SEF14 fimbriae are only found in some strains of serogroup-D Salmonella such as S. enteritidis, suggesting that SEF14 fimbriae may affect serovar-specific virulence traits. In this study, we found that prevalence of sefA, sefD and sefR genes in S. dublin and S. enteritidis was 100%. In 18 isolates of S. pullorum, the prevalence of sefA gene was 100%, while the prevalence of sefD and sefR genes was 38.9% (7/18), and 11 strains isolated after 1980s did not contain any gene sefD or sefR. Interestingly, among the 7 strains of S. pullorum before 1980s, the sefD sequence has a missing base pair at position 196 and caused open reading frame (ORF) shift, resulting in a stop codon (TAG) at position 71 amino acid residual (Leu of TTA at position 214-216 shift into stop codon of TAG at position 215-217). Unlike S. pullorum, all S. enteritidis and S. dublin tested could express SEF14 fimbriae in vitro.