Elucidation of Listeria monocytogenes Contamination Routes in Cold-Smoked Salmon Processing Plants Detected by DNA-Based Typing Methods

The contamination routes of Listeria monocytogenes in cold-smoked salmon processing plants were investigated by analyzing 3,585 samples from products (produced in 1995, 1996, 1998, and 1999) and processing environments (samples obtained in 1998 and 1999) of two Danish smokehouses. The level of product contamination in plant I varied from 31 to 85%, and no L. monocytogenes was found on raw fish (30 fish were sampled). In plant II, the levels of both raw fish and product contamination varied from 0 to 25% (16 of 185 raw fish samples and 59 of 1,000 product samples were positive for L. monocytogenes). A total of 429 strains of L. monocytogenes were subsequently compared by random amplified polymorphic DNA (RAPD) profiling, and 55 different RAPD types were found. The RAPD types detected on the products were identical to types found on the processing equipment and in the processing environment, suggesting that contamination of the final product (cold-smoked salmon) in both plants (but primarily in plant I) was due to contamination during processing rather than to contamination from raw fish. However, the possibility that raw fish was an important source of contamination of the processing equipment and environment could not be excluded. Contamination of the product occurred in specific areas (the brining and slicing areas). In plant I, the same RAPD type (RAPD type 12) was found over a 4-year period, indicating that an established in-house flora persisted and was not eliminated by routine hygienic procedures. In plant II, where the prevalence of L. monocytogenes was much lower, no RAPD type persisted over long periods of time, and several different L. monocytogenes RAPD types were isolated. This indicates that persistent strains may be avoided by rigorous cleaning and sanitation; however, due to the ubiquitous nature of the organism, sporadic contamination occurred. A subset of strains was also typed by using pulsed-field gel electrophoresis and amplified fragment length polymorphism profiling, and these methods confirmed the type division obtained by RAPD profiling.     

Characterization of Listeria monocytogenes isolated from production lines of fresh and cold-smoked fish

AIMS: The aims of this study were to characterize strains of Listeria monocytogenes isolated from cold-smoking fish plants to establish possible routes of contamination through the processing chain. METHODS AND RESULTS: Listeria monocytogenes from fresh fish suppliers, raw materials, factory sites and finished products isolated in Portugal (162 isolates) and England (28 isolates) were characterized by serotyping, phage typing, tetracycline, cadmium and arsenic resistance, and plasmid profiling. On the basis of serotyping and phage typing, the isolates were categorized into eight groups. Although cultures within some of the groups could be further differentiated on the basis of plasmid profiling and cadmium and arsenite typing, consideration of all typing data predominantly clustered together isolates from a single location. L. monocytogenes strains: from fresh salmon suppliers were not found in the processing lines; from fresh salmon from different locations differed; and from the water where salmon trout were farmed differed from those isolated from the fish samples. SIGNIFICANCE AND IMPACT OF THE STUDY: No clear source or route of contamination in the cold-smoked processing chain could be established; however, these results highlight the complexity in tracking this bacterium through food chains.     

One group of genetically similar Listeria monocytogenes strains frequently dominates and persists in several fish slaughter- and smokehouses

Contamination of foods with the human pathogen Listeria monocytogenes may occur during processing, and the purpose of this study was to determine whether genetically similar strains colonize different processing plants or whether specific persistent strains are unique to each processing plant. We hypothesized that specific L. monocytogenes strains may be better adapted to specific environmental niches in the processing environment. L. monocytogenes contamination patterns were identified by the collection of 686 and 267 samples from the processing environments: raw fish and products of four fish smokehouses and four fish slaughterhouses, respectively. Samples were collected both during production and after cleaning and disinfection. Typically, these samplings were separated by 1 to 3 months. Sampling sites were targeted toward areas likely to harbor the bacterium. L. monocytogenes was isolated from 213 samples, and one strain from each positive sample was typed by RAPD (random amplified polymorphic DNA) analysis with four different primers. The 213 strains were divided into 37 RAPD types. One RAPD type was predominant; 86 of 213 strains belonged to this type. This type was found in three smokehouses and two slaughterhouses and was predominant in three of these plants. A subset of 35 strains was also analyzed by amplified fragment length polymorphism typing, which confirmed the genetic similarity of the groups. Moreover, strains of the dominant RAPD type were indistinguishable from strains isolated frequently from smoked fish products 10 years ago. One smokehouse was surveyed for a year and a half, and the dominant RAPD type persisted throughout the survey period and accounted for 94 of 118 isolates. Our study indicates that strains of L. monocytogenes that are genetically very closely related may be especially adapted to colonizing the processing equipment or especially resistant to cleaning and disinfection.     

