Dataset Information

Microfluidic droplet application for bacterial surveillance in fresh-cut produce wash waters.

ABSTRACT: Foodborne contamination and associated illness in the United States is responsible for an estimated 48 million cases per year. Increased food demand, global commerce of perishable foods, and the growing threat of antibiotic resistance are driving factors elevating concern for food safety. Foodborne illness is often associated with fresh-cut, ready-to-eat produce commodities due to the perishable nature of the product and relatively minimal processing from farm to the consumer. The research presented here optimizes and evaluates the utility of microfluidic droplets, also termed ultra-miniaturized bioreactors, for rapid detection of viable Salmonella enterica ser. Typhimurium in a shredded lettuce wash water acquired from a major Mid-Atlantic produce processing facility (denoted as Producer) in the U.S. Using a fluorescently-labeled anti-S. Typhimurium antibody and relative fluorescence intensities, paired with in-droplet incubation, S. Typhimurium was detected and identified with 100% specificity in less than 5 h. In initial optimization experiments using S. Typhimurium-spiked sterile water, the relative fluorescence intensity of S. Typhimurium was approximately two times that of the observed relative intensities of five non-S. Typhimurium negative controls at 4-h incubation in droplets containing Rappaport-Vasiliadis (RV) broth at 37°C: relative fluorescence intensity for S. Typhimurium = 2.36 (95% CI: 2.15-2.58), Enterobacter aerogens 1.12 (95% CI: 1.09-1.16), Escherichia coli 700609 = 1.13 (95% CI: 1.09-1.17), E. coli 13706 1.13 (95% CI: 1.07-1.19), E. coli 700891 1.05 (95% CI: 1.03-1.07) and Citrobacter freundii 1.04 (95% CI: 1.03-1.05). S. Typhimurium- and E. aerogens-spiked shredded lettuce wash waters acquired from the Producer were then incubated 4 h in-droplet at 37°C with RV broth. The observed relative fluorescence of S. Typhimurium was significantly higher than that of E. aerogens, 1.56 (95% CI: 1.42-1.71) and 1.10 (95% CI: 1.08-1.12), respectively. While further optimization focusing on compatible concentration methodologies for highly-dilute produce water samples is needed, this application of droplet microfluidics shows great promise in dramatically shortening the time necessary-from days to hours-to confirm viable bacterial contamination in ready-to-eat produce wash waters used throughout the domestic and international food industry.

SUBMITTER: Harmon JB

PROVIDER: S-EPMC7282644 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Microfluidic droplet application for bacterial surveillance in fresh-cut produce wash waters.

Harmon J Brian JB Gray Hannah K HK Young Charles C CC Schwab Kellogg J KJ

PloS one 20200609 6

Foodborne contamination and associated illness in the United States is responsible for an estimated 48 million cases per year. Increased food demand, global commerce of perishable foods, and the growing threat of antibiotic resistance are driving factors elevating concern for food safety. Foodborne illness is often associated with fresh-cut, ready-to-eat produce commodities due to the perishable nature of the product and relatively minimal processing from farm to the consumer. The research prese ...[more]

PMID: 32516315

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Project description:Whole and fresh-cut produce are minimally processed and susceptible to microbial contamination. This study evaluated the survival or growth of L. monocytogenes on peeled rinds, and fresh-cut produce at different storage temperatures. Fresh-cut fruits and vegetables, including cantaloupe, watermelon, pear, papaya, pineapple, broccoli, cauliflower, lettuce, bell pepper, and kale (25 g pieces) were spot inoculated with 4 log CFU/g of L. monocytogenes and stored at 4 or 13°C for 6 days. Cantaloupe and bell pepper rind disks (20 cm2), mimicking whole produce were inoculated with low inoculum level (4 log CFU/mL) and high inoculum level (6 log CFU/mL) and stored at 24°C up to 8 days and 4°C up to 14 days, respectively. L. monocytogenes counts on fresh-cut pear samples stored at 4°C increased significantly by 0.27 log CFU/g. However, Listeria levels on kale (day 4), cauliflower (day 6), and broccoli (day 2) were significantly reduced by 0.73, 1.18, and 0.80 log CFU/g, respectively, at 4°C. At 13°C, the bacterial counts increased significantly after a day of storage on fresh-cut watermelons (increasing by 1.10 log CFU/g) and cantaloupes (increasing by 1.52 log CFU/g). Similar increases were observed on pears (1.00 log CFU/g), papayas (1.65 log CFU/g), and green bell peppers (1.72 log CFU/g). Pineapple samples did not support the growth of L. monocytogenes at 13°C with a significant reduction of 1.80 log CFU/g by day 6. L. monocytogenes levels significantly increased in fresh-cut lettuce at 13°C but remained stable on kale, cauliflower, and broccoli after 6 days of storage. Stable population was observed also on cantaloupe rinds up to 8 days at 24°C. While on the outer surface of bell peppers, the population level decreased below the detectable limit of the test (10 CFU/20 cm2) after 14 days of storage at 4°C. The results demonstrated variable survival behavior of L. monocytogenes on fresh-cut produce with produce type and storage temperature.

Dataset Information

Microfluidic droplet application for bacterial surveillance in fresh-cut produce wash waters.

Publications

Microfluidic droplet application for bacterial surveillance in fresh-cut produce wash waters.

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