There may not have been a correlation between serotype and RAPD because only a small number of genes is involved in serotyping while the entire genome is analyzed with the RAPD LY333531 purchase technique [22]. Our SDS-PAGE results agree with those of Oliviera and Pijoan [30] who reported that isolates from systemic sites were usually virulent
and clustered together as shown by using a computer-based analysis of protein profiles from serovars 1, 2, 4, 5, 7, 12, 13, 14 and nontypeable (NT) isolates. Their results are similar to protein profiles described in our study for field isolates and their isolation sites and pathogenesis as selleck chemicals shown in the WCP lysate dendrogram of Figure 5 and Table 2. The field strains clustered in Subclade A1 and Clades B and C were primarily systemic. Ruiz et al. [33] found different OMP profiles between isolates AZD5363 price from healthy pigs and those from diseased pigs. However, they concluded that respiratory isolates were more heterogeneous than systemic isolates. Four studies have stated that a protein of approximately 36–38.5 kDa may be associated with Glässer’s disease [29, 30, 33, 56]. In this work, a protein band was
observed at approximately 40 kDa in all of the field isolates and thirteen of fifteen of the reference strains (Figure 4). The results shown for the WCP lysate dendrogram (Figure 5) imply that protein expression may be related to age or number of passages of the isolatein vitro, because reference strains clustered together, as did the “old” field strains (26–29) isolated in 1999 (Figure mTOR inhibitor 5, Subclades A2 (C-G, J-O), A3 (A-B, H-I), and A1 (26–29), respectively). The phenotypic change of an isolate after serial passage was also reported by Rapp-Gabrielson and Gabrielson and Oliviera et al. [12, 57]. Although we had only seven samples from North Carolina, three isolates (27–29) from 1999 grouped together in Subclade A1 of the SDS-PAGE neighbor joining dendrogram (Figure 5). Our WCP lysate patterns
easily discriminated between A. pleuropneumoniae serotype 1 and H. parasuis as well as the other three outgroup strains (Figure 2B). Identical H. parasuis field isolates (H. parasuis IA84-29755 and 31) (Figure 5), bands did not match sufficiently to obtain identity in the protein profile computer analysis. This may have been because the bands were not fully “matched” in the Gel Compar II program. They were, however, in the same clonal branch of Subclade A3. Oliviera and Pijoan [30], Kielstein and Rapp-Gabrielson [5], Rosner et al. [58] and Blackall et al. [59] did not find any correlation between virulence and serotype of the isolate. However, the results reported in this study seem to indicate an association of virulence with isolates of Clade C in the WCP lysate analysis. There also seemed to be more serotypeable isolates among the recent field isolates of Clade C.
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