Molecular studies on the ecology of Listeria monocytogenes in the smoked fish processing industry

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 < or = 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 food-borne pathogen in the food industry.     

Sources of Listeria monocytogenes Contamination in a Cold-Smoked Rainbow Trout Processing Plant Detected by Pulsed-Field Gel Electrophoresis Typing

Sites of Listeria monocytogenes contamination in a cold-smoked rainbow trout (Oncorhynchus mykiss) processing plant were detected by sampling the production line, environment, and fish at different production stages. Two lots were monitored. The frequency of raw fish samples containing L. monocytogenes was low. During processing, the frequency of fish contaminated with L. monocytogenes clearly rose after brining, and the most contaminated sites of the processing plant were the brining and postbrining areas. A total of 303 isolates from the raw fish, product, and the environment were characterized by pulsed-field gel electrophoresis (PFGE). PFGE yielded nine pulsotypes, which formed four clusters. The predominating L. monocytogenes pulsotypes of the final product were associated with brining and slicing, whereas contaminants of raw fish were not detected in the final product. Air-mediated contamination in the plant could not be proved. In accordance with these results, an L. monocytogenes eradication program was planned. The use of hot steam, hot air, and hot water seemed to be useful in eliminating L. monocytogenes. None of the control samples taken in the 5 months after the eradication program was implemented contained L. monocytogenes.     

One Group of Genetically Similar Listeria monocytogenes Strains Frequently Dominates and Persists in Several Fish Slaughter- and Smokehouses

Contamination of foods with the human pathogen Listeria monocytogenes may occur during processing, and the purpose of this study was to determine whether genetically similar strains colonize different processing plants or whether specific persistent strains are unique to each processing plant. We hypothesized that specific L. monocytogenes strains may be better adapted to specific environmental niches in the processing environment. L. monocytogenes contamination patterns were identified by the collection of 686 and 267 samples from the processing environments: raw fish and products of four fish smokehouses and four fish slaughterhouses, respectively. Samples were collected both during production and after cleaning and disinfection. Typically, these samplings were separated by 1 to 3 months. Sampling sites were targeted toward areas likely to harbor the bacterium. L. monocytogenes was isolated from 213 samples, and one strain from each positive sample was typed by RAPD (random amplified polymorphic DNA) analysis with four different primers. The 213 strains were divided into 37 RAPD types. One RAPD type was predominant; 86 of 213 strains belonged to this type. This type was found in three smokehouses and two slaughterhouses and was predominant in three of these plants. A subset of 35 strains was also analyzed by amplified fragment length polymorphism typing, which confirmed the genetic similarity of the groups. Moreover, strains of the dominant RAPD type were indistinguishable from strains isolated frequently from smoked fish products 10 years ago. One smokehouse was surveyed for a year and a half, and the dominant RAPD type persisted throughout the survey period and accounted for 94 of 118 isolates. Our study indicates that strains of L. monocytogenes that are genetically very closely related may be especially adapted to colonizing the processing equipment or especially resistant to cleaning and disinfection.     

Effects of high-pressure processing on Listeria monocytogenes, spoilage microflora and multiple compound quality indices in chilled cold-smoked salmon

AIMS: To evaluate the effect of high-pressure processing (HPP) on Listeria monocytogenes, microbial and chemical changes and shelf-life in chilled cold-smoked salmon (CSS). METHODS AND RESULTS: First, challenge tests with L. monocytogenes were carried out using HPP of the product at 0.1 (control), 150, 200 and 250 MPa. Secondly, storage trials with the naturally contaminated product and HPP at 0.1 (control) and 200 MPa were realized. Shelf-life, microbial changes and chemical changes were determined and existing predictive models and multiple compound quality indices evaluated. HPP with 250 MPa did not inactivate L. monocytogenes but significant lag phases of 17 and 10 days were observed at ca 5 and 10 degrees C, respectively. HPP with 200 MPa had a marked effect on both colour and texture of CSS. CONCLUSIONS: High-pressure processing was unable to prevent growth of L. monocytogenes or spoilage of chilled CSS. Existing mathematical models allowed growth rates of L. monocytogenes and shelf-life of samples without high-pressure treatments to be predicted. SIGNIFICANCE AND IMPACT OF THE STUDY: High-pressure processing seems more appropriate for new types of salmon products than for a classical product like CSS where consumers expect specific quality attributes.     

