Two Gram-positive, rod-shaped, alkali-tolerant (pH 10.5), endospore-forming bacteria (strains KSC SF8bT and KSC SF10a) were isolated from surfaces within the Payload Hazardous Servicing Facility, where robotic spacecraft are assembled and tested before launch, at the Kennedy Space Center at Cape Canaveral. Based on 16S rRNA gene sequence similarities, these strains were shown to belong to the family Bacillaceae and the genus Bacillus. The highest 16S rRNA gene sequence similarity was ∼97.5 %, observed between the novel strains and Bacillusselenatarsenatis SF-1T. Several phenotypic characteristics, such as growth with 10 % NaCl and assimilation of melibiose and lactose, were useful in the discrimination of this novel species from the closely related alkali-tolerant species Bacillus firmus and B. selenatarsenatis. DNA–DNA hybridization studies revealed reassociation values of less than 45 % between strain KSC SF8bT and its closest genotypic neighbours. The combination of unique phenotypic and genotypic characteristics allowed the differentiation of these alkali- and halotolerant spore-forming strains from related Bacillus species, and a novel species, Bacillus canaveralius sp. nov., is proposed. The type strain is KSC SF8bT (=ATCC BAA-1493T=MTCC 8908T).
Claus, D. & Berkeley, R. C. W.(1986). Genus Bacillus Cohn, 1872. In Bergey's Manual of Systematic Bacteriology, vol. 2, pp. 1105–1139. Edited by P. H. A. Sneath, N. S. Mair, M. E. Sharpe & J. G. Holt. Baltimore: Williams & Wilkins.
Heyrman, J., Vanparys, B., Logan, N. A., Balcaen, A., Rodriguez-Diaz, M., Felske, A. & De Vos, P.(2004).Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands. Int J Syst Evol Microbiol54, 47–57.[CrossRef][Google Scholar]
Horneck, G.(1993). Responses of Bacillus subtilis spores to space environment: results from experiments in space. Orig Life Evol Biosph23, 37–52.[CrossRef][Google Scholar]
Kempf, M. J., Chen, F., Kern, R. & Venkateswaran, K.(2005). Recurrent isolation of hydrogen peroxide-resistant spores of Bacillus pumilus from a spacecraft assembly facility. Astrobiology5, 391–405.[CrossRef][Google Scholar]
Klieve, A. V., Plumb, J. J. & Blackall, L. L.(1999). Isolation and characterization of an unusual bacterium, allied to the soil bacterium Bacillus benzoevorans, from feedlot manure pads in Australia. Lett Appl Microbiol29, 71–75.[CrossRef][Google Scholar]
La Duc, M. T., Dekas, A. E., Osman, S., Moissl, C., Newcombe, D. & Venkateswaran, K.(2007). Isolation and characterization of bacteria capable of tolerating the extreme conditions of clean-room environments. Appl Environ Microbiol73, 2600–2611.[CrossRef][Google Scholar]
Lane, D. J.(1991). 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics, pp. 115–175. Edited by E. Stackebrandt & M. Goodfellow. Chichester: Wiley.
Mandel, M. & Marmur, J.(1968). Use of ultraviolet absorbance-temperature profile for determining the guanine plus cytosine content of DNA. Methods Enzymol12B, 195–206.
[Google Scholar]
NASA(1980). Standard procedures for the microbiological examination of space hardware, NHB 5340.1B, Rev. B. In National Aeronautics and Space Administration. Washington, DC: NASA.
Newcombe, D. A., Schuerger, A. C., Benardini, J. N., Dickinson, D., Tanner, R. & Venkateswaran, K.(2005). Survival of spacecraft-associated microorganisms under simulated martian UV irradiation. Appl Environ Microbiol71, 8147–8156.[CrossRef][Google Scholar]
Newcombe, D. A., La Duc, M. T., Vaishampayan, P. & Venkateswaran, K.(2008). Impact of assembly, testing and launch operations on the airborne bacterial diversity within a spacecraft assembly facility clean-room. Int J Astrobiol7, 223–236.[CrossRef][Google Scholar]
Pichinoty, F., Asselineau, J. & Mandel, M.(1984). Biochemical characterization of Bacillus benzoevorans sp. nov., a new filamentous, sheathed mesophilic species, degrading various aromatic acids and phenols. Ann Microbiol (Paris)135B, 209–217.
[Google Scholar]
Priest, F. G. & Alexander, B.(1988). A frequency matrix for probabilistic identification of some bacilli. J Gen Microbiol134, 3011–3018.
[Google Scholar]
Ruimy, R., Breittmayer, V., Elbaze, P., Lafay, B., Boussemart, O., Gauthier, M. & Christen, R.(1994). Phylogenetic analysis and assessment of the genera Vibrio, Photobacterium, Aeromonas, and Plesiomonas deduced from small-subunit rRNA sequences. Int J Syst Bacteriol44, 416–426.[CrossRef][Google Scholar]
Rummel, J. D.(2001). Planetary exploration in the time of astrobiology: protecting against biological contamination. Proc Natl Acad Sci U S A98, 2128–2131.[CrossRef][Google Scholar]
Tamura, K., Dudley, J., Nei, M. & Kumar, S.(2007). MEGA4: Molecular Evolutionary Genetics Analysis (mega) software version 4.0. Mol Biol Evol24, 1596–1599.[CrossRef][Google Scholar]
Tiago, I., Pires, C., Mendes, V., Morais, P. V., da Costa, M. S. & Verissimo, A.(2006).Bacillus foraminis sp. nov., isolated from a non-saline alkaline groundwater. Int J Syst Evol Microbiol56, 2571–2574.[CrossRef][Google Scholar]
Tourova, T. P. & Antonov, A. S.(1987). Identification of microorganisms by rapid DNA-DNA hybridization. Methods in Microbiology19, 333–355.
[Google Scholar]
UN Treaty(1967).Treaty on Principles Governing the Activities of States in the Exploration and Use of Outer Space, Including the Moon and Other Celestial Bodies. Article IX, UN Doc. A/RES/222/(XXI), TIAS No. 6347.
Venkateswaran, K., Kempf, M., Chen, F., Satomi, M., Nicholson, W. & Kern, R.(2003).Bacillus nealsonii sp. nov., isolated from a spacecraft-assembly facility, whose spores are gamma-radiation resistant. Int J Syst Evol Microbiol53, 165–172.[CrossRef][Google Scholar]
Wayne, L. G.(1988). International Committee on Systematic Bacteriology: announcement of the report of the ad hoc Committee on Reconciliation of Approaches to Bacterial Systematics. Zentralbl Bakteriol Mikrobiol Hyg [A]268, 433–434.
[Google Scholar]
Yamamura, S., Yamashita, M., Fujimoto, N., Kuroda, M., Kashiwa, M., Sei, K., Fujita, M. & Ike, M.(2007).Bacillus selenatarsenatis sp. nov., a selenate- and arsenate-reducing bacterium isolated from the effluent drain of a glass-manufacturing plant. Int J Syst Evol Microbiol57, 1060–1064.[CrossRef][Google Scholar]
Zhang, Z., Schwartz, S., Wagner, L. & Miller, W.(2000). A greedy algorithm for aligning DNA sequences. J Comput Biol7, 203–214.[CrossRef][Google Scholar]