Bacillus Globigii

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Tuan Vodinh - One of the best experts on this subject based on the ideXlab platform.

  • intensified biochip system using chemiluminescence for the detection of Bacillus Globigii spores
    Analytical and Bioanalytical Chemistry, 2008
    Co-Authors: Dimitra N Stratiscullum, Guy D Griffin, Joel Mobley, Tuan Vodinh
    Abstract:

    This paper reports the first intensified biochip system forchemiluminescence detectionandthefeasibilityof using this system for the analysis of biological warfare agents is demonstrated. An enzyme-linked immunosorbent assay targeting Bacillus Globigii spores, a surrogate species for Bacillus anthracis, using a chemiluminescent alkaline phosphatase substrate is combined with a compact intensi- fied biochip detection system. The enzymatic amplification was found to be an attractive method for detection of low spore concentrations when combined with the intensified biochip device. This system was capable of detecting ap- proximately 1×10 5 Bacillus Globigii spores. Moreover, the chemiluminescence method, combined with the self- contained biochip design, allows for a simple, compact sys- tem that does not require laser excitation and is readily adaptable to field use.

  • surface enhanced raman scattering detection of chemical and biological agents using a portable raman integrated tunable sensor
    Sensors and Actuators B-chemical, 2007
    Co-Authors: Fei Yan, Tuan Vodinh
    Abstract:

    Abstract This paper describes a compact and rugged Raman integrated tunable sensor coupled with surface-enhanced Raman scattering substrates for the screening of a wide variety of chemical and biological agents for homeland defense applications. The field-deployable instrument, consisting of an 830-nm diode laser for excitation and an avalanche photodiode (APD) for detection, is a fully integrated, tunable, “point-and-shoot” Raman device based on solid-state acousto-optic tunable filter (AOTF) technology. It can provide direct identification of chemical and biological samples in a few seconds under field conditions. This paper illustrates some applications of this portable device for the detection of various compounds of particular interest for homeland defense applications. These include methyl parathion (a nerve agent simulant) and dipicolinic acid (a biomarker for Bacillus endospore), and other chemical warfare simulants such as dimethyl methylphosphonate, pinacolyl methylphosphonate, diethyl phosphoramidate, and 2-chloroethyl ethylsulfide, which are simulants for sarin (GB), soman (GD), tabun (GA), and sulfur mustard (HD), respectively, and intact bacteria such as Bacillus Globigii, Erwinia herbicola, and Bacillus thuringiensis, which are simulants for biological warfare agents.

  • a compact cmos biochip immunosensor towards the detection of a single bacteria
    Biosensors and Bioelectronics, 2005
    Co-Authors: Joon Myong Song, Guy D Griffin, Mustafa Culha, Paul M Kasili, Tuan Vodinh
    Abstract:

    Recent use of biological warfare (BW) agents has led to a growing interest in the rapid and sensitive detection of pathogens. Therefore, the development of field-usable detection devices for sensitive and selective detection of BW agents is an important issue. In this work, we report a portable biochip system based on complementary metal oxide semiconductor (CMOS) technology that has great potential as a device for single-bacteria detection. The possibility of single-bacteria detection is reported using an immunoassay coupled to laser-induced fluorescence (LIF) detection. Bacillus Globigii spores, which are a surrogate species for B. anthracis spores, were used as the test sample. Enzymatic amplification following immunocomplex formation allowed remarkably sensitive detection of B. Globigii spores, and could preclude a complicated optical and instrumental system usually required for high-sensitive detection. Atomic force microscopy (AFM) was employed to investigate whether B. Globigii spores detected in the portable biochip system exist in single-cell or multicellular form. It was found that B. Globigii spores mostly exist in multicellular form with a small minority of single-cell form. The results showed that the portable biochip system has great potential as a device for single-particle or possibly even single-organism detection.

  • a miniature biochip system for detection of aerosolized Bacillus Globigii spores
    Analytical Chemistry, 2003
    Co-Authors: Dimitra N Stratiscullum, Guy D Griffin, Joel Mobley, And Arpad A Vass, Tuan Vodinh
    Abstract:

    The feasibility of using a novel detection scheme for the analysis of biological warfare agents is demonstrated using Bacillus Globigii spores, a surrogate species for Bacillus anthracis. In this paper, a sensitive and selective enzyme-linked immunosorbent assay using a novel fluorogenic alkaline phosphatase substrate (dimethylacridinone phosphate) is combined with a compact biochip detection system, which includes a miniature diode laser for excitation. Detection of aerosolized spores was achieved by coupling the miniature system to a portable bioaerosol sampler, and the performance of the antibody-based recognition and enzyme amplification method was evaluated. The bioassay performance was found to be compatible with the air sampling device, and the enzymatic amplification was found to be an attractive amplification method for detection of low spore concentrations. The combined portable bioaerosol sampler and miniature biochip system detected 100 B. Globigii spores, corresponding to 17 aerosolized spore...

