Figure 3 Dendrogram grouping based on the composite RAPD electrophoretic band patterns of representative H. parasuis strains and outgroup strains. Band patterns from all three single-primer experiments were combined to obtain a composite-primer RAPD dendrogram. Reference strains are designated A-O (Table 1), field isolates are designated 1–31 (Table 2), and outgroups are Pasteurella multocida (PM), Mannheimia haemolytica (MH), Pasteurella trehalosi (PT) and Actinobacillus pleuropneumoniae (AP). Three

clade designations are shown. PND-1186 order Reference strains were obtained between 1978 and 1990. Field strains 1–24, 25–29, 30–31 were obtained in 2004, 1999, and 1984, respectively. Numbers at the nodes indicate percentages of AZD0530 bootstrap values after 1000 replicates. Both of the recent

field isolates in Clade A (7 and 9) could be serotyped and 79% of the recent field isolates in Clade C (6, 14, 10–11, 16–22) were typeable, whereas 72% of the recent field isolates (1–2, 12–13, 15, 24) in Clade B were classified as “Unk”. Three isolates (20–22) from the same animal but with two different serotypes (4 and 5) clustered in the same clonal grouping (Figure 3). Comparison of SDS-PAGE protein profiles and pattern analysis Protein bands between 8 and 180 kilodalton (kDa) were present in all of the reference strains and field isolates (Figure 4), as well as a few bands higher than 180 kDa in four of the reference strains C, F, H, and I, respectively. The latter reference strains corresponded to serovars 3, 6, 8, and 9, respectively, Tanespimycin which all designated as avirulent. Isolates gave identical patterns when the analysis was performed in triplicate. Each serovar showed unique band patterns, but there were also common protein bands why among the reference serovars (lanes A-O) and field isolates (lanes 1–31). For example, reference strains C and F showed a common protein at 253 kDa; and reference strains H and I showed a common band at 217 kDa. All reference strains (lanes A-O) and field isolates 25–31 expressed prominent

bands at 140 kDa and 70 kDa and all strains except reference strains B and H (serovars 2 and 8, respectively) showed prominent bands at approximately 40 kDa. Visual inspection of the protein profiles of the field strains 25–31 (Figure 4) showed that these were similar to but not identical to reference strains K and L. Field strains 1–24 protein profiles were more heterogeneous than the reference strains or field isolates 25–31 protein profiles. Field isolates 3, 6, 13, 20, and 29 all had major protein bands at approximately 50 kDa, which were not apparent in the other protein profiles. Outgroup strains (Figure 2B) had unique WCP lysate patterns, which differed from the H. parasuis pattern, on an SDS-PAGE gel. Figure 4 SDS-PAGE profiles of representative H. parasuis strains.