Application of Sample6 DETECT™ HT/L Kit for the Detection of Listeria in Mixed Leafy Greens.

BACKGROUND: Early and accurate detection of Listeria in foods is vital. Current methods require 24 h enrichment for detection. OBJECTIVE: This study aimed to demonstrate enhanced early detection of Listeria in mixed leafy greens using Sample6 DETECT™ HT/L, a phage-based detection system. METHODS: A method comparison between the reference method (U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 10) for the detection of Listeria spp. and the Sample6 DETECT HT/L using a new proprietary R2 Medium was performed in mixed leafy greens. RESULTS: Using the R2 Medium enrichment with Sample6 DETECT HT/L, detection of L. innocua was possible at 12 h (with a centrifugation step), and L. monocytogenes was detected by 18 h, with equivalent performance as the 24 h enrichments using the reference method detection. The Sample6 DETECT HT/L gave an equivalent performance for L. innocua at 15 h as the reference method at 24 h. Detection was accomplished by the addition of reagents in the kit, following the package insert, and reading results in a detection chamber using a 96-well plate reader that detects a fluorescent signal. CONCLUSIONS: Results indicate the new R2 Medium and Sample6 DETECT HT/L allowed for earlier detection of Listeria spp. in mixed leafy greens. HIGHLIGHTS: Sample6 DETECT HT/L with the new R2 Medium allowed the early detection of Listeria spp. and may be applied in other food matrices and environmental samples.

Thermal and Chemical Treatments To Reduce Salmonella on Alfalfa (Medicago sativa) and Broccoli (Brassica oleracea var. italica) Seeds before and during the Sprouting Process: A Hurdle Approach.

Sprouts are vehicles of foodborne diseases caused by pathogens such as Salmonella. The aim of this study was to evaluate thermal and chemical treatments applied as a hurdle approach to reduce Salmonella in alfalfa (Medicago sativa) and broccoli (Brassica oleracea var. italica) seeds before and during their germination. Seeds, inoculated and then dried at 55°C for 48 h, were subjected to a chemical treatment and a thermal shock with (i) 75 mM caprylic acid at 70°C for 5 s, (ii) 0.04% CaO at 70°C for 5 s, or (iii) 1% H2O2 at 70°C for 5 s. After each treatment, seeds were immersed in water at 3°C for 5 s. Next, the imbibition process was carried out with 0.016% H2O2 at pH 3.0. Finally, the seeds were transferred to a rotary drum-type germinator and were sprayed with the same chemical solution that was applied before the imbibition process, for 20 s at intervals of 5 min for 40 min at 3 rpm. All chemical treatments reduced Salmonella at least 5 log CFU/g on both seeds. Germination rates between 90 and 93% were obtained after application of thermal and chemical treatments. Salmonella was not detected after the imbibition stage when caprylic acid and H2O2 treatments were applied. However, during the germination process of both seeds, Salmonella counts of >6 log CFU/g were obtained despite all treatments being applied at different stages of the sprouting process. These results demonstrated that thermal and chemical treatments used as a hurdle approach to control Salmonella on alfalfa and broccoli seeds significantly reduced the pathogen concentration on seeds >5 log but were ineffective to eliminate Salmonella and to control its growth during the sprouting process. The production of safe sprouts continues to be a major challenge for industry.

Application of an Environmental Phage-Based Assay (Sample6 Detect HT/L) for the Detection of Listeria spp. in Ice Cream.

Background: Dairy products are common sources of Listeria outbreaks, and early detection of the pathogen is critical to prevent outbreaks of illnesses and financial losses for dairy producers. Objective: This study aimed to evaluate Sample6 Detect HT/L for effective detection of Listeria monocytogenes and L. innocua in ice cream. Methods: Performance of the Sample6 DETECT HT/L was compared with U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) Chapter 10 method for detection of Listeria spp. in ice cream using an unpaired study design. Results: R2-enriched samples tested with Sample6 Detect HT/L performed as well as the reference method at all time points tested from 15 to 24 h. R2 is a proprietary blend for use with the test kit that helps with early detection. All the dPODC values (Sample6 Detect HT/L presumptive and confirmed results) equaled zero, indicating 100% concordance between the methods. Both Sample6 Detect HT/L and FDA BAM results showed low dPODC values, with confidence intervals indicating no significant differences between Sample6 Detect HT/L and reference method results. Conclusions: Sample6 Detect HT/L is suitable to detect Listeria spp. in ice cream, even with a 12 h enrichment. Sample6 Detect HT/L demonstrated equivalent detection of L. monocytogenes and L. innocua from R2-enriched samples as expected with 15 and 18 h enrichment when compared with the 24 h FDA BAM method for L. monocytogenes. Highlights: These results indicate that Sample6 Detect HT/L, primarily developed for environmental samples, can be used to detect Listeria spp. in ice cream with less incubation time, resulting in faster detection.

Rapid Detection of Listeria in Ice Cream in 13 Hours Using the Roka Listeria Detection Assay.

Background: Listeria contamination is a major concern in the ice cream industry; therefore, early and accurate detection is vital. Current detection methods require about a 24 h enrichment period for detection. Objective: Enhance the early detection of Listeria in ice cream using the highly sensitive isothermal ribosomal RNA-based Roka/Atlas Listeria Detection Assay. Methods: The R2 Medium was developed for Listeria enrichment by Molecular Epidemiology, Inc. (Seattle, WA). Comparative growth curve studies were performed on the new R2 Medium for Listeria and the currently validated media for the Roka Listeria Detection Assay. Subsequently, a method comparison between the Roka Listeria Detection Assay and the U.S. Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM) Chapter 10 reference method on ice cream was carried out. Results: The R2 Medium supports the growth of L. monocytogenes better than Buffered Listeria Enrichment Broth, Demi-Fraser broth, and Modified University of Vermont Broth, as indicated by the faster growth rate of the organism. When used as an enrichment medium in a method comparison study of ice cream, the results showed that R2 Medium-enriched samples tested with the Roka Listeria Detection Assay gave an equivalent performance compared with the 24 h FDA-BAM reference method at 10 and 18 h post-enrichment for Listeria. Conclusions: The results from this study indicate that the new R2 Medium and the highly sensitive Roka Listeria Detection Assay allowed for the rapid detection of Listeria species in ice cream in 13 h. Highlights: The Roka Listeria Detection Assay, in conjunction with a new media formulation (R2 Medium), allowed for the early detection of Listeria in ice cream and may be applied in other food matrixes and environmental samples.