Unveiling contamination sources and dissemination routes of Salmonella sp. in pigs at a Portuguese slaughterhouse through macrorestriction profiling by pulsed-field gel electrophoresis

Sixty-nine isolates of Salmonella sp. isolated from the ileum, tonsils, carcass and mandibular and ileocolic lymph nodes of individual pigs slaughtered for consumption in one abattoir were analyzed using serotyping and macrorestriction profiling by Pulsed-Field Gel Electrophoresis (RFLP-PFGE), in order to identify clonal relationships. XbaI macrorestriction was able to distinguish 18 genotypes among the eight identified serotypes: Salmonella Typhimurium (4 genotypes), Salmonella Rissen (3), Salmonella Tennessee (2), Salmonella Enteritidis (2), Salmonella 4,[5],12:i:- (4), Salmonella Give (1), Salmonella Anatum (1), and Salmonella Derby (1). Except for one sample, the serotype and the genotype identified in the samples from the same pork were always the same, allowing to unravel possible dissemination routes of Salmonella sp. through these pork tissues and equate presumptive sources of contamination or infection. Highly significant associations (p < 0.001) were observed for the presence of Salmonella sp. in the ileum and in the ileocolic lymph nodes, as well as between the carcass contamination and the presence of Salmonella sp. in others samples of the correspondent slaughtered pig, such as the ileum, the ileocolic and mandibular lymph nodes and the tonsils. Moreover, 80% of the pigs with ileum and ileocolic lymph nodes positive samples also presented the same salmonella genotype in the correspondent tonsils and, among pigs with positive tonsils, 70% also carried the same genotype in the corresponding mandibular lymph nodes. The occurrence of cross-contamination was also detected, since a genotype identified in other pigs slaughtered in the same day was found in 31% of positive carcasses. The global analysis of the genotypes suggested three different sources of pig infection: the farm of origin, the transportation and the lairage. A particular attention should be paid to the last one, since the majority of the isolates from pig samples were related to infection in the lairage. Since the presence of Salmonella sp. in the ileum of pigs and faeces ingestion promotes tonsils infection and internal dissemination of the agent through the mandibular lymph nodes, as well as drainage to the ileocolic lymph nodes, a potential risk exists at slaughter for Salmonella sp. contamination in the carcasses during pork processing. This risk may be increased by incorrect evisceration techniques and by hygienically inappropriate meat inspection procedures, especially those concerned to the mandibular lymph nodes incisions.