  • a miniature biochip system for detection of aerosolized Bacillus Globigii spores
    Analytical Chemistry, 2003
    Co-Authors: Dimitra N Stratiscullum, Guy D Griffin, Joel Mobley, And Arpad A Vass, Tuan Vodinh
    Abstract:

    The feasibility of using a novel detection scheme for the analysis of biological warfare agents is demonstrated using Bacillus Globigii spores, a surrogate species for Bacillus anthracis. In this paper, a sensitive and selective enzyme-linked immunosorbent assay using a novel fluorogenic alkaline phosphatase substrate (dimethylacridinone phosphate) is combined with a compact biochip detection system, which includes a miniature diode laser for excitation. Detection of aerosolized spores was achieved by coupling the miniature system to a portable bioaerosol sampler, and the performance of the antibody-based recognition and enzyme amplification method was evaluated. The bioassay performance was found to be compatible with the air sampling device, and the enzymatic amplification was found to be an attractive amplification method for detection of low spore concentrations. The combined portable bioaerosol sampler and miniature biochip system detected 100 B. Globigii spores, corresponding to 17 aerosolized spores/L of air. Moreover, the incorporation of the miniature diode laser with the self-contained biochip design allows for a compact system that is readily adaptable to field use. In addition, these studies have included investigations into the tradeoff between assay time and sensitivity.

Paul J Jackson - One of the best experts on this subject based on the ideXlab platform.

  • small scale dna sample preparation method for field pcr detection of microbial cells and spores in soil
    Applied and Environmental Microbiology, 1998
    Co-Authors: Cheryl R Kuske, Kaysie L Banton, Dante L Adorada, Peter C Stark, Karen K Hill, Paul J Jackson
    Abstract:

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus Globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent-bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. Globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. Globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR.

  • small scale dna sample preparation method for field pcr detection of microbial cells and spores in soil
    Applied and Environmental Microbiology, 1998
    Co-Authors: Cheryl R Kuske, Kaysie L Banton, Dante L Adorada, Peter C Stark, Karen K Hill, Paul J Jackson
    Abstract:

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus Globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent‐bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. Globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. Globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR. PCR analysis provides a sensitive and specific means to detect and monitor microorganisms in complex environmental samples. Successful detection and characterization of microbial DNA in the environment require efficient extraction of the DNA from environmental samples and adequate purification from the coextracted contaminants that inhibit PCR. Soils and sediments vary greatly in chemical and organic composition. They also contain abundant humic and fulvic acids that are inhibitory to Taq DNA polymerase and other enzymes (24, 26, 28; for a recent review, see reference 29). Soils are therefore one of the most challenging environmental matrices from which to obtain microbial DNA that will support PCR. Two applications in environmental microbial assessment require simultaneous extraction of the DNA from a wide range of microorganisms in a single sample. For analysis of the diversity and dynamics of natural microbial communities, a broad-based, nonselective DNA extraction procedure is desirable to obtain unbiased representation of community members. For forensic and other investigative analyses, a simple, small-scale procedure is needed to provide rapid, sensitive detection of a wide variety of potentially released organisms, including several medically important bacterial and fungal pathogens, for in-the-field analysis of environmental samples. Direct comparisons of the relative effectiveness of different extraction and purification procedures for simultaneous preparation of both bacterial and fungal propagules have not been made. Most studies describing recovery of microbial DNA from soils or sediments have focused on extraction of DNA from a single introduced microorganism, usually vegetative cells of a gram-negative organism, or have examined only a

Catherine Fenselau - One of the best experts on this subject based on the ideXlab platform.

