Effect of supplementary manganese on the sporulation of <i>Bacillus</i> endospores analysed by Raman spectroscopy

Stöckel, Stephan; Meisel, Susann; Böhme, René; Elschner, Mandy C.; Rösch, Petra; Popp, Jürgen

doi:10.1002/jrs.2292

Cited by 39 publications

(32 citation statements)

References 63 publications

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“…21 Nevertheless, for this databases a huge variety of all essential environmental parameters has to be taken into account. 8,9 Sampling of microorganisms An additional difference between the conventional identification schemes and the application of micro-Raman spectroscopy is the different isolation approach. For Raman spectroscopy the single cells should not be destroyed during isolation.…”

Section: Resultsmentioning

confidence: 99%

“…7 Hence, all kind of environmental parameters relevant for the specific habitat under investigation have to be included in an identification database. [8][9][10] In addition, also the influence of antibiotics on the Raman spectra of different bacteria have to be taken into account. 11,12 Some bacteria exhibit high amounts of storage material like e.g.…”

Section: Introductionmentioning

confidence: 99%

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Bacterial identification in real samples by means of micro-Raman spectroscopy

et al. 2011

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Pathogen detection is essential without time delay especially for severe diseases like sepsis. Here, the survival rate is dependent on a prompt antibiosis. For sepsis three hours after the onset of shock the survival rate of the patient drops below 60 %. Unfortunately, the results from standard diagnosis methods like PCR or microbiology can normally be received after 12 or 36 h, respectively. Therefore diagnosis methods which require less cultivation or even no cultivation at all have to be established for medical diagnosis. Here, Raman spectroscopy, as a vibrational spectroscopic method, is a very sensitive and selective approach and monitors the biochemical composition of the investigated sample. Applying micro-Raman spectroscopy allows for a spatial resolution below 1 µm and is therefore in the size range of bacteria. Raman spectra of bacteria depend on the physiological status. Therefore, the databases require the inclusion of the necessary environmental parameters such as temperature, pH, nutrition, etc. Such large databases therefore require a specialized chemometric approach, since the variation between different strains is small. In this contribution we will demonstrate the capability of Raman spectroscopy to identify pathogens without cultivation even from real environmental or medical samples.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Bacterial identification in real samples by means of micro-Raman spectroscopy

et al. 2011

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“…Due to the changed nutrient and incubation conditions by adaptation to the new matrix, it is probable that the bacterial growth behavior was modified (16,45). Changes caused by these factors become manifest as variations in the bacterial Raman spectra.…”

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confidence: 99%

Raman Spectroscopy as a Potential Tool for Detection of Brucella spp. in Milk

Meisel

Stöckel

Elschner

et al. 2012

Appl Environ Microbiol

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Detection of Brucella, causing brucellosis, is very challenging, since the applied techniques are mostly time-demanding and not standardized. While the common detection system relies on the cultivation of the bacteria, further classical typing up to the biotype level is mostly based on phenotypic or genotypic characteristics. The results of genotyping do not always fit the existing taxonomy, and misidentifications between genetically closely related genera cannot be avoided. This situation gets even worse, when detection from complex matrices, such as milk, is necessary. For these reasons, the availability of a method that allows early and reliable identification of possible Brucella isolates for both clinical and epidemiological reasons would be extremely useful. We evaluated micro-Raman spectroscopy in combination with chemometric analysis to identify Brucella from agar plates and directly from milk: prior to these studies, the samples were inactivated via formaldehyde treatment to ensure a higher working safety. The single-cell Raman spectra of different Brucella, Escherichia, Ochrobactrum, Pseudomonas, and Yersinia spp. were measured to create two independent databases for detection in media and milk. Identification accuracies of 92% for Brucella from medium and 94% for Brucella from milk were obtained while analyzing the single-cell Raman spectra via support vector machine. Even the identification of the other genera yielded sufficient results, with accuracies of >90%. In summary, micro-Raman spectroscopy is a promising alternative for detecting Brucella. The measurements we performed at the single-cell level thus allow fast identification within a few hours without a demanding process for sample preparation.

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“…2 the spore composition of the particular culture, emerging, e.g., from fluctuating CaDPA contents in single spores among a population (19). The band positions, on the other hand, remain stable among the spectra; this is not necessarily the case, since the use of divalent manganese as a sporulation accelerator can cause some minor CaDPA band shifts (53).…”

Section: Resultsmentioning

confidence: 99%

Raman Spectroscopy-Compatible Inactivation Method for Pathogenic Endospores

Stöckel

Schumacher

Meisel

et al. 2010

Appl Environ Microbiol

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Micro-Raman spectroscopy is a fast and sensitive tool for the detection, classification, and identification of biological organisms. The vibrational spectrum inherently serves as a fingerprint of the biochemical composition of each bacterium and thus makes identification at the species level, or even the subspecies level, possible. Therefore, microorganisms in areas susceptible to bacterial contamination, e.g., clinical environments or food-processing technology, can be sensed. Within the scope of point-of-care-testing also, detection of intentionally released biosafety level 3 (BSL-3) agents, such as Bacillus anthracis endospores, or their products is attainable. However, no Raman spectroscopy-compatible inactivation method for the notoriously resistant Bacillus endospores has been elaborated so far. In this work we present an inactivation protocol for endospores that permits, on the one hand, sufficient microbial inactivation and, on the other hand, the recording of Raman spectroscopic signatures of single endospores, making species-specific identification by means of highly sophisticated chemometrical methods possible. Several physical and chemical inactivation methods were assessed, and eventually treatment with 20% formaldehyde proved to be superior to the other methods in terms of sporicidal capacity and information conservation in the Raman spectra. The latter fact has been verified by successfully using self-learning machines (such as support vector machines or artificial neural networks) to identify inactivated B. anthracis-related endospores with adequate accuracies within the range of the limited model database employed.The detection of biological warfare agents requires methods for detecting and rapidly identifying bacterial endosporessuch as Bacillus anthracis, the etiological agent of the acute fatal zoonosis anthrax in mammals-that are released in buildings or distributed in the environment. A great number of different technologies and combinations, such as DNA detection by PCR or DNA sequencing (42), are applied for genetic analysis. In addition, different microscopic approaches, such as atomic force microscopy (64) or fluorescence microscopy (26, 27), mass spectroscopy, and infrared (23,34,50) and Raman spectroscopy (15), have been used. With these optical detection methods, preparation and analysis time can be considerably shortened relative to that required for currently established methods based on, e.g., microbiology, immunoassays, and genetic and molecularly based approaches for identification.

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Effect of supplementary manganese on the sporulation of Bacillus endospores analysed by Raman spectroscopy

Cited by 39 publications

References 63 publications

Bacterial identification in real samples by means of micro-Raman spectroscopy

Bacterial identification in real samples by means of micro-Raman spectroscopy

Raman Spectroscopy as a Potential Tool for Detection of Brucella spp. in Milk

Raman Spectroscopy-Compatible Inactivation Method for Pathogenic Endospores

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