Evaluation of Listeria monocytogenes biofilms attachment and formation on different surfaces using a CDC biofilm reactor.

Listeria monocytogenes can adapt, persist, and form biofilms on food premises surfaces, representing a challenge for food safety, since they led to disease transmission, food contamination and spoilage during production. Physical interventions (scrubbing and wiping) can help controlling formation, nevertheless when biofilms are formed, they are usually very resistant to current control strategies used in the food industry. Biofilm attachment and formation is influenced by environment characteristics, substrate properties and microbial motility. The purpose of this study was to evaluate the ability of L. monocytogenes to attach and form biofilms on different surfaces (wood, nylon, and polycarbonate) representative of the materials used during produce harvesting and storage. Multi-strain L. monocytogenes biofilms were grown in a CDC Biofilm reactor at 20 ± 2 °C up to 96-h and characterized for: a) attachment strength by enumerating cells after rinsing; b) hydrophobicity and interfacial tension by contact angle measurements; c) biofilm architecture by Laser Scanning Confocal Microscopy. All experiments were done in triplicate. Material, incubation, and solvent significantly affected the hydrophobicity and wetting properties of L. monocytogenes biofilms (P < 0.05). The type of material and incubation time significantly influenced hydrophobicity and wetting properties of L. monocytogenes biofilms (P < 0.05). Highest contact angle and lowest interfacial tension were observed on polycarbonate coupons. The data presented contributes to understanding Listeria biofilms grow on different surfaces commonly used in produce harvesting and storage. The data obtained in this study can be used when evaluating intervention strategies to control this pathogen in food premises.

Potential risk-factors affecting Salmonella sp. and Escherichia coli occurrence and distribution in Midwestern United States swine feed mills.

AIM: This study aimed to evaluate the patterns and potential risk factors associated with the occurrence of Salmonella sp. and Escherichia coli in selected United States swine feed mills. METHODS AND RESULTS: A total of 405 samples were collected during fall 2018, spring and summer 2019 from selected sites including floors, equipment, shoes and feed in six feed mills in the US Midwest region. Each sample was analysed for the presence of Salmonella and E. coli with culture methods and confirmed by PCR. A survey regarding production volumes, hygiene practices and microbial testing capabilities was conducted in each facility All mills had at least one sampling site positive for either Salmonella or E. coli. Of the 405 samples, 4·7, and 14·1% were positive for Salmonella sp., and E. coli respectively. Sites with higher percentages of positive samples were the receiving, manufacturing, and control area floors. The survey responses indicated that the age of the mill might be a risk factor for bacterial contamination: the older the facility, the higher the number of positive samples. Other risk factors evaluated, such as the production capacity, did not appear to relate to bacterial prevalence. CONCLUSION: The data documents the presence of E. coli and Salmonella in selected US swine feed mills, and an association between E. coli occurrence and number of ingredient suppliers to feed mill. SIGNIFICANCE AND IMPACT OF THE STUDY: This information could be used to understand risk factors affecting the occurrence of Salmonella sp. and E. coli in feed mills and help implement monitoring and mitigation strategies for public health.

The application of high-concentration short-time chlorine dioxide treatment for selected specialty crops including Roma tomatoes (Lycopersicon esculentum), cantaloupes (Cucumis melo ssp. melo var. cantaloupensis) and strawberries (Fragaria×ananassa).

The effects of high-concentration short-time chlorine dioxide (ClO2) gas treatment on food-borne pathogens inoculated onto the surface of tomatoes, cantaloupes, and strawberries were studied. Produce were spot-inoculated with a mixture of Salmonella enterica (serotypes Montevideo, Javiana and Baildon), Escherichia coli O157:H7 (serotypes 204 P, EDL 933 and C792) or Listeria monocytogenes (serotypes Scott A, F 5069 and LCDC 81-861), and treated with ClO2 gas at 10 mg/l for 180 s. After ClO2 gas treatment, surviving populations were determined and shelf-life studies were conducted (microbial spoilage population, change in color and overall appearance). Significant microbial reduction (p < 0.05) was observed for all treated samples. Nearly a 5LogCFU/cm(2)Salmonella reduction was found on tomatoes, cantaloupe and strawberries, while a ~3LogCFU/cm(2) reduction was observed for E. coli and Listeria on all produce surfaces. E. coli and Listeria appeared to be more resistant to ClO2 gas as compared to Salmonella spp. Treatments significantly (p < 0.05) reduced initial microflora population, while produce color surface was not significantly influenced, as compared to the control (p > 0.05). Results obtained suggest the potential use of high-concentration short-time ClO2 gas treatment as an effective online pathogen inactivation technology for specialty crops in large-scale produce packing operations.

A comparative study on the effectiveness of chlorine dioxide gas, ozone gas and e-beam irradiation treatments for inactivation of pathogens inoculated onto tomato, cantaloupe and lettuce seeds.

