Pyrrole-2-carboxylic acid inhibits biofilm formation and suppresses the virulence of Listeria monocytogenes.

Bacterial adhesion and biofilm formation of Listeria monocytogenes on food-contact surfaces result in serious safety concerns. This study aimed to explore the antibiofilm efficacy of pyrrole-2-carboxylic acid (PCA) against L. monocytogenes. Crystal violet staining assay demonstrated that PCA reduced the biofilm biomass of L. monocytogenes. The 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide reduction and flow cytometric assays indicated that PCA attenuated the metabolic activity of L. monocytogenes biofilm together with a decrease in viability. Morphologic observations revealed that PCA exposure collapsed the biofilm architecture. PCA administration of 0.75 mg ml-1 decreased the excretion of extracellular DNA, protein and polysaccharide by 48.58%, 61.60% and 75.63%, respectively. PCA failed to disperse the mature biofilm, even at 1.5 mg ml-1. However, PCA suppressed L. monocytogenes adhesion on common food-contact surfaces. Additionally, PCA exposure suppressed the hemolytic activity of L. monocytogenes. These findings suggested that PCA might serve as an alternative antibiofilm agent to control L. monocytogenes contamination.

Antibacterial Mechanism of 2R,3R-Dihydromyricetin Against Staphylococcus aureus: Deciphering Inhibitory Effect on Biofilm and Virulence Based on Transcriptomic and Proteomic Analyses.

Staphylococcus aureus is a major foodborne pathogen that leads to various diseases due to its biofilm and virulence factors. This study aimed to investigate the inhibitory effect of 2R,3R-dihydromyricetin (DMY), a natural flavonoid compound, on the biofilm formation and virulence of S. aureus, and to explore the mode of action using transcriptomic and proteomic analyses. Microscopic observation revealed that DMY could remarkably inhibit the biofilm formation by S. aureus, leading to a collapse on the biofilm architecture and a decrease in viability of biofilm cell. Moreover, the hemolytic activity of S. aureus was reduced to 32.7% after treatment with subinhibitory concentration of DMY (p < 0.01). Bioinformation analysis based on RNA-sequencing and proteomic profiling revealed that DMY induced 262 differentially expressed genes and 669 differentially expressed proteins (p < 0.05). Many downregulated genes and proteins related to surface proteins were involved in biofilm formation, including clumping factor A (ClfA), iron-regulated surface determinants (IsdA, IsdB, and IsdC), fibrinogen-binding proteins (FnbA, FnbB), and serine protease. Meanwhile, DMY regulated a wide range of genes and proteins enriched in bacterial pathogenesis, cell envelope, amino acid metabolism, purine and pyrimidine metabolism, and pyruvate metabolism. These findings suggest that DMY targets S. aureus through multifarious mechanisms, and especially prompt that interference of surface proteins in cell envelope would lead to attenuation of biofilm and virulence.

Ratiometric-enhanced G-Quadruplex Probes for Amplified and Mix-to-Read Detection of Mercury Pollution in Aquatic Products.

Mercury, as a global toxic pollutant, is easy to be accumulated in aquatic products and poses a great threat to human health. In this work, we proposed a mix-to-read, label-free, and robust assay for detecting mercury pollution in aquatic products by engineering a ratiometric-enhanced G-quadruplex probe. The transformation from the G-quadruplex to a hairpin-like structure allows us to confer a ratiometric and leveraged response to Hg2+, amplifying the signal-to-background ratio for Hg2+ detection. Hg2+ response was further improved by screening parallel- and antiparallel-, single-, and multiple-stranded G-quadruplex structures. Compared to the common aptamer probes, the ratiometric-enhanced G-quadruplex probe increased the sensitivity for Hg2+ detection by 4.7 times. This proposed sensing system allowed a simple and one-tube homogenous detection of Hg2+ at room temperature using a single unlabeled DNA sequence. Its application for Hg2+ detection in fish and shrimp conferred satisfactory recovery rates ranging from 98.5 to 105.9%. The label-free and mix-to-read assay is promising for the onsite detection of mercury pollution and facilitating food safety of aquatic products.