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The direct immunofluorescent detection of pathogens on pork skin is evaluated. Calibrate contamination of pork skin with Salmonella Typhimurium (ST) and Listeria monocytogenes (Lm) is developed in 2 h at 4 degrees C. Then a specific indirect immunofluorescent staining protocol is optimized in order to obtain specific and intensive signals able to be detected by electronic cameras (deported microscopy). Despite the individual staining of ST and Lm is possible on pork skin and is specific and bright, the deported microscopy failed to detect these particles. After respectively 3 and 6 h, we obtain micro-colonies of ST and Lm. Due to the limited power of the video camera used, only the microscope permits the detection on the skin. However, our work gives standard conditions to mime the pathogens contamination and staining directly on a biological matrix such as pork skin. This work is a first step in the development of direct and rapid detection of pathogens on biological matrix.

Aims: The origin of Salmonella contamination of pork products is not well established. In order to further this knowledge, the transmission of Salmonella spp. from live pigs to pork cuts was investigated in two pork slaughter and cutting plants., Methods and Results: Salmonella spp. were isolated from both pork (pigs, carcasses, cuts) and the environment before and during slaughterhouse activities. Eight serotypes were identified. XbaI and SpeI macrorestriction distinguished 20 genotypes of Salmonella Typhimurium and 16 genotypes of Salmonella Derby. A major cluster of Salmonella Typhimurium genotypes was common to both plants and all pig-related genotypes, while a predominant pig-related Salmonella Derby genotype was common to both plants., Conclusion: None of the Salmonella strains persisted for long periods in the pork-processing environments., Significance and Impact of the Study: This work shows that contaminated live pigs, because of bacterial spread due to the process and ineffective cleaning procedures, are involved in Salmonella contamination.

In order to determine the origin of pork cuts contamination by Listeria monocytogenes, 287 isolates, collected from five French pork slaughtering and cutting plants, from live pigs to pork cuts, were characterised using three molecular typing methods: random amplification of polymorphic DNA (RAPD) carried out with five different primers, genomic macrorestriction using ApaI with pulsed-field gel electrophoresis (PFGE) and a PCR-restriction enzyme analysis (PCR-REA) based on the polymorphism existing within the inlA and inlB genes. Results obtained from RAPD and PFGE were closely related and distinguished respectively 17 RAPD types (r1-r17) and 17 PFGE types (a1-a17) among the 287 isolates, whereas the PCR-REA analysis only yielded two profiles (p1 and p2). Considering the combined results obtained with the three molecular typing methods, 19 Listeria monocytogenes genotypes (1-19) were distinguished. Serotyping led at least four serotypes being distinguished: 1/2a, 3a, 1/2c and 3c. The application of genotyping identified the predominance of a Listeria monocytogenes strain of type (1) and other very closely related ones (5, 9, 10, 12, 13, 14, 16 and 19) which were present on pork as well as in the environment within the five investigated plants. This study also pointed out the presence of these closely related Listeria monocytogenes strains over a 1-year period in the environments of two plants, even after cleaning and disinfection procedures. This highlights the possibility for some Listeria monocytogenes strains to persist in pork processing environments and raises the problem of the efficiency of cleaning and disinfection procedures used in pork slaughterhouses, chilling and cutting rooms.