Evaluation and interlaboratory validation of a selective agar for phosphatidylinositol-specific phospholipase C activity using a chromogenic substrate to detect Listeria monocytogenes from foods

Phosphatidylinositol-specific phospholipase C (PI-PLC) activity is a potential virulence factor and is exhibited only by the Listeria species Listeria monocytogenes and Listeria ivanovii. A chromogenic substrate for the direct detection of PI-PLC activity is available in a new medium (BCM L. monocytogenes plating agar). The use of a chromogenic substrate offers a mechanism with which to directly screen for L. monocytogenes and L. ivanovii other than the esculin used in Oxford (OXF) and Palcam (PAL) agars, which screen for all Listeria species. The specificity levels of BCM plating agar and of BCM confirmation and rhamnose agars were evaluated with 107 Listeria and 10 Bacillus species isolates. In addition, BCM L. monocytogenes plating agar was compared with standard Listeria selective agars (OXF and PAL agars) with regard to the recovery of L. monocytogenes from 2,000 food and environmental samples obtained from eight participating laboratories. A Listeria species was isolated from at least one of the agars in 209 analyses, and L. monocytogenes was isolated in 135 of these analyses. In 27 of the analyses in which L. monocytogenes was isolated, one or more of the selective differential agars used failed to isolate L. monocytogenes, and therefore the results of these analyses were discrepant. Relative to a reference method involving the use of all three agars (OXF, PAL, and BCM agars), the OXF-BCM, PAL-BCM, and OXF-PAL combinations had sensitivities of 99.3, 99.2, and 90.2%, respectively. In statistical analyses of the different combinations of agars, the OXF-BCM and BCM-PAL combinations were found to be superior to the OXF-PAL combination for the detection of L. monocytogenes.     

Modelling the fate of Listeria monocytogenes during manufacture and ripening of smeared cheese made with pasteurised or raw milk

The dynamics of the physicochemical characteristics of foods help to determine the fate of pathogens throughout processing. The aim of this study was to assess the behaviour of Listeria monocytogenes during cheesesmaking and ripening and to model the growth observed under the dynamic conditions of the cheese. A laboratory scale cheese was made in 4 independent replicates from pasteurised or raw cow's milk, artificially contaminated with L. monocytogenes. No growth of L. monocytogenes occurred during raw milk cheese-making, whereas growth did occur in pasteurised milk. During ripening, growth occurred in raw milk cheese, but inactivation occurred in pasteurised milk cheese. The behaviour observed for L. monocytogenes was modelled using a logistic primary model coupled with a secondary cardinal model, taking into account the effect of physicochemical conditions (temperature, pH, water activity and lactate). A novel statistical approach was proposed to assess the optimal growth rate of a microorganism from experiments performed in dynamic conditions. This complex model had an acceptable quality of fit on the experimental data. The estimated optimum growth rates can be used to predict the fate of L. monocytogenes during cheese manufacture in raw or pasteurized milk in different physicochemical conditions. The data obtained contributes to a better understanding of the potential risk that L. monocytogenes presents to cheese producers (growth on the product, if it is contaminated) and consumers (the presence of high numbers) and constitutes a very useful set of data for the completion of chain-based modelling studies.     

Prevalence of Listeria spp. in feces and carcasses at a lamb packing plant in Brazil

The objective of this work was to study the occurrence of Listeria species in feces and on dressed and cooled carcasses of lambs at a packing plant in Brazil. Listeria spp. were recovered on Oxford and Palcam agars. The 35 fecal samples yielded Listeria welshimeri (20%) and Listeria innocua (8.6%). The 69 carcass samples yielded L. innocua (34.8%), Listeria monocytogenes (4.3%), and Listeria ivanovii (1.5%). More Listeria spp. were recovered with two selective agars than with either agar alone.     

Recovery of Listeria monocytogenes on selective agar media in a collaborative study using reference samples

Sixteen laboratories compared counts of Listeria monocytogenes in reference samples using Blood agar, Palcam(y) agar and Oxford agar. Significant differences were found between laboratories. The mean counts on Blood agar were significantly higher than on Palcam(y) or Oxford agar. The mean counts on Palcamy agar were somewhat higher than on Oxford agar (only after 48 h incubation), but no significant difference was found. Addition of egg yolk to Palcam agar seems to be beneficial for the recovery of sublethally injured cells. Recovery of L. monocytogenes was higher after 48 h incubation for all media tested.     

