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Title: Following the Biochemical and Morphological Changes of Bacillus atrophaeus during Sporulation using Bioaerosol Mass Spectrometry

Abstract

The overall objective of this report is to develop a real-time single-particle mass spectrometry technique called Bio-Aerosol Mass Spectrometry (BAMS) in order to efficiently screen and identify bioaerosols and single cells of national security and public health concern.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
899381
Report Number(s):
UCRL-CONF-221679
TRN: US200708%%258
DOE Contract Number:
W-7405-ENG-48
Resource Type:
Conference
Resource Relation:
Conference: Presented at: 54th ASMS Conference, Seattle , WA, United States, May 27 - Jun 01, 2006
Country of Publication:
United States
Language:
English
Subject:
45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEFENSE; 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; BACILLUS; MASS SPECTROSCOPY; MORPHOLOGICAL CHANGES; NATIONAL SECURITY; PUBLIC HEALTH; SCREENS

Citation Formats

Tobias, H J, Pitesky, M E, Fergenson, D P, Horn, J, Frank, M, and Gard, E E. Following the Biochemical and Morphological Changes of Bacillus atrophaeus during Sporulation using Bioaerosol Mass Spectrometry. United States: N. p., 2006. Web.
Tobias, H J, Pitesky, M E, Fergenson, D P, Horn, J, Frank, M, & Gard, E E. Following the Biochemical and Morphological Changes of Bacillus atrophaeus during Sporulation using Bioaerosol Mass Spectrometry. United States.
Tobias, H J, Pitesky, M E, Fergenson, D P, Horn, J, Frank, M, and Gard, E E. Wed . "Following the Biochemical and Morphological Changes of Bacillus atrophaeus during Sporulation using Bioaerosol Mass Spectrometry". United States. doi:. https://www.osti.gov/servlets/purl/899381.
@article{osti_899381,
title = {Following the Biochemical and Morphological Changes of Bacillus atrophaeus during Sporulation using Bioaerosol Mass Spectrometry},
author = {Tobias, H J and Pitesky, M E and Fergenson, D P and Horn, J and Frank, M and Gard, E E},
abstractNote = {The overall objective of this report is to develop a real-time single-particle mass spectrometry technique called Bio-Aerosol Mass Spectrometry (BAMS) in order to efficiently screen and identify bioaerosols and single cells of national security and public health concern.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Wed May 03 00:00:00 EDT 2006},
month = {Wed May 03 00:00:00 EDT 2006}
}

Conference:
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  • We have conducted studies to fully characterize the mass spectral signature of individual Bacillus atrophaeus, previously known as Bacillus subtilis var niger or Bacillus globigii, spores obtained in matrix-free bioaerosol mass spectrometry (BAMS). Mass spectra of spores grown in unlabeled, {sup 13}C-labeled and {sup 15}N-labeled growth media are used to determine the number of carbon and nitrogen atoms associated with each mass peak. To determine the parent ion structure associated with fragment ions present in the spore spectra, the mass-to-charge (m/z) fragmentation pattern of several chemical standards was obtained. Our results agree with prior assignments of dipicolinic acid, amino acidsmore » and calcium complex ions made in the spore mass spectra. Identity of several previously unidentified mass peaks, key to recognition of Bacillus spore by matrix-free BAMS, is revealed. Specifically, a set of fragment peaks in the negative polarity is shown to be consistent with the fragmentation pattern of purine nucleobase containing compounds. The identity of m/z=+74, a marker peak that helps discriminate Bacillus atrophaeus from Bacillus thuringiensis spores grown in rich medium, is surprisingly a non-description, viz. [N{sub 1}C{sub 4}H{sub 12}]{sup +}. A probable precursor molecule for the [N{sub 1}C{sub 4}H{sub 12}]{sup +} ion observed in spore spectra is trimethyl glycine ({sup +}N(CH{sub 3}){sub 3}CH{sub 2}COOH) that produces a m/z=74 peak in presence of dipicolinic acid.« less
  • No abstract prepared.
  • Bioaerosol mass spectrometry (BAMS) is being developed to analyze and identify biological aerosols in real-time. Mass spectra of individual Bacillus endospores were measured here with a bipolar aerosol time-of-flight mass spectrometer in which molecular desorption and ionization were produced using a single laser pulse from a Q-switched, frequency-quadrupled Nd:YAG laser that was modified to have an approximately flattop profile. The flattened laser profile allowed the minimum fluence required to desorb and ionize significant numbers of ions from single aerosol particles to be determined. For Bacillus spores this threshold had a mean value of approximately 1 nJ/{micro}m{sup 2} (0.1 J/cm{sup 2}).more » Thresholds for individual spores, however, could apparently deviate by 20% or more from the mean. Threshold distributions for clumps of MS2 bacteriophage and bovine serum albumin were subsequently determined. Finally, the flattened profile was observed to increase the reproducibility of single spore mass spectra. This is consistent with the general conclusions of our earlier paper on the fluence dependence of single spore mass spectra and is particularly significant because it is expected to enable more robust differentiation and identification of single bioaerosol particles.« less
  • The BioAerosol Mass Spectrometry (BAMS) system is a rapidly fieldable, fully autonomous instrument that can perform correlated measurements of multiple orthogonal properties of individual aerosol particles. The BAMS front end uses optical techniques to nondestructively measure a particle's aerodynamic diameter and fluorescence properties. Fluorescence can be excited at 266nm or 355nm and is detected in two broad wavelength bands. Individual particles with appropriate size and fluorescence properties can then be analyzed more thoroughly in a dual-polarity time-of-flight mass spectrometer. Over the course of two deployments to the San Francisco International Airport, more than 6.5 million individual aerosol particles were fullymore » analyzed by the system. Analysis of the resulting data has provided a number of important insights relevant to rapid bioaerosol detection, which are described here.« less