MALDI-TOFMS compared with other polyphasic taxonomy approaches for the identification and classification of Bacillus pumilus spores

Dickinson, Danielle; Duc, Myron T. La; Satomi, Masataka; Winefordner, J. D.; Powell, David H.; Venkateswaran, Kasthuri

doi:10.1016/j.mimet.2004.02.011

Cited by 47 publications

(37 citation statements)

References 26 publications

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“…S1 available in IJSEM Online), all of the novel isolates are grouped together in their own cluster, separate from the distinct cluster formed by previously described B. pumilus strains, supporting the gyrB analysis and previous MALDI-TOF MS results (Dickinson et al, 2004). The most notable difference between the protein profiles of the B. pumilus type strain (ATCC 7061 T ) and the proposed FO36b T strain group is the presence of a peak at 7620 Da, found only in the FO-36b T group strains (Dickinson et al, 2004). Another difference between the two groups is that the FO36b T group forms a tight cluster in terms of MALDI-TOF MS protein profiling.…”

supporting

confidence: 84%

“…It has been used in recent taxonomic studies to evaluate DNA relatedness among bacterial strains (Thompson et al, 2003). The rep-PCR results generated in our work clearly distinguish the novel isolates from B. pumilus and, when coupled with supporting MALDI-TOF MS profiles (Dickinson et al, 2004), strongly suggest that these isolates should be recognized as belonging to a distinct genotype. To confirm the conclusions generated from the results of the phylogenetic analyses, DNA-DNA hybridization was performed (Supplementary Table S1 available in IJSEM Online).…”

supporting

confidence: 69%

“…The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA and gyrB gene sequences referred to in this communication are given in to assess similarities amongst isolates, two very distinct and consistent groups of B. pumilus were revealed (Dickinson et al, 2004). A detailed taxonomic characterization of this group of bacteria was deemed necessary to clarify the distribution of these species in spacecraft-assembly facilities.…”

mentioning

confidence: 99%

“…The novel isolates were Gram-positive, spore-forming rods and were aerobic, motile and oxidase-and catalase-positive and so demonstrated several morphological and physiological characteristics typical of members of the genus Bacillus (Claus & Berkeley, 1986); this indicated that all of these isolates belonged to this genus or related genera. The Biolog identification system indicated that the novel isolates were most similar to B. pumilus (Dickinson et al, 2004). Additionally, 174 phenotypic tests were performed to study the phenotype of the novel isolates.…”

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confidence: 99%

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Bacillus safensis sp. nov., isolated from spacecraft and assembly-facility surfaces

Satomi

Duc

Venkateswaran

2006

International Journal of Systematic and Evolutionary Microbiology

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187

119

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Thirteen strains of a novel spore-forming, Gram-positive, mesophilic heterotrophic bacterium were isolated from spacecraft surfaces (Mars Odyssey Orbiter) and assembly-facility surfaces at the Jet Propulsion Laboratory in California and the Kennedy Space Center in Florida. Phylogenetic analysis of 16S rRNA gene sequences has placed these novel isolates within the genus Bacillus, the greatest sequence similarity (99?9 %) being found with Bacillus pumilus. However, these isolates share a mere 91?2 % gyrB sequence similarity with Bacillus pumilus, rendering their 16S rRNA gene-derived relatedness suspect. Furthermore, DNA-DNA hybridization showed only 54-66 % DNA relatedness between the novel isolates and strains of B. pumilus. rep-PCR fingerprinting and previously reported matrix-assisted laser desorption/ionization time-of-flight mass spectrometry protein profiling clearly distinguished these isolates from B. pumilus. Phenotypic analyses also showed some differentiation between the two genotypic groups, although the fatty acid compositions were almost identical. The polyphasic taxonomic studies revealed distinct clustering of the tested strains into two distinct species. On the basis of phenotypic characteristics and the results of phylogenetic analyses of 16S rRNA and gyrB gene sequences, repetitive element primer-PCR fingerprinting and DNA-DNA hybridization, the 13 isolates represent a novel species of the genus Bacillus, for which the name Bacillus safensis sp. nov. is proposed. The type strain is FO-36b T (=ATCC BAA-1126 T =NBRC 100820 T ).Spacecraft and associated clean-room assembly-facility surfaces harbour an extremely low biomass (La Duc et al., 2003;Venkateswaran et al., 2001), because of stringent maintenance. However, colonization by micro-organisms specifically adapted to such facility conditions, especially those yet to be cultured and/or characterized, is of major concern to those commissioning modern-day space-related experimentation. The search for extraterrestrial life will rely heavily on validated cleaning and bioreduction strategies to ensure that terrestrial microbial contamination does not compromise the scientific integrity of such missions. It is crucial both to minimize and eradicate such microbial contaminants and to identify and characterize the recurring, prevalent micro-organisms associated with the surfaces of spacecraft and associated environments.Studies have repeatedly shown that extremely resilient, spore-forming members of the genus Bacillus are the most strongly represented micro-organisms in samples collected from spacecraft and facility surfaces (La Duc et al., 2003; Puleo et al., 1977). The extremely oligotrophic, lowhumidity, temperature-controlled conditions of spacecraftassembly facilities appear to select for micro-organisms able to withstand such unfavourable surroundings. During monitoring of the microbial diversity of spacecraftassociated environments over a period of 5 years (1999)(2000)(2001)(2002)(2003)(2004), Bacillus pumilus was found to be the second most domi...