  • complete sequences of small acid soluble proteins from Bacillus Globigii
    Journal of Mass Spectrometry, 2004
    Co-Authors: Jeffrey R Whiteaker, Yetrib Hathout, Bettina Warscheid, Partick Pribil, Catherine Fenselau
    Abstract:

    Three abundant small acid-soluble proteins (SASPs) from spores of Bacillus Globigii were sequenced using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with post-source decay and nanoelectrospray collision-induced dissociation tandem mass spectrometry. The proteins were extracted from spores with 1 M HCl. Scanning electron micrographs of spores before and after acid extraction show that the spores retain their overall structure but have a shriveled texture following the acid treatment. Extracted SASPs were purified by high-performance liquid chromatography and molecular masses of the SASPs were identified at 7068 (SASP-1), 7332 (SASP-2), and 8889 (γ-SASP). De novo peptide sequencing was used to determine the protein sequences. The correct ordering of peptide sequences was aided by mapping overlapping enzymatic digests and by comparison with homologous SASPs from Bacillus stearothermophilus. B. Globigii is used in many field tests as a surrogate for B. anthracis. Thus complete SASP sequences from B. Globigii will facilitate the development of methods for rapid identification of bacteria based on mass spectrometry and the examination of taxonomic relationships between Bacillus species. Copyright © 2004 John Wiley & Sons, Ltd.

  • structural characterization of lipopeptide biomarkers isolated from Bacillus Globigii
    Journal of Mass Spectrometry, 2002
    Co-Authors: Bruce H Williams, Yetrib Hathout, Catherine Fenselau
    Abstract:

    Spectra obtained using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry of Bacillus Globigii (Bacillus subtilis niger) spores, vegetative cells and the culture supernatant show a cluster of biomarkers centered at a molecular mass of 1478 Da. Three biomarkers were isolated from the cell-free culture supernatant by solid-phase extraction and reversed-phase high-performance liquid chromatography, and characterized using various kinds of mass spectrometry. A Fourier transform mass spectrometer with a MALDI source was used to determine the monoisotopic protonated masses at 1463.8, 1477.8, and 1505.8 Da in order of elution. The mass differences of 14 and 28 Da suggest that they are homologous molecules. Alkaline hydrolysis of each species showed that it contained a lactone linkage. Strong acid hydrolysis released a fatty acid from an amide bond, consistent with a lipopeptide. A quadrupole time-of-flight instrument with a nanospray source was used to sequence the hydrolyzed forms of the three biomarkers. The cyclic lipopeptides were found to have amino acid sequences identical with those in fengycins and plipastatins, antimicrobial compounds with phospholipase inhibitor activity, previously identified in related species of Bacillus subtilis and Bacillus cereus.

  • rapid characterization of spores of Bacillus cereus group bacteria by matrix assisted laser desorption ionization time of flight mass spectrometry
    Applied and Environmental Microbiology, 2000
    Co-Authors: Victor Ryzhov, Yetrib Hathout, Catherine Fenselau
    Abstract:

    Matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry was used to characterize the spores of 14 microorganisms of the Bacillus cereus group. This group includes the four Bacillus species B. anthracis, B. cereus, B. mycoides, and B. thuringiensis. MALDI mass spectra obtained from whole bacterial spores showed many similarities between the species, except for B. mycoides. At the same time, unique mass spectra could be obtained for the different B. cereus and B. thuringiensis strains, allowing for differentiation at the strain level. To increase the number of detectable biomarkers in the usually peak-poor MALDI spectra of spores, the spores were treated by corona plasma discharge (CPD) or sonicated prior to MALDI analysis. Spectra of sonicated or CPD-treated spores displayed an ensemble of biomarkers common for B. cereus group bacteria. Based on the spectra available, these biomarkers differentiate B. cereus group spores from those of Bacillus subtilis and Bacillus Globigii. The effect of growth medium on MALDI spectra of spores was also explored.

Cheryl R Kuske - One of the best experts on this subject based on the ideXlab platform.

  • small scale dna sample preparation method for field pcr detection of microbial cells and spores in soil
    Applied and Environmental Microbiology, 1998
    Co-Authors: Cheryl R Kuske, Kaysie L Banton, Dante L Adorada, Peter C Stark, Karen K Hill, Paul J Jackson
    Abstract:

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus Globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent-bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. Globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. Globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR.

