To evaluate DNA fingerprinting as an epidemiologic tool, pulsed-field gel electrophoresis (PFGE) was performed on isolates of Salmonella, including S. typhimurium, S. thompson, and S. enteritidis. Chromosomal DNA was digested with the restriction endonucleases Bln I and Xba I. The patterns of S. thompson and S. typhimurium isolates from various sources were different from one another. There was no correlation between the phage type and the digestion pattern of S. enteritidis isolates. Some strains belonging to one phage type were distinguished by their PFGE pattern in this study. These results suggest that the Bln I and Xba I digestion patterns of chromosomal DNA are useful for epidemiological analysis of an outbreak of Salmonella infection or food poisoning.
Pulsed-field gel electrophoresis (PFGE) of XbaI-digested DNA fragments of enterohemorrhagic Escherichia coli (EHEC) O157:H7 strains showed disappearance of a 70- or 80-kb fragment in their patterns associated with loss of Shiga toxin genes during maintenance or subcultivation. Hybridization experiments with a DNA probe complementary to Shiga toxin sequences revealed that the Shiga toxin genes in the parental strain were located on fragments the same size as the lost fragments from the toxin-negative derivatives. The evidence indicates that PFGE pattern of EHEC O157:H7 may change due to loss of Shiga toxin genes, which is likely to be associated with curing of Shiga toxin gene carrying phages in vitro.
Eggs that harbor Salmonella in their edible contents pose a significant risk of transmitting disease to consumers. Although Salmonella deposition inside yolks does not usually occur at a high frequency in naturally contaminated eggs, bacterial penetration through the vitelline membrane could lead to rapid and extensive multiplication in the nutrient-rich yolk contents. The present study used an in vitro egg contamination model to assess the ability of Salmonella strains to penetrate the vitelline membrane and multiply inside yolks. An S. enteritidis strain and 2 Salmonella heidelberg strains, initially inoculated onto the outside of the vitelline membrane, were able to enter the yolk contents (at frequencies ranging from 10 to 25% of experimentally contaminated eggs) during 24 h of incubation at 30 degrees C. Variants of these parent strains, obtained by in vivo passage into eggs laid by infected hens, penetrated the yolk membrane at significantly higher frequencies. These results demonstrate that pathogens such as S. enteritidis and S. heidelberg can penetrate into and begin to multiply inside the yolks of contaminated eggs during the first day of storage at warm temperatures.
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