CTC staining and counting of actively respiring bacteria in natural stone using confocal laser scanning microscopy

Bartosch, Sabine; Mansch, R.; Knötzsch, K; Bock, Eberhard

doi:10.1016/s0167-7012(02)00133-1

Cited by 42 publications

(26 citation statements)

References 30 publications

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“…Because GFP is not suitable as a viability marker since the protein remains detectable even in dead cells (Tsien, 1998), the redox dye CTC was employed as a viability stain (Bartosch et al, 2003) for microscopic detection of metabolizing L-forms. Co-localization of red (CTC) and green (GFP) signals strongly suggests viability of the ICL (Figure 3).…”

Section: Resultsmentioning

confidence: 99%

Cell-wall deficient L. monocytogenes L-forms feature abrogated pathogenicity

Schnell¹,

Staubli²,

Harris

et al. 2014

Front. Cell. Infect. Microbiol

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Stable L-forms are cell wall-deficient bacteria which are able to multiply and propagate indefinitely, despite the absence of a rigid peptidoglycan cell wall. We investigated whether L-forms of the intracellular pathogen L. monocytogenes possibly retain pathogenicity, and if they could trigger an innate immune response. While phagocytosis of L. monocytogenes L-forms by non-activated macrophages sometimes resulted in an unexpected persistence of the bacteria in the phagocytes, they were effectively eliminated by IFN-γ preactivated or bone marrow-derived macrophages (BMM). These findings were in line with the observed down-regulation of virulence factors in the cell-wall deficient L. monocytogenes. Absence of Interferon-β (IFN-β) triggering indicated inability of L-forms to escape from the phagosome into the cytosol. Moreover, abrogated cytokine response in MyD88-deficient dendritic cells (DC) challenged with L. monocytogenes L-forms suggested an exclusive TLR-dependent host response. Taken together, our data demonstrate a strong attenuation of Listeria monocytogenes L-form pathogenicity, due to diminished expression of virulence factors and innate immunity recognition, eventually resulting in elimination of L-form bacteria from phagocytes.

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Section: Resultsmentioning

confidence: 99%

Cell-wall deficient L. monocytogenes L-forms feature abrogated pathogenicity

Schnell¹,

Staubli²,

Harris

et al. 2014

Front. Cell. Infect. Microbiol

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“…For the epifluorescent analysis, MRSA and EMRSA CFU on inoculated coupons were stained with 5-cyano-2,3-ditolyl tetrazolium (CTC) which detects actively respiring bacteria. 25 Coupons were sterilized and inoculated with 20 mL of culture as described in the alloy testing protocol. Based on findings from the previous tests, metal sample C19700 was analysed after an exposure period of 45, 60 and 90 min for MRSA (10442), EMRSA-1 and EMRSA-16, respectively.…”

Section: Edic and Epifluorescent Microscopy Analysismentioning

confidence: 99%

Potential use of copper surfaces to reduce survival of epidemic meticillin-resistant Staphylococcus aureus in the healthcare environment

Noyce

Michels

Keevil

2006

Journal of Hospital Infection

341

229

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“…Optimal CTC staining conditions should be changed in different environments. For example, the optimal CTC concentration for staining microorganisms in stone material was found to be 15 mM 3) .…”

Section: Discussionmentioning

confidence: 99%

An Improved Redox Dye-Staining Method Using 5-Cyano-2,3-Ditoryl Tetrazolium Chloride for Detection of Metabolically Active Bacteria in Activated Sludge

Yoshida

Hiraishi

2004

Microb. Environ.

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A fluorochrome-staining method using 5-cyano-2,3-ditoryl tetrazolium chloride (CTC) was improved to enhance the sensitivity of detection of metabolically active bacteria in activated sludge. The highest efficiency of CTC staining was obtained with 6 mM CTC in MOPS buffer at pH 6.5. CTC staining efficiency was also increased by adding a substrate mixture (0.05% peptone, 0.05% yeast extract and 1 mM glucose), Meldola's Blue as an electron transfer mediator and KCN. The bacterial counts detected by the CTC staining method thus improved accounted for 41-76% of the total counts in activated sludge with the average value being 54%. This value was approximately two-fold higher than those obtained by the traditional CTC staining assay which has been performed in phosphate buffer or phosphate-buffered saline without any addition of external substrate or electron transfer mediator. There was a high positive correlation between CTC-stained direct counts and the plate counts of aerobic chemoorganotrophic bacteria or viable cell counts measured using a LIVE/DEAD BacLight Bacterial Viability kit. The viable population in the activated sludges as measured by BacLight staining and plate counting accounted for 73±7.6% and 8.7±4.7% of the total population, respectively. These data suggested that most of the "viable but nonculturable" bacteria (i.e., BacLight-positive but unculturable population) potentially existing in the sludge system were so metabolically active as to be detectable by the improved CTC staining method.

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CTC staining and counting of actively respiring bacteria in natural stone using confocal laser scanning microscopy

Cited by 42 publications

References 30 publications

Cell-wall deficient L. monocytogenes L-forms feature abrogated pathogenicity

Cell-wall deficient L. monocytogenes L-forms feature abrogated pathogenicity

Potential use of copper surfaces to reduce survival of epidemic meticillin-resistant Staphylococcus aureus in the healthcare environment

An Improved Redox Dye-Staining Method Using 5-Cyano-2,3-Ditoryl Tetrazolium Chloride for Detection of Metabolically Active Bacteria in Activated Sludge

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