XTT assay for evaluating the effect of alcohols, hydrogen peroxide and benzalkonium chloride on biofilm formation of Staphylococcus epidermidis

Chaieb, Kamel; Zmantar, Tarek; Souiden, Yosra; Mahdouani, Kacem; Bakhrouf, Amina

doi:10.1016/j.micpath.2010.11.004

Cited by 38 publications

(19 citation statements)

References 29 publications

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“…The XTT assay has been frequently used for the quantification of bacteria and bacterial biofilms [42], [60] where it detects the presence of cells that are viable and metabolically active [42] regardless of being culturable. The viability of P. aeruginosa biofilms treated with a 20 kHz plasma discharge was evaluated using this method.…”

Section: Resultsmentioning

confidence: 99%

Eradication of Pseudomonas aeruginosa Biofilms by Atmospheric Pressure Non-Thermal Plasma

et al. 2012

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Bacteria exist, in most environments, as complex, organised communities of sessile cells embedded within a matrix of self-produced, hydrated extracellular polymeric substances known as biofilms. Bacterial biofilms represent a ubiquitous and predominant cause of both chronic infections and infections associated with the use of indwelling medical devices such as catheters and prostheses. Such infections typically exhibit significantly enhanced tolerance to antimicrobial, biocidal and immunological challenge. This renders them difficult, sometimes impossible, to treat using conventional chemotherapeutic agents. Effective alternative approaches for prevention and eradication of biofilm associated chronic and device-associated infections are therefore urgently required. Atmospheric pressure non-thermal plasmas are gaining increasing attention as a potential approach for the eradication and control of bacterial infection and contamination. To date, however, the majority of studies have been conducted with reference to planktonic bacteria and rather less attention has been directed towards bacteria in the biofilm mode of growth. In this study, the activity of a kilohertz-driven atmospheric pressure non-thermal plasma jet, operated in a helium oxygen mixture, against Pseudomonas aeruginosa in vitro biofilms was evaluated. Pseudomonas aeruginosa biofilms exhibit marked susceptibility to exposure of the plasma jet effluent, following even relatively short (∼10′s s) exposure times. Manipulation of plasma operating conditions, for example, plasma operating frequency, had a significant effect on the bacterial inactivation rate. Survival curves exhibit a rapid decline in the number of surviving cells in the first 60 seconds followed by slower rate of cell number reduction. Excellent anti-biofilm activity of the plasma jet was also demonstrated by both confocal scanning laser microscopy and metabolism of the tetrazolium salt, XTT, a measure of bactericidal activity.

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Section: Resultsmentioning

confidence: 99%

Eradication of Pseudomonas aeruginosa Biofilms by Atmospheric Pressure Non-Thermal Plasma

et al. 2012

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“…) and 2,3‐bis (2‐methoxy‐4‐nitro‐5‐sulfophenyl)‐5‐[(phenylamino)carbonyl]‐2 H ‐tetrazolium hydroxide (XTT) quantification (Chaieb et al . ) assays, respectively, on two randomly selected prewashed containers. Additionally, two sterile containers were treated with each assay as blanks.…”

Section: Methodsmentioning

confidence: 99%

“…Before treatments of L. monocytogenes monoculture biofilm, levels of biofilm biomass and viable cells within biofilm were quantified by crystal violet (CV) staining (Harvey et al 2007) and 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) quantification (Chaieb et al 2011) assays, respectively, on two randomly selected prewashed containers. Additionally, two sterile containers were treated with each assay as blanks.…”

Section: Quantification Of Initial Biofilm Levelmentioning

confidence: 99%

Combined effect of benzalkonium chloride and ultrasound againstListeria monocytogenesbiofilm on plastic surface

