Fluorescent in situ hybridization and flow cytometry as tools to evaluate the treatments for the control of slime-forming enterobacteria in paper mills

Torres, Claudia; Gibello, Alicia; Nande, Mar; Martín, Margarita; Blanco, Ángeles

doi:10.1007/s00253-008-1369-6

Cited by 13 publications

(12 citation statements)

References 29 publications

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“…Additionally, the SYTO ® -9 stain served as the donor dye for the downstream specific FRET-ISH assay. Stained bacteria were then diluted in 10 mL distilled water for a final concentration of 1 …”

Section: Fluorescent Staining Of Cellsmentioning

confidence: 99%

“…This approach dramatically reduces the time required for pan-enterobacterial detection to less than thirty minutes from receipt of sample (Table 1), compared to multiple hours needed for traditional RT-PCR and its requisite sample preparation. A DNA probe specific to a region of the enterobacterial repetitive intergenic consensus (ERIC) probe was employed by Torres, et al [1] to detect multiple species of enterobacterial contaminants in environmental slime samples by fluorescent in situ hybridization (FISH). That approach was successfully used to detect a range of enterobacteria, specifically in the presence of an environmental matrix, and in the present work that method has been modified to drastically decrease assay time.…”

Section: Introductionmentioning

confidence: 99%

“…Further study is required for validation of this approach with more complex sample matrices and other species of bacteria. FISH was first introduced in the 1980s and has since found widespread application in bacterial identification [1][2][3][4][5]. Although capable of species-specific microbiological detection, FISH traditionally requires fixation, permeabilization, denaturation, probe hybridization, washing, and detection.…”

Section: Introductionmentioning

confidence: 99%

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Novel, rapid DNA-based on-chip bacterial identification system combining dielectrophoresis and amplification-free fluorescent resonance energy transfer assisted in-situ hybridization (FRET-ISH)

2011

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Although real-time PCR (RT-PCR) has become a diagnostic standard for rapid identification of bacterial species, typical methods remain time-intensive due to sample preparation and amplification cycle times. The assay described in this work incorporates on-chip dielectrophoretic capture and concentration of bacterial cells, thermal lysis, cell permeabilization, and nucleic acid denaturation and fluorescence resonance energy transfer assisted in situ hybridization (FRET-ISH) species identification. Combining these techniques leverages the benefits of all of them, allowing identification to be accomplished completely on chip less than thirty minutes after receipt of sample, compared to multiple hours required by traditional RT-PCR and its requisite sample preparation. OPEN ACCESS

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Section: Fluorescent Staining Of Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Novel, rapid DNA-based on-chip bacterial identification system combining dielectrophoresis and amplification-free fluorescent resonance energy transfer assisted in-situ hybridization (FRET-ISH)

2011

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“…Combined with fluorescence in situ hybridization (FISH), flow cytometry (flow-FISH) has become a powerful tool that has been used to quantify rare bacterial cells from mixtures (3,7), enrich for subpopulations from complex environmental matrices (15,44), monitor the effectiveness of antimicrobial treatments (38), identify physiological and ecological diversity (16), measure the modulation of microbiota in response to nutrient shifts (5), and enable the detection of pathogenic microbes (12). Flow-FISH probes are typically designed to target rRNA due to the relative cellular abundance of rRNA molecules and their accepted use in species identification.…”

mentioning

confidence: 99%

Locked Nucleic Acid and Flow Cytometry-Fluorescence In Situ Hybridization for the Detection of Bacterial Small Noncoding RNAs

Robertson

Vora

2012

Appl Environ Microbiol

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We describe the development and testing of a high-throughput method that enables the detection of small noncoding RNAs (ncRNAs) from single bacterial cells using locked nucleic acid probes (LNA) and flow cytometry-fluorescence in situ hybridization (flow-FISH). The LNA flow-FISH method and quantitative reverse transcription-PCR (qRT-PCR) were used to monitor the expression of three ncRNAs (6S, CsrB, and TPP-2) in Vibrio campbellii ATCC BAA-1116 cultures during lag phase, mid-log phase, and stationary phase. Both LNA flow-FISH and qRT-PCR revealed that CsrB and TPP-2 were highly expressed during lag phase but markedly reduced in mid-log phase and stationary phase, whereas 6S demonstrated no to little expression during lag phase but increased thereafter. Importantly, while LNA flow-FISH and qRT-PCR demonstrated similar overall expression trends, only LNA flow-FISH, which enabled the detection of ncRNAs in individual cells as opposed to the lysate-based ensemble measurements generated by qRT-PCR, was able to capture the cell-to-cell heterogeneity in ncRNA expression. As such, this study demonstrates a new method that simultaneously enables the in situ detection of ncRNAs and the determination of gene expression heterogeneity within an isogenic bacterial population.

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“…Paper manufacturers have traditionally used biocides such as chlorine, bromine, isothiazolones, glutaraldehyde and others at different points of the process for various purposes, for example to preserve raw materials and filler slurries, to prevent the formation of biofilm deposits and bad odours or to avoid the corrosion of machine components (Torres et al 2008). …”

mentioning

confidence: 99%

Enzymatic treatment for preventing biofilm formation in the paper industry

Torres

Lenon

Craperi

et al. 2011

Appl Microbiol Biotechnol

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Microbiological control programmes at industrial level should aim at reducing both the detrimental effects of micro-organisms on the process and the environmental impact associated to the use of biocides as microbiological control products. To achieve this target, new efficient and environmentally friendly products are required. In this paper, seventeen non-specific, commercial enzymatic mixtures were tested to assess their efficacy for biofilm prevention and control at laboratory and pilot plant scale. Pectin methylesterase, an enzyme found in the formulation of two of the mixtures tested, was identified as an active compound able to reduce biofilm formation by 71% compared to control tests.

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Fluorescent in situ hybridization and flow cytometry as tools to evaluate the treatments for the control of slime-forming enterobacteria in paper mills

Abstract: Running title, Control of slime-forming enterobacteria using fluorescent in situ hybridization and flow cytometry.

Cited by 13 publications

References 29 publications

Novel, rapid DNA-based on-chip bacterial identification system combining dielectrophoresis and amplification-free fluorescent resonance energy transfer assisted in-situ hybridization (FRET-ISH)

Novel, rapid DNA-based on-chip bacterial identification system combining dielectrophoresis and amplification-free fluorescent resonance energy transfer assisted in-situ hybridization (FRET-ISH)

Locked Nucleic Acid and Flow Cytometry-Fluorescence In Situ Hybridization for the Detection of Bacterial Small Noncoding RNAs

Enzymatic treatment for preventing biofilm formation in the paper industry

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