Comparative evaluation of culture- and BAX polymerase chain reaction-based detection methods for Listeria spp. and Listeria monocytogenes in environmental and raw fish samples

Two commercial polymerase chain reaction (PCR)-based Listeria detection systems, the BAX for Screening/Listeria monocytogenes and the BAX for Screening/Genus Listeria, and a culture-based detection system, the Biosynth L. monocytogenes Detection System (LMDS), were evaluated for their ability to detect L. monocytogenes and Listeria spp. in raw ingredients and the processing environment. For detection of L. monocytogenes from raw fish, enrichment was performed in Listeria enrichment broth (LEB), followed by plating on both Oxford agar and LMDS L. monocytogenes plating medium (LMPM). Detection of Listeria and L. monocytogenes from environmental samples was performed using LMDS enrichment medium, followed by plating on both Oxford agar and LMPM. A total of 512 environmental samples and 315 raw fish were taken from two smoked fish processing facilities and screened using these molecular and cultural Listeria detection methods. The BAX for Screening/L monocytogenes was used to screen raw fish and was 84.8% sensitive and 100% specific. The BAX for Screening/Genus Listeria was evaluated on environmental samples and had 94.7% sensitivity and 97.4% specificity. In conjunction with enrichment in LEB, LMPM had a sensitivity and specificity for detection of L. monocytogenes from raw fish of 97.8 and 100%, respectively. Use of LMDS enrichment medium followed by plating on LMPM allowed for sensitivity and specificity rates of 94.8 and 100%, respectively, for detection of L. monocytogenes from environmental samples. We conclude that both the BAX systems and the use of LMPM allow for reliable and rapid detection of Listeria spp. and L. monocytogenes. While the BAX systems provide screening results in about 3 days, the use of LMPM allows for L. monocytogenes isolation in 4 to 5 days.     

Important vectors for Listeria monocytogenes transmission at farm dairies manufacturing fresh sheep and goat cheese from raw milk

The aim of this study was to determine the transmission routs of Listeria spp. in dairy farms manufacturing fresh cheese made from ovine and caprine raw milk and to evaluate the impact of Listeria monocytogenes mastitis on raw milk contamination. Overall, 5,799 samples, including 835 environmental samples, 230 milk and milk product samples, and 4,734 aseptic half-udder foremilk samples were collected from 53 dairy farms in the dairy intensive area of Lower Austria. Farms were selected for the study because raw milk was processed to cheese that was sold directly to consumers. A total of 153 samples were positive for Listeria spp., yielding an overall prevalence of 2.6%; L. monocytogenes was found in 0.9% of the samples. Bulk tank milk, cheese, and half-udder samples were negative for Listeria spp. Because none of the sheep and goats tested positive from udder samples, L. monocytogenes mastitis was excluded as a significant source of raw milk contamination. L. monocytogenes was detected at 30.2% of all inspected farms. Swab samples from working boots and fecal samples had a significantly higher overall prevalence (P < 0.001) of L. monocytogenes (15.7 and 13.0%, respectively) than did swab samples from the milk processing environment (7.9%). A significant correlation was found between the prevalence of L. monocytogenes in the animal and in the milk processing environment and the silage feeding practices. Isolation of L. monocytogenes was three to seven times more likely from farms where silage was fed to animals throughout the year than from farms where silage was not fed to the animals.     

Comparison of media and sampling locations for isolation of Listeria monocytogenes in queso fresco cheese

Listeriosis associated with Hispanic-style soft cheese is an ongoing public health concern. Although rapid detection methods based on molecular and immunological technologies have been applied successfully for detecting Listeria monocytogenes in foods, obtaining isolates of the pathogen is a critical procedure for epidemiologic studies and regulatory analysis. Oxford agar, a medium recommended by the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) to isolate L. monocytogenes from cheese, is unable to differentiate L. monocytogenes from other Listeria species. Hence, two selective isolation media, L. monocytogenes blood agar (LMBA) and Rapid 'L. mono agar (RLMA), were compared with Oxford agar for isolating L. monocytogenes from cheese. Queso fresco cheese was inoculated at 10(0) or 10(1) CFU/g with a five-strain mixture of L. monocytogenes or with the five-strain L. monocytogenes mixture and Listeria innocua. Cheese samples were stored at 21, 12, and 4 degrees C and Listeria counts were determined at 3, 7, and 10 days; 7, 10, 14, 21 days; and 2, 4, 8, and 12 weeks postinoculation, respectively. Surface and interior cheese samples as well as liquid exudate produced during storage were assayed individually to determine differences in Listeria contamination at different sampling locations. L. monocytogenes was more easily differentiated from L. innocua on RLMA than LMBA and Oxford agar. Similar L. monocytogenes counts (ca. 10(4) CFU/g) were obtained on the last sampling day on the surface and interior of cheese samples (P > 0.05) for all storage temperatures and both initial inoculation levels, but smaller cell numbers were detected in the exudate produced during storage. In addition, simultaneous inoculation of L. innocua with L. monocytogenes did not affect the final L. monocytogenes counts in the cheese. The amount of exudate released from the cheese and decrease of pH correlated with storage temperature. More exudate was produced and a greater decrease of pH occurred at 21 degrees C than at 12 or 4 degrees C. Our results indicate that RLMA is a suitable medium for isolating L. monocytogenes from queso fresco cheese. Higher counts of L. monocytogenes were obtained from surface and interior samples of cheese than from the exudate of the cheese during storage. In addition, pH may be a useful indicator of improperly stored queso fresco cheese.     

