Antigens of Eubacterium species including E. alactolyticum, E. brachy, E. nodatum, E. saburreum, E. timidum, E. yurii subsp. yurii and E. yurii subsp. margaretiae, which have been isolated frequently from periodontal pockets and associated with periodontal diseases, were extracted by ultrasonication from whole bacterial cells. Antigens were also prepared from E. aerofaciens, E. lentum and E. rectale, which have been found in intestinal tracts and infected abscesses in human oral cavities. The antigens of the oral Eubacterium species were compared with antigens from E. limosum, the type species of the genus Eubacterium, by using SDS-PAGE and Western immunoblot assays. SDS-PAGE gels stained with Coomassie brilliant blue indicated that no major peptide bands were common among the Eubacterium species examined. The protein profile patterns were distinctly different from each other. Western immunoblotting reactions with rabbit antisera showed that the Eubacterium species could be clearly distinguished serologically, and that the species-specific antigens were peptide components of ultrasonic extracts from the whole bacterial cells. The present study demonstrates that these Eubacterium species show great heterogeneity in their peptide components and immunological reactions, which may be useful for identification of the Eubacterium species from human oral specimens.
BurnetteW. N.1981; Western blotting electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodilied nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Analytical Biochemistry 112:195
FayeL. M. J.,
ChrispeelsM. L.1985; Characterization of N-linked oligosaccharides by affinoblotting with concanavalin A peroxidase and treatment of the blots with glycosidases. Analytical Biochemistry 149:218–224
GunsolleyJ. C.,
TewJ. G.,
GoossC.,
MarshallD. R.,
BurmeisterJ. A.,
SchenkeinH. A.1990; Serum antibodies to periodontal bacteria. Journal of Periodontology 61:412–419
HanN.,
XiaoX.,
ZhangL.,
RiX.,
ZhangJ.,
TongY.,
YangM.,
XiaoZ.1991; Bacteriological study of juvenile periodontitis in China. Journal of Periodontal Research 26:409–414
HillG. B.,
AyersO. M.,
KohanA. P.1987; Characteristics and sites of infection of Eubacterium nodatum, Eubacterium timidum, Eubacterium brachy, and other asaccharolytic eubacteria. Journal of Clinical Microbiology 25:1540–1545
HoldemanL. V.,
CatoE. P.,
MooreW. E. C.1977; Media. In Anaerobe Laboratory Manual, 4th edn. pp. 141–147HoldemanL. V.,
CatoE. P.,
MooreW. E. C.
Edited by Blacksburg: Virginia Polytechnic Institute and State University;
HoldemanL. V.,
CatoE. P.,
BurmeisterJ. A.,
MooreW. E. C.1980; Descriptions of Eubacterium timidum sp. nov., Eubacterium brachy sp. nov., and Eubacterium nodatum sp. nov isolated from human periodontitis. International Journal of Systematic Bacteriology 30:163–169
KondoW.,
NakazawaF.,
SatoM.,
ItoT.1983; Structural studies of the antigenic polysaccharide of Eubacterium saburreum, strain T27. Carbohydrate Research 117:125–131
MartinS. A.,
FalklerW. A.JrVincentJ. W.,
MacklerB. F.,
SuzukiJ. B.1986; A comparison of the reactivity of Eubacterium species with localized and serum immunoglobulins from rapidly progressive and adult periodontitis patients. Journal of Periodontology 59:32–39
MashimoP. A.,
MurayamaY.,
ReynoldsH.,
MoutonC.,
EllisonS. A.,
GencoR. J.1981; Eubacterium saburreum and Veillonella parvula. Journal of Periodontology 52:374–379
MooreW. E. C.,
MooreL. V. H.1986; Genus Eubacterium. In Bergey’s Manual of Systematic Bacteriology pp. 1353–1372SneathP. H. A.,
MairN. S.,
SharpeM. E.,
HoltJ. G.
Edited Baltimore: Williams & Wilkins;
MooreW. E. C.,
HoldemanL. V.,
SmibertR. M.,
HashD. E.,
BurmeisterJ. A.,
RanneyR. R.1982a; Bacteriology of severe periodontitis in young adult humans. Infection and Immunity 38:1137–1148
MooreW. E. C.,
RanneyR. R.,
HoldemanL. V.1982b; Subgingival microflora in periodontal disease: culture studies. In Host Parasite Interaction in Periodontal Disease pp. 13–26GencoR. J.,
MergenhagenS. E.
Edited by Washington DC: American Society for Microbiology;
MooreW. E. C.,
HoldemanL. V.,
CatoE. P.,
SmibertR. M.,
BurmeisterJ. A.,
RanneyR. R.1983; Bacteriology of moderate (chronic) periodontitis in mature adult humans. Infection and Immunity 42:510–515
MooreW. E. C.,
HoldemanL. V.,
CatoE. P.,
SmibertR. M.,
BurmeisterJ. A.,
PalcanisK. G.,
RanneyR. R.1985; Comparative bacteriology of juvenile periodontitis. Infection and Immunity 48:507–519
NakazawaF.,
HoshinoE.1992; Immunochemical and structural characterization of the antigenic polysaccharide from Eubacterium saburreum T18. Oral Microbiology and Immunology 7:182–186
NakazawaF.,
ItoT.,
SatoN.,
SatoM.,
HoshinoE.1987; Chemical composition and immunological characterization of polysaccharide antigen from Eubacterium saburreum T18. Infection and Immunity 55:871–876
TannerA. C. R.,
SocranskyS. S.,
GoodsonJ. M.1984; Microbiota of periodontal pockets losing crestal alveolar bone. Journal of Periodontal Research 19:279–291
TewJ. G.,
MareshallD. R.,
MooreW. E. C.,
BestA. M.,
PalcanisK. G.,
RanneyR. R.1985; Serum antibody reactive with predominant organisms in the subgingival flora of young adults with generalized severe periodontitis. Infection and Immunity 48:303–311
ToloK.,
JorkjendL.1990; Serum antibodies and loss of periodontal bone in patients with rheumatoid arthritis. Journal of Clinical Periodontology 17:288–291
ToloK.,
SchenckK.1985; Activity Of serum immunoglobulins G, A and M to six anaerobic, oral bacteria in diagnosis of periodontitis. Journal of Periodontal Research 20:113–121
VincentJ. K.,
FalklerW. A.JrDalessandroN. F.,
MillerR. A.,
HeathJ. R.III1985; Reaction of human sera with Eubacterium brachy : isolation and characterization of an extracellular antigen. Infection and Immunity 47:592–597
WadeW. G.,
MoranJ.,
MorganJ. R.,
NewcombeR.,
AddyM.1992; The effects of antibacterial acrylic strips on the subgingival microflora in chronic periodontitis. Journal of Clinical Periodontology 19:127–134