Effect of Sub-Lethal Concentrations of Biocides on the Structural Parameters and Viability of the Biofilms Formed by <i>Salmonella</i> Typhimurium

Capita, Rosa; Buzón-Durán, Laura; Riesco-Peláez, Félix; Alonso‐Calleja, Carlos

doi:10.1089/fpd.2016.2241

Cited by 39 publications

(29 citation statements)

References 12 publications

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“…Structural parameter values in the present work are also similar to those previously observed for biofilms formed on polystyrene by a S . Typhimurium isolate from food (S175), as recorded by Capita et al [ 2 ]. It should be noted that in both the above-mentioned papers only fluorescent dyes for staining nucleic acids were used, and the EPS matrix was not studied.…”

Section: Discussionmentioning

confidence: 99%

“…Salmonella has the ability to form biofilms, which have been identified as an important factor for the persistence of food-borne pathogens in food-processing environments [ 2 , 3 , 4 ]. Salmonella Agona strains have been classified as strong biofilm producers [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

“…Thus, the quantitative three-dimensional results recorded by CLSM are an important basis for understanding, controlling and modelling of biofilms formed by pathogenic bacteria. In previous work, the structural parameters and cellular viability of 48-hour-old Salmonella Typhimurium biofilms were studied using CLSM [ 2 ]. However, it would appear that the individual cellular and EPS matrix components of biofilms formed by Salmonella at different stages of development have not hitherto been visualized or quantitatively characterized.…”

Section: Introductionmentioning

confidence: 99%

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Visualization and quantification of the cellular and extracellular components of Salmonella Agona biofilms at different stages of development

et al. 2018

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Salmonella is a major food-borne pathogen able to persist in food processing environments because of its ability to form biofilms. A Salmonella enterica serotype Agona isolate from poultry (S24) was grown at 37°C in biofilms for up to 144 hours (H144) in attachment to polystyrene surfaces. Biofilm structures were examined at different stages in their development (H3, H24, H48, H72, H96 and H144) using confocal laser scanning microscopy (CLSM) in conjunction with fluorescent dyes for live cells (SYTO 9), dead cells (propidium iodide), proteins (fluorescein isothiocyanate isomer I), lipids (DiD’oil), α-polysaccharides (concanavalin A, tetramethylrhodamine conjugate), and β-polysaccharides (calcofluor white M2R). Strain S24 developed a robust biofilm at H72 (biovolume of 166,852.5 ± 13,681.8 μm3 in the observation field of 16,078.2 μm2). The largest biovolume of live cells was also detected at H72 (128,110.3 ± 4,969.1 μm3), decreasing thereafter, which was probably owing to the detachment of cells prior to a new phase of colonization. The percentage of dead cells with regard to total cells in the biofilms increased throughout the incubation, ranging from 2.3 ± 1.1% (H24) to 44.2 ± 11.0% (H144). Proteins showed the greatest biovolume among the extracellular components within the biofilms, with values ranging from 1,295.1 ± 1,294.9 μm3 (H3) to 19,186.2 ± 8,536.0 μm3 (H96). Maximum biovolume values of 15,171.9 ± 660.7 μm3 (H48), 7,055.3 ± 4,415.2 μm3 (H144), and 2,548.6 ± 1,597.5 μm3 (H72) were observed for β-polysaccharides, α-polysaccharides and lipids, respectively. A strong (P < 0.01) positive correlation was found between the total biovolume of biofilm and the biovolume of live cells, proteins and β-polysaccharides, which may serve as useful markers of biofilm formation. The present work provides new insights into the formation of S. Agona biofilms. Our findings may contribute to the designing of reliable strategies for preventing and removing these bacterial communities.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Visualization and quantification of the cellular and extracellular components of Salmonella Agona biofilms at different stages of development

et al. 2018

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“…The formation of biofilms is a survival strategy adopted by bacterial cells in response to the damage caused by environmental stressors, such as sub-lethal concentrations of antimicrobial and oxidizing agents [ 146 , 147 , 148 ]. The molecular mechanisms involved in biofilm stimulation by these stressors have not been elucidated.…”

Section: Adaptive Response Of Gram-negative Cells To Hoclmentioning

confidence: 99%

Surviving Reactive Chlorine Stress: Responses of Gram-Negative Bacteria to Hypochlorous Acid

