Use of PFGE typing for tracing contamination with Listeria monocytogenes in three cold-smoked salmon processing plants

The sites of Listeria monocytogenes contamination in three cold-smoked salmon (Salmo salar) processing plants were detected by sampling salmon and the plant's environment and equipment at different production stages. Of the 141 samples collected from three processing plants, 59 (42%) were contaminated with L. monocytogenes. The rates of contamination varied as to the plant and the sample source. L. monocytogenes isolates from 17 various contaminated seafood products (fresh, frozen and smoked fishes, cooked mussels) were also studied. A total of 155 isolates from the three plants and the various seafoods were characterized by genomic macrorestriction using ApaI and SmaI with pulsed-field gel electrophoresis (PFGE) and 82 isolates were serotyped. Macrorestriction yielded 20 pulsotypes and serotyping yielded four serovars: 1/2a, 1/2b, 1/2c, 4b (or e), with 77 (93%) belonging to serovar 1/2a. One clone of L. monocvtogenes predominated and persisted in plant I and was the only pulsotype detected in the final product although it was not isolated from raw salmon. No L. monocytogenes was detected in the smoked skinned salmon processed in plant II, even though 87% of the raw salmon was contaminated. All the smoked salmon samples collected in plant III were contaminated with a unique clone of L. monocytogenes, which may have occurred during slicing. In the three plants, the contamination of final products did not seem to originate from the L. monocytogenes present on raw salmon, but from the processing environment.     

Tracing Listeria monocytogenes isolates from cold-smoked salmon and its processing environment in Iceland using pulsed-field gel electrophoresis

Listeria spp. and Listeria monocytogenes contamination of cold-smoked salmon (n=125) and its processing environment (n=522) were evaluated during surveys conducted in 1997-1998 and 2001 as well as in samples of final products analysed in 2001. The overall frequencies of Listeria spp. and L. monocytogenes in samples from all sources were 15.1% and 11.3%, respectively, but the incidence of L. monocytogenes in cold-smoked salmon final products was only 4%. A total of 201 L. monocytogenes isolates were characterised by Pulsed-Field Gel Electrophoresis (PFGE) in order to trace L. monocytogenes contamination in the processing plants. The combination of AscI and ApaI macrorestriction patterns yielded 24 different pulsotypes in 6 plants. One pulsotype observed by AscI restriction digestion comprised 148 of the 167 typed isolates from two processing plants. Two other pulsotypes predominated in samples from raw material, processing environments and final products. The results indicate that raw material, floors, and drains are potential sources of the L. monocytogenes found on cold-smoked salmon products. This highlights the need to readdress the design and cleaning of processing plants and equipment, and staff behavior. Hindering the introduction into and spread of the organism through the processing environment is necessary to avoid jeopardizing safety of the final product.     

Evaluation of the international reference methods NF EN ISO 11290-1 and 11290-2 and an in-house method for the isolation of Listeria monocytogenes from retail seafood products in france

Retail seafood products were analyzed on their use-by date using the international reference methods NF EN ISO 11290-1 and 11290-2 (collectively method R) or an in-house method (method B) for the isolation of Listeria monocytogenes. The sensitivity of the methods was about 78%. Method R detected more positive samples of smoked salmon and herb-flavored slices of smoked salmon than did method B, whereas the reverse was true for samples of carpaccio-like salmon, herb-flavored slices of raw salmon, and smoked trout. Most products produced a positive result after the first of two enrichments, and little difference was observed after changing the isolation medium (Listeria selective agar, L. monocytogenes blood agar, agar for Listeria according to Ottaviani and Agosti, Oxford agar, and Palcam agar). L. monocytogenes was isolated from 151 (27.8%) of the 543 samples, with concentrations mostly below 100 CFU/g. The pathogen prevalence and concentration in these seafood products varied greatly depending on the producer and the nature of the product. In certain cases, these differences could be explained by problems in cleaning and disinfection operations in the food-processing environment. The identities of L. monocytogenes isolates were confirmed by PCR, and isolates were characterized by random amplification of polymorphic DNA and pulsed-field gel electrophoresis (PFGE). PFGE patterns obtained with the enzymes Apal and AscI produced 26 different pulsotypes. In general, different pulsotypes were present in the different categories of seafood products and were not specific to one producer. The genetic diversity observed in the products was not related to the prevalence found at the manufacturing site. It is therefore important for producers to determine the source(s) of contamination of their product so the risks linked to the presence of L. monocytogenes can be reduced.     

