Field detection of bacillus spore aerosols with stand-alone pyrolysis-gas chromatography-ion mobility spectrometry

Snyder, A. Peter; Maswadeh, Waleed M.; Parsons, J. A.; Tripathi, Ashish; Meuzelaar, Henk L. C.; Dworzański, Jacek P.; Kim, Man-Goo

doi:10.1002/(sici)1520-6521(1999)3:4/5<315::aid-fact10>3.0.co;2-2

Cited by 38 publications

(12 citation statements)

References 15 publications

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“…Atmospheric pressure IMS is well suited in outdoors, field applications. ,,− Field utility of IMS is attractive, because it has a high degree of portability and low logistics and consumable requirements.…”

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“…Outdoor bioaerosols consisting of Gram-positive Bacillus atrophaeus (formerly Bacillus subtilis var. globigii ) spores, Gram-negative Pantoea agglomerans (formerly Erwinia herbicola ) cells, ovalbumin protein (OV), and MS-2 coliphage virus were released during formal trials at Western desert and prairie test sites in the United States and Canada. − Differentiation of the four bioaerosols was possible by visual 18-20 and multivariate factor analysis determinations (unpublished data) of the dispersion of pyrolysate peaks in the GC−IMS data space. These series of tests documented the first successful detection and differentiation of outdoor-released bioaerosols with an ambient temperature and pressure IMS detector interfaced to a biological sample Py-GC processing system.…”

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Correlation of Mass Spectrometry Identified Bacterial Biomarkers from a Fielded Pyrolysis-Gas Chromatography-Ion Mobility Spectrometry Biodetector with the Microbiological Gram Stain Classification Scheme

et al. 2004

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Public reporting burden for fthi collection of Informaiaon is estimated to, average I hour per response, including the time for reviewing Instructions. searchilng erristing data sources, gatheing and maintsteng the data needed, and completing and review"n this collection of Information. Send cofmment regarding this burden estimate or any othier aspect of this collection of Infomuation, Indluding suggestions for reducing fti burden to Departmrent PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORTDIR ECBC, ATTN: AMSRD-ECBC-RT-II, APG, MD 21010-5424 NUMBER GEO-CENTERS, Inc., Gunpowder Branch, APG, MD 21010-5424 ECBC-TR-415 SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSORIMONITOR'S ACRONYM(S) SPONSORIMONITOR'S REPORT NUMBER(S) DISTRIBUTION I AVAILABILITY STATEMENTApproved for public release; distribution is unlimited. SUPPLEMENTARY NOTES ABSTRACTThis work discusses the significance of the pyrolyzate peaks observed in the gas chromatography-ion mobility spectrometry (GC-IMS) dataspace of the pyrolysis-GC-IMS (Py-GC-IMS) briefcase system. This system has the ability to detect and classifyi deliberately released bioaerosols in outdoor field scenarios. The bioaerosols include Gram-positive spores and Gram-negative bacteria, the MS-2 Escherichia coliphage virus, and ovalbumnin (OV) protein species. The work suggests certain improvements that can be made to the IMS detection System. A pyrolysis-gas chromatography-ion mobility spectrometry (Py-GC-IMS) briefcase system can detect and classify deliberately released bioaerosols in outdoor field scenarios. The bioaerosols include Gram-positive spores and Gram-negative bacteria, the MS-2 Escherichia coliphage virus, and ovalbumin (OV) protein species. However, the origin and structural identities of the pyrolyzate peaks observed in the GC-IMS dataspace, their microbiological information content, and taxonomic importance with respect to biodetection have not been determined. SUBJECT TERMS PyrolysisThe present work interrogates the identities of the peaks by inserting a time-offlight (TOF) mass spectrometry (MS) system in parallel with the IMS detector through a Tee connection in the GC module. Biological substances, producing ion mobility peaks from the pyrolysis of microorganisms, have been identified by their GC retention times, by matching their electron ionization mass spectra with authentic standards, and by the National Institute of Standards and Technology (NIST) mass spectral database.Strong signals from 2-pyridinecarboxamide were identified in Bacillus samples, including Bacillus anthracis, and their origins were traced to the cell wall peptidoglycan macromolecule. 3-Hydroxymyristic acid is a component of lipopolysaccharides (LPS) in the cell walls of Gram-negative organisms. The Gram-negative E. coli organism showed significant amounts of 3-Hydroxymyristic acid derivatives and degradation products in Py-GC-MS analyses. Some of the fatty acid derivatives were observed in very low abundance in the ion mobi...