Modification of a virulence-associated phenotype after growth of Listeria monocytogenes on food

AIM: To assess the effect of different foods, which have been implicated or not in cases of listeriosis, on the in vitro virulence-associated phenotype level of different Listeria monocytogenes strains. METHODS AND RESULTS: The virulence-associated phenotype level of L. monocytogenes was studied with the in vitro cell test based on a plaque-forming assay with a human adenocarcinoma cell line (HT-29) monolayer. Three strains of L. monocytogenes were grown in preparations (homogenate, 1-mum filtrate or 0.2-mum filtrate) of different food extracts ['rillettes' (potted minced pork), milk, raw salmon and cold-smoked salmon] or in a control medium, brain heart infusion (BHI). The bacterial suspensions grown in food extracts or in BHI at 37 degrees C were diluted with their growth medium (food extract or BHI) or with minimum essential medium before seeding on confluent HT-29 cell monolayers. Filtration of food extracts had no significant effect on the plaque numbers formed by the bacteria. A significant decrease in the plaque numbers was noted for the three strains when they grew in the rillettes extracts, compared with the other food extracts and BHI. The levels of in vitro virulence-associated phenotype of the strains after growth in the rillettes extract were similar to or lower than that of the hypovirulent internal reference strain L. monocytogenes 442. After growth in milk and cold-smoked salmon, the impact on virulence-associated phenotype depended on the strain. In contrast, plaque-forming assay indicated increased virulence-associated phenotype when the strains were switched from a nutrient-rich medium (food extract or BHI) to a minimum essential medium. CONCLUSIONS: In vitro virulence-associated phenotype level of the studied strains grown in BHI or cold-smoked salmon was the same as the control virulent strain EGD. In contrast, the nutrients present in rillettes may therefore substantially reduce the number of plaques but not the growth of L. monocytogenes. The utilization of minimum essential medium as diluent attenuates changes the effect of the food extract on virulence-associated phenotype in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: In the experimental design of this study, we showed that the nature of the food could affect the in vitro virulence-associated phenotype level of L. monocytogenes.     

Use of molecular typing methods to trace the dissemination of Listeria monocytogenes in a shrimp processing plant.

Molecular typing of bacteria has been widely used in epidemiological studies but not as extensively for tracing the transmission of pathogenic bacteria in food plants. This study was conducted to examine the potential use of two molecular typing methods, random amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE), to trace Listeria monocytogenes contamination in a shrimp processing plant. Ribotyping and phase typing were also performed on a select number of strains. One hundred fifteen strains of L. monocytogenes collected in different areas of a shrimp processing plant were first serotyped and then subtyped by molecular typing. RAPD and PFGE showed great promise for typing L. monocytogenes isolates since distinguishable and reproducible DNA polymorphisms were obtained. When the composite profile from both (RAPD and PFGE) methods was generated, there was an increase in the discriminatory power to discern differences between strains of L. monocytogenes. The results indicated that environmental strains all fell into composite profile groupings unique to the environment, while strains from both water and utensils shared another composite profile group. L. monocytogenes fresh shrimp isolates belonging to one profile group were found in different areas of the processing line. This same profile group was also present in food handlers from the processing and packaging areas of the plant.     

Characterization of Listeria monocytogenes isolated from poultry products and from the poultry-processing environment by random amplification of polymorphic DNA and multilocus enzyme electrophoresis.