Occurrence and typing of Listeria monocytogenes strains in retail vacuum-packed fish products and in a production plant.

One hundred and ten samples of ready-to-eat, vacuum-packed, smoked and cold-salted fish products were collected from retail outlets in southern Finland during 1996 for examination of the occurrence and level of Listeria monocytogenes. The samples originated from 12 producers. Positive samples with levels exceeding 100 CFU/g were encountered mainly in one of the producers (no. 8). Therefore, 200 samples from the plant and the products of this producer were studied during August-September 1996 and May-September 1997, as well as 55 samples from the six fish farms providing raw material fish to this plant, during September 1997-January 1998. The isolates were characterised by serotyping and pulsed-field gel electrophoresis (PFGE). L. monocytogenes was isolated in 20% (22/110) of the samples from the retail market, originating from 6 producers. Ten of these positive samples contained L. monocytogenes at > 100 CFU/g (maximum 1.37 X 10(4) CFU/g). Seventeen percent (5/30) of cold-smoked and 50% (16/32) of cold-salted rainbow trout samples were contaminated. Only one hot-smoked fish product (2%) was found to be positive by enrichment. Nineteen (86%) of the strains isolated from the retail samples belonged to serovar 1/2a and three (14%) to serovar 4b. In further studies the production line of plant no. 8 was found to be contaminated. All of isolates from up until autumn, 1997 both the products and the production plant were serovar 1/2a; thereafter one strain of 4b and one of 1/2 (H-antigen untypeable) were isolated from the plant. The samples from raw material fish were all negative for L. monocytogenes. The samples from retail market fell into seven PFGE types. Five and nine PFGE types, respectively, were found from the products and the plant of producer no. 8. PFGE type A was detected from the retail products of four producers and was also dominant among the isolates from production plant no. 8. PFGE type A was the only one found repeatedly from skinning, salting and slicing units as well as from products throughout the whole period. PFGE proved to be a powerful tool for studying contamination points and routes in the production plant. The measures based on hazard analysis critical control points (HACCP) program resulted in L. monocytogenes negative samples at production plant no. 8 from the beginning of January 1998.     

Contamination patterns of Listeria monocytogenes in cold-smoked pork processing

Contamination patterns of Listeria monocytogenes were studied in a cold-smoked pork processing plant to identify the sources and possible reasons for the contamination. Environmental sampling combined with pulsed-field gel electrophoresis (PFGE) subtyping and serotyping were applied to investigate the genetic diversity of L. monocytogenes in the plant environment and ready-to-eat (RTE) cold-smoked pork products. A total of 183 samples were collected for contamination analyses, including samples of the product at different stages during manufacture (n = 136) and environmental samples (n = 47) in 2009. L. monocytogenes isolates, previously recovered from 73 RTE cold-smoked pork samples and collected from the same meat processing plant in 2004, were included in this study. The brining machine and personnel working with brining procedures were the most contaminated places with L. monocytogenes. The overall prevalence of L. monocytogenes in raw pork (18%) increased to 60% after the brining injections. The brining machine harbored six different PFGE types belonging to serotypes 1/2a, 1/2c, 4b, and 4d, which were found on the feeding teeth, smooth surfaces, and spaces of the machine, thus potentially facilitating dissemination of L. monocytogenes contamination. Two PFGE types (2 and 8) belonging to serotypes 1/2a and 1/2c were recovered from RTE cold-smoked pork collected in 2004, and from surfaces of the brining machine sampled in 2009, and may indicate the presence of persistent L. monocytogenes strains in the plant. Due to poor hygiene design, removal of the brining machine from the production of cold-smoked meat products should be considered to reduce L. monocytogenes contamination in the finished products.     

Prevalence of Listeria monocytogenes in broilers at the abattoir, processing plant, and retail level.

The environment and products from two broiler abattoirs and processing plants and raw broiler pieces at the retail level were sampled for Listeria monocytogenes in order to evaluate the contamination level of the broiler carcasses and products. Sampling started in the slaughtering process and finished with raw broiler meat or ready-to-eat cooked product. Sampling sites positive for L. monocytogenes at the broiler abattoir were the air chiller, the skin-removing machine, and the conveyor belt leading to the packaging area. The L monocytogenes contamination rate varied from 1 to 19% between the two plants studied. Furthermore, 62% (38 of 61) of the raw broiler pieces, bought from retail stores, were positive for L. monocytogenes. Altogether, 136 L. monocytogenes isolates were obtained for serotyping and pulsed-field gel electrophoresis (PFGE) characterization performed with two rare-cutting enzymes (ApaI and AscI). Altogether three serotypes (1/2a, 1/2c, and 4b) and 14 different PFGE types were obtained using information provided from both ApaI and AscI patterns for discrimination basis. The two broiler abattoirs studied did not share the same PFGE types. However, the same PFGE types found in the raw broiler pieces at the retail level were also found in the broiler abattoirs where the broilers had been slaughtered.     

