Using three reference strains of Brachyspira hyodysenteriae (B204, B234, B169), one B. pilosicoli (P43/6/78), one B. murdochii (56-150), one B. intermedia (PWS/A), one B. innocens (B256) and ten Korean isolates, PCR-RFLP analysis of DNA encoding 23S rRNA was performed to establish a rapid and accurate method for characterizing porcine intestinal spirochetes. Consequently, B. hyodysenteriae and B. pilosicoli revealed different restriction patterns; however, the other three species shared the same pattern. These findings are not consistent with a prior report. Differences in 23S rRNA gene sequences, between two B. murdochii strains, 56-150 and 155-20, were observed. These results indicate that 23S rRNA PCR-RFLP could be used as an identification method for pathogenic Brachyspira spp. (B. hyodysenteriae and B. pilosicoli) as well as an epidemiological tool for characterizing spirochetes isolated from swine.
Animals ; DNA, Bacterial/genetics ; Dysentery, Bacillary/diagnosis/microbiology/*veterinary ; Korea ; Phylogeny ; Polymerase Chain Reaction/methods/*veterinary ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 23S/chemistry/*genetics ; Spirochaetales/*genetics/*isolation&purification ; Spirochaetales Infections/diagnosis/microbiology/*veterinary ; Swine ; Swine Diseases/diagnosis/*microbiology

This study was done to characterize diversity in 10 Brachyspira hyodysenteriae isolates in Korea. The isolates were compared with 14 well-characterized non-Korean strains of various Brachyspira species. All Korean isolates showed strong beta haemolysis and had blunt cell ends with 7~14 periplasmic flagella. They produced indole, and did not ferment fructose. They were alpha-glucosidase positive and alpha-galatosidase negative using the APIZYM kit. Using polyclonal antisera raised in rabbits against recognized serotypes, all isolates showed a strong reaction to B. hyodysenteriae antisera E, A and B. Using multilocus enzyme electrophoresis (MLEE) with 15 enzymes and 5 buffer systems, the Korean and non-Korean isolates were divided into 22 electrophoretic types (ETs) and 5 divisions (A, B, C, D and E). Division A corresponded to B. hyodysenteriae, B to B. innocens, C to B. intermedia, D to B. murdochii and E to B. pilosicoli. The 10 Korean isolates of B. hyodysenteriae were relatively diverse, being divided into 9 ETs within MLEE division A. They were all distinct from the non-Korean strains.
Animals ; Electrophoresis ; Genes, Bacterial ; Korea/epidemiology ; Rabbits ; Serotyping ; Serpulina hyodysenteriae/classification/genetics/*isolation&purification ; Spirochaetales Infections/*microbiology ; Swine/microbiology ; Swine Diseases/*microbiology ; Variation (Genetics)