Saracatinib outgroups were included to compare the presence or absence of bands in these isolates to the bands in the more closely related H. parasuis isolates. The only monophyletic ingroup with the four “outgroups” was the SDS-PAGE dendrogram as determined by the neighbor

joining Selleckchem ABT-263 analysis (Figure 5, Clade A3). The results suggest that the four outgroup species selected may have been too closely related to H. parasuis to act as a true outgroup. Dijkman et al. [20] were also unable to discriminate A. minor and A. porcinus strains from H. parasuis strains in an ERIC-PCR technique. Additionally, Olvera et al. [18] could not demonstrate that A. indolicus and A. minor strains were outgroups to H. parasuis strains when they used the variation of the partial hsp60 sequence of H. parasuis as a classification tool. Others have shown that the geographic distribution or age of the isolate may cause the “outgroup” to act as an ingroup [38] and that if the isolates in the study were too closely related, then the outgroups could be rerooted to locations within phylogenetic trees [39]. A fourth possibility

for the lack of outgroup observance in the dendrograms could be that horizontal gene transfer has occurred between the outgroup species and H. parasuis[40], which would cause unexpected similarities and unusual phyletic patterns [18]. This theory is supported by the presence of bacteriophages in H. parasuis[41–43], E. coli[44], P. multocida[45], M. haemolytica[46], and P. trehalosi[47], plasmids in H. parasuis[48] and A. pleuropneumoniae[49], and a DNA uptake sequence in H. parasuis[50]. Although isolates from known systemic AZD2014 order sites [51] (lung in an animal with polyserositis, joint, brain, heart, or septicemia) were able to be separated into groups by the RAPD

technique described here, the composite diagram of the three individual primers ultimately showed a limited degree of relatedness based on pathogenicity among the reference strains and the 31 field strains. The strains showed high heterogeneity with the RAPD method which indicated possible horizontal transfer of genes or chromosomal recombination between unrelated and potentially transient Benzatropine strains. Wang et al. [25] compared RAPD and MEE and found that RAPD data that combined five primers was more discriminatory than MEE tests that used 34 enzymes. The ERIC-PCR technique is a comparable method to RAPD. Zulkifli et al. [52] found RAPD to be more discriminatory than ERIC-PCR. Some H. parasuis isolates were not able to be assayed by using the ERIC-PCR [20] because they gave no or very poor results. Recent studies have found a high diversity of H. parasuis strains isolated in various geographic areas but have not been able to assign a clear correlation between virulence or serovar and ERIC-PCR clusters [19–21]. This conclusion agrees with other H. parasuis ERIC-PCR studies [12, 18]. Macedo et al.