Mouse virulence and the ability to adhere to, and invade cultured MDCK cells were investigated in 38 phage type reference strains of Salmonella enterica serotype Enteritidis and correlated with genomic lineage. The genomic lineage of 11 of the strains was determined in the present study; one IS200 and one ribotype pattern that had not been reported previously were observed. Log c.f.u. in the spleen 10 days post intraperitoneal (i.p.) infection with 3×103 bacteria (logVC10) varied between 2·9 and 8·7. The reference strains of PT7 and PT23 were found to be semi-rough and were of low virulence. All other strains possessed smooth LPS. Within each of the two major clonal lines, as well as among phage types outside these, both highly virulent and moderate to low virulent strains were present. While all strains of PT1, PT2 and PT8 were highly virulent, low virulent strains were detected in PT4 and PT13. The ability to adhere to, and invade MDCK cells varied between phage types (adherence between 13 and 61% of the inocula and invasion between 4 and 151% of the adherent cells). The results of the cell culture experiments did not correlate with the results of mouse virulence tests. No correlation between clonal lineage and virulence was found within S. Enteritidis. It seems most likely that some strains have lost some of the essential factors enabling this serotype to cause successful systemic infection.