The increase in reported food-borne outbreaks linked with consumption of raw fruits and vegetables has motivated new research focusing on prevention of pre-harvest produce contamination. This study evaluates and compares the effectiveness of three non-thermal technologies, chlorine dioxide gas, ozone gas and e-beam irradiation, for inactivation of Salmonella enterica and Escherichia coli O157:H7 on pre-inoculated tomato, lettuce and cantaloupe seeds, and also their corresponding effect on seeds germination percentage after treatments. Samples were treated with 10mg/l ClO(2) gas for 3 min at 75% relative humidity, with 4.3mg/l ozone gas for 5 min and with a dose of 7 kGy electron beam for 1 min. Initial load of pathogenic bacteria on seeds was ~6 log CFU/g. Results demonstrate that all treatments significantly reduce the initial load of pathogenic bacteria on seeds (p<0.05). In particular, after ozone gas treatments 4 log CFU/g reduction was always observed, despite the seeds and/or microorganisms treated. ClO(2) and e-beam treatments were noticeably more effective against Salmonella on contaminated tomato seeds, where 5.3 and 4.4 log CFU/g reduction were respectively observed. Germination percentage was not affected, except for cantaloupe seeds, where the ratio was significantly lowered after ClO(2) treatments. Overall, the results obtained show the great applicability of these non-thermal inactivation techniques to control and reduce pathogenic bacteria contamination of seeds.

Use of high-concentration-short-time chlorine dioxide gas treatments for the inactivation of Salmonella enterica spp. inoculated onto Roma tomatoes.

Salmonella outbreaks have been recently linked to the consumption of fresh tomatoes. Thus, there is a need to develop systems that reduce the risk of microbial contamination to increase product shelf-life and keep fresh fruit attributes. The objectives of this study were to evaluate high-concentration-short-time chlorine dioxide gas treatments effects on Salmonella-inoculated Roma tomatoes and determine the optimal treatment conditions for microbial inactivation and shelf-life extension. Effects of ClO(2) concentration (2, 5, 8 and 10mg/l) and exposure time (10, 30, 60, 120 and 180 s) on inoculated Roma tomatoes were studied. Salmonella enterica strains, serotype Montevideo, Javiana and Baildon, were used to experimentally inoculate the food product. After ClO(2) treatments, tomatoes were stored at room temperature for 28 days. Inherent microbial population, change in tomato color, and chlorine dioxide gas residuals were evaluated. ANOVA analysis showed that both ClO(2) concentration and treatment time were significant (p<0.01) for Salmonella inactivation. Surviving Salmonella populations of 3.09, 2.17 and 1.16 logCFU/cm(2) were obtained treating tomatoes with 8 mg/l ClO(2) for 60 s, 10 mg/l ClO(2) for 120 s, and 10 mg/l for 180 s, respectively (initial Salmonella population: 6.03±0.11 log CFU/cm(2)). The selected treatments significantly reduced background microflora (p<0.05), while fruit color and residual contents were not significantly different (p>0.05), as compared to the control. Results suggest the potential for high-concentration-short-time treatments ClO(2) gas as an effective pathogen inactivation technology for large-scale produce packing operations.

Isolation, taxonomic identification and hydrogen peroxide production by Lactobacillus delbrueckii subsp. lactis T31, isolated from Mongolian yoghurt: inhibitory activity on food-borne pathogens.

AIMS: The aim of this work was to isolate lactic acid bacteria (LAB) strains from Mongolian tarag (a traditionally homemade yoghurt) displaying antimicrobial activities against food-borne pathogens, identify inhibitory substances and study the kinetics of their production. METHODS AND RESULTS: Inhibitory substance-producing bacterial strains were isolated from tarag. From 300 bacterial clones, 31 were able to inhibit the growth of the indicator strain Lactobacillus bulgaricus 340. One of the most active strains was identified as Lactobacillus delbrueckii subsp. lactis strain T31 by using cluster analysis of amplified fragment length polymorphism (AFLP) DNA fingerprints. The antimicrobial substance was inactivated by catalase, demonstrating the production of hydrogen peroxide (H(2)O(2)). Production of H(2)O(2) was studied under aerated and nonaerated culture conditions. The amount of H(2)O(2) in the culture supernatant increased during bacterial growth and reached a maximum (5.12 mmol l(-1)) at the early stationary phase under aerated conditions (agitated cultures). H(2)O(2) was not detected in the culture performed without agitation. In mixed cultures performed in milk with either Lact. delbrueckii subsp. lactis T31 in the presence of Escherichia coli, or Lact. delbrueckii subsp. lactis T31 in the presence of Listeria innocua under aerated and nonaerated conditions, a significant decrease in pathogen count was observed in aerated cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: The significant decrease in Listeria viability observed in aerated mixed cultures of Lact. delbrueckii subsp. lactis T31 is mainly because of H(2)O(2) production. Lactobacillus delbrueckii subsp. lactis T31 could be used as a protective culture in food industries or as a probiotic to prevent intestinal and urogenital infections.