The Predominant Actinomyces spp. Isolated from Infected Dentin of Active Root Caries Lesions

Abstract

Actinomyces are Gram-positive pleomorphic rods (GPPR) which form a large proportion of the oral microflora of all mammals. They have been implicated in root caries, although their role in dental caries initiation and progression is not well-understood. Many studies have focused on Actinomyces naeslundii, but few reports have documented other members of the GPPR. Therefore, we investigated the GPPRs isolated from infected dentin of active root caries lesions (n = 9) to determine which species were the most frequently isolated. The GPPR were isolated under both aerobic and anaerobic conditions and identified by biochemical and physiological tests to the species level according to the new taxonomy. Of 654 GPPR isolates investigated, 607 were identified as belonging to the genus Actinomyces. Of these, 242 were identified as A. israelii, 225 as A. gerencseriae, 109 as A. naeslundii, 15 as A. odontolyticus, and 13 as A. georgiae. Individual strains of A. israelii (n = 56) and A. gerencseriae (n = 46) were also investigated at the DNA level by means of Repetitive Extragenic Palindromic polymerase chain-reactions (REP-PCR) for the study of clonal diversity. Although only a small number of isolates was investigated, REP-PCR showed that the genotypes of both A. gerencseriae and A. israelii populations were heterogeneous within individual root caries lesions. A. gerencseriae and A. israelii strains from the same lesions did not share the same REP-PCR patterns, showing the robustness of the identification scheme. A significantly greater proportion of A. gerencseriae was isolated from the aerobic plates (p < 0.05), while the proportion of A. israelii was significantly (p < 0.05) greater from anaerobic plates. The role of individual Actinomyces spp. in the root caries process remains unclear, since various populations of GPPRs were isolated from individual active root caries lesions.

Keywords

Actinomyces spp.root caries taxonomy REP-PCR typing

Get full access to this article

View all access options for this article.

References

Aamdal-Scheie A. , Luan WM , Dahlen G. , Fejerskov O (1996). Plaque pH and microflora of dental plaque on sound and carious root surfaces. J Dent Res 75:1901-1908.

Barrow GI , Feltham Rka (1993). Cowan and Steel's manual for the identification of medical bacteria. 3rd rev. ed. Cambridge: Cambridge University Press.

Barsotti O , Morrier JJ , Decoret D. , Benay G. , Rocca JP (1993). An investigation into the use of restriction endonuclease analysis for the study of the transmission of Actinomyces. J Clin Periodontol 20:436-442.

Beighton D. , Lynch E. (1995). Comparison of selected microflora of plaque and underlying carious dentin associated with primary root caries lesions. Caries Res 29:154-158.

Beighton D. , Miller WA (1977). A microbiological study of normal flora of macropod dental plaque. J Dent Res 56 :995-1000.

Beighton D. , Hellyer PH , Lynch EJ , Heath MR (1991). Salivary levels of mutans streptococci, lactobacilli, yeasts, and root caries prevalence in non-institutionalized elderly dental patients. Community Dent Oral Epidemiol 19:302-307.

Beighton D. , Lynch E. , Heath MR (1993). A microbiological study of primary root-caries lesions with different treatment needs. J Dent Res 72:623-629.

Bowden Ghw (1990). Microbiology of root surface caries in humans . J Dent Res 69:205-1210.

Bowden GH , Ekstrand J. , McNaughton B. , Challacombe SJ (1991). The association of selected bacteria with the lesions of root surface caries. Oral Microbiol Immunol 5:346-351.

10.

Bowden G. , Johnson J. , Schachtele C. (1993). Characterization of Actinomyces with genomic DNA fingerprints and rRNA gene probes. J Dent Res 72:1171-1179.

11.

Brailsford SR , Lynch EJ , Beighton D. (1998). The isolation of Actinomyces naeslundii from sound root surfaces and root carious lesions. Caries Res 32:100-106.

12.

Brown LR , Billings RJ , Kaster AG (1986). Quantitative comparisons of potentially cariogenic microorganisms cultured from noncarious and carious root and coronal tooth surfaces. Infect Immun 51:765-770.

13.

Dent VE , Marsh PD (1981). Evidence for a basic plaque microbial community on the tooth surface in animals. Arch Oral Biol 26:171-179.

14.

Ellen RP , Banting DW , Fillery ED (1985). Longitudinal microbiological investigation of a hospitalized population of older adults with a high root surface caries risk. J Dent Res 64:1377-1381.

15.

Gold OG , Jordan HV , van Houte J. (1973). A selective medium for Streptococcus mutans. Arch Oral Biol 18:1357-1364.

16.

Holdemann LV , Cato EP , Moore Wec (1977). Anaerobe laboratory manual. Blacksburg, VA: VA Polytechnic Institute and State University, P.O. Box 49, 24060.

17.

Holmberg K. , Nord CE ( 1975). Numerical taxonomy and laboratory identification of Actinomyces and Arachnia and related bacteria. J Gen Micro 91:17-44.

18.

