Abstract
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.
Highlights
Biofilms are defined as complex microbial communities embedded in a self-produced extracellular polymeric matrix that attach to surfaces and are the predominant mode of microbial growth in nature (Steenackers et al, 2012)
It has been estimated that the minimum concentration of biocide required to kill bacteria within a biofilm may be up to 100–1000 times higher compared to planktonic cells (Costerton et al, 1995; Stoodley et al, 2002; Høiby et al, 2010)
We describe the development of a robust and validated experimental approach using a High-Throughput Screening (HTS) platform to identify natural compounds that either can inhibit the formation or can eliminate or disperse S. enterica preformed biofilms
Summary
Biofilms are defined as complex microbial communities embedded in a self-produced extracellular polymeric matrix that attach to surfaces and are the predominant mode of microbial growth in nature (Steenackers et al, 2012). Natural Antibiofilm Molecules against Salmonella might provide a reservoir of pathogenic bacteria, increasing the risk of microbial contamination, and leading to critical problems in terms of public health and economical lost (Shi and Zhu, 2009). They are recalcitrant, highly resistant to antimicrobials and disinfectants and are extremely difficult to eradicate (O’Toole et al, 2000; Bridier et al, 2011). Since most antimicrobials currently in use show a poor activity against bacteria embedded in biofilms (Høiby et al, 2010), search for new molecules with antibiofilm properties is a thrilling challenge
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