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Correlation of Mass Spectrometry Identified Bacterial Biomarkers from a Fielded Pyrolysis-Gas Chromatography-Ion Mobility Spectrometry Biodetector with the Microbiological Gram Stain Classification Scheme

et al. 2004

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“…Bacterial spores are challenging to detect and enumerate by conventional methods (Gould 2006). Several techniques such as pyrolysis/mass spectrometry (Ryzhov et al 2000), enzymelinked immunosorbent assays (Smith and Ulrich 1983), pyrolysis-gas chromatography-ion mobility spectrometry (Snyder et al 1999) and molecular approaches (Fasanella et al 2003;Guidi et al 2010) have been tested, but these procedures require special personnel training and may not be easily adapted to routine testing. Therefore, there is a need to develop rapid, simple and cost-effective methods to detect them.…”

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Use of Attenuated Total Reflectance Infrared Microspectroscopy to Discriminate Bacillus Spores

Lamo-Castellví

Rodríguez-Saona

2011

Journal of Food Safety

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In the present study, the potential of Fourier‐transform infrared microspectroscopy to differentiate and discriminate Bacillus spp. spores that sporulated in two nonselective media were evaluated. Spore crops of Bacillus subtilis (ATCC 11779), Bacillus thuringiensis (ATCC 13368, ATCC 35866 and ATCC 55177), Bacillus amyloliquefacies (ATCC 496763) and Bacillus stearothermophilus (ATCC 7953) were obtained with Nutrient and Trypticase Soy Agar supplemented with 10 ppm of MnSO4.H2O. Spores were sonicated, washed with deionized water, and pellets (108 cfu/mL) were deposited onto the grids of hydrophobic membrane filters and dried to produce a uniform and thin film. Spectra were collected in the attenuated total reflectance mode in the mid‐infrared region (4,000–700 cm−1). Our classification models, soft independent modeling of class analogy obtained from derivatized infrared spectra (1,800–900 cm−1), showed that the discrimination among spore crops was mainly attributed to infrared frequencies of spore coat proteins. PRACTICAL APPLICATIONS Despite regulatory efforts and emergence of new processing technologies, food‐related illnesses remain a major concern for consumers and producers. Detection of bacterial spores using conventional methods is time consuming and challenging. Therefore, there is a need to develop rapid, simple and cost‐effective methods to detect Bacillus spores. Fourier‐transform infrared microspectroscopy is a viable technology to meet these demands.

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“…Miniaturization of detectors and gas chromatography (GC) systems for real-time detection of high-priority chemicals such as explosives, toxic industrial compounds, and chemical warfare agents (CWA) is a field of increasing interest over the past few years. − The motivations behind miniaturization of analytical systems are to reduce weight and resource consumption while providing sensitive and reliable detection devices that can be used for on-site analysis. GC separations can be accelerated by using short columns, fast temperature programming, and increased carrier gas flow rates.…”

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Ultrafast Gas Chromatographic Separation of Organophosphor and Organosulfur Compounds Utilizing a Microcountercurrent Flame Photometric Detector

et al. 2006

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A microcountercurrent flame photometric detector (microcc-FPD) was adapted and optimized for ultrafast gas chromatographic (GC) separation and detection of organophosphor (OP) and organosulfur (OS) compounds on short chromatographic columns. Air and hydrogen are introduced to the microcc-FPD from opposite directions, creating a hydrogen-rich flame. In this microcc-FPD, combustion takes place between the burner tips without touching them. The separation between the tips and the flame reduces heat loss from the flame to the surrounding environment, resulting in low hydrogen consumption and a compact flame. The microcc-FPD is capable of detecting very narrow (13 ms) chromatographic peaks. An ultrafast GC separation of a group of six OP and OS compounds is achieved within less than 5 s using fast temperature programming of a 0.5-m-long microbore column. Very fast separations are also demonstrated on a 1-m-long microfabricated column consisting of 150-microm-wide, 240-microm-deep channels, etched in a 1.9-cm square silicon chip, covered with a Pyrex wafer, and statically coated with dimethyl polysiloxane. With a hydrogen flow rate of 10 mL/min, the detection limit for OP is 12 pg of P/s and 3 ng of S/s for OS compounds at a signal-to-noise ratio of 2. The coupling of a microfabricated column and a miniature FPD is an important step toward the development of a miniaturized GC-FPD capable of ultrafast detection of low levels of OP and OS compounds.

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Field detection of bacillus spore aerosols with stand-alone pyrolysis-gas chromatography-ion mobility spectrometry

Cited by 38 publications

References 15 publications

Correlation of Mass Spectrometry Identified Bacterial Biomarkers from a Fielded Pyrolysis-Gas Chromatography-Ion Mobility Spectrometry Biodetector with the Microbiological Gram Stain Classification Scheme

Correlation of Mass Spectrometry Identified Bacterial Biomarkers from a Fielded Pyrolysis-Gas Chromatography-Ion Mobility Spectrometry Biodetector with the Microbiological Gram Stain Classification Scheme

Use of Attenuated Total Reflectance Infrared Microspectroscopy to Discriminate Bacillus Spores

Ultrafast Gas Chromatographic Separation of Organophosphor and Organosulfur Compounds Utilizing a Microcountercurrent Flame Photometric Detector

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