Howell A. Jr , Jordan HV , Georg LK , Pine L. ( 1957). Odontomyces viscosus, gen. nov., spec. nov., a filamentous microorganism isolated from periodontal plaque in hamsters. J Dent Res 35:65-68.

19.

Irving JT , Socransky SS , Heeley JD (1974). Histological changes in experimental periodontal disease in gnotobiotic rats and conventional hamsters. J Periodont Res 9:73-80.

20.

Johnson JL , Moore LV , Kaneko B. , Moore WE (1990). Actinomyces georgiae sp. nov., Actinomyces gerencseriae sp. nov., designation of two genospecies of Actinomyces naeslundii and inclusion of A . naeslundii serotypes II and III and Actinomyces viscosus serotype II in A. naeslundii genospecies 2. Int J Syst Bacteriol 40:273-286.

21.

Jordan HV , Hammond BF (1972). Filamentous bacteria isolated from human root surface caries. Arch Oral Biol 17:1333-1342.

22.

Liu T. , Gibbons RJ , Hay DI , Skobe Z. (1991). Binding of A. viscosus to collagen. Association with the type I fimbrial adhesin. Oral Microbiol Immunol 6:1-5.

23.

Nyvad B. (1993). Microbial colonisation of human tooth surfaces . APMIS 32(Suppl):1-45.

24.

Nyvad B. , Kilian M. (1986). Microbiology of the early colonization of human enamel and root surfaces in vivo. Scand J Dent Res 95:369-380.

25.

Nyvad B. , Kilian M. (1990). Microflora associated with experimental root surfaces caries in humans. Infect Immun 58 :1628-1633.

26.

Ozaki K. , Matsuo T. , Nakae H. , Noiri Y. , Yoshigama M. , Ebisu S. (1994). A quantitative comparison of selected bacteria in human carious dentine by microbiological counts. Caries Res 28:137-145.

27.

Putnins EE , Bowden GH (1993). Antigenic relationships among oral Actinomyces isolates, Actinomyces naeslundii genospecies 1 and 2, Actinomyces howellii, Actinomyces denticolens and Actinomyces slackii. J Dent Res 72:1374-1385.

28.

Rodriguez-Barradas MC , Hamill RJ , Houston ED , Georghiou PR , Clarridge JE , Regnery RL , et al. (1995). Genomic fingerprinting of Bartonella species by repetitive element PCR for distinguishing species and isolates. J Clin Microbiol 33:1089-1093.

29.

Schaal KP (1986). Actinomyces spp. In: Bergey's manual of systematic bacteriology. Vol. 2. 8th ed. Sneath PHA , Mair NS , Sharpe ME , Holt JG , editors. Baltimore, MD: Williams and Wilkins, pp. 1383-1418.

30.

Schupbach P. , Osterwalder V. , Guggenheim B. (1995). Human root caries: microbiota in plaque covering sound, carious and arrested root surfaces. Caries Res 29:382-395.

31.

Schupbach P. , Osterwalder V. , Guggenheim B. (1996). Human root caries: microbiota of a limited number of root caries lesions. Caries Res 30:52-64.

32.

Slack JM , Landfried S. , Gerencser MA (1969). Morphological, biochemical and serological studies on 64 strains of Actinomyces israelii. J Bacteriol 97:873-884.

33.

Socransky SS , Manganiello SD (1971). The oral microbiota of man from birth to senility . J Periodontol 42:485-496.

34.

Sumney DL , Jordan HV (1974). Characterization of bacteria isolated from human root surface carious lesions. J Dent Res 53:343-351.

35.

Syed SA , Loesche WJ , Pape HL Jr , Grenier E. (1975). Predominant cultivable flora isolated from human root surface caries plaque. Infect Immun 11:727-731.

36.

ter Steeg PF , van der Hoeven JS , de Jong MH , van Munster PJ , Jansen MJ (1987). Enrichment of subgingival microflora on human serum leading to accumulation of Bacteroides species, Peptostreptococci and Fusobacteria. Antonie Van Leeuwenhoek 53:261-272.

37.

van der Hoeven JS , Mikx FH , Konig KG , Plasschaert AJ (1974). Plaque formation and dental caries in gnotobiotic and SPF Osborne-Mendel rats associated with Actinomyces viscosus. Caries Res 8:211-223.

38.

van Houte J. , Lopman J. , Kent R. ( 1994). The predominant cultivable flora of sound and carious human root surfaces. J Dent Res 73:1727-1734.

39.

Versalovic J. , Koeuth T. , Lupski JR (1991). Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res 19:6823-6831.

40.

Versalovic J. , Kapur V. , Mason EO Jr , Shah U. , Koeuth T. , Lupski JR , et al. (1993). Penicillin-resistant Streptococcus pneumoniae strains recovered in Houston: identification and molecular characterization of multiple clones. J Infect Dis 167:850-856.

41.

Woods CR Jr , Versalovic J. , Koeuth T. , Lupski JR (1992). Analysis of relationships among isolates of Citrobacter diversus by using DNA fingerprints generated by repetitive sequence-based primers in the polymerase chain reaction. J Clin Microbiol 30:2921-2129.