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other·research methods, gastroenterology, food science, preclinical research·PMC10030034

Evaluating the efficacy of non-thermal microbial load reduction treatments of heat labile food components for in vitro fermentation experiments

PLOS ONE · 2 authors, 2 centres

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TL;DR

Pulse flours from four sources were treated with bleach, alcohol, or hydrogen peroxide at varying concentrations to reduce microbial contamination for in vitro simulated colon fermentation. All treatments significantly reduced microbial loads, and bleach and hydrogen peroxide treated flours showed minimal differences from untreated controls in microbial composition and short chain fatty acid production.

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Liquid chemical sterilant alternatives to autoclaving were evaluated for treating pulse flours from green lentil, field pea, chickpea, and sprouted green lentile prior to in vitro simulated colon fermentation. Bleach, alcohol, and hydrogen peroxide were tested at varying concentrations, with microbial reduction assessed by plate counts, treated substrate integrity evaluated by scanning electron microscopy, and fermentation impact measured by microbial community composition and short chain fatty acid production. All treatments significantly reduced microbial loads, though 70% alcohol did not reduce counts below detectable levels. Simulated colon fermentations of bleach- and hydrogen peroxide-treated pulses showed minimal differences from untreated controls in both microbial composition and short chain fatty acid production. SEM showed minimal impact on gross flour structure. Alcohol did not fully eliminate microbes, and some sterilization treatments exhibited a minor shift in community structure. These results suggest bleach and hydrogen peroxide are effective nonthermal methods for eliminating contaminating microbes without significantly damaging fermentable substrates.

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other

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research methods

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ORIGINAL PAPER

PMC10030034

Evaluating the efficacy of non-thermal microbial load reduction treatments of heat labile food components for in vitro fermentation experiments

PLOS ONE

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ABSTRACT

Increasingly, in vitro simulated colon fermentations are being used as a pre-clinical step to assess the impacts of foods and drugs on the gut microbiota in a cost-effective manner. One challenge in such systems is that they are potentially susceptible to the influences of contaminating microbes in test materials. Simulated gastric and intestinal digestion can relieve some of these concerns, however, live microbes may remain that can confound analysis. Autoclave treatment of test materials is the surest way to eliminate these microbes but presents problems when using heat labile components such as resistant starch. In this study, liquid chemical sterilant alternatives to moist heat sterilization were explored for treating pulse flours for use during in vitro simulated colon fermentation. Key attributes considered in chemical selection were accessibility, impact on treated food components, and effectiveness of the treatments for reducing microbial load. Three chemicals were selected for evaluation, bleach, alcohol, and hydrogen peroxide, at varying concentrations. Flours chosen for testing were from green lentil, field pea, chickpea, or sprouted green lentil. All treatments significantly reduced microbial loads, though there were still detectable levels of microbes after alcohol treatments. Furthermore, in vitro simulated colon fermentations of the treated pulses showed minimal difference from the untreated control both in terms of microbial composition and short chain fatty acid production. Scanning electron microscopy showed minimal impact of sterilization treatments on the gross structure of the pulse flours. Together these results suggest that bleach and hydrogen peroxide treatments can be effective nonthermal treatments to eliminate contaminating microbes in pulse flours without causing significant damage to starch and other fermentable substrates. This is thus also a promising treatment method for other starchy food substrates, though further testing is required.

CITATION EXPORT

Andrew Paff, Darrell W. Cockburn. Evaluating the efficacy of non-thermal microbial load reduction treatments of heat labile food components for in vitro fermentation experiments. PLOS ONE. (2023). https://doi.org/10.1371/journal.pone.0283287

AUTHORS

2 · 2 CENTRES

Andrew Paff, Darrell W. Cockburn

Department of Food Science, College of Agricultural Sciences, The Pennsylvania State University, University Park, Pennsylvania, United States of America
University of Illinois Urbana-Champaign, UNITED STATES

EDITOR / REVIEWER

Brenda A. Wilson