Abstract

Foodborne pathogens are known to adhere strongly to surfaces and can form biofilms in food processing facilities; therefore, their potential to contaminate manufactured foods underscores the importance of sanitation. The objectives of this study were to (1) examine the efficacy of a new-generation sanitizer (Decon7) on Staphylococcus and Pseudomonas biofilms, (2) identify biofilm bacteria from workers’ boots in relation to previous sanitizer chemistry, (3) validate the efficacy of Decon7 on biofilm from workers’ boots from an abattoir/food processing environment, and (4) compare the sensitivity of isolated boot biofilm bacteria to new- and early (Bi-Quat)-generation QAC sanitizers. Decon7 was applied at two concentrations (5%, 10%) and was shown to be effective within 1 min of exposure against enhanced biofilms of Staphylococcus spp. and Pseudomonas spp. in 96-well microplates. Decon7 was also used to treat workers’ boots that had accumulated high levels of biofilm bacteria due to ineffective sanitization. Bacteria isolated before enzyme/sanitizer treatment were identified through 16S rRNA PCR and DNA sequencing. All treatments were carried out in triplicate and analyzed by one-way RM-ANOVA or ANOVA using the Holm–Sidak test for pairwise multiple comparisons to determine significant differences (p < 0.05). The data show a significant difference between Decon7 sanitizer treatment and untreated control groups. There was a ~4–5 log reduction in Staphylococcus spp. and Pseudomonas spp. (microplate assay) within the first 1 min of treatment and also a > 3-log reduction in the bacterial population observed in the biofilms from workers’ boots. The new next-generation QAC sanitizers are more effective than prior QAC sanitizers, and enzyme pre-treatment can facilitate biofilm sanitizer penetration on food contact surfaces. The rotation of sanitizer chemistries may prevent the selective retention of chemistry-tolerant microorganisms in processing facilities.

Highlights

  • IntroductionBiofilms are a mixture of bacterial cells and organic material attached to surfaces [1]

  • Biofilms are a mixture of bacterial cells and organic material attached to surfaces [1].Most biofilms evolve through a series of steps, involving initial microbial attachment, biofilm development and proliferation, followed by detachment and dispersal

  • In our prior work, untreated microplates were used to examine the innate ability of strains of Listeria monocytogenes isolated from food processing plants to form biofilms [19,32,33]

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Summary

Introduction

Biofilms are a mixture of bacterial cells and organic material attached to surfaces [1]. Most biofilms evolve through a series of steps, involving initial microbial attachment, biofilm development and proliferation, followed by detachment and dispersal. Any microorganism can initiate attachment to surfaces via Van der Waals interactions and electrostatic charges from numerous chemical constituents on its surface, including pili, fimbriae, flagella, lipopolysaccharides, and other proteins and carbohydrates [2]. The most important factor is the adherence provided by extracellular polysaccharides (EPS), which acts as a ‘glue’ for the biofilm matrix and may even trap other microorganisms that are not actively involved in biofilm formation [3,4]. The bacterial EPS may limit the transfer of nutrients and chemicals into the biofilm, which must diffuse through the 3-dimensional matrix to supply the embedded cells [5,6]. The disparity in the nutrient availability of cells on/near

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