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.     

Raw and processed fish show identical Listeria monocytogenes genotypes with pulsed-field gel electrophoresis

A total of 257 raw fish samples at two different sites were examined for the presence of Listeria monocytogenes. The prevalence of L. monocytogenes was 4%. From 11 positive samples, nine different L. monocytogenes pulsed-field gel electrophoresis genotypes were recovered. From nine pulsotypes recovered from raw fish and 32 pulsotypes shown by 101 fish product isolates, two raw fish and fish product pulsotypes were indistinguishable from each other. Although the prevalence of L. monocytogenes in raw fish is low, the range of L. monocytogenes strains entering the processing plant in large amounts of raw material is wide. This indicates that the raw material is an important initial contamination source of L. monocytogenes in fish processing plants. This postulation is supported by the identical pulsotypes recovered from both raw and processed fish. Some L. monocytogenes strains entering a plant may thus contaminate and persist in the processing environment, causing recurrent contamination of the final products via processing machines.     

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.     

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.     

Prevalence of Salmonellae, Listeria monocytogenes, and fecal coliforms in bulk tank milk on US dairies

The objective of this study was to determine the prevalence of Salmonella, Listeria monocytogenes, and fecal coliforms in bulk tank milk in the United States. As part of the NAHMS Dairy 2002 survey, 861 bulk tank milk samples were collected from farms in 21 states. Milk was directly plated on selective agars for direct bacterial enumeration and was enriched in selective broths to increase detection sensitivity. Somatic cell counts (SCC) and standard plate counts (SPC) were also determined. Coliforms were detected in 95% (818 of 860) of the samples, and the average SCC was 295,000 cells/mL. Twenty-two samples (2.6%) were culture-positive for Salmonella, and 9 serotypes were identified: Montevideo (n = 7), Newport (n = 4), Muenster (n = 2), Meleagridis (n = 2), Cerro (n = 2), 44:Z36 (Z38) (n = 2), Dublin (n = 1), Anatum (n = 1), and 9, 12:nonmo-tile (n = 1). Listeria monocytogenes was isolated from 56 (6.5%) samples, and serotyping of these isolates yielded 5 serotypes (1/2a, 1/2b, 3b, 4b, and 4c). Of the L. monocytogenes isolates, 93% were serotypes 1/2a, 1/2b, and 4b, the most common human clinical isolates. Regional differences in L. monocytogenes and Salmonella prevalence were observed, but more studies are needed to determine the validity of these differences. There were no apparent relationships between SCC or SPC and incidence of Salmonella or L. monocytogenes. Although the prevalence of L. monocytogenes and Salmonella was low, these pathogens represent a potential risk to consumers of raw milk and raw milk products.     

Distribution of Listeria monocytogenes subtypes within a poultry further processing plant

Samples from environmental sites and raw product in a chicken further processing plant were collected every 6 weeks for 12 months. Each sample site was examined before and after a complete production shift. All samples were examined for the presence of Listeria monocytogenes, which was detected in floor drains on the raw product side of the plant preoperation and in drains on both raw and cooked sides following 8 h of processing operation. L. monocytogenes also was detected in raw product and once in fully cooked product but never on cooked product contact surfaces. One hundred sixty-one isolates were collected from 75 positive samples. All isolates were subtyped using a sequence-based method, and 14 unique subtypes were detected through the course of the study. Four of these types were found repeatedly and appeared to be resident in the plant. Three of the four resident strains were detected on raw product at some point during the year-long study, suggesting that raw product may be one source of L. monocytogenes in the processing plant environment. These data highlight the need for research to investigate why some types of L. monocytogenes persist in a processing plant environment but others do not.     

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.     

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.     

Estimation of low bacterial concentration: Listeria monocytogenes in raw milk

A time-series bacteriological analysis has been carried out on milk collected on farms from 1997 to 2001 by a plant producing raw milk soft cheese, with the purpose of assessing the time course of the presence/absence of Listeria monocytogenes. A standard data collection procedure was used, in which farms were tested on a monthly or biweekly basis and 2-3 days after the detection of milk tanker contamination. This procedure yielded low figures for contamination frequencies. The average value and the median of the monthly prevalence of farms detected positive for L. monocytogenes were 2.4 and 0%, respectively. A seasonal effect (with peaks in winter) was observed. Between 1997 and 2001, there was no significant decrease of contamination rates, in spite of the efforts on the contaminated farms. Over the last year of the study (from March 2000 to February 2001), a new data collection procedure was implemented that allowed much better detection of sporadic occurrences. Milk samples were collected from the bulk tank of each participating farm just before pick-up, then stored and subsequently analysed whenever the milk tanker was found contaminated. The average value and the median of the monthly prevalence of positive farms were found equal to 7.7 and 0%, respectively (for a mean prevalence of L. monocytogenes in the milk tanker of 3.2%). These results confirm that farm milk contamination is, most often, a sporadic event In addition to this prevalence study, contamination levels were quantified by enumerating L. monocytogenes using direct plating of small volumes of farm milk previously tested positive. Most often, these levels were extremely low. A simple simulation model shows that, when milk tankers were found positive, contamination levels in the corresponding bulk-tank milk are themselves very low (typically, below 3 L. monocytogenes per millilitre with most probable concentration 0.1 Colony Forming Unit (CFU)/ml and median ranging from 5.10(-2) to 0.1 CFU/ml). Such low levels are very likely to be due to environmental contamination.     

Incidence of Listeria monocytogenes in different types of meat products on the Belgian retail market

A survey was undertaken to determine the incidence and numbers of L. monocytogenes in a variety of meat products (cooked meat products, raw cured meat products (dried or not), mayonnaise based salads and prepared meals). As expected, raw cured meat products were significantly higher contaminated with L. monocytogenes than cooked meat products, 13.71% (113/824) and 4.90% (167/3405), respectively. Also a larger proportion of raw cured meat product samples contained a high initial level of the pathogen ( > 10 cfu/g). Higher incidence rates were obtained for whole cooked meat products (e.g. cooked ham, bacon) after slicing than before slicing, 6.65 and 1.56%, respectively, indicating cross-contamination. Due to multiple handling and processing steps, the incidence rate of the pathogen was higher for cooked minced meat products than for whole cooked meat products, 6.14 and 3.96%, respectively. No significant differences were obtained in the incidence of L. monocytogenes in whole cured meat products (e.g., raw ham) and minced cured meat products (e.g., dry fermented sausage), 14.92 and 11.69%, respectively. Lower incidence rates of L. monocytogenes were obtained for raw, cured meat products using beef or horse meat, 4.65 and 5.88%, respectively, A high incidence rate of L. monocytogenes was noted for the mayonnaise based salads (21.28% (186/874)) as well as for prepared meals (11.70% (92/786)), the latter especially due to contamination of vegetarian meals.     

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.     

Incidence of Listeria monocytogenes in different food products commercialized in Portugal

Several types of food products on sale in Portugal, were examined for the presence of Listeria monocytogenes. Secondary enrichments, in Fraser broth, were analysed by the mini-Vidas LMO, enzyme-linked fluorescent immunoassay technique. Positive samples were confirmed by isolation on Oxford and PALCAM selective agars followed by biochemical characterization. Of 1035 samples, 72 (7.0%) were positive for L. monocytogenes, the majority being from raw products (milk, meat, fish, flour) although some heat-processed or fermented foods (ready-to-eat) were also positive. In Portugal, a predilection for fresh cheese was indicated as a potential risk for consumers.     

Longitudinal study on the sources of Listeria monocytogenes contamination in cold-smoked salmon and its processing environment in Italy

The aim of the present study was to investigate the sources of Listeria monocytogenes contamination in a cold smoked salmon processing environment over a period of six years (2003-2008). A total of 170 samples of raw material, semi-processed, final product and processing surfaces at different production stages were tested for the presence of L. monocytogenes. The L. monocytogenes isolates were characterized by multiplex PCR for the analysis of virulence factors and for serogrouping. The routes of contamination over the six year period were traced by PFGE. L. monocytogenes was isolated from 24% of the raw salmon samples, 14% of the semi-processed products and 12% of the final products. Among the environmental samples, 16% were positive for L. monocytogenes. Serotyping yielded three serovars: 1/2a, 1/2b, 4b, with the majority belonging to serovars 1/2a (46%) and 1/2b (39%). PFGE yielded 14 profiles: two of them were repeatedly isolated in 2005-2006 and in 2007-2008 mainly from the processing environment and final products but also from raw materials. The results of this longitudinal study highlighted that contamination of smoked salmon occurs mainly during processing rather than originating from raw materials, even if raw fish can be a contamination source of the working environment. Molecular subtyping is critical for the identification of the contamination routes of L. monocytogenes and its niches into the production plant when control strategies must be implemented with the aim to reduce its prevalence during manufacturing.