Prevalence of Listeria Species and Listeria monocytogenes on Raw Produce Arriving at Frozen Food Manufacturing Facilities.

ABSTRACT: The ubiquity of Listeria monocytogenes in the environment affects the food industry and presents concerns for frozen food facilities. This study determined the prevalence and numbers of Listeria species and L. monocytogenes on raw produce arriving at frozen food facilities. Raw produce was collected using multilevel blinding protocols to ensure anonymity of participants and avoid traceback. Five raw vegetables were selected: corn, carrots, green beans, peas, and spinach. Raw products were collected after arrival at the facilities but before cleaning or other preprocessing steps that are typically performed inside the facility. The U.S. Food and Drug Administration's Bacteriological Analytical Manual method for detection of Listeria spp. and L. monocytogenes was followed, with PCR screening followed by selective plating methods. Listeria numbers were estimated from positive samples using the most-probable-number (MPN) methodology. A total of 290 samples were collected, with 96 and 17 samples positive for Listeria spp. (33.1%) and L. monocytogenes (5.9%), respectively. Enumeration data for the 96 Listeria spp. samples indicated 82 samples had greater than 100 MPN of Listeria spp. per g and 14 samples had less than 100 MPN Listeria spp. per g. The prevalence of Listeria spp. varied by commodity: spinach (66.7%), peas (50%), corn (32.2%), green beans (22.2%), and carrots (13%). L. monocytogenes prevalence was determined in corn (13.6%), peas (6.3%), and green beans (4.2%) arriving at processing facilities. Such data were previously unavailable to frozen vegetable processors and are valuable in implementing process control standards. The prevalence and pathogen concentration data from raw commodities found in this study can provide the industry with information to conduct more accurate quantitative risk assessments and a baseline to model and target appropriate pathogen reduction steps during processing.

Thermal Inactivation of Listeria monocytogenes and Salmonella during Water and Steam Blanching of Vegetables.

Thermal inactivation of Listeria monocytogenes and Salmonella was evaluated on peas, spinach, broccoli, potatoes, and carrots that were treated with hot water and steam. One gram-positive bacterium, L. monocytogenes, and one gram-negative bacterium, Salmonella, were selected as pertinent human pathogens for evaluation. Samples were inoculated with a composite of five strains each of L. monocytogenes and Salmonella to achieve approximately 108 to 109 CFU/g. Inoculated samples were treated with hot water at 85 and 87.8°C and with steam at 85 and 96.7°C for up to 3.5 min. A greater than 5-log reduction of L. monocytogenes and Salmonella was achieved on all products within 0.5 min by hot water blanching at 85 and 87.8°C. Steam blanching at 85°C reduced Salmonella populations by greater than 5 log on spinach and peas within 2 min and on carrots and broccoli within 3.5 min. Populations of Salmonella were reduced by more than 5 log within 1 min on carrot, spinach, and broccoli and within 2 min on peas by steam blanching at 96.7°C. Steam blanching at 85°C reduced L. monocytogenes populations by more than 5 log on carrots and spinach within 2 min and on broccoli and peas within 3.5 min. L. monocytogenes populations were reduced more than 5 log within 1 min on carrot, spinach, peas and broccoli by steam blanching at 96.7°C. Longer treatment times and higher temperatures were required for steam-blanched samples than for samples blanched with hot water. Results suggest that hot water and steam blanching practices commonly used by the frozen vegetable industry will achieve the desired 5-log lethality of L. monocytogenes and Salmonella and will enhance microbiological safety prior to freezing.

Retention of Acid Tolerance and Acid Shock Responses of Escherichia coli O157:H7 and Non-O157:H7 Isolates.

Escherichia coli O157:H7 and non-O157:H7 survival due to induced acid tolerance or shock responses when exposed to lactic acid over time was studied. Induced acid tolerance or shock responses could allow pathogens, like E. coli O157:H7, to survive acidic conditions in foods during storage. Escherichia coli O157:H7 isolates 932 and E009 and a non-Ol57:H7 strain, 23716, were grown to stationary phase at 32°C and exposed to one of two treatments: acid shock or acid adaption. Acid-shocked cells were exposed to lactic acid at pH 3.5 or 4.0. Acid-adapted cells were exposed to pH 5.5 for an adaptation period and then exposed to an acid challenge of pH 3.5 or 4.0. Samples were incubated at either 25 or 32°C and survival of the isolates at 0, 3, 24, 48, 72, and 168 h (7 days), 336 h (14 days), and 504 h (21 days) was determined. All three isolates survived longer with larger populations at pH 4.0 and 25°C compared to the other treatments. In cases where a difference was observed in the two responses, acid-shocked cells had a higher survival rate (typically less than 2 logs) than acid-adapted cells in most cases. Isolate differences were observed at the two pH and temperature levels. Isolate 932 was the most resistant to the acidic conditions during the incubation period, E009 intermediate, and strain 23716 was the most sensitive.

Growth and Production of Toxin of Clostridium botulinum Type E in Rainbow Trout under Various Storage Conditions.

Rainbow trout ( Oncorhynchus mykiss ) were inoculated with 3 to 4 1og10 spores per g of fish of a mixed pool of four strains of Clostridium botulinum type E (Beluga, Minnesota, G21-5, and 070). The trout were vacuum-skin packaged with either oxygen-barrier or oxygen-permeable films. Trout packaged with oxygen-permeable film were stored at 4°C for 21 days, while trout packaged with oxygen-barrier film were stored either at 4°C for 21 days or at 10°C for 15 days. Storage at 10°C was used to simulate commercial temperature abuse. Clostridium botulinum outgrowth was determined by a most probable-number (MPN) method using (tryptone peptone yeast extract glucose trypsin) anaerobic broth. Toxin production was evaluated using a mouse bioassay. Psychrotrophic and anaerobic populations increased with time regardless of packaging type. After 6 days at l0°C, botulinum toxin was detected in the packaged trout; however, the fish was noticeably spoiled before that time. No botulinum toxin was detected in trout packaged with either barrier or permeable films and stored at 4°C for 21 days, although the product was considered spoiled by day 12.