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1.
J Appl Microbiol ; 111(4): 997-1005, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21794036

ABSTRACT

AIMS: The chromosomal ail gene (attachment and invasion locus) is commonly used as target gene for the detection of pathogenic Y. enterocolitica strains in food testing. The ail PCR does not detect strains of biotype 1A (BT1A), which are regarded as non-pathogenic because BT1A strains lack the virulence plasmid and chromosomally encoded virulence genes. In some recent reports, however, BT1A strains were discovered that harboured the ail gene. We isolated an ail-positive strain and characterized this strain with phenotypic and genotypic methods to study its possible relation to pathogenic Y. enterocolitica strains. METHODS AND RESULTS: The ail region of the BT1A strain was sequenced and compared with the corresponding region of nonpathogenic BT1A strains and pathogenic strains. Pulsed field gel electrophoresis (PFGE) analysis was applied revealing no similarity of the PFGE pattern of this strain to the patterns of pathogenic strains. Virulence-gene-based PCR analyses showed the strain to be positive for ystB, but negative for virulence genes ystA, virF and yadA. Whole-cell MALDI-TOF MS combined with a shrinkage discriminant analysis approach was applied and clearly classified the ail-positive biotype 1A strain within the cluster of BT1A strains. CONCLUSIONS: PCR detection of ail sequences in food matrices should be followed by the isolation of the responsible strain and its characterization using phenotypic or genotypic methods. SIGNIFICANCE AND IMPACT OF THE STUDY: The ail gene may be present in Y. enterocolitica BT1A strains, which are commonly considered as nonpathogenic. Efficient methods such as PCR typing of other virulence genes or rapid MALDI-TOF MS-based bacterial profiling allow a more comprehensive assessment of the pathogenicity potential of Yersinia strains.


Subject(s)
Bacterial Outer Membrane Proteins/genetics , Virulence Factors/genetics , Yersinia enterocolitica/genetics , Cluster Analysis , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Genes, Bacterial , Genotype , Plasmids , Polymerase Chain Reaction , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Virulence , Yersinia enterocolitica/isolation & purification , Yersinia enterocolitica/pathogenicity
2.
Article in English | MEDLINE | ID: mdl-16000112

ABSTRACT

The efficacy of enterocoliticin, a phage tail-like bacteriocin, as antimicrobial compound against infections with pathogenic Yersinia enterocolitica serotype O3 strains was assessed. In cell cultures, which were infected with the Y. enterocolitica strains 13 169 or 6471/76, bactericidal activity of enterocoliticin was found for bacteria adhering to the surface of eukaryotic cells, whereas bacteria, which had invaded the eukaryotic cells, were not accessible to the bacteriocin. The interaction of enterocoliticin with Y. enterocolitica was further examined in animals. Female BALB/c mice were experimentally infected with the two Y. enterocolitica strains and enterocoliticin was applied as antimicrobial compound by the oral route. Experimental variations concerning the infectious doses of the Y. enterocolitica strains and the time points of application of the bacteriocin were investigated. The increase of the Yersinia CFU titre in animals was retarded at time points shortly after the application of enterocoliticin indicating that the particles were effective on recently introduced Yersinia. The repeated application of enterocoliticin, however, did not prevent the colonization of the gastrointestinal tract by Yersinia.


Subject(s)
Bacteriocins/pharmacology , Yersinia Infections/veterinary , Yersinia enterocolitica/drug effects , Animals , Bacterial Adhesion/drug effects , Bacteriocins/therapeutic use , Bacteriophages , Cells, Cultured/microbiology , Duodenum/cytology , Female , Mice , Mice, Inbred BALB C , Microbial Sensitivity Tests , Specific Pathogen-Free Organisms , Stomach/cytology , Swine , Yersinia Infections/drug therapy , Yersinia enterocolitica/classification , Yersinia enterocolitica/physiology
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