2020

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Sodium hypochlorite (NaOCl) and its active ingredient, hypochlorous acid (HOCl), are the most commonly used chlorine-based disinfectants. HOCl is a fast-acting and potent antimicrobial agent that interacts with several biomolecules, such as sulfur-containing amino acids, lipids, nucleic acids, and membrane components, causing severe cellular damage. It is also produced by the immune system as a first-line of defense against invading pathogens. In this review, we summarize the adaptive responses of Gram-negative bacteria to HOCl-induced stress and highlight the role of chaperone holdases (Hsp33, RidA, Cnox, and polyP) as an immediate response to HOCl stress. We also describe the three identified transcriptional regulators (HypT, RclR, and NemR) that specifically respond to HOCl. Besides the activation of chaperones and transcriptional regulators, the formation of biofilms has been described as an important adaptive response to several stressors, including HOCl. Although the knowledge on the molecular mechanisms involved in HOCl biofilm stimulation is limited, studies have shown that HOCl induces the formation of biofilms by causing conformational changes in membrane properties, overproducing the extracellular polymeric substance (EPS) matrix, and increasing the intracellular concentration of cyclic-di-GMP. In addition, acquisition and expression of antibiotic resistance genes, secretion of virulence factors and induction of the viable but nonculturable (VBNC) state has also been described as an adaptive response to HOCl. In general, the knowledge of how bacteria respond to HOCl stress has increased over time; however, the molecular mechanisms involved in this stress response is still in its infancy. A better understanding of these mechanisms could help understand host-pathogen interactions and target specific genes and molecules to control bacterial spread and colonization.

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“…It was observed that contact between biofilms and BZK increased the amount of biomass (markedly so for concentrations of 2500 ppm). An enhanced ability to form a biofilm in the presence of BZK has previously been demonstrated for strains of Escherichia coli [72], methicillin-resistant Staphylococcus aureus (MRSA) [73], Salmonella [71,74], and L. monocytogenes [14]. In the present study, the biofilms were exposed to BZK for five minutes and the biocide was then eliminated, although it is probable that residual quantities remained in the wells of the microtiter plate.…”

Section: The Effect Of Disinfectants On Biofilmsmentioning

confidence: 70%

Persistent Listeria monocytogenes Isolates from a Poultry-Processing Facility Form More Biofilm but Do Not Have a Greater Resistance to Disinfectants than Sporadic Strains

et al. 2019

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Some strains of Listeria monocytogenes can persist in food-processing environments, increasing the likelihood of the contamination of foodstuffs. To identify traits that contribute to bacterial persistence, a selection of persistent and sporadic L. monocytogenes isolates from a poultry-processing facility was investigated for biofilm-forming ability (crystal violet assay). The susceptibility of sessile cells to treatments (five minutes) with sodium hypochlorite having 10% active chlorine (SHY: 10,000 ppm, 25,000 ppm, and 50,000 ppm) and benzalkonium chloride (BZK: 2500 ppm, 10,000 ppm, and 25,000 ppm) was also studied. All isolates exhibited biofilm formation on polystyrene. Persistent strains showed larger (p < 0.001) biofilm formation (OD580 = 0.301 ± 0.097) than sporadic strains (OD580 = 0.188 ± 0.082). A greater susceptibility to disinfectants was observed for biofilms of persistent strains than for those of sporadic strains. The application of SHY reduced biofilms only for persistent strains. BZK increased OD580 in persistent strains (2500 ppm) and in sporadic strains (all concentrations). These results indicate that the use of BZK at the concentrations tested could represent a public health risk. Findings in this work suggest a link between persistence and biofilm formation, but do not support a relationship between persistence and the resistance of sessile cells to disinfectants.

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Effect of Sub-Lethal Concentrations of Biocides on the Structural Parameters and Viability of the Biofilms Formed by Salmonella Typhimurium

Cited by 39 publications

References 12 publications

Visualization and quantification of the cellular and extracellular components of Salmonella Agona biofilms at different stages of development

Visualization and quantification of the cellular and extracellular components of Salmonella Agona biofilms at different stages of development

Surviving Reactive Chlorine Stress: Responses of Gram-Negative Bacteria to Hypochlorous Acid

Persistent Listeria monocytogenes Isolates from a Poultry-Processing Facility Form More Biofilm but Do Not Have a Greater Resistance to Disinfectants than Sporadic Strains

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