Listeria monocytogenes contamination patterns for the smoked fish processing environment and for raw fish

Reliable data on the sources of Listeria monocytogenes contamination in cold-smoked fish processing are crucial in designing effective intervention strategies. Environmental samples (n = 512) and raw fish samples (n = 315) from two smoked fish processing facilities were screened for L. monocytogenes, and all isolates were subtyped by automated ribotyping to examine the relationship between L. monocytogenes contamination from raw materials and that from environmental sites. Samples were collected over two 8-week periods in early spring and summer. The five types of raw fish tested included lake whitefish, sablefish, farm-raised Norwegian salmon, farm-raised Chilean salmon, and feral (wild-caught) salmon from the U.S. West Coast. One hundred fifteen environmental samples and 46 raw fish samples tested positive for L. monocytogenes. Prevalence values for environmental samples varied significantly (P < 0.0001) between the two plants; plant A had a prevalence value of 43.8% (112 of 256 samples), and plant B had a value of 1.2% (3 of 256 samples). For plant A, 62.5% of drain samples tested positive for L. monocytogenes, compared with 32.3% of samples collected from other environmental sites and 3.1% of samples collected from food contact surfaces. Ribotyping identified 11 subtypes present in the plant environments. Multiple subtypes, including four subtypes not found on any raw fish, were found to persist in plant A throughout the study. Contamination prevalence values for raw fish varied from 3.6% (sablefish) to 29.5% (U.S. West Coast salmon), with an average overall prevalence of 14.6%. Sixteen separate L. monocytogenes subtypes were present on raw fish, including nine that were not found in the plant environment. Our results indicate a disparity between the subtypes found on raw fish and those found in the processing environment. We thus conclude that environmental contamination is largely separate from that of incoming raw materials and includes strains persisting, possibly for years, within the plant. Operational and sanitation procedures appear to have a significant impact on environmental contamination, with both plants having similar prevalence values for raw materials but disparate contamination prevalence values for the environmental sites. We also conclude that regular L. monocyrogenes testing of drains, combined with molecular subtyping of the isolates obtained, allows for efficient monitoring of persistent L. monocytogenes contamination in a processing plant.     

Identification and molecular characterization of Listeria monocytogenes isolated in raw milk in the region of Algiers (Algeria)

Listeria monocytogenes was isolated from raw milk, whey and curdled milk produced and collected in the region of Algiers and Blida between September 2003 and July 2004. Four out of 153 (2.61%) farm milk samples and 6 out of 80 (7.50%) tankers' samples tested positive for L. monocytogenes. All samples of whey and curdled milk (n=12) tested negative for L. monocytogenes, but 2 of 22 (9%) samples of whey were contaminated by L. innocua. L. monocytogenes isolates were grouped by a multiplex PCR assay; all isolates belonged to the PCR-group IVb, which corresponds to serovars 4b, 4d and 4e. L. monocytogenes isolates were characterized by Pulsed-Field Gel Electrophoresis (PFGE). The combination of AscI and ApaI macrorestriction patterns yielded five different pulsovars (I to V). The results indicate that raw milk, and raw milk products are potential sources of the L. monocytogenes and represent a potential risk for consumers.     

Serological and molecular ecology of Listeria monocytogenes isolates collected from 13 French pork meat salting-curing plants and their products

The purpose of this study was dual: 1. to evaluate the serotype distribution of 1028 Listeria monocytogenes isolates collected in 13 French salting factories and their products and 2. to identify sources of L. monocytogenes contamination in these factories and trace the routes of spread by PFGE (Pulsed-Field Gel Electrophoresis) typing. Serotypes 1/2a, 1/2b, 1/2c, 4b and 4e occurred. Pulsotype diversity was high among strains collected in plants and products. Furthermore, strains showing similar pulsotypes occurred on the same surfaces after an interval of at least two weeks and in unrelated factories. Forty five strains were genetically closely related to a 4b serotype L. monocytogenes strain isolated from a human clinical case of listeriosis. Our results highlighted the fact that L. monocytogenes is introduced into meat processing plants through raw meat. To overcome such contamination, suppliers of raw material should adhere to specific microbiological control measures. In addition, more attention should be focused on the appropriateness and compliance with procedures of cleaning and disinfection.     