A total of 289 Listeria monocytogenes strains isolated from a poultry-processing environment and poultry products over a 6-month period were characterized by random amplification of polymorphic DNA, (RAPD) to pinpoint sources of contamination within the plant and gain some measure of the persistence of individual genotypes within this environment. Eighteen RAPD profiles (A through R) were identified within this group, with 64% (184 of 289) of all strains displaying a single RAPD profile, RAPD type A. This genotype was more prevalent in the raw-poultry-processing environment, where, although its origin within this environment appeared to be the incoming birds, it was also widespread on food contact surfaces, floors, and drains. This was the only genotype which persisted throughout the entire 6-month period, and it and RAPD type B were the only two genotypes found in both the raw- and cooked-poultry-processing environments. L. monocytogenes strains isolated from cooked poultry products and the cooked-poultry-processing environment up to 1 year later (17 strains) contained only RAPD types A and B, highlighting the potential which exists for persistent strains to cross-contaminate foods processed in that environment. The other genotypes (C through R) occurred more sporadically, suggesting varied sources of contamination. These were confined to either the raw- or the cooked-poultry-processing environment and were relatively short-lived. Further characterization of a selection of RAPD type A strains, together with strains of RAPD types B through R, was carried out by multilocus enzyme electrophoresis. Strains of RAPD type A contained two electrophoretic types, one of which was serotype 1/2a and the other was 1/2c.(ABSTRACT TRUNCATED AT 250 WORDS)     

A novel real-time PCR for Listeria monocytogenes that monitors analytical performance via an internal amplification control

We describe a novel quantitative real-time (Q)-PCR assay for Listeria monocytogenes based on the coamplification of a target hly gene fragment and an internal amplification control (IAC). The IAC is a chimeric double-stranded DNA containing a fragment of the rapeseed BnACCg8 gene flanked by the hly-specific target sequences. This IAC is detected using a second TaqMan probe labeled with a different fluorophore, enabling the simultaneous monitoring of the hly and IAC signals. The hly-IAC assay had a specificity and sensitivity of 100%, as assessed using 49 L. monocytogenes isolates of different serotypes and 96 strains of nontarget bacteria, including 51 Listeria isolates. The detection and quantification limits were 8 and 30 genome equivalents, and the coefficients for PCR linearity (R2) and efficiency (E) were 0.997 and 0.80, respectively. We tested the performance of the hly-IAC Q-PCR assay using various broth media and food matrices. Fraser and half-Fraser media, raw pork, and raw or cold-smoked salmon were strongly PCR-inhibitory. This Q-PCR assay for L. monocytogenes, the first incorporating an IAC to be described for quantitative detection of a food-borne pathogen, is a simple and robust tool facilitating the identification of false negatives or underestimations of contamination loads due to PCR failure.     

The incidence and level of Listeria monocytogenes contamination of food sources at primary production and initial processing

Listeria monocytogenes was isolated in low numbers from a variety of environmental samples associated with the primary production of food, including vegetation, faeces and meat. The organism was rarely detected on growing grass and vegetables prior to processing. The excretion of L. monocytogenes by farm animals was linked to their diet, with animals fed entirely on hay or manufactured diets not excreting detectable levels of Listeria (i.e. absence in 25 g). However, animals fed on silage, which is frequently contaminated with L. monocytogenes , commonly excreted the organism. Transport of live animals over long distances (> 100 km) significantly increased the level of excretion of Listeria , but the contamination of carcasses of sheep and cattle was not high. Pigs and poultry faeces were free of Listeria prior to slaughter and pig carcasses were not found to have Listeria present. Frozen and chilled chicken did show detectable levels reflecting the greater potential for contamination during poultry processing. Samples of minced beef were tested and 21 of 23 samples were positive for L. monocytogenes , demonstrating that processing significantly increases the level of contamination compared to whole carcasses. Multilocus enzyme electrophoresis of a representative selection of the isolates showed that there was a wide range of electrophoretic types present in the primary production environment, relatively few of which have been linked to cases of human listeriosis. However, these types do arise on farms and occasional contamination of food raw material by potentially virulent strains may be sufficient to allow adaptable strains to become established in the processing environment and thus be responsible for more widespread contamination of the food available to the consumer.     

