Real-time monitoring of Bacillus subtilis endospore components by attenuated total reflection Fourier-transform infrared spectroscopy during germination

Cheung, Him; Cui, Jianxun; Sun, Su-Qin

doi:10.1099/13500872-145-5-1043

Cited by 42 publications

(27 citation statements)

References 18 publications

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“…The kinetics of germination of a spore population is usually monitored by measuring the optical density at 600 nm (OD 600 ) of spore cultures, which falls ∼60% upon completion of Stage II of germination (13), or by measuring DPA release (15–18). However, the kinetics of germination of individual spores cannot be determined from these population measurements due to significant heterogeneity in the germination of individual spores (19–21).…”

Section: Introductionmentioning

confidence: 99%

Elastic and Inelastic Light Scattering from Single Bacterial Spores in an Optical Trap Allows the Monitoring of Spore Germination Dynamics

et al. 2009

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Raman scattering spectroscopy and elastic light scattering intensity (ESLI) were used to simultaneously measure levels of Ca-dipicolinic acid (CaDPA) and changes in spore morphology and refractive index during germination of individual B. subtilis spores with and without the two redundant enzymes (CLEs), CwlJ and SleB, that degrade spores' peptidoglycan cortex. Conclusions from these measurements include: 1) CaDPA release from individual wild-type germinating spores was biphasic; in a first heterogeneous slow phase, T lag , CaDPA levels decreased ∼15% and in the second phase ending at T release , remaining CaDPA was released rapidly; 2) in L-alanine germination of wild-type spores and spores lacking SleB: a) the ESLI rose ∼2-fold shortly before T lag at T 1 ; b) following T lag , the ESLI again rose ∼2-fold at T 2 when CaDPA levels had decreased ∼50%; and c) the ESLI reached its maximum value at ∼T release and then decreased; 3) in CaDPA germination of wild-type spores: a) T lag increased and the first increase in ESLI occurred well before T lag , consistent with different pathways for CaDPA and L-alanine germination; b) at T release the ESLI again reached its maximum value; 4) in L-alanine germination of spores lacking both CLEs and unable to degrade their cortex, the time ΔT release (T release -T lag ) for excretion of ≥75% of CaDPA was ∼15-fold higher than that for wild-type or sleB spores; and 5) spores lacking only CwlJ exhibited a similar, but not identical ESLI pattern during L-alanine germination to that seen with cwlJ sleB spores, and the high value for ΔT release .

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Section: Introductionmentioning

confidence: 99%

Elastic and Inelastic Light Scattering from Single Bacterial Spores in an Optical Trap Allows the Monitoring of Spore Germination Dynamics

et al. 2009

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“…The initial events, breaking of endospore dormancy and start of metabolic functions, are termed germination and outgrowth and occur rapidly after spore association with host phagocytes, or potentially, within certain bodily fluids (5). These processes remain incompletely defined at nearly all molecular levels but entail, among other events, dramatic changes in the bacterial cell surface as the structural and chemical compositions of the surfaces differ greatly between endospore and bacillus morphotypes (2,7,26,31). While the cell surface of the endospore contributes greatly to its resistance properties in the environment (6, 7), this surface is deconstructed and replaced by vegetative cell-specific structures during germination and outgrowth (26,31).…”

mentioning

confidence: 99%

The dltABCD Operon of Bacillus anthracis Sterne Is Required for Virulence and Resistance to Peptide, Enzymatic, and Cellular Mediators of Innate Immunity

Fisher¹,

Shetron-Rama²,

Herring-Palmer³

et al. 2006

J Bacteriol

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In the environment, the gram-positive bacterium Bacillus anthracis persists as a metabolically dormant endospore. Upon inoculation into the host the endospores germinate and outgrow into vegetative bacilli able to cause disease. The dramatic morphogenic changes to the bacterium during germination and outgrowth are numerous and include major rearrangement of and modifications to the bacterial surface. Such modifications occur during a time in the B. anthracis infectious cycle when the bacterium must guard against a multitude of innate immune mediators. The dltABCD locus of B. anthracis encodes a cell wall D-alanine esterification system that is initiated by transcriptional activation during endospore outgrowth. The level of transcription from the dltABCD operon determined B. anthracis resistance to cationic antibacterial peptides during vegetative growth and cationic peptide, enzymatic, and cellular mediators of innate immunity during outgrowth. Mutation of dltABCD was also attenuating in a mouse model of infection. We propose that the dltABCD locus is important for protection of endosporeforming bacteria from environmental assault during outgrowth and that such protection may be critical during the establishment phase of anthrax.

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“…Chemical changes in Bacillus subtilis spore components during germination on a real-time basis was studied by attenuated total reflection-FTIR. 7 Johnsen and Nielsen 8 showed that the classification of various Pseudomonas species by the FTIR method was comparable to that obtained by repetitive extragenic palindromic-polymerase chain reaction, and that FTIR proved to be rapid and good for subspecies level identification. FTIR spectroscopy was also used for rapid identification of 10 group isolates of Bacillus cereus.…”

Section: Introductionmentioning

confidence: 93%

Use of FTIR spectroscopy to distinguish between capsular types and capsular quantities in Streptococcus pneumoniae

et al. 2006

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Fourier transform infrared (FTIR) spectroscopy has shown remarkable ability in distinguishing between bacterial species and identifying bacterial colony structures, when used in tandem with methods such as cluster analysis, principal component analysis, or linear discriminant analysis. The present work was aimed to evaluate the potential of FTIR-microscopy (FTIR-MSP) to distinguish between different serotypes and capsular quantities of Streptococcus pneumoniae. In general, the results obtained have consistently proven that the spectral information at the region 900-1,185 cm(-1) was sufficient to distinguish between various pneumococcal serotypes. Moreover, the method was able to differentiate between S. pneumoniae phase variants on the basis of their relative carbohydrate content. The unsupervised cluster analysis of the samples showed differences, not only in the carbohydrate content, but also in the region 1,350-1,480 cm(-1), which is dominated by absorptions due to lipids and phospholipids. This approach proved to be useful for the distinction between S. pneumoniae serotypes and between phase variants, which were shown to acquire different pathogenic capacity.

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Real-time monitoring of Bacillus subtilis endospore components by attenuated total reflection Fourier-transform infrared spectroscopy during germination

Cited by 42 publications

References 18 publications

Elastic and Inelastic Light Scattering from Single Bacterial Spores in an Optical Trap Allows the Monitoring of Spore Germination Dynamics

Elastic and Inelastic Light Scattering from Single Bacterial Spores in an Optical Trap Allows the Monitoring of Spore Germination Dynamics

The dltABCD Operon of Bacillus anthracis Sterne Is Required for Virulence and Resistance to Peptide, Enzymatic, and Cellular Mediators of Innate Immunity

Use of FTIR spectroscopy to distinguish between capsular types and capsular quantities in Streptococcus pneumoniae

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