Persistent and nonpersistent Listeria monocytogenes contamination in meat and poultry processing plants

Contamination analysis of persistent and nonpersistent Listeria monocytogenes strains in three meat processing plants and one poultry processing plant were performed in order to identify factors predisposing to or sustaining persistent plant contamination. A total of 596 L. monocytogenes isolates were divided into 47 pulsed-field gel electrophoresis (PFGE) types by combining the restriction enzyme patterns of AscI (42 patterns) and ApaI (38 patterns). Persistent and nonpersistent strains were found in all plants. Nonpersistent PFGE types were found mostly at one sampling site, with the processing environment being the most common location, whereas the persistent strains were found at several sampling sites in most cases. The processing machines were frequently contaminated with persistent L. monocytogenes PFGE types, and it was of concern that surfaces having direct contact with the products were contaminated. The role of the processing machines in sustaining contamination and in contaminating the products appeared to be important because the final product of several processing lines was contaminated with the same L. monocytogenes PFGE type as that found in the processing machine. The proportion of persistent PFGE types in heat-treated products was eight times higher than in the raw products, showing the importance of the persistent PFGE types as contaminants of the final heat-treated products. The contamination status of the processing lines and machines appeared to be influenced by the compartmentalization of the processing line, with poor compartmentalization increasing L. monocytogenes contamination. The separation of raw and post-heat treatment areas seemed especially important in the contamination status of post-heat treatment lines.     

Ecology of Listeria spp. in a fish farm and molecular typing of Listeria monocytogenes from fish farming and processing companies

This study focused on the ecology of Listeria monocytogenes in a fish farm by following the changes in its occurrence in different types of samples for a three year period. In addition, L. monocytogenes isolates from different seafood industry areas were compared with pulsed field gel electrophoresis (PFGE) typing to discover possible associations between primary production, further processing and final products. Weather conditions were found to have a strong influence on the probability of finding Listeria spp. in a fish farm environment. The number of samples contaminated with Listeria spp. was typically bigger after rainy periods. Brook and river waters as well as other runoff waters seemed to be the main contamination source at the farm studied. The farmed fish originally found to carry L. monocytogenes become gradually Listeria free. The time needed for the purification of the fish was several months. The sea bottom soil samples were the ones that preserved the L. monocytogenes contamination the longest time. It can be stated that the fish and fish farm equipment studied did not spread listeria contamination. On the contrary, they were found to suffer from listeria contamination coming from outside sources like the brook water. There was a wide range of different L. monocytogenes PFGE-pulsotypes (30) found at 15 Finnish fish farms and fish processing factories. L. monocytogenes isolates from the final products often belonged to the same pulsotypes as did the isolates from the processing environment as well as from the raw fish. This suggests that, in addition to the fish processing factory environment, the fish raw materials are important sources of L. monocytogenes contamination in final products.     

Listeria monocytogenes in foods in Norway

Three-hundred-and-eighty-two samples of different retail food items in Norway (imported soft cheese, raw chicken, minced meat, fermented sausages, vacuum-packed processed meat products, smoked salmon, peeled shrimps, raw minced fish) and 78 carcass samples (sheep, pig, cattle), were screened for Listeria monocytogenes. Of the 460 samples investigated, 78 were found to contain L. monocytogenes. Five of these contained greater than 10(3) cfu/g, four greater than 10(2) cfu/g, while the remainder were shown to contain L. monocytogenes only after enrichment. L. monocytogenes was isolated most frequently from raw chicken, sporadically from soft cheese, shrimps, processed meat products and smoked salmon, and not at all from carcasses and fermented sausages.     

Listeria monocytogenes in the smoked salmon industry

Smoked salmon is sporadically contaminated with Listerial monocytogenes. Contamination levels are normally low and consumers are probably seldom exposed to risk concentrations. No clones of L. monocytogenes seem to be specific to smoked salmon, some clones found in smoked salmon having been isolated from several sources, including patients. Cold-smoking has been shown to eliminate L. monocytogenes in challenge tests at temperatures from 17.1 to 21.1 degrees C, while from 22.2 to 30 degrees C the bacteria survived. Under natural cold-smoking conditions (19 to 22 degrees C) the frequency and level of L. monocytogenes seems to decrease. Hot-smoking seems to eliminate the bacteria when smoke is applied during the whole heating process. The prevention of recontamination of both cold-smoked and hot-smoked salmon is therefore of great importance. L. monocytogenes multiply considerably in smoked salmon during storage. Growth is faster in challenge tests than in naturally-contaminated smoked salmon. The declared shelf-life under refrigeration should be shorter than that customarily stipulated by many producers. While the sources of L. monocytogenes in smoked salmon processing plants have still to be determined, raw salmon does not seem to be an important source. The main issue for producers is to prevent colonization of the processing environment and spread of the bacteria to products. This should be achieved by the systemic implementation of hygienic measures, including the HACCP approach.     