Listeria monocytogenes occurrence and characterization in meat-producing plants

AIMS: The prevalence, level of contamination and epidemiological profile of Listeria monocytogenes were investigated in two meat-producing plants during a 20-month period. METHODS AND RESULTS: Sampling for L. monocytogenes was carried out in a cattle slaughterhouse (n = 72) and a swine meat-processing plant (n = 68) during a 20-month period. Swabs and food samples were analysed with the most probable number (MPN) technique for L. monocytogenes and the isolated strains were characterized by AscI-restriction analysis pulsed-field gel electrophoresis (REA-PFGE). Contamination of meat and meat products was always at low level (below 50 MPN per gram). The seven L. monocytogenes positive samples isolated in the bovine slaughterhouse yielded strains with the same REA-PFGE profile. However, the seven strains isolated in the swine meat processing plant showed six different profiles. Two of them showed indistinguishable profiles with L. monocytogenes strains collected from other meat processing facilities located in the same area. SIGNIFICANCE AND IMPACT OF THE STUDY: The genotyping method is a valuable tool to investigate contamination sources. The study of REA-PFGE profiles indicated that environmental contamination was probably responsible for the persistence of over 16 months of one strain of L. monocytogenes in the cattle slaughterhouse. Several meat suppliers could be responsible for the contamination in the pig meat processing facility, and this is confirmed by the finding of some identical strain in other meat processing facilities located in the same area.     

Random amplification of polymorphic bacterial DNA: evaluation of 11 oligonucleotides and application to food contaminated with Listeria monocytogenes

CH. NIEDERHAUSER, CH. HÖFELEIN, M. ALLMANN, P. BURKHALTER, J. LÜTHY AND U. CANDRIAN. 1994. The polymerase chain reaction was used to obtain randomly-amplified polymorphic DNA (RAPD) profiles from Listeria spp. and enterobacteria. Eleven different oligonucleotides were evaluated. Only one, HR4 (19mer), generated reproducible and specific profiles for Listeria spp., while results for enterobacteria were controversial. A total of 57 different Listeria strains were subjected to the RAPD analysis and 27 different profiles were recognized. RAPD typing allowed strains of the same serotype to be distinguished but the same profile was obtained from different serotypes of L. monocytogenes in three cases and in one case two different serotypes of L. innocua yielded the same profile. RAPD-typing with HR4 allowed L. monocytogenes contamination in several food outlets to be traced back to a food processing plant. In additional experiments, the general utility of this RAPD system in typing Yersinia enterocolitica, verotoxigenic Escherichia coli and Salmonella enteritidis was evaluated.     

Behavior of Listeria monocytogenes during fabrication and storage of experimentally contaminated smoked salmon

Experiments were carried out to examine the behavior of Listeria monocytogenes in the course of fabrication and storage of smoked salmon. In three trials, raw salmon fillets were surface inoculated with L. monocytogenes, marinated, smoked at 26 to 30 degrees C, and stored at 4 or 10 degrees C for up to 30 days. At different times during the fabrication and storage, samples were taken and, by means of the three-tube most probable number (MPN) method, quantitatively analyzed for the concentration of L. monocytogenes. The initial Listeria levels in the raw fillets were 10(4) MPN/g in trial 1, 10(1) MPN/g in trial 2, and 10(2) MPN/g in trial 3. During the fabrication, neither an increase nor a decrease of the inoculated quantities was observed. During the storage, however, a significant growth was measured in two of three trials; in trial 1, a 2.5 log10 MPN/g increase and in trial 3, an increase of even 4.5 log10 MPN/g. In the second trial, the Listeria level remained about the same. The results indicate the importance of preventing pre- and postprocessing contamination of L. monocytogenes in raw and smoked salmon. Because a significant increase of L. monocytogenes was measured during storage, there might be an increasing risk of infection for the consumer by storing such fish for a long time.     

Behavior of Listeria monocytogenes during fabrication and storage of experimentally contaminated smoked salmon.

