Abrasive treatment of microtiter plates improves the reproducibility of bacterial biofilm assays

Bordeleau, Emily; Mazinani, Sina Atrin; Nguyen, David; Betancourt, Frank; Yan, Hongbin

doi:10.1039/c8ra06352d

Cited by 9 publications

(4 citation statements)

References 34 publications

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“…While a majority of other in vitro biofilm-oriented tests apply various types of polished polystyrene/polypropylene inserts or plugs for this purpose [36], we hypothesized that a porous agar surface used in the AntiBioVol test may be more appropriate for enabling the cells' adhesion. The correlation between higher biofilm adherence and surface porosity has already been demonstrated by other research teams [37]. Indeed, as shown in Figure 6, the microorganisms used agar pores to anchor to the surface.…”

Section: Discussionsupporting

confidence: 70%

The Novel Quantitative Assay for Measuring the Antibiofilm Activity of Volatile Compounds (AntiBioVol)

et al. 2020

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Herein, we present a new test, dubbed AntiBioVol, to be used for the quantitative evaluation of antibiofilm activity of volatile compounds in vitro. AntiBioVol is performed in two 24-well plates using a basic microbiological laboratory equipment. To demonstrate AntiBioVol usability, we have scrutinized the activity of volatilized eucalyptus, tea tree, thyme essential oils, and ethanol (used for method suitability testing) against biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. We have also compared AntiBioVol with the standard disc volatilization method, placing a special stress on evaluating the impact of various technical parameters on the outcomes of the latter method. The obtained results indicate that AntiBioVol allows analyzing the antibiofilm activity of volatile compounds in a high number of repeats and provides semi-quantitative or quantitative results of high repeatability. In comparison to disc volatilization, AntiBioVol is a more space- and cost-effective method that allows analyzing various types of microbial aggregates. Moreover, we have indicated that the possible reasons for the discrepancies in the results obtained by means of the standard disc volatilization method may be related to various parameters of the testing dishes used (height, volume, diameter) and to various volumes of the agar medium applied. In turn, the application of a 24-well plate and a strictly defined AntiBioVol protocol provide a higher control of experimental conditions. Therefore, the application of AntiBioVol may enable an optimization of and introduction of volatile compounds to the fight against infective biofilms.

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Section: Discussionsupporting

confidence: 70%

The Novel Quantitative Assay for Measuring the Antibiofilm Activity of Volatile Compounds (AntiBioVol)

et al. 2020

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“…Such procedure may affect the different physiology of H. pylori cells, while it guarantees obtaining high cell biomass, which is difficult to obtain for H. pylori using classical methods. The porosity of surface (enabled in the experimental setting by application of agar-based medium) is a known factor increasing adherence of settled communities of microbes [ 50 ] and it allows to obtain more repeatable results than in case when smooth polystyrene and polypropylene surfaces are used for in vitro testing [ 51 ]. The proper surface porosity correlates with a higher adherence of cells, which in turn decreases the effect of aggressive pipette-based washing, i.e., the random de-attachment of large amounts of biofilm and/or multicellular aggregates [ 52 ].…”

Section: Discussionmentioning

confidence: 99%

Potential of Bacterial Cellulose Chemisorbed with Anti-Metabolites, 3-Bromopyruvate or Sertraline, to Fight against Helicobacter pylori Lawn Biofilm

Krzyżek

Gościniak

Fijałkowski

et al. 2020

IJMS

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Helicobacter pylori is a bacterium known mainly of its ability to cause persistent inflammations of the human stomach, resulting in peptic ulcer diseases and gastric cancers. Continuous exposure of this bacterium to antibiotics has resulted in high detection of multidrug-resistant strains and difficulties in obtaining a therapeutic effect. The purpose of the present study was to determine the usability of bacterial cellulose (BC) chemisorbed with 3-bromopyruvate (3-BP) or sertraline (SER) to act against lawn H. pylori biofilms. The characterization of BC carriers was made using a N2 adsorption/desorption analysis, tensile strength test, and scanning electron microscopy (SEM) observations. Determination of an antimicrobial activity was performed using a modified disk-diffusion method and a self-designed method of testing antibacterial activity against biofilm microbial forms. In addition, bacterial morphology was checked by SEM. It was found that BC disks were characterized by a high cross-linking and shear/stretch resistance. Growth inhibition zones for BC disks chemisorbed with 2 mg of SER or 3-BP were equal to 26.5–27.5 mm and 27–30 mm, respectively. The viability of lawn biofilm H. pylori cells after a 4-h incubation with 2 mg SER or 3-BP chemisorbed on BC disks was ≥4 log lower, suggesting their antibacterial effect. SEM observations showed a number of morphostructural changes in H. pylori cells exposed to these substances. Concluding, SER and 3-BP chemisorbed on BC carriers presented a promising antibacterial activity against biofilm H. pylori cells in in vitro conditions.

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“…The bio lm-forming ability (BFA) of the soil was determined by the method of Golby et al [11] with few modi cations. The at-bottom polystyrene 48-well plates were grinded [12] with autoclaved gravel (0.8-1.2mm) on a rotary shaker at 400rpm for 30 minutes, rinsed with 96% ethanol, and dried before use. The growth medium consisted of 2ml BG11 0 medium, 10µl MgSO 4 1M, 50µl glucose 22.2mM, 10µl nutrient broth, 50µl 100mM arginine, and 2.88ml of sterile distilled water.…”

Section: Full Textmentioning

confidence: 99%

Sound and rhythm impact on soil biofilm-forming ability

Fičko

Palijan

2023

Preprint

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Modern civilization is producing alarming levels of noise which often is developed in soil proximity having an opportunity to influence soil organisms. Yet, its effects on soil microbial communities are unexplored. To investigate the effects of different sound types we tested classical music of Mozart and Beethoven, construction, and road works noise and rhythm played on drums at the environmentally relevant sound levels and time intervals. All tested sounds impacted soil biofilm forming ability but differently between soil and land use types. Agricultural fields were less impacted by noise. Interestingly only regular rhythm stimulated all tested samples. Such results are the first evidence that noise can change the functional properties of soil microbial communities and that its effects are dependent on the type of sound.

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Abrasive treatment of microtiter plates improves the reproducibility of bacterial biofilm assays

Abstract: The reproducibility of bacterial biofilm assays is improved using abrasively-treated microtiter plates.

Cited by 9 publications

References 34 publications

The Novel Quantitative Assay for Measuring the Antibiofilm Activity of Volatile Compounds (AntiBioVol)

The Novel Quantitative Assay for Measuring the Antibiofilm Activity of Volatile Compounds (AntiBioVol)

Potential of Bacterial Cellulose Chemisorbed with Anti-Metabolites, 3-Bromopyruvate or Sertraline, to Fight against Helicobacter pylori Lawn Biofilm

Sound and rhythm impact on soil biofilm-forming ability

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