suis, B. melitensis, and B. abortus isolates were passaged
in vitro 14 times over 270 days, that only the B. abortus isolate showed an increase in one TRs copy number at one locus (VNTR 12B) towards the end of this time course [27]. This locus that showed a change was hypervariable to DI 0.88. The clinical isolates would, however, prior to routine, undergo the MLVA assay, which indicates that in-vitro cultivation will not lead to significant changes in the MLVA profiles [27]. To measure the stability of 17 loci via in-vivo passage, native Korean cattle and ICR mice were experimentally infected with the B. abortus strains. The B. abortus RB51 vaccine strains inoculated in the Korean native cattle were not found to have undergone any change in 17 loci, but some of the B. abortus 2308 strains that were isolated NSC 683864 in vitro in the mice were found to have increased TRs copy numbers at Hoof-3 (Figure 5).
Although this difference was naturally caused, it may be generated in the course of the adaptation to the changes in the host. If brucella isolates are Roscovitine manufacturer transferred GS-9973 to the non-preference hosts, there may be changed to TRs copy numbers in some of 17 loci. As the B. abortus strain has infected various animals besides the Bovidae, there seems to be a need for these changes to be further investigated in using the MLVA assay as an epidemiological trace-back tool for transmissions between natural and heterogeneous hosts. Conclusion Korean B. abortus isolates were clustered into nine clusters and 23 genotypes, although they were not highly divided and had low DI values. The MLVA assay showed enough discrimination power in the Brucella species level and could thus be utilized as a tool for epidemiological trace-back in a restricted area. Moreover, it must be considered that even in the farm that was contaminated by one source, the Brucella isolates were able to undergo minor changes at C59 mouse some loci with high DI values especially. The stability studies performed via the in-vivo and in-vitro passages showed that although further investigation may
be needed to determine the stability of marker by changes of the host, 17 loci in this study are sufficiently stable markers for the identification of the original inoculation strain. The MLVA assay can also be applied to determine the relationship between the Brucella isolates from animals and from humans. Methods B. abortus isolates and DNA template preparation A total of 177 isolate that originated from 105 cattle farms (including one elk farm) for the period 1996 to 2008 were selected as representatives for the nine provinces of Korea, namely: Chungbuk (CB), Chungnam (CN), Gyeongbuk (GB), Gyeongnam (GN), Gyeonggi (GG), Jeonbuk (JB), Jeonnam (JN), Jeju (JJ), and Kangwon (KW) [see Additional file 1].
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