Experiments were carried out to examine the behavior of Listeria monocytogenes in the course of fabrication and storage of smoked salmon. In three trials, raw salmon fillets were surface inoculated with L. monocytogenes, marinated, smoked at 26 to 30 degrees C, and stored at 4 or 10 degrees C for up to 30 days. At different times during the fabrication and storage, samples were taken and, by means of the three-tube most probable number (MPN) method, quantitatively analyzed for the concentration of L. monocytogenes. The initial Listeria levels in the raw fillets were 10(4) MPN/g in trial 1, 10(1) MPN/g in trial 2, and 10(2) MPN/g in trial 3. During the fabrication, neither an increase nor a decrease of the inoculated quantities was observed. During the storage, however, a significant growth was measured in two of three trials; in trial 1, a 2.5 log10 MPN/g increase and in trial 3, an increase of even 4.5 log10 MPN/g. In the second trial, the Listeria level remained about the same. The results indicate the importance of preventing pre- and postprocessing contamination of L. monocytogenes in raw and smoked salmon. Because a significant increase of L. monocytogenes was measured during storage, there might be an increasing risk of infection for the consumer by storing such fish for a long time.     

Biodiversity of Listeria monocytogenes sensitivity to bacteriocin-producing Carnobacterium strains and application in sterile cold-smoked salmon

AIMS: The aim of this study was to demonstrate the inhibitory capacity of Carnobacterium strains against a collection of Listeria monocytogenes strains in cold-smoked salmon (CSS). METHODS AND RESULTS: Three bacteriocin-producing strains, Carnobacterium divergens V41, C. piscicola V1 and C. piscicola SF668, were screened for their antilisterial activity against a collection of 57 L. monocytogenes strains selected from the French smoked salmon industry, using an agar spot test. All the Listeria strains were inhibited but three different groups could be distinguished differing in sensitivity to the three Carnobacterium strains. However, C. divergens V41 always had the highest inhibitory effect. The antilisterial capacity was then tested in sterile CSS blocks co-inoculated with Carnobacterium spp. and mixtures of L. monocytogenes strains. C. divergens V41 was the most efficient strain, maintaining the level of L. monocytogenes at <50 CFU g(-1) during the 4 weeks of vacuum storage at 4 and 8 degrees C, whatever the sensitivity of the set of L. monocytogenes strains. CONCLUSIONS: C. divergens V41 may be a good candidate for biopreservation in CSS. SIGNIFICANCE AND IMPACT OF THE STUDY: A biopreservation strategy for CSS against the risk of L. monocytogenes was investigated using bacteriocin-producing lactic acid bacteria.     

Prevalence and growth of Listeria monocytogenes in naturally contaminated cold-smoked salmon

AIM: To investigate Listeria monocytogenes contamination and behaviour in naturally contaminated French cold-smoked salmon (CSS). METHOD AND RESULTS: Between 2001 and 2004, L. monocytogenes was detected in 104 of 1010 CSS packs, produced by nine French plants, with different prevalence (from 0% to 41%). The initial contamination, measured with a sensitive filtration method, was low (92% of contaminated products below 1 CFU g(-1)) and growth was limited. CONCLUSION: Growth was consistent with results of a predictive model including microbial competition. SIGNIFICANCE AND IMPACT OF THE STUDY: To be included in a quantitative risk assessment.     

Incidence of Listeria spp. and Listeria monocytogenes in a poultry processing environment and in poultry products and their rapid confirmation by multiplex PCR.

The incidence of Listeria spp. and Listeria monocytogenes in a poultry processing plant and in raw and cooked poultry products was determined over a 6-month period. Within the raw and cooked poultry processing environments, 46% (36 of 79) and 29% (51 of 173) of the samples contained Listeria spp. while 26% (21 of 79) and 15% (27 of 173) contained L. monocytogenes, respectively. Various sites within the processing environment were found to be consistently positive for L. monocytogenes throughout the entire sampling period. Of the raw and cooked products tested, 91% (53 of 58) and 8% (8 of 96) were found to contain Listeria spp. while 59% (34 of 58) and 0% (0 of 96) contained L. monocytogenes, respectively. Although L. monocytogenes was not detected in the cooked products examined, the presence of other Listeria spp. highlights the potential which exists for postprocessing contamination. Multiplex PCR proved to be a convenient and time-saving technique for rapid confirmation of Listeria spp. and L. monocytogenes in a single reaction.     

Incidence of Listeria in hot- and cold-smoked fish

Twenty-three samples of hot-smoked fish and 58 samples of cold-smoked fish were tested for the presence of Listeria. Listeria spp. were isolated from two (8.7%) and five (8.6%) of the respective samples tested. Listeria monocytogenes was only isolated from 2 samples (3.4%) of the cold-smoked fish.