Torlak

Sert

2013

Lett Appl Microbiol

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Significance and Impact of the Study: Listeria monocytogenes is capable of adhering and forming biofilm on inert surfaces such as plastics. Biofilms are more resistant to disinfectants compared to planktonic cells, and this makes their elimination from food processing facilities a big challenge. Moreover, the emergence of resistant to disinfectants bacteria clearly shows that new biofilm control strategies are required. Among new strategies, the ultrasound treatment attracts attention as a nonchemical and environmentally friendly technology. This study demonstrated that the combination of ultrasound and benzalkonium chloride is a feasible strategy for removal of L. monocytogenes biofilms from plastic food-contact surfaces. AbstractThis study was performed to evaluate the effectiveness of benzalkonium chloride combined with ultrasound in eliminating Listeria monocytogenes biofilm from polystyrene surface. The test strain of L. monocytogenes, previously classified as strong biofilm producer, was grown to form biofilm in tryptic soy broth at 20°C for 6 days in polystyrene specimen containers. The biofilms formed on surface of containers were treated with ultrasound alone, benzalkonium chloride (100 and 400 mg l À1 ) alone and their combination for three different exposure times (1, 5 and 15 min) at room temperature. Sonication was performed using an ultrasound bath at a constant ultrasound frequency of 35 kHz. After treatments, levels of biofilm biomass and viable cells in biofilm were determined using crystal violet staining and XTT assays, respectively. The combined treatment of ultrasound and benzalkonium chloride resulted in significant (P < 0Á05) more decrease in the level of viable cells in the L. monocytogenes biofilm compared to individual treatments of benzalkonium chloride. Our results suggest that the combination of benzalkonium chloride with ultrasound is useful approach for the elimination of L. monocytogenes biofilms from plastic surfaces.

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“…Biofilm formation was interpreted as follows: Highly positive (OD570 ≥ 1), moderately to weakly positive (0.1 ≤ OD570 < 1), or negative (OD570 ≤ 0.1) [24].…”

Section: Adherence Assaymentioning

confidence: 99%

Interaction of Vibrio to Biotic and Abiotic Surfaces: Relationship between Hydrophobicity, Cell Adherence, Biofilm Production, and Cytotoxic Activity

2018

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Vibrio parahaemolyticus and Vibrio alginolyticus are important pathogenic agents for both humans and aquatic animals. Twenty-five bacterial strains were isolated from infected sea bass (Dicentrarchus labrax) on thiosulfate citrate bile salts sucrose (TCBS) agar plates. For the species-specific detection of V. alginolyticus and V. parahaemolyticus, a multiplex PCR assay using two collagenase-targeted primer pairs allows the detection of four strains of V. parahaemolyticus and three strains of V. alginolyticus. The seven identified isolates were partitioned for capsule production, hydrophobicity, adherence, biofilm formation, invasion, and cytotoxicity against Hep-2 cells. Two V. parahaemolyticus (Spa2 and Spa3) and one V. alginolyticus (Va01) were capsule producers developing almost black colonies on CRA, they showed a strong hydrophobicity using bacterial adhesion to hydrocarbons test (BATH), and were able to produce high biofilm. Isolates were able to adhere and invade Hep-2 cells and exhibited dissimilar levels of cytotoxicity in epithelial cells. This study shows the strong relationship between adhesion, biofilm formation, invasion and the cytotoxicity of Vibrio strains. Thus, we found a strong and significant positive correlation between different virulence properties of these isolates. The present study shows that bacterial contact with the cells as well as adhesion and invasion are essential steps to induce cytotoxicity. However, the invasion is seen to be a post adherence event.

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XTT assay for evaluating the effect of alcohols, hydrogen peroxide and benzalkonium chloride on biofilm formation of Staphylococcus epidermidis

Cited by 38 publications

References 29 publications

Eradication of Pseudomonas aeruginosa Biofilms by Atmospheric Pressure Non-Thermal Plasma

Eradication of Pseudomonas aeruginosa Biofilms by Atmospheric Pressure Non-Thermal Plasma

Combined effect of benzalkonium chloride and ultrasound againstListeria monocytogenesbiofilm on plastic surface

Interaction of Vibrio to Biotic and Abiotic Surfaces: Relationship between Hydrophobicity, Cell Adherence, Biofilm Production, and Cytotoxic Activity

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