Previous research involving internalin A (inlA) and internalin B (inlB) single-and double-knockout mutants of Listeria monocytogenes has suggested the involvement of two surface proteins, InlA and InlB, in the adherence of the cells to a glass surface. This phenomenon was further investigated with a larger number (n=27) of L. monocytogenes wild-type strains that were isolated from catfish processing plants and catfish fillets, in addition to internal controls, one ATCC 7644 strain, and L. monocytogenes EGDe strain. Of the wild-type strains, three were shown to produce truncated forms of InlA protein. A blot succession method was used to measure the ease of detachment of sessile L. monocytogenes from a glass surface after attachment at 4°C for 8 h. Real-time reverse transcriptase polymerase chain reaction was used to quantitate mRNA levels of inlA and inlB in L. monocytogenes strains after the cells were incubated at 4°C for 8 h. An inverse relationship between the ease of cell removal from glass surface and the relative inlA and inlB mRNA levels with R 2 value of 0.664 and 0.431, respectively, was observed. This suggests that the attachment strength of L. monocytogenes on glass surface is positively correlated with inlA and inlB expression. There were no differences (p>0.05) in attachment strength among serotypes. These results suggest that L. monocytogenes InlA and InlB proteins play a role in adherence to a glass surface at low temperature, and the attachment ability is independent of serotype.
Incidence of Listeria spp. in whole raw catfish, catfish fillets, and processing environments from two catfish processing facilities was determined in August 2008 and August 2009. Thirty-nine (18.4%) of 212 samples collected in August 2008 were positive for Listeria monocytogenes. Prevalences of Listeria species L. innocua and L. seeligeri-L. welshimeri-L. ivanovii were 11.3 and 23.6%, respectively. Of 209 samples collected in August 2009, 12.4% were positive for L. monocytogenes, 11% for L. innocua, and 19.6% for L. seeligeri-L. welshimeri-L. ivanovii. No Listeria grayi was detected in any of the samples. L. monocytogenes was not found in catfish skins and intestines, but was detected in catfish fillets, on food contact surfaces, and on non-food contact surfaces with frequencies of 45.0, 12.0, and 11.1%, respectively. In August 2008 isolates, serotypes 1/2b (62.2%) and 3b (15.6%) were frequently isolated, whereas the majority of the August 2009 isolates (92.3%) were serotype 1/2b. Genotyping analyses revealed that some genotypes of L. monocytogenes isolates were detected in one facility even after a year, but no persistence of L. monocytogenes was observed in the other facility. In addition, some L. monocytogenes isolates from fresh fillets showed genotypes that were either identical, or more than 90% similar, to those of L. monocytogenes isolates from food contact surfaces in the processing lines. The results of this study suggest that processing environment rather than whole raw catfish is an important source of L. monocytogenes contamination in the catfish fillets. These results should assist the catfish industry to develop better control and prevention strategies for L. monocytogenes.
A 2-year study was performed at two ready-to-eat tilapia sashimi processing plants (A and B) to identify possible routes of contamination with Listeria monocytogenes during processing. Samples were collected from the aquaculture environments, transportation tanks, processing plants, and final products. Seventy-nine L. monocytogenes isolates were found in the processing environments and final products; 3.96% (50 of 1,264 samples) and 3.86% (29 of 752 samples) of the samples from plants A and B, respectively, were positive for L. monocytogenes . No L. monocytogenes was detected in the aquaculture environments or transportation tanks. The predominant L. monocytogenes serotypes were 1/2b (55.70%) and 4b (37.97%); serotypes 3b and 4e were detected at much lower percentages. At both plants, most processing sections were contaminated with L. monocytogenes before the start of processing, which indicated that the cleaning and sanitizing methods did not achieve adequate pathogen removal. Eleven seropulsotypes were revealed by pulsed-field gel electrophoresis and serotyping. Analysis of seropulsotype distribution revealed that the contamination was disseminated by the processing work; the same seropulsotypes were repeatedly found along the work flow line and in the final products. Specific seropulsotypes were persistently found during different sampling periods, which suggests that the sanitation procedures or equipment used at these plants were inadequate. Plant staff should improve the sanitation procedures and equipment to reduce the risk of L. monocytogenes cross-contamination and ensure the safety of ready-to-eat tilapia products.
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