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

2006 ◽  
Vol 72 (6) ◽  
pp. 4313-4322 ◽  
Author(s):  
Gitte Wulff ◽  
Lone Gram ◽  
Peter Ahrens ◽  
Birte Fonnesbech Vogel

ABSTRACT 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.

2003 ◽  
Vol 66 (1) ◽  
pp. 52-60 ◽  
Author(s):  
ADAM D. HOFFMAN ◽  
KENNETH L. GALL ◽  
DAWN M. NORTON ◽  
MARTIN WIEDMANN

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. monocytogenes testing of drains, combined with molecular subtyping of the isolates obtained, allows for efficient monitoring of persistent L. monocytogenes contamination in a processing plant.


1983 ◽  
Vol 91 (3) ◽  
pp. 479-490 ◽  
Author(s):  
B. W. Adams ◽  
G. C. Mead

SUMMARYThe incidence ofStaphylococcus aureuson turkeys sampled at various stages of processing and further-processing was determined on four occasions at each of three different processing plants. For freshly-slaughtered birds, counts from neck skin varied from plant to plant over the range < 102to > 105/g but in all cases the corresponding counts obtained from carcasses sampled after chilling rarely exceeded 103/g and the same was true for samples of mechanically recovered meat (MRM), the final raw product examined.Despite the limited susceptibility of isolates from the different factories to typing by means of either standard human or poultry bacteriophages (55–94% untypable), evidence was obtained with the aid of biotyping for the presence of both human and animal-derived strains. However, some biotypes isolated from MRM were not detected at earlier stages of processing.At one processing plant, an ‘indigenous’ type ofS. aureuswas clearly demonstrated. It occurred in high numbers in the defeathering machines (up to 105/swab), was found on carcasses at all subsequent stages of processing over the survey period and was shown to survive routine cleaning and disinfection procedures. Isolates of this type produced unusually large amounts of extracellular ‘slime’ in artificial culture.Two of the three processing plants yielded isolates which were enterotoxigenic. Of 55 strains from Plant 1, 60% produced enterotoxin C and all were of the ‘indigenous’ type. In the case of Plant 2, only two type D-and one type F-producing strain were found.


2004 ◽  
Vol 67 (2) ◽  
pp. 328-341 ◽  
Author(s):  
JOANNE THIMOTHE ◽  
KENDRA KERR NIGHTINGALE ◽  
KEN GALL ◽  
VIRGINIA N. SCOTT ◽  
MARTIN WIEDMANN

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 &lt; 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.


2008 ◽  
Vol 71 (5) ◽  
pp. 1028-1034 ◽  
Author(s):  
ANNE JENSEN ◽  
DENITA WILLIAMS ◽  
ELIZABETH A. IRVIN ◽  
LONE GRAM ◽  
MARY ALICE SMITH

The foodborne pathogen Listeria monocytogenes can cause infection in immunocompromised humans and in the fetuses of pregnant women. We have demonstrated that one group of genetically similar L. monocytogenes strains (random amplified polymorphic DNA [RAPD] type 9) dominate and persist in several independent fish processing plants. The purpose of the present study was to determine the virulence potential of one RAPD type 9 strain (La111), one human clinical strain (Scott A), and one monkey clinical strain (12443) in a pregnant guinea pig model. Animals were orally exposed to 108 CFU of L. monocytogenes in whipping cream on gestation day (GD) 36 and euthanized on GD 42, 45, or 56. Strains 12443 and Scott A were shed from treated animals for 20 days, whereas La111 was shed only in the first 10 days. Strains 12443 and Scott A were recovered from maternal liver, spleen, and gallbladder on all 3 days of euthanization, whereas La111 was recovered only at GD 45 and 56. Scott A was not isolated from any placentas or fetuses. For dams treated with 12443, 22% of the fetuses were positive for L. monocytogenes, and surprisingly, treatment of dams with La111 resulted in 56% infected fetuses. L. monocytogenes was isolated from 16 and 20% of placentas for 12443 and La111, respectively. The study demonstrates that a food processing plant persistent strain of L. monocytogenes is able to cross the fetoplacental barrier in pregnant guinea pigs. Furthermore, we demonstrate that although information can be gained from model virulence assays, assessment of the virulence potential of a strain may require more complex hosts.


2005 ◽  
Vol 68 (6) ◽  
pp. 1228-1231 ◽  
Author(s):  
ANNUKKA MARKKULA ◽  
TIINA AUTIO ◽  
JANNE LUNDÉN ◽  
HANNU KORKEALA

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.


2006 ◽  
Vol 69 (4) ◽  
pp. 835-841 ◽  
Author(s):  
HIROMI NAKAMURA ◽  
YUKA TOKUDA ◽  
AYUMI SONO ◽  
TOMOKA KOYAMA ◽  
JUN OGASAWARA ◽  
...  