Rapid quantitative detection of, Listeria monocytogenes in salmon products: evaluation of pre-real-time PCR strategies

The spread and persistence of Listeria monocytogenes in smoked fish products and seafood processing factories are big concerns. Thus, the corresponding quality assurance programs must include adequate microbiological control measures. We evaluated eight different pre-PCR sample processing strategies to be coupled with a previously developed real-time PCR assay for the quantitative detection of L. monocytogenes in salmon products. The optimal pre-PCR procedure involved filtration and DNA purification with the use of a commercial kit. This strategy could detect 10 CFU of L. monocytogenes per g of smoked salmon and could quantify 1,000 CFU/g with excellent accuracy compared with the standard plate count method. Thus, this method could be a promising alternative for the quantitative detection of L. monocytogenes in smoked fish products and processing factories. This method could also detect the bacterium in raw salmon.     

Low prevalence of Listeria monocytogenes in foods from Italy

Listeria monocytogenes is an important foodborne pathogen that causes gastrointestinal disorders, and, especially in immunocompromised people, serious extraintestinal diseases, such as septicemia and meningitis, as well as abortion in pregnant women. Many foods, from both plant and animal origin, have been involved in listeriosis outbreaks. This article reports the results of a 12-year survey (1993 through 2004) on the presence of L. monocytogenes in several kinds of food marketed in Italy. Of 5,788 analyzed samples, 121 (2.1%) were contaminated with L. monocytogenes. The highest prevalence was found in smoked salmon (10.6%) and in poultry meat samples (8.5%) and the lowest in red meat (0.3%). L. monocytogenes was not found in 154 samples of fresh seafood products. Fifty-two isolates were also serotyped by the agglutination method. The most common serotypes detected in the 52 strains tested were 1/2a (36.5%), followed by 1/2c (32.8%), 1/2b (13.5%), 4b (11.5%), 3a (3.8%), and 3b (1.9%). The results of the present study showed low levels of L. monocytogenes in the analyzed samples. A total of 61.5% of the 52 L. monocytogenes strains analyzed belonged to serotypes 1/2a, 1/2b, and 4b, namely the serovars that are most commonly involved in extraintestinal human listeriosis outbreaks. In the ready-to-eat samples, these three serotypes were 40.0% (1/2a), 17.1% (1/2b), and 14.3% (4b). This finding highlights the need to implement strict hygienic measures during the production, distribution, and sale of foods to reduce the risk of foodborne listeriosis in humans to an acceptable level.     

Longitudinal studies on Listeria in smoked fish plants: impact of intervention strategies on contamination patterns

Four ready-to-eat smoked fish plants were monitored for 2 years to study Listeria contamination patterns and the impact of plant-specific Listeria control strategies, including employee training and targeted sanitation procedures, on Listeria contamination patterns. Samples from the processing plant environment and from raw and finished product were collected monthly and tested for Listeria spp. and Listeria monocytogenes. Before implementation of intervention strategies, 19.2% of raw product samples (n = 276), 8.7% of finished product samples (n = 275), and 26.1% of environmental samples (n = 617) tested positive for Listeria spp. During and after implementation of Listeria control strategies, 19.0% of raw product samples (n = 242), 7.0% of finished product samples (n = 244), and 19.5% of environmental samples (n = 527) were positive for Listeria spp. In one of the four fish plants (plant 4), no environmental samples were positive for L. monocytogenes, and this plant was thus excluded from statistical analyses. Based on data pooled from plants 1, 2, and 3, environmental Listeria spp. prevalence was significantly lower (P < 0.05) for nonfood contact surfaces and the finished product area and for the overall core environmental samples after implementation of control strategies. Listeria prevalence for floor drains was similar before and after implementation of controls (49.6 and 54.2%, respectively). Regression analysis revealed a significant positive relationship (P < 0.05) between L. monocytogenes prevalence in the environment and in finished products before implementation of control strategies; however, this relationship was absolved by implementation of Listeria control strategies. Molecular subtyping (EcoRI ribotyping) revealed that specific L. monocytogenes ribotypes persisted in three processing plants over time. These persistent ribotypes were responsible for all six finished product contamination events detected in plant 1. Ribotype data also indicated that incoming raw material is only rarely a direct source of finished product contamination. While these data indicate that plant-specific Listeria control strategies can reduce cross-contamination and prevalence of Listeria spp. and L. monocytogenes in the plant environment, elimination of persistent L. monocytogenes strains remains a considerable challenge.     

Tracking of Listeria monocytogenes in smoked fish processing plants

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.     

Mathematical model of Listeria monocytogenes cross-contamination in a fish processing plant

Listeriosis is a foodborne disease caused by the bacterium Listeria monocytogenes. The food industry and government agencies devote considerable resources to reducing contamination of ready-to-eat foods with L. monocytogenes. Because inactivation treatments can effectively eliminate L. monocytogenes present on raw materials, postprocessing cross-contamination from the processing plant environment appears to be responsible for most L. monocytogenes food contamination events. An improved understanding of cross-contamination pathways is critical to preventing L. monocytogenes contamination. Therefore, a plant-specific mathematical model of L. monocytogenes cross-contamination was developed, which described the transmission of L. monocytogenes contamination among food, food contact surfaces, employees' gloves, and the environment. A smoked fish processing plant was used as a model system. The model estimated that 10.7% (5th and 95th percentile, 0.05% and 22.3%, respectively) of food products in a lot are likely to be contaminated with L. monocytogenes. Sensitivity analysis identified the most significant input parameters as the frequency with which employees' gloves contact food and food contact surfaces, and the frequency of changing gloves. Scenario analysis indicated that the greatest reduction of the within-lot prevalence of contaminated food products can be achieved if the raw material entering the plant is free of contamination. Zero contamination of food products in a lot was possible but rare. This model could be used in a risk assessment to quantify the potential public health benefits of in-plant control strategies to reduce cross-contamination.     

Traceback identification of an ingredient (pork dewlap) as the possible source of Listeria monocytogenes serotype 4b contamination in raw chicken products

In surveys conducted on finished product samples from a single poultry processing plant in Spain, Listeria monocytogenes was found in 14 different uncooked products. To track contamination patterns, 77 L. monocytogenes isolates were characterized by PCR-based serotyping, pulsed-field gel electrophoresis (PFGE) restriction analysis, and PCR-based allelic analysis of the virulence gene actA. Serotyping revealed that 12 isolates (15.6%) were of the L. monocytogenes serotype 4b complex (serotype 4b or the closely related serotypes 4d and 4e). A combination of endonucleases AscI and ApaI PFGE patterns yielded 15 different pulsotypes among all 77 tested isolates. All the serotype 4b isolates belonged to one pulsotype. Sequencing of the actA gene confirmed that all serotype 4b isolates corresponded to the same allelic subtype. The subtype was recovered from five product types, but its presence was not correlated with the production line or the date of isolation, suggesting a possible association of this strain with a common ingredient. This traceback investigation established that pork dewlap, an ingredient common to all the products contaminated with this strain, was the most probable source of L. monocytogenes 4b. The same 4b strain was isolated from four samples of pork dewlap from one specific supplier. After replacement of this contaminated ingredient in the fresh products, this strain of L. monocytogenes serotype 4b was not detected. This study confirms the effectiveness of molecular subtyping to control contamination by specific strains of L. monocytogenes and the importance of testing the different ingredients added to the food products.     

Prevalence and contamination levels of Listeria monocytogenes in retail foods in Japan

Retail foods in Japan were surveyed for the presence and contamination levels of L. monocytogenes. It was isolated from 12.2, 20.6, 37.0 and 25.0% of 41 minced beef, 34 minced pork, 46 minced chicken and 16 minced pork-beef mixture samples, respectively. MPN values were higher than 100/g in five (10.9%) minced chicken samples, but lower than 100/g in all minced beef, pork and pork-beef mixture samples. The organism was also isolated from 5.4% of the 92 smoked salmon samples at MPN values lower than 10/g, and from 3.3% of 213 ready-to-eat raw seafood samples at MPN values from lower than 0.3 to higher than 100/g. None of the 285 vegetable samples were contaminated with L. monocytogenes. These findings indicate that ready-to-eat raw seafoods are relatively high risk among the foods surveyed in this study.