A total of 120 minimally processed, cut and packaged lettuce samples were purchased from retail supermarkets or provided by a salad production facility over an 8‐month period. The samples were tested for total aerobic plate counts and for the presence of potentially pathogenic species belonging to the genera of Listeria, Aeromonas and Yersinia. The aerobic plate counts ranged from 103 to 109 colony forming units (cfu) g−1. Most samples (76%) contained between 105 and 107 cfu g−1 total aerobic bacteria. Listeria monocytogenes was isolated from three samples, Aeromonas hydrophila or Aeromonas caviae from 66 samples, and Yersinia enterocolitica from 71 samples. The pathogenic potential of Y. enterocolitica isolates was determined by screening for an array of biochemical, serological and genetic traits (heat‐stable enterotoxin gene, the attachment and invasion gene locus, the invasin gene locus and the virulence plasmid). The Y. enterocolitica isolates lacked many of the phenotypic and genetic markers associated with virulence in primary pathogenic strains. As the roles of the reputed virulence factors of Aeromonas spp. in human infection are uncertain, the pathogenic potential of the Aeromonas isolates in lettuce remains unclear.
The polymerase chain reaction (PCR) was used as the basis for the development of highly sensitive and specific diagnostic tests for organisms harboring botulinum neurotoxin type A through E genes. Synthetic DNA primers were selected from nucleic acid sequence data for Clostridium botulinum neurotoxins. Individual components of the PCR for each serotype (serotypes A through E) were adjusted for optimal amplification of the target fragment. Each PCR assay was tested with organisms expressing each of the botulinum neurotoxin types (types A through G), Clostridium tetani, genetically related nontoxigenic organisms, and unrelated strains. Each assay was specific for the intended target. The PCR reliably identified multiple strains having the same neurotoxin type. The sensitivity of the test was determined with different concentrations of genomic DNA from strains producing each toxin type. As little as 10 fg of DNA (approximately three clostridial cells) was detected. C. botulinum neurotoxin types A, B, and E, which are most commonly associated with human botulism, could be amplified from crude DNA extracts, from vegetative cells, and from spore preparations. This suggests that there is great potential for the PCR in the identification and detection of botulinum neurotoxin-producing strains.
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