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supporting

confidence: 84%

supporting

confidence: 69%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Bacillus safensis sp. nov., isolated from spacecraft and assembly-facility surfaces

Satomi

Duc

Venkateswaran

2006

International Journal of Systematic and Evolutionary Microbiology

Self Cite

187

119

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“…(41). The B. anthracis, B. cereus sensu stricto, and B. thuringiensis strains were also selected to completely cover the previously described SASP diversity found within each of these species (8)(9)(10)(11)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39). A dry powder formulation of B. atrophaeus spores was obtained from Dugway Proving Grounds (Dugway, UT), and a dry powder pesticide formulation (Turex WP 50) containing B. thuringiensis spores was obtained from Certis Europe (Utrecht, The Netherlands).…”

mentioning

confidence: 99%

Rapid Identification of Bacillus anthracis Spores in Suspicious Powder Samples by Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

Dybwad

Laaken²,

Blatny

et al. 2013

Appl Environ Microbiol

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cRapid and reliable identification of Bacillus anthracis spores in suspicious powders is important to mitigate the safety risks and economic burdens associated with such incidents. The aim of this study was to develop and validate a rapid and reliable laboratory-based matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis method for identifying B. anthracis spores in suspicious powder samples. A reference library containing 22 different Bacillus sp. strains or hoax materials was constructed and coupled with a novel classification algorithm and standardized processing protocol for various powder samples. The method's limit of B. anthracis detection was determined to be 2.5 ؋ 10 6 spores, equivalent to a 55-g sample size of the crudest B. anthracis-containing powder discovered during the 2001 Amerithrax incidents. The end-to-end analysis method was able to successfully discriminate among samples containing B. anthracis spores, closely related Bacillus sp. spores, and commonly encountered hoax materials. No false-positive or -negative classifications of B. anthracis spores were observed, even when the analysis method was challenged with a wide range of other bacterial agents. The robustness of the method was demonstrated by analyzing samples (i) at an external facility using a different MALDI-TOF MS instrument, (ii) using an untrained operator, and (iii) using mixtures of Bacillus sp. spores and hoax materials. Taken together, the observed performance of the analysis method developed demonstrates its potential applicability as a rapid, specific, sensitive, robust, and cost-effective laboratory-based analysis tool for resolving incidents involving suspicious powders in less than 30 min.

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MALDI‐TOF Mass Spectrometry of Intact Bacteria

Lay¹,

Liyanage²

2005

Identification of Microorganisms by Mass Spectrometry

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MALDI-TOFMS compared with other polyphasic taxonomy approaches for the identification and classification of Bacillus pumilus spores

Cited by 47 publications

References 26 publications

Bacillus safensis sp. nov., isolated from spacecraft and assembly-facility surfaces

Bacillus safensis sp. nov., isolated from spacecraft and assembly-facility surfaces

Rapid Identification of Bacillus anthracis Spores in Suspicious Powder Samples by Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

MALDI‐TOF Mass Spectrometry of Intact Bacteria

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