In this study, Listeria monocytogenes contamination in a cold-smoked fish processing plant in Osaka, Japan, was examined from 2002 to 2004. A total of 430 samples were collected and divided into five categories: raw fish, materials during processing, processing equipment, environment, and finished products. A total of 59 finished products were examined throughout this study. L. monocytogenes was isolated from four of these samples during summer and autumn but was not found during winter or spring. During the warmer seasons, L. monocytogenes was more prevalent on processing equipment, especially slicing machines (8 of 54 samples in summer and autumn versus 1 of 50 samples in winter and spring). L. monocytogenes was not detected on whole skins removed from 23 frozen raw fish. L. monocytogenes strains isolated from 56 samples were characterized by serotyping, pulsed-field gel electrophoresis, and three PCR-based methods. Seventy-seven L. monocytogenes strains were recognized as contaminants of the samples: 2 distinguishable strains were identified in each of 13 samples, 3 strains were identified in 2 samples, 5 strains were identified in 1 sample, and the other 40 strains were identified in 40 samples. Combining the results from these techniques, 77 strains were classified into 13 different types. Three of these types prevailed throughout the plant, and two of the three were also isolated from final products. The DNA subtype found in the product was also found on the slicing machines. Our findings suggest that the slicing machines at this plant were the source of the product contamination. Implementing an appropriate cleaning regime for the slicing machines was effective in preventing contamination.


1999 ◽  
Vol 65 (1) ◽  
pp. 150-155 ◽  
Author(s):  
Tiina Autio ◽  
Sebastian Hielm ◽  
Maria Miettinen ◽  
Anna-Maija Sjöberg ◽  
Kaarina Aarnisalo ◽  
...  

ABSTRACT 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. monocytogenespulsotypes 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 eliminatingL. monocytogenes. None of the control samples taken in the 5 months after the eradication program was implemented containedL. monocytogenes.


2005 ◽  
Vol 68 (10) ◽  
pp. 2068-2077 ◽  
Author(s):  
VIRGÍNIA F. ALVES ◽  
ELAINE C. P. DE MARTINIS ◽  
MARIA TERESA DESTRO ◽  
BIRTE FONNESBECH VOGEL ◽  
LONE GRAM

Data on the prevalence and growth of Listeria monocytogenes in lightly preserved fish products from subtropical and tropical regions are very scarce. Our research describes L. monocytogenes that was detected in 5% of the packages of cold-smoked surubim, a native Brazilian freshwater fish that we analyzed, and shows that the strains isolated were of the same random amplified polymorphic DNA subtype as the strains that were isolated from the same factory 4 years earlier. A bacteriocinogenic strain of Carnobacterium piscicola (strain C2), isolated from vacuum-packed cold-smoked surubim, and two C. piscicola strains, isolated from vacuum-packed, cold-smoked salmon, were capable of limiting or completely inhibiting the growth of an L. monocytogenes (strain V2) isolated from surubim in fish peptone model systems incubated at 10°C. Mono-cultures of L. monocytogenes reached 108 CFU/ml (g), whereas the growth of L. monocytogenes was completely inhibited by C. piscicola C2. The bacteriocinogenic C. piscicola A9b+ and its nonbacteriocinogenic mutant A9b− reduced maximum Listeria levels by 2 to 3 log units. Both bacteriocinogenic C. piscicola strains prevented listerial growth in cold-smoked fish juices (surubim and salmon). Although the carnobacteria grew poorly on cold-smoked surubim at 10°C, the strains were able to reduce maximum Listeria counts by 1 to 3 log units in an artificially inoculated product (surubim). We conclude that Brazilian smoked fish products harbor L. monocytogenes and should be stabilized against the growth of the organism. C. piscicola C2 has the potential for use as a bioprotective culture in surubim and other lightly preserved fish, but further studies are required to optimize its effect.


2001 ◽  
Vol 64 (7) ◽  
pp. 994-999 ◽  
Author(s):  
MARIA K. MIETTINEN ◽  
LIISA PALMU ◽  
K. JOHANNA BJÖRKROTH ◽  
HANNU KORKEALA

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.


2006 ◽  
Vol 69 (4) ◽  
pp. 815-819 ◽  
Author(s):  
CHUNG-HSI CHOU ◽  
JUAN L. SILVA ◽  
CHINLING WANG

Raw channel catfish fillets collected from three processing plants during four time periods were tested for the presence of Listeria species. Listeria monocytogenes was the predominant Listeria species found in these catfish fillets, with 25 to 47% prevalence. Other Listeria species, such as L. welshimeri, L. innocua, L. ivanovii, L. grayi, and L. seeligeri, were also found. L. monocytogenes isolates were further fingerprinted by a repetitive element PCR. Forty distinctive electrophoretic types (ETs) and three genetic clusters were determined by Dice coefficient analysis and UPGMA (unweighted pair group method using arithmetic averages). Twenty of 40 ETs were represented by a single isolate, and the other 20 ETs were represented by 2 to 11 isolates. Thirty-five ETs, represented by 76 isolates, were found in processing plant A, B, or C and designated plant-specific types. The remaining five ETs, represented by 21 isolates, were found in multiple plants and designated nonplant-specific types. In addition, 10 ETs from 52 isolates were found repeatedly during different seasons. Plant-specific and nonplant-specific L. monocytogenes coexisted in processed catfish fillets. Some isolates were persistently found in processed fillets, suggesting that either the current sanitation procedures used by these plants are inadequate or that these isolates originated from the natural habitats of the catfish. The results also suggest that the repetitive element PCR is a useful tool for differentiating L. monocytogenes subtypes and can be used for tracing the source of a contamination.


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