  • small scale dna sample preparation method for field pcr detection of microbial cells and spores in soil
    Applied and Environmental Microbiology, 1998
    Co-Authors: Cheryl R Kuske, Kaysie L Banton, Dante L Adorada, Peter C Stark, Karen K Hill, Paul J Jackson
    Abstract:

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus Globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent‐bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. Globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. Globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR. PCR analysis provides a sensitive and specific means to detect and monitor microorganisms in complex environmental samples. Successful detection and characterization of microbial DNA in the environment require efficient extraction of the DNA from environmental samples and adequate purification from the coextracted contaminants that inhibit PCR. Soils and sediments vary greatly in chemical and organic composition. They also contain abundant humic and fulvic acids that are inhibitory to Taq DNA polymerase and other enzymes (24, 26, 28; for a recent review, see reference 29). Soils are therefore one of the most challenging environmental matrices from which to obtain microbial DNA that will support PCR. Two applications in environmental microbial assessment require simultaneous extraction of the DNA from a wide range of microorganisms in a single sample. For analysis of the diversity and dynamics of natural microbial communities, a broad-based, nonselective DNA extraction procedure is desirable to obtain unbiased representation of community members. For forensic and other investigative analyses, a simple, small-scale procedure is needed to provide rapid, sensitive detection of a wide variety of potentially released organisms, including several medically important bacterial and fungal pathogens, for in-the-field analysis of environmental samples. Direct comparisons of the relative effectiveness of different extraction and purification procedures for simultaneous preparation of both bacterial and fungal propagules have not been made. Most studies describing recovery of microbial DNA from soils or sediments have focused on extraction of DNA from a single introduced microorganism, usually vegetative cells of a gram-negative organism, or have examined only a

Karen K Hill - One of the best experts on this subject based on the ideXlab platform.

  • small scale dna sample preparation method for field pcr detection of microbial cells and spores in soil
    Applied and Environmental Microbiology, 1998
    Co-Authors: Cheryl R Kuske, Kaysie L Banton, Dante L Adorada, Peter C Stark, Karen K Hill, Paul J Jackson
    Abstract:

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus Globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent-bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. Globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. Globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR.

  • small scale dna sample preparation method for field pcr detection of microbial cells and spores in soil
    Applied and Environmental Microbiology, 1998
    Co-Authors: Cheryl R Kuske, Kaysie L Banton, Dante L Adorada, Peter C Stark, Karen K Hill, Paul J Jackson
    Abstract:

    Efficient, nonselective methods to obtain DNA from the environment are needed for rapid and thorough analysis of introduced microorganisms in environmental samples and for analysis of microbial community diversity in soil. A small-scale procedure to rapidly extract and purify DNA from soils was developed for in-the-field use. Amounts of DNA released from bacterial vegetative cells, bacterial endospores, and fungal conidia were compared by using hot-detergent treatment, freeze-thaw cycles, and bead mill homogenization. Combining a hot-detergent treatment with bead mill homogenization gave the highest DNA yields from all three microbial cell types and provided DNA from the broadest range of microbial groups in a natural soil community. Only the bead mill homogenization step was effective for DNA extraction from Bacillus Globigii (B. subtilis subsp. niger) endospores or Fusarium moniliforme conidia. The hot-detergent‐bead mill procedure was simplified and miniaturized. By using this procedure and small-scale, field-adapted purification and quantification procedures, DNA was prepared from four different soils seeded with Pseudomonas putida cells or B. Globigii spores. In a New Mexico soil, seeded bacterial targets were detected with the same sensitivity as when assaying pure bacterial DNA (2 to 20 target gene copies in a PCR mixture). The detection limit of P. putida cells and B. Globigii spores in different soils was affected by the amount of background DNA in the soil samples, the physical condition of the DNA, and the amount of DNA template used in the PCR. PCR analysis provides a sensitive and specific means to detect and monitor microorganisms in complex environmental samples. Successful detection and characterization of microbial DNA in the environment require efficient extraction of the DNA from environmental samples and adequate purification from the coextracted contaminants that inhibit PCR. Soils and sediments vary greatly in chemical and organic composition. They also contain abundant humic and fulvic acids that are inhibitory to Taq DNA polymerase and other enzymes (24, 26, 28; for a recent review, see reference 29). Soils are therefore one of the most challenging environmental matrices from which to obtain microbial DNA that will support PCR. Two applications in environmental microbial assessment require simultaneous extraction of the DNA from a wide range of microorganisms in a single sample. For analysis of the diversity and dynamics of natural microbial communities, a broad-based, nonselective DNA extraction procedure is desirable to obtain unbiased representation of community members. For forensic and other investigative analyses, a simple, small-scale procedure is needed to provide rapid, sensitive detection of a wide variety of potentially released organisms, including several medically important bacterial and fungal pathogens, for in-the-field analysis of environmental samples. Direct comparisons of the relative effectiveness of different extraction and purification procedures for simultaneous preparation of both bacterial and fungal propagules have not been made. Most studies describing recovery of microbial DNA from soils or sediments have focused on extraction of DNA from a single introduced microorganism, usually vegetative cells of a gram-negative organism, or have examined only a