Biofilm Specific Activity: A Measure to Quantify Microbial Biofilm

Corte, Laura Della; Pierantoni, Debora Casagrande; Tascini, Carlo; Roscini, Luca; Cardinali, Gianluigi

doi:10.3390/microorganisms7030073

Cited by 47 publications

(25 citation statements)

References 45 publications

(59 reference statements)

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“…One‐day‐old biofilm mass was quantified with the crystal violet assay; metabolic activity was examined with the XTT assay as previously described by O'Toole (2011) and Hawser et al (1996), respectively. The crystal violet and XTT measures of biofilm formation do not necessarily correlate and can be considered two independent characteristics of a biofilm . Therefore, isolates were categorised either based on their level of biomass or based on metabolic activity distribution as low, intermediate or high biofilm producers according to a study published by Rajendran et al (2016) .…”

Section: Methodsmentioning

confidence: 99%

“…The crystal violet and XTT measures of biofilm formation do not necessarily correlate and can be considered two independent characteristics of a biofilm. 13,17 Therefore, isolates were categorised either based on their level of biomass or based on metabolic activity distribution as low, intermediate or high biofilm producers according to a study published by Rajendran et al (2016). 10 Isolates within the first quartile (Q1) were classified as low biofilm producers, isolates with a biomass higher than the third quartile (Q3) were classified as high biofilm formers and those in between were classified as intermediate biofilm producers (Q2).…”

Section: Biofilm Formationmentioning

confidence: 99%

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Candida biofilm production is associated with higher mortality in patients with candidaemia

Vitális

Nagy

Tóth

et al. 2020

Mycoses

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Summary Background Candidaemia is a common life‐threatening disease among hospitalised patients, but the effect of the Candida biofilm‐forming ability on the clinical outcome remains controversial. Objective The aim was to determine the impact of biofilms, specifically focusing on biofilm mass and metabolic activity, on the mortality in candidaemia. Patients/Methods The clinical data of patients (n = 127) treated at the University of Debrecen, Clinical Centre, between January 2013 and December 2018, were investigated retrospectively. Biofilm formation was assessed using the crystal violet and XTT assays, measuring the biofilm mass and metabolic activity, respectively. Isolates were classified as low, intermediate and high biofilm producers both regarding biofilm mass and metabolic activity. The susceptibility of one‐day‐old biofilms to fluconazole, amphotericin B, anidulafungin, caspofungin and micafungin was evaluated and compared to planktonic susceptibility. Results Intermediate/high biofilm mass was associated with significantly higher mortality (61%). All Candida tropicalis, Candida parapsilosis and Candida glabrata isolates originating from fatal infections were intermediate/high biofilm producers, whereas this ratio was 85% for Candida albicans. Solid malignancy was associated with intermediate/high biofilm producers (P = .043). The mortality was significantly higher in infections caused by Candida strains producing biofilms with intermediate/high metabolic activity (62% vs. 33%, P = .010). The ratio of concomitant bacteraemia was higher for isolates forming biofilms with low metabolic activity (53% vs 28%, P = .015). Conclusions This study provides evidence that the Candida biofilms especially with intermediate/high metabolic activity are related to higher mortality in candidaemia.

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Section: Methodsmentioning

confidence: 99%

Section: Biofilm Formationmentioning

confidence: 99%

Candida biofilm production is associated with higher mortality in patients with candidaemia

Vitális

Nagy

Tóth

et al. 2020

Mycoses

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“…Crystal violet (CV) and safranin (SAF) assays are commonly used for biomass quantification in biofilm-based research of Staphylococcus spp. [83][84][85]. Both methods are based on the use of specific dyes that penetrate through the biomass of pre-grown biofilms.…”

Section: Minimal Biofilm Eradication/eliminating Concentration Assaysmentioning

confidence: 99%

Methods Used for the Eradication of Staphylococcal Biofilms

et al. 2019

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Staphylococcus aureus is considered one of the leading pathogens responsible for community and healthcare-associated infections. Among them, infections caused by methicillin-resistant strains (MRSA) are connected with ineffective or prolonged treatment. The therapy of staphylococcal infections faces many difficulties, not only because of the bacteria’s resistance to antibiotics and the multiplicity of virulence factors it produces, but also due to its ability to form a biofilm. The present review focuses on several approaches used for the assessment of staphylococcal biofilm eradication. The methods described here are successfully applied in research on the prevention of biofilm-associated infections, as well as in their management. They include not only the evaluation of the antimicrobial activity of novel compounds, but also the methods for biomaterial functionalization. Moreover, the advantages and limitations of different dyes and techniques used for biofilm characterization are discussed. Therefore, this review may be helpful for those scientists who work on the development of new antistaphylococcal compounds.

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“…Non-invasive measurements of activity in biofilms are not trivial, as many current methods rely on disrupting the biofilm and altering the chemical microenvironment, though some genetic (Poulsen et al, 1993;Kragh et al, 2014) and chemical reporters do exist (Corte et al, 2019). Most of these methods, however, only cover part of the metabolic output linked to, e.g., growth or production of molecules produced under certain conditions (Whooley and McLoughlin, 1982;Poulsen et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Metabolic flux fingerprinting differentiates planktonic and biofilm states ofPseudomonas aeruginosaandStaphylococcus aureus

Lichtenberg¹,

Kragh

Fritz³

et al. 2020

Preprint

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The challenges of defining the biofilm phenotype has been clear for decades. Many biomarkers for biofilm are known, but methods for identifying these are often invasive and/or complicated. These methods often rely on disrupting the biofilm matrix or examining virulence factors and compounds, which may only be expressed under certain conditions.We used microcalorimetric measurements of metabolic energy release to investigate whether unchallenged, planktonic Pseudomonas aeruginosa displayed differences in metabolism compared to surface-bound and non-attached biofilms.The pattern of energy release observed in the recorded microcalorimetric thermograms clearly depended on growth state, though the total energy expenditure was not different between growth states. To characterize these differences, we developed a classification pipeline utilizing machine learning algorithms to classify growth state, based on the observed patterns of energy release. With this approach, we could with high accuracy detect the growth form of blinded samples. To challenge the algorithm, we attempted to limit the amount of training data. By training the algorithm with only a single data point from each growth form, we obtained a mean accuracy of 90.5% using two principal components. Further validation of the classification pipeline showed that the approach was not limited to P. aeruginosa but could also be used for detection of gram-positive Staphylococcus aureus biofilm. We propose that microcalorimetric measurements, in combination with this new quantitative framework, can be used as a non-invasive biomarker to detect the presence of biofilm.These results could have a significant potential in clinical settings where the detection of biofilms in infections often means a different outcome and treatment regime for the patient.

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Biofilm Specific Activity: A Measure to Quantify Microbial Biofilm

Cited by 47 publications

References 45 publications

Candida biofilm production is associated with higher mortality in patients with candidaemia

Candida biofilm production is associated with higher mortality in patients with candidaemia

Methods Used for the Eradication of Staphylococcal Biofilms

Metabolic flux fingerprinting differentiates planktonic and biofilm states ofPseudomonas aeruginosaandStaphylococcus aureus

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