Evaluation of iron dextran and mucin for enhancement of the virulence of Yersinia enterocolitica serotype O:3 in mice

The pathogenic role of Yersinia enterocolitica serotypes O:3, O:8, and O:9 in human infections is well documented. Whereas the virulence of the O:8 strains can be readily demonstrated in mice by 50% lethal dose determinations, the O:3 and O:9 strains have no lethal effect on mice by any route of inoculation. A mouse virulence test for the O:3 and O:9 strains is described. Y. enterocolitica strains were first tested for the presence of virulence-associated plasmid characteristics by auto-agglutination and gel electrophoresis procedures before mouse virulence determinations. The 50% lethal dose of the O:3 strains injected intraperitoneally with 2.5% mucin was about 10(7) colony-forming units. However, histological examinations showed that mucin allowed the growth of Y enterocolitica on the surface of the livers and spleens of the mice without internal lesions. The 50% lethal dose of the same O:3 strains injected intraperitoneally with 1 ml of 10% iron dextran in saline was about 10(5) to 10(6) colony-forming units, and the nonlethal infective dose with typical lesion development was 20 to 200 colony-forming units. The infected mice developed symptoms and extensive liver and spleen lesions which differed from those in mice infected intraperitoneally with the virulent O:8 strains. These results showed that the virulence of the O:3 Y. enterocolitica strains can be measured by intraperitoneal injection with iron dextran. This procedure was used to test the virulence of food isolates, plasmidless strains, and the effect of growth temperatures.