An improved and versatile methodology to quantify biofilms formed on solid surfaces and exposed to the air–liquid interphase

Paytubi, Sònia; Guirado, Pedro; Balsalobre, Carlos; Madrid, Cristina

doi:10.1016/j.mimet.2014.05.020

Cited by 4 publications

(3 citation statements)

References 7 publications

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“…CV binds to negatively charged molecules and can be used to stain and quantify total biomass comprising bacteria and extracellular polymeric substances (EPS) within a biofilm (Stepanovic et al, 2000 ). CV staining protocol was performed as described by Paytubi et al ( 2014 ) with minor modifications. Briefly, plates were rinsed twice with distilled water and biofilms were fixed by heating at 80°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

A High-Throughput Screening Platform of Microbial Natural Products for the Discovery of Molecules with Antibiofilm Properties against Salmonella

et al. 2017

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In this report, we describe a High-Throughput Screening (HTS) to identify compounds that inhibit biofilm formation or cause the disintegration of an already formed biofilm using the Salmonella Enteritidis 3934 strain. Initially, we developed a new methodology for growing Salmonella biofilms suitable for HTS platforms. The biomass associated with biofilm at the solid-liquid interface was quantified by staining both with resazurin and crystal violet, to detect living cells and total biofilm mass, respectively. For a pilot project, a subset of 1120 extracts from the Fundación MEDINA's collection was examined to identify molecules with antibiofilm activity. This is the first validated HTS assay of microbial natural product extracts which allows for the detection of four types of activities which are not mutually exclusive: inhibition of biofilm formation, detachment of the preformed biofilm and antimicrobial activity against planktonic cells or biofilm embedded cells. Currently, several extracts have been selected for further fractionation and purification of the active compounds. In one of the natural extracts patulin has been identified as a potent molecule with antimicrobial activity against both, planktonic cells and cells within the biofilm. These findings provide a proof of concept that the developed HTS can lead to the discovery of new natural compounds with antibiofilm activity against Salmonella and its possible use as an alternative to antimicrobial therapies and traditional disinfectants.

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Section: Methodsmentioning

confidence: 99%

A High-Throughput Screening Platform of Microbial Natural Products for the Discovery of Molecules with Antibiofilm Properties against Salmonella

et al. 2017

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“…For biofilm on glass, qualitative assays were performed as described [ 27 ]. For quantitative assays on plastic, CFA cultures were grown using flat-bottomed 96-well polystyrene plates (Nalge Nunc International, Roskilde, Denmark).…”

Section: Methodsmentioning

confidence: 99%

Gre Factors Are Required for Biofilm Formation in Salmonella enterica Serovar Typhimurium by Targeting Transcription of the csgD Gene

et al. 2022

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Rdar biofilm formation of Salmonella typhimurium and Escherichia coli is a common ancient multicellular behavior relevant in cell–cell and inter-organism interactions equally, as in interaction with biotic and abiotic surfaces. With the expression of the characteristic extracellular matrix components amyloid curli fimbriae and the exopolysaccharide cellulose, the central hub for the delicate regulation of rdar morphotype expression is the orphan transcriptional regulator CsgD. Gre factors are ubiquitously interacting with RNA polymerase to selectively overcome transcriptional pausing. In this work, we found that GreA/GreB are required for expression of the csgD operon and consequently the rdar morphotype. The ability of the Gre factors to suppress transcriptional pausing and the 147 bp 5′-UTR of csgD are required for the stimulatory effect of the Gre factors on csgD expression. These novel mechanism(s) of regulation for the csgD operon might be relevant under specific stress conditions.

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“…Crystal violet assay was performed as previously described, [ 51,52 ] with minor modifications. Overnight cultures were diluted 1:100 in M9 medium with 0.4 or 4% glucose.…”

Section: Methodsmentioning

confidence: 99%

An Organic Electrochemical Transistor to Monitor Salmonella Growth in Real‐Time

Butina

Filipović

Richter‐Dahlfors

et al. 2021

Adv Materials Inter

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Organic electrochemical transistors (OECTs) are used in research and diagnostic applications due to their facile manufacture, scalability, and biocompatibility. In these devices, the source–drain current upon gate voltage application depends on ion concentration in the electrolyte. This study investigates whether an OECT can be employed to monitor bacterial growth since it is known that the concentration of charged species increases in bacterial cultures during growth. A poly(3,4‐ethylenedioxythiophene):polystyrene sulfonate‐based single‐well OECT, compatible with long‐term incubation of bacterial cultures, is fabricated. It is shown that the growth of Salmonella alters the transfer characteristics of the device and demonstrates how it can be applied to monitor growth in real‐time by recording the source–drain current at gate voltage +0.5 V. The signal can also be measured in filtrates of bacterial cultures, devoid of bacterial cells. This suggests that the signal originates from charged metabolic products. Bacterial biofilm formation does not alter the device response. This proof‐of‐principle study presents OECT recordings as an alternative to optical methods, allowing bacterial growth to be monitored in transparent and opaque media alike. By measuring metabolic products rather than bacterial cell multiplication, insight into the stationary phase and other nondividing states may be obtained in the future.

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An improved and versatile methodology to quantify biofilms formed on solid surfaces and exposed to the air–liquid interphase

Cited by 4 publications

References 7 publications

A High-Throughput Screening Platform of Microbial Natural Products for the Discovery of Molecules with Antibiofilm Properties against Salmonella

A High-Throughput Screening Platform of Microbial Natural Products for the Discovery of Molecules with Antibiofilm Properties against Salmonella

Gre Factors Are Required for Biofilm Formation in Salmonella enterica Serovar Typhimurium by Targeting Transcription of the csgD Gene

An Organic Electrochemical Transistor to Monitor Salmonella Growth in Real‐Time

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