Application of Bacteriophage in Food Manufacturing Facilities for the Control of Listeria monocytogenes and Listeria sp
Reinhard, Robert Gordon
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The purpose of this research was to determine if bacteriophage (phage) could be used to treat and reduce the incidence of Listeria in food manufacturing facilities, and thereby reduce the risk of food products being cross-contaminated with Listeria monocytogenes. Listeria incidence in food manufacturing ready-to-eat environments was surveyed at 31 ready-to-eat (RTE) food plants. A total 4,829 samples were collected from all locations. Nine (29%) facilities had zero samples positive for Listeria spp., whereas 22 (71%) had one or more samples positive. The total incidence of Listeria spp. in all RTE food plants was 4.5%. The effectiveness of phage against Listeria was determined when applied to stainless steel, polyurethane thermoplastic, and epoxy. Each material was inoculated with a cocktail containing L. monocytogenes and L. innocua (4 to 5-log10 CFU/cm2) and treated with two different concentrations of phage (2x10^7 and 1x10^8 PFU/cm2). Treated samples were held at 4 or 20°C for 1 and 3h. After treatment with phage, Listeria reductions ranged from 1.27–3.33 log10 CFU/cm2 on stainless steel, 1.17–2.76 log10 CFU/cm2 on polyurethane thermoplastic, and 1.19–1.76 log10 CFU/cm2 on epoxy. Listeria reduction occurred on all materials tested, under all conditions. Higher phage concentration, longer time, and higher environmental temperatures led to significantly (P<0.05) greater reduction of Listeria on stainless-steel and polyurethane thermoplastic. The effectiveness of a phage against Listeria spp. was evaluated in two food manufacturing facilities, operating at either 4°C or 20°C. First, a moderate application of a 2x10^7 PFU/mL phage was applied once per day over three days and samples were collected and analyzed for Listeria at 0, 24, 48 and 72 h. This phage treatment led to a decrease in the incidence of Listeria by 67%. A second application method was studied with phage being applied in the food manufacturing environment in an intensified manner (3 times in 18 hours) at a higher concentration of phage (1x10^8 pfu/mL). This intensified application led to a 32% overall reduction in the incidence of Listeria in the production environment. Applications of Listeria specific phage can be an additional intervention strategy for controlling pathogenic Listeria organisms in food production facilities.
General Audience Abstract
Listeriosis is a serious illness caused by the bacterium Listeria monocytogenes. Annually in the United States it is estimated that 95.7% of all listeriosis illnesses are caused by the consumption of contaminated food, and it is generally recognized that L. monocytogenes is caused by cross contamination of ready-to-eat foods from an environmental source. The purpose of this research was to determine if food manufacturing facilities could use bacteriophage (phage) to treat and reduce the incidence of Listeria in food manufacturing plant, and thereby reduce the risk of food products being cross-contaminated with L. monocytogenes. The incident rate of Listeria was surveyed by collecting 4,829 samples from 31 ready-to-eat (RTE) food plants across the United States. Nine (29%) facilities had zero samples positive for Listeria spp., whereas 22 (71%) plants had one or more samples positive. The total incidence of Listeria spp. in all RTE food plant samples was 4.5%. Second, research was completed to determine if phage at different levels reduced Listeria on three common food manufacturing plant materials (stainless steel, polyurethane belting and epoxy flooring). After Listeria was attached to each material (4 to 5-log10 CFU/cm2), they were treated with two different levels of phage (2x10^7 and 1x10^8 PFU/cm2) at two temperatures (4 or 20°C) for 1 and 3 hours. After treatment with phage, Listeria reduction of 93.2 to 99.9% occurred on all materials tested, under all conditions. Finally, two different methods of applying phage (moderate and intensified) in a food manufacturing plant was studied. The moderate application was a single treatment with lower concentration of phage (2x10^7 PFU/mL) once per day for three days, while the intensified treatment was the application of high phage concentrations (1x10^8 pfu/mL) three different times, all in a single day. Both application methods reduced the incidence of Listeria in the food manufacturing plant. The total reduction across all trials was 67% using the moderate application method and 32% using the intensified application of phage.
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