are shown in Supplementary Fig. S3. The analysis confirmed that the two strains showed identical patterns and they can be considered identical.

The two collections have attempted to obtain a subculture of the original strain from the depositor, but the strain is no longer available in the depositor's laboratory. According to Rule 18c of the International Code of Nomenclature of Bacteria (Lapage et al., 1992Go), it is proposed that the Judicial Commission of the International Committee on Systematics of Prokaryotes places the name Bacillus aeolius Gugliandolo et al. 2003 on the list of rejected specific and subspecific epithets in names of species and subspecies of bacteria if a suitable replacement for the type strain or a neotype cannot be found within 2 years of publication of this Request.

ACKNOWLEDGEMENTS

We wish to thank Dr J. P. Euzéby for his help with the taxonomic commentaries.

REFERENCES

Ash, C., Farrow, J. A. E., Wallbanks, S. & Collins, M. D. (1991). Phylogenetic heterogeneity of the genus Bacillus revealed by comparative analysis of small-subunit-ribosomal RNA. Lett Appl Microbiol 13, 202–206.

Gugliandolo, C., Maugeri, T. L., Caccamo, D. & Stackebrandt, E. (2003a). Bacillus aeolius sp. nov. A novel thermophilic, halophilic marine Bacillus species from Eolian Islands (Italy). Syst Appl Microbiol 26, 172–176.[CrossRef][Medline]

Gugliandolo, C., Maugeri, T. L., Caccamo, D. & Stackebrandt, E. (2003b). Bacillus aeolius sp. nov. In Validation of Publication of New Names and New Combinations Previously Effectively Published Outside the IJSEM, List no. 94. Int J Syst Evol Microbiol 53, 1701–1702.[Abstract/Free Full Text]

Lapage, S. P., Sneath, P. H. A., Lessel, E. F., Skerman, V. B. D., Seeliger, H. P. R. & Clark, W. A. (editors) (1992). International Code of Nomenclature of Bacteria (1990 revision). Bacteriological Code. Washington, DC: American Society for Microbiology.

Maugeri, T. L., Gugliandolo, C., Caccamo, D. & Stackebrandt, E. (2001). A polyphasic study of thermophilic Bacilli from shallow, marine vents. Syst Appl Microbiol 24, 572–587.[CrossRef][Medline]

Schumann, P. (2007). MALDI-TOF mass spectrometry versus Riboprinting in the quality control of bacterial strains. In Connections between Collections. Proceedings of the 11th International Conference on Culture Collections, Goslar, Germany, 7–11 October 2007, pp. 82–86. Edited by E. Stackebrandt, M. Wozniczka, V. Weihs & J. Sikorsky. Braunschweig: DSMZ and the World Federation of Culture Collections.





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