We have applied molecular approaches, including PCR-based detection strategies and DNA fingerprinting methods, to study the ecology of Listeria monocytogenes in food processing environments. A total of 531 samples, including raw fish, fish during the cold-smoking process, finished product, and environmental samples, were collected from three smoked fish processing facilities during five visits to each facility. A total of 95 (17.9%) of the samples tested positive for L. monocytogenes using a commercial PCR system (BAX for Screening/Listeria monocytogenes), including 57 (27.7%) environmental samples (n ‫؍‬ 206), 8 (7.8%) raw material samples (n ‫؍‬ 102), 23 (18.1%) samples from fish in various stages of processing(n ‫؍‬ 127), and 7 (7.3%) finished product samples (n ‫؍‬ 96). L. monocytogenes was isolated from 85 samples (16.0%) using culture methods. Used in conjunction with a 48-h enrichment in Listeria Enrichment Broth, the PCR system had a sensitivity of 91.8% and a specificity of 96.2%. To track the origin and spread of L. monocytogenes, isolates were fingerprinted by automated ribotyping. Fifteen different ribotypes were identified among 85 isolates tested. Ribotyping data established possible contamination patterns, implicating raw materials and the processing environment as potential sources of finished product contamination. Analysis of the distribution of ribotypes revealed that each processing facility had a unique contamination pattern and that specific ribotypes persisted in the environments of two facilities over time (P < 0.0006). We conclude that application of molecular approaches can provide critical information on the ecology of different L. monocytogenes strains in food processing environments. This information can be used to develop practical recommendations for improved control of this important foodborne pathogen in the food industry.The advent of molecular methodology has revolutionized our ability to investigate and understand microbial ecology, offering new and unique opportunities to explore the ecology of food-borne pathogens, including Listeria monocytogenes, throughout the food chain and in the food processing environment. Highly discriminatory molecular typing methods, including multilocus enzyme electrophoresis, pulsed-field gel electrophoresis (PFGE), random amplification of polymorphic DNA, ribotyping, and phage typing, have been successfully applied to investigations of contamination patterns in foods and in the food processing environment and are increasingly used for surveillance of human disease cases and for tracking of outbreak sources (2-4, 7, 12, 26, 34, 36, 37, 39). While each method provides discriminatory differentiation of L. monocytogenes subtypes, highly automated and standardized methods provide a simplified approach to molecular subtyping and data analysis. The RiboPrinter Microbial Characterization System (Qualicon, Inc., Wilmington, Del.) is one example of such an approach. This system is based on ribotyping, a subtyping method based upon scoring restriction poly...

The effects of baking, broiling, deep frying and cooking in a microwave oven on the proximate, mineral and fatty acid composition of grouper (Epinephelus morio), red snapper (LuGanus campechanus), Florida pompano (Trachinotus carolinus) and Spanish mackerel (Scomberomorus macuhtus) were determined. The lipid content of low fat species was not significantly changed by cooking, but lipid was lost from fatty fillets during cooking. The fatty acid composition of all fillets was not significantly changed by baking, broiling or microwave cooking. Deep fried fiets absorbed the major fatty acids in the cooking medium, and as the filet lipid content increased the extent of absorption of fatty acids from the cooking medium decreased. Sodium, potassium and magnesium levels were decreased when low fat species were cooked, but these minerals were not lost when raw fillets containing higher lipid levels were cooked. Cooking did not significantly affect the concentration of the microelements, zinc, copper, iron and manganese.

Four smoked fish processing plants were used as a model system to characterize Listeria monocytogenes contamination patterns in ready-to-eat food production environments. Each of the four plants was sampled monthly for approximately 1 year. At each sampling, four to six raw fish and four to six finished product samples were collected from corresponding lots. In addition, 12 to 14 environmental sponge samples were collected several hours after the start of production at sites selected as being likely contamination sources. A total of 234 raw fish, 233 finished products, and 553 environmental samples were tested. Presumptive Listeria spp. were isolated from 16.7% of the raw fish samples, 9.0% of the finished product samples, and 27.3% of the environmental samples. L. monocytogenes was isolated from 3.8% of the raw fish samples (0 to 10%, depending on the plant), 1.3% of the finished product samples (0 to 3.3%), and 12.8% of the environmental samples (0 to 29.8%). Among the environmental samples, L. monocytogenes was found in 23.7% of the samples taken from drains, 4.8% of the samples taken from food contact surfaces, 10.4% of the samples taken from employee contact surfaces (aprons, hands, and door handles), and 12.3% of the samples taken from other nonfood contact surfaces. Listeria spp. were isolated from environmental samples in each of the four plants, whereas L. monocytogenes was not found in any of the environmental samples from one plant. Overall, the L. monocytogenes prevalence in the plant environment showed a statistically significant (P < 0.0001) positive relationship with the prevalence of this organism in finished product samples. Automated EcoRI ribotyping differentiated 15 ribotypes among the 83 L. monocytogenes isolates. For each of the three plants with L. monocytogenes-positive environmental samples, one or two ribotypes seemed to persist in the plant environment during the study period. In one plant, a specific L. monocytogenes ribotype represented 44% of the L. monocytogenes-positive environmental samples and was also responsible for one of the two finished product positives found in this plant. In another plant, a specific L. monocytogenes ribotype persisted in the raw fish handling area. However, this ribotype was never isolated from the finished product area in this plant, indicating that this operation has achieved effective separation of raw and finished product areas. Molecular subtyping methods can help identify plant-specific L. monocytogenes contamination routes and thus provide the knowledge needed to implement